WO2023208005A1

WO2023208005A1 - Cyclic compounds, preparation methods and medicinal uses thereof

Info

Publication number: WO2023208005A1
Application number: PCT/CN2023/090706
Authority: WO
Inventors: Avinash KHANNA; Hugh Y. Zhu; Weidong Pan
Original assignee: Hansoh Bio Llc; Shanghai Hansoh Biomedical Co., Ltd.; Jiangsu Hansoh Pharmaceutical Group Co., Ltd.
Priority date: 2022-04-25
Filing date: 2023-04-25
Publication date: 2023-11-02
Also published as: TW202400607A

Abstract

Compounds of formula (I), formula (II) or formula (III), the preparation method thereof, pharmaceutical compositions comprising the compounds, and the pharmaceutical uses for the treatment of diseases or disorders are provided.

Description

CYCLIC COMPOUNDS, PREPARATION METHODS AND MEDICINAL USES THEREOF

FIELD OF THE INVENTION

The present invention belongs to the field of medicine, and relates to nitrogen-containing tetracyclic compounds, preparation methods thereof, pharmaceutical compositions comprising the compounds, and medical uses thereof.

BACKGROUD OF THE INVENTION

The most notable members of the RAS subfamily are HRAS, KRAS and NRAS, mainly for being implicated in many types of cancer.

Kirsten Rat Sarcoma 2 Viral Oncogene Homolog ( “KRas” ) is a small GTPase and a member of the Ras family of oncogenes. KRas serves as a molecular switch cycling between inactive (GDP-bound) and active (GTP-bound) states to transduce upstream cellular signals received from multiple tyrosine kinases to downstream effectors to regulate a wide variety of processes, including cellular proliferation (e.g., see Alamgeer et al., (2013) Current Opin Pharmcol. 13: 394-401) .

The role of activated KRas in malignancy was observed over thirty years ago. Aberrant expression of KRas accounts for up to 20%of all cancers and oncogenic KRas mutations that stabilize GTP binding and lead to constitutive activation of KRas and downstream signaling have been reported in 25 -30%of lung adenocarcinomas, (e.g., see Samatar and Poulikakos (2014) Nat Rev Drug Disc 13 (12) : 928-942) . Single nucleotide substitutions that result in missense mutations at codons 12 and 13 of the KRas primary amino acid sequence comprise approximately 40%of these KRas driver mutations in lung adenocarcinoma. KRAS G12D mutation is present in 25.0%of all pancreatic ductal adenocarcinoma patients, 13.3%of all colorectal carcinoma patients, 10.1%of all rectal carcinoma patients, 4.1%of all non-small cell lung carcinoma patients and 1.7%of all small cell lung carcinoma patients (e.g., see The AACRProject GENIE Consortium, (2017) Cancer Discovery; 7 (8) : 818-831. Dataset Version 4) .

The well-known role of KRas in malignancy and the discovery of these frequent mutations in KRas in various tumor types made KRas a highly attractive target of the pharmaceutical industry for cancer therapy. Notwithstanding thirty years of large-scale discovery efforts to patent of WO 2021/041671 develop inhibitors of KRas for treating cancer, no KRas inhibitor has yet demonstrated sufficient safety and/or efficacy to obtain regulatory approval.

Compounds that inhibit KRas activity are still highly desirable and under investigation, including those that disrupt effectors such as guanine nucleotide exchange factors (e.g., see Sun etal., (2012) Agnew Chem IntEd Engl. 51 (25) : 6140-6143) as well recent advances in the covalent targeting of an allosteric pocket ofKRas G12C (e.g., see Ostrem et ah, (2013) Nature 503: 548-551 and Fell et ah, (2018) ACS Med. Chem. Lett. 9: 1230-1234) .

While progress has been made in this field, there remains a need in the art for improved compounds and methods for treatment of cancer, for example by inhibition of KRAS, HRAS or NRAS. The present invention fulfills this need and provides further related advantages.

SUMMARY OF THE INVENTION

The present invention, in one aspect, provides a compound of formula (I) , formula (II) , or formula (III) , or a pharmaceutically acceptable salt, solvate, or prodrug thereof, including tautomers, cis-or trans-isomers, mesomers, racemates, enantiomers, diastereomers, and mixtures thereof:

or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof,

wherein,

ring A and ring B are each independently selected from the group consisting of cycloalkyl, heterocyclyl, aryl and heteroaryl;

R₁ is selected from the group consisting of cycloalkyl, heterocyclyl, aryl and heteroaryl,

wherein each of the cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl, C_3-12 cycloalkyl, C_3-12 heterocyclyl, wherein the C_3-12 heterocyclyl is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl, heterocyclyl;

L and M are each independently selected from the group consisting of alkyl, alkoxy, alkyl-O-C (O) -, alkoxy-O-C (O) -, -alkyl-cycloalkyl-alkyl-O-C (O) -;

R₂, R₃, R₄, R₅, R₆, R₇ and R₈ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl, cycloalkyl and heterocyclyl;

or, R₄ and R₅ together with the carbon atom to which they are bound form cycloalkyl, heterocyclyl, wherein each of the cycloalkyl, heterocyclyl is optionally substituted with one or more substituents selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl;

n is 1, 2 or 3;

G iswherein, R_a and R_b are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl;

or, R_a and R_b together with the carbon atom to which they are bound form cycloalkyl, heterocyclyl, aryl and heteroaryl;

K is selected from the group consisting of vinyl ketone, vinyl sulfone, ynone, alkynyl sulfone, -S (O) ₂ and alkyl, -C (O) -C (O) -alkyl;

J is selected from the group consisting of cycloalkyl, heterocyclyl, aryl and heteroaryl and NR_cR_d;

R_c and R_d are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkenyl, alkynyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl;

or J is absent with the proviso that K is selected from alkyl, -C (O) -C (O) -alkyl.

In some embodiments, the compound of formula (I) or formula (II) being a compound of formula (Ia) or formula (IIa) , or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof:

wherein,

R₁₃ is selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl, heteroaryl;

m is 1, 2 or 3.

In some embodiments, the compound of formula (I) or formula (II) being a compound of formula (Ib) or formula (IIb) , or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof:

wherein,

X₁ is absent, C or N;

X₂ is C, O, N;

R₉, R₁₀, R₁₁ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl, cycloalkyl and heterocyclyl.

In some embodiments, ring A and ring B are each independently selected from the group consisting of C_3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-12 aryl and C_3-12 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S.

In some embodiments, ring B is each independently selected from the group consisting of C_3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-12 aryl and C_3-12 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S.

In some embodiments, R1 is selected from the group consisting of C3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-12 aryl and C_3-12 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S, wherein each of the C_3-12 cycloalkyl, C_3-12 heterocyclyl, C_3-12 aryl and C_3-12 heteroaryl is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-10 cycloalkyl, C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_7-10 bridged heterocyclyl containing 1 to 3 heteroatoms independently selected from N, O or S, wherein each of the C_3-6 heterocyclyl and the C_7-10 bridged heterocyclyl is optionally substituted with one or more substituents selected from C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_3-5 heterocyclyl.

In some embodiments, L and M are each independently selected from the group consisting of bond, C_1-8 alkyl, C_1-8 alkoxy, C_1-8 alkyl-O-C (O) -, C_1-8 alkoxy-O-C (O) -, -C_1-6 alkyl-C_3-6 cycloalkyl-C_1-6 alkyl-O-C (O) -.

In some embodiments, R₂, R₃, R₄, R₅, R₆, R₇, R₈ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-12 cycloalkyl and C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S;

or, R₄ and R₅ together with the carbon atom to which they are bound form C_3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, wherein each of the C_3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S is optionally substituted with one or more substituents selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl.

In some embodiments, R₉, R₁₀, R₁₁ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-12 cycloalkyl and C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S.

In some embodiments, wherein, n is 1.

In some embodiments, G iswherein, R_a and R_b are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl;

or, R_a and R_b together with the carbon atom to which they are bound form C_3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-12 aryl and C_3-12 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S.

In some embodiments, K is selected from the group consisting of C_1-6 vinyl ketone, C_1-6 vinyl sulfone, C_1-6 ynone, C_1-6 alkynyl sulfone, C_1-3 alkyl and -C (O) -C (O) -C_1-3 alkyl.

In some embodiments, J is selected from the group consisting of C_3-12 cycloalkyl, C_3- ₁₂ heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-12 aryl and C_3-12 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S, and NR_cR_d;

R_c and R_d are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-3 alkyl, C_1-3 alkenyl, C_1-3 alkynyl, C_1-3 alkoxy, hydroxyl, C_1-3 hydroxyalkyl, C_1-3 haloalkyl;

or J is absent, with the proviso that K is C_1-3 alkyl.

In some embodiments, ring A and ring B are each independently selected from the group consisting of C_5-10 cycloalkyl, C_5-10 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_5-10 aryl and C_5-10 heteroaryl containing 1 to to 2 heteroatoms independently selected from N, O or S, C_5-10 aryl and C_5-10 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S.

In some embodiments, ring B is each independently selected from the group consisting of C_5-10 cycloalkyl, C_5-10 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_5-10 aryl and C_5-10 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S.

R₁ is selected from the group consisting of C_5-8 cycloalkyl, C_5-8 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_5-8 aryl and C_5- ₈ heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S, wherein each of the C_5-8 cycloalkyl, C_5-8 heterocyclyl, C_5-8 aryl and C_5-8 heteroaryl is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl, C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, C_7-10 bridged heterocyclyl containing 1 to 3 heteroatoms independently selected from N or O, wherein each of the C_3-6 heterocyclyl and the C_7-10 bridged heterocyclyl is optionally substituted with one or more substituents selected from C_1-6 alkyl, C_3-5 heteroaryl containing a heteroatom selected from O.

In some embodiments, L is independently selected from the group consisting of linear or branched C_3-6 alkyl, linear or branched C_3-6 alkoxy, linear or branched C_3-6 alkyl-O-C (O) -, linear or branched C_3-6 alkoxy-O-C (O) -and -C_1-3 alkyl-C_3-6 cycloalkyl-C_1-3 alkyl-O-C (O) -.

In some embodiments, M is independently selected from the group consisting of bond, C_1-3 alkyl, C_1-3 alkoxy.

In some embodiments, R₂, R₃, R₄, R₅, R₆, R₇, R₈ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl, C_3-6 cycloalkyl and C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S;

or, R₄ and R₅ together with the carbon atom to which they are bound form C_3-6 cycloalkyl, C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, wherein each of the C_3-6 cycloalkyl, C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S is optionally substituted with one or two substituents selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl.

In some embodiments, R₉, R₁₀, R₁₁ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl, C_3-6 cycloalkyl and C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S.

In some embodiments, G iswherein, R_a and R_b are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl;

or, R_a and R_b together with the carbon atom to which they are bound form C_3-8 cycloalkyl, C_3-8 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-8 aryl and C_3-8 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S.

In some embodiments, K is selected from the group consisting of C_3-6 vinyl ketone, C_3-6 vinyl sulfone, C_3-6 ynone, C_3-6 alkynyl sulfone, -C (O) -C (O) -methyl, -C (O) -C (O) -ethyl, -C (O) -C (O) -propyl.

In some embodiments, J is selected from the group consisting of C_3-8 cycloalkyl, C_3-8 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3- ₈ aryl and C_3-8 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S and NR_cR_d;

R_c and R_d are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-3 alkyl, C_1-3 alkenyl, C_1-3 alkynyl, C_1-3 alkoxy, hydroxyl, C_1-3 hydroxyalkyl, C_1-3 haloalkyl.

In some embodiments, ring A and ring B are each independently selected from the group consisting of:

In some embodiments, R₁ is selected from the group consisting of phenyl, pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, wherein each of the phenyl, pyridinyl, pyrimidinyl, pyrazolyl and imidazolyl is optionally substituted with one or two substituents selected from hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy,

hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl,

In some embodiments, G is selected from the group consisting of pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, piperidyl, piperazinyl, pyrrolidyl, oxazolidinyl, thiazolidinyl.

In some embodiments, K iswherein, R₁₂ is C_1-6 alkyl.

In some embodiments, R₁₂ is C_1-3 alkyl.

In some embodiments, J is selected from the group consisting of pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, piperidyl, piperazinyl, pyrrolidyl, oxazolidinyl, thiazolidinyl, morpholinyl, thiomorpholinyl.

In some embodiments, the compound of formula (I) being a compound of formula (Ic) , or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof:

wherein,

X₂ is N, CH;

R₁₁ is each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkenyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-8 cycloalkyl and C_3-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, wherein the C_3-8 cycloalkyl and C_3-8 heterocyclyl are optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, C_1-6 alkyl;

preferably, the C_3-12 heterocyclyl is C₃-₇ mono-heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, or fused C_8-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S;

R₁₀ is ndependently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl;

R₃ is independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl;

or two of R₃ together with ring B to which they are attached form a bridged C_3-8 cycloalkyl.;

r is 0, 1, 2 or 3;

s is 0, 1, 2 or 3;

In some embodiments, the compound of formula (III) being a compound of formula (IIIc) or (IIId) , or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof:

wherein,

X₃ is N, CH and S;

r is 0, 1, 2 or 3;

s is 0, 1, 2 or 3;

In some embodiments, the compound is a compound of formula (IV) , or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof:

R₁₄ is each independently selected from the group consisting of hydrogen, deuterium, C_1-6 alkyl, C_1-6 alkenyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-8 cycloalkyl and C_3-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, wherein the C_3-8 cycloalkyl and C_3-8 heterocyclyl are optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, C_1-6 alkyl and C_4-6 heterocyclyl containing 1, 2 or 3 heteroatoms selected form N;

u is 0, 1, 2 or 3;

R₄, R₅, R₆, R₁₃ and m are as definded above.

In some embodiments, the compound is a compound of formula (V) , or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof:

u is 0, 1, 2 or 3;

R₄, R₅, R₆, R₇, R₈, R₁₃ and m are as definded above.

In some embodiments, the compounds of formula (I) , or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is being a compound of formula (Id)

wherein,

X₂ is N, CH;

X₃ is N, CH and S;

or two of R₃ together with ring B to which they are attached form a bridged C_3-8 cycloalkyl;

r is 0, 1, 2 or 3;

In preferred embodiments, ring B is selected from the group consisting of phenyl or

In more preferred embodiments, ring B is selected from the group consisting of

phenyl or

In some embodiments, R₁₁ is selected from the group consisting of linear or branched C_1-3 alkoxy, C_2-4 haloalkenyl, C_5-8 mono heterocyclyl containing 1, 2 or 3 heteroatoms selected from N, O, S, C_5-12 fused heterocyclyl containing 1, 2 or 3 heteroatoms selected from N, O, S, wherein the mono heterocyclyl and the fused heterocyclyl are optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, C_1-3 alkyl.

In preferred embodiments, R₁₁ is selected from the group consisting of branched C_1-3 alkoxy, C_2-4 fluorine-alkenyl,

r is 1 or 2;

In some embodiments, R₁₃ is selected from the group consisting of C5-7 heteroaryl comprising 1, 2 or 3 heteroatoms selected from N, O or S;

In some embodiments, the compounds of formula (I) , or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is being a compound of formula (VIII)

wherein,

X₂ is N, CH;

L is independently selected from the group consisting of linear or branched C_3-6 alkyl, linear or branched C_3-6 alkoxy, linear or branched C_3-6 alkyl-O-C (O) -, linear or branched C_3-6 alkoxy-O-C (O) -, -C_1-3 alkyl-C_3-6 cycloalkyl-C_1-3 alkyl-O-C (O) -;

M is independently selected from the group consisting of bond, C1-3 alkyl, branched or linear C1-3 alkoxy;

R₆, R₇ and R₈ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl, C_3-6 cycloalkyl and C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S;

R₁₀ is ndependently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl, C_3-8 cycloalkyl, 4-8 membered heterocyclyl containing one or moreheteroatoms independently selected from N, O or S;

R₁₁ is each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkenyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-8 cycloalkyl and C_3-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, wherein the C_3-8 cycloalkyl and C_3-8 heterocyclyl are optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, C_1-6 alkyl; preferably, the C_3-12 heterocyclyl is C₃-₇ mono-heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, or fused C_8-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S;

R₁₅ and R₁₆ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C1-4 alkoxy, C_2-4 alkenyl, C_2-4 alkynyl, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl, C_2-4 haloalkenyl, C_3-6 cycloalkyl and 3-6 membered heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S;

ring B is 7-12 membered fused heterocyclyl comprising 1, 2 or 3 heteroatoms selected from N, O, S;

ring C is selected from the group consisting of 5-10 membered mono heterocyclyl comprising 1, 2 or 3 heteroatoms selected from N, O, S, 7-12 membered bi-heterocyclyl comprising 1, 2 or 3 heteroatoms selected from N, O, S;

r is 0, 1, 2 or 3;

t is 0, 1, 2 or 3.

In some embodiments, ring B is selected from the group consisting of

In a preferred embodiment, ring B is selected from the group consisting of

In some embodiments, ring C is selected from the group consisting of

In some embodiments, the compounds of formula (I) , or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, is being a compound of formula (VIIIa)

In another aspect, the present invention provides following compounds, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt solvate, or prodrug thereof,

The present invention also provides a pharmaceutical composition, comprising a therapeutically effective amount of a compound of formulas as defined above, or a tautomer, cis-or trans isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, together with one or more pharmaceutically acceptable carriers, diluents or excipients.

In some embodiments, in above stated pharmaceutical composition, the amount of the compound, tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salts thereof is about 0.1-95%by weight of free base; preferably, is about 5-70%, e.g. 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%.

In some embodiment, above stated pharmaceutical composition is formulated as a tablet, capsule, liquid form or injection form.

In some embodiment, in above stated pharmaceutical composition, the amount of the compound, tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salts thereof is about 1-1000mg; preferably, is about 1-500mg, more preferably, is about 1mg, 2mg, 3mg, 5mg, 10mg, 20mg, 40mg, 50mg, 60mg, 80mg, 100mg, 200mg, 300mg, 400m or 500mg. In some embodiment, the compound, tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salts thereof is can be administered by any suitable route of administration, e.g. oral, parenteral, buccal, sublingual, nasal, rectal, intrathecal or transdermal administration, and the pharmaceutical compositions adapted accordingly.

In some embodiment, the compound, tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salts is formulated as a soild or liquid form, e.g. syrups, suspension, emulsion, tablets, capsules, powders, granules or lozenges.

In another aspect, the present invention relates to a method for treatment of disease mediated by RAS mutation, comprising administering to a subject in need thereof an effective amount of a compound of formula (I) to formula (VIIIa) as defined above, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt solvate, or prodrug thereof, or the pharmaceutical composition comprising the same.

In some embodiments, the RAS mutation is seleted from the grouop consisting of KRAS, HRAS or NRAS mutation.

In a preferred embodiment, the RAS mutation is KRAS mutation selected from the group consisting of KRAS G12C, KRAS G12D, KRAS G12V or KRAS G13D mutation. In another aspect, the present invention relates to a method for treating cancer comprising administering to a subject in need thereof an effective amount of a compound of formula (I) to formula (VIIIa) as defined above, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt solvate, or prodrug thereof, or the pharmaceutical composition comprising the same.

In an embodiment, the cancer is associated with KRAS G12C, KRAS G12D, KRAS G12V or KRAS G13D mutation.

In a preferred embodiment, the cancer is selected from the group consisting of Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma) , myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma) , alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma) , stomach (carcinoma, lymphoma, leiomyosarcoma) , pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma) , small bowel (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma) , large bowel (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma) ; Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor (nephroblastoma) , lymphoma, leukemia) , bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma) , prostate (adenocarcinoma, sarcoma) , testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma) ; Liver: hepatoma (hepatocellular carcinoma) , cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: gall bladder carcinoma, ampullary carcinoma, cholangiocarcinoma; Bone: osteogenic sarcoma (osteosarcoma) , fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma) , multiple myeloma, malignant giant cell tumor chordoma, osteochronfroma (osteocartilaginous exostoses) , benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans) , meninges (meningioma, meningiosarcoma, gliomatosis) , brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma) , glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors) , spinal cord neurofibroma, meningioma, glioma, sarcoma) ; Gynecological: uterus (endometrial 'carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma) , granulosa-thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma) , vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma) , vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma) , fallopian tubes (carcinoma) ; Hematologic: blood (myeloid leukemia (acute and chronic) , acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome) , Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma) ; Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma.

DETAILED DESCRIPTION OF THE INVENTION

Given below are definitions of terms used in this application. Any term not defined herein takes the normal meaning as the skilled person would understand the term. “Alkyl” refers to a saturated aliphatic hydrocarbon group including C₁-C₂₀ straight chain and branched chain groups. Preferably an alkyl group is an alkyl having 1 to 12, sometimes preferably 1 to 6, sometimes more preferably 1 to 4, carbon atoms. Representative examples include, but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1, 1-dimethyl propyl, 1, 2-dimethyl propyl, 2, 2-dimethyl propyl, 1-ethyl propyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 1, 2-trimethylpropyl, 1, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-dimethylpentyl, 2, 4-dimethylpentyl, 2, 2-dimethylpentyl, 3, 3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2, 3-dimethylhexyl, 2, 4-dimethylhexyl, 2, 5-dimethylhexyl, 2, 2-dimethylhexyl, 3, 3- dimethylhexyl, 4, 4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2, 2-diethylpentyl, n-decyl, 3, 3-diethylhexyl, 2, 2-diethylhexyl, and the isomers of branched chain thereof. More preferably an alkyl group is a lower alkyl having 1 to 6 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 1, 2-trimethylpropyl, 1, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, etc. The alkyl group can be substituted or unsubstituted. When substituted, the substituent group (s) can be substituted at any available connection point, preferably the substituent group (s) is one or more substituents independently selected from the group consisting of alkyl, halogen, alkoxy, alkenyl, alkynyl, alkylsulfo, alkylamino, thiol, hydroxy, nitro, cyano, amino, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic, cycloalkylthio, heterocylic alkylthio and oxo group.

“Alkenyl” refers to an alkyl defined as above that has at least two carbon atoms and at least one carbon-carbon double bond, for example, vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl, etc., preferably C_2-20 alkenyl, more preferably C_2-12 alkenyl, and most preferably C_2-6 alkenyl. The alkenyl group can be substituted or unsubstituted. When substituted, the substituent group (s) is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, group (s) independently selected from the group consisting of alkyl, halogen, alkoxy, alkenyl, alkynyl, alkylsulfo, alkylamino, thiol, hydroxy, nitro, cyano, amino, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic, cycloalkylthio, heterocylic alkylthio and oxo group.

“Alkynyl” refers to an alkyl defined as above that has at least two carbon atoms and at least one carbon-carbon triple bond, for example, ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl etc., preferably C_2-20 alkynyl, more preferably C_2-12 alkynyl, and most preferably C_2-6 alkynyl. The alkynyl group can be substituted or unsubstituted. When substituted, the substituent group (s) is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, group (s) independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfo, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic alkoxyl, cycloalkylthio and heterocylic alkylthio.

“Alkylene” refers to a saturated linear or branched aliphatic hydrocarbon group, wherein having 2 residues derived by removing two hydrogen atoms from the same carbon atom of the parent alkane or two different carbon atoms. The straight or branched chain group containing 1 to 20 carbon atoms, preferably has 1 to 12 carbon atoms, more preferably 1 to 6 carbon atoms. Non-limiting examples of alkylene groups include, but are not limited to, methylene (-CH₂-) , 1, 1-ethylene (-CH (CH₃) -) , 1, 2-ethylene (-CH₂CH₂) -, 1, 1-propylene (-CH (CH₂CH₃) -) , 1, 2-propylene (-CH₂CH (CH₃) -) , 1, 3-propylene (-CH₂CH₂CH₂-) , 1, 4-butylidene (-CH₂CH₂CH₂CH₂-) etc. The alkylene group can be substituted or unsubstituted. When substituted, the substituent group (s) is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, group (s) independently selected from the group consisting of selected from alkyl, alkenyl, alkynyl, alkoxy, alkylsulfo, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic alkoxyl, cycloalkylthio and heterocylic alkylthio.

“Alkenylene” refers to an alkylene defined as above that has at least two carbon atoms and at least one carbon-carbon double bond, preferably C_2-20 alkenylene, more preferably C_2-12 alkenylene, and most preferably C_2-6 alkenylene. Non-limiting examples of alkenylene groups include, but are not limited to, -CH=CH-, -CH=CHCH₂-, -CH=CHCH₂CH₂-, -CH₂CH=CHCH₂-etc. The alkenylene group can be substituted or unsubstituted. When substituted, the substituent group (s) is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, group (s) independently selected from the group consisting of selected from alkyl, alkenyl, alkynyl, alkoxy, alkylsulfo, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic alkoxyl, cycloalkylthio and heterocylic alkylthio.

“Alkynylene” refers to an alkynyl defined as above that has at least two carbon atoms and at least one carbon-carbon triple bond, preferably C_2-20 alkynylene, more preferably C_2-12 alkynylene, and most preferably C_2-6 alkynylene. Non-limiting examples of alkenylene groups include, but are not limited to, -CH≡CH-, -CH≡CHCH₂-, -CH≡CHCH₂CH₂-, -CH₂CH≡CHCH₂-etc. The alkynylene group can be substituted or unsubstituted. When substituted, the substituent group (s) is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, group (s) independently selected from the group consisting of selected from alkyl, alkenyl, alkynyl, alkoxy, alkylsulfo, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic alkoxyl, cycloalkylthio and heterocylic alkylthio.

“Cycloalkyl” refers to a saturated and/or partially unsaturated monocyclic or polycyclic hydrocarbon group having 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 8 carbon atoms or 3 to 6 carbon atoms. Representative examples of monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc. Polycyclic cycloalkyl includes a cycloalkyl having a spiro ring, fused ring or bridged ring.

“Spiro Cycloalkyl” refers to a 5 to 20 membered polycyclic group with rings connected through one common carbon atom (called a spiro atom) , wherein one or more rings can contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Preferably a spiro cycloalkyl is 6 to 14 membered, more preferably 7 to 10 membered, and most preferably . 7 to 8 membered. According to the number of common spiro atoms, a spiro cycloalkyl is divided into mono-spiro cycloalkyl, di-spiro cycloalkyl, or poly-spiro cycloalkyl, and preferably refers to a mono-spiro cycloalkyl or di-spiro cycloalkyl, more preferably 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered, or 5-membered/6- membered mono-spiro cycloalkyl. Representative examples of spiro cycloalkyl include, but are not limited to the following substituents:

“Fused Cycloalkyl” refers to a 5 to 20 membered polycyclic hydrocarbon group, wherein each ring in the system shares an adjacent pair of carbon atoms with another ring, wherein one or more rings can contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Preferably, a fused cycloalkyl group is 6 to 14 membered, more preferably 7 to 10 membered, and most preferably . 7 to 8 membered. According to the number of membered rings, fused cycloalkyl is divided into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl, and preferably refers to a bicyclic or tricyclic fused cycloalkyl, more preferably 5-membered/5-membered, or 5-membered/6-membered bicyclic fused cycloalkyl. Representative examples of fused cycloalkyls include, but are not limited to, the following substituents:

“Bridged Cycloalkyl” refers to a 5 to 20 membered polycyclic hydrocarbon group, wherein every two rings in the system share two disconnected carbon atoms. The rings can have one or more double bonds, but have no completely conjugated pi-electron system. Preferably, a bridged cycloalkyl is 6 to 14 membered, more preferably 7 to 10 membered, and most preferably 7 to 8 membered. According to the number of membered rings, bridged cycloalkyl is divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl, and preferably refers to a bicyclic, tricyclic or tetracyclic bridged cycloalkyl, more preferably a bicyclic or tricyclic bridged cycloalkyl. Representative examples of bridged cycloalkyls include, but are not limited to, the following substituents:

The cycloalkyl can be fused to the ring of an aryl, heteroaryl or heterocyclic alkyl, wherein the ring bound to the parent structure is cycloalkyl. Representative examples include, but are not limited to indanylacetic, tetrahydronaphthalene, benzocycloheptyl and so on. The cycloalkyl is optionally substituted or unsubstituted. When substituted, the substituent group (s) is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, substituents independently selected from the group consisting of alkyl, halogen, alkoxy, alkenyl, alkynyl, alkylsulfo, alkylamino, thiol, hydroxy, nitro, cyano, amino, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic, cycloalkylthio, heterocylic alkylthio and oxo group. Representative examples include, but are not limited to, the following substituents:

“Heterocyclyl” refers to a 3 to 20 membered saturated and/or partially unsaturated monocyclic or polycyclic hydrocarbon group having one or more, sometimes preferably one to five, sometimes more preferably one to three, heteroatoms selected from the group consisting of N, O, and S (O) _m (wherein m is 0, 1, or 2) as ring atoms, but excluding -O-O-, -O-S-or -S-S-in the ring, the remaining ring atoms being C. Preferably, heterocyclyl is a 3 to 12 membered having 1 to 4 heteroatoms; more preferably a 3 to 10 membered having 1 to 3 heteroatoms; most preferably a 5 to 6 membered having 1 to 2 heteroatoms. Representative examples of monocyclic heterocyclyls include, but are not limited to, pyrrolidyl, piperidyl, piperazinyl, morpholinyl, sulfo-morpholinyl, homopiperazinyl, and so on. Polycyclic heterocyclyl includes the heterocyclyl having a spiro ring, fused ring or bridged ring.

“Spiro heterocyclyl” refers to a 5 to 20 membered polycyclic heterocyclyl with rings connected through one common carbon atom (called a spiro atom) , wherein said rings have one or more, sometimes preferably one to five, sometimes more preferably one to three, heteroatoms selected from the group consisting of N, O, and S (O) _m (wherein m is 0, 1 or 2) as ring atoms, the remaining ring atoms being C, wherein one or more rings can contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Preferably a spiro heterocyclyl is 6 to 14 membered, more preferably 7 to 10 membered, and most preferably 7 to 8 membered. According to the number of common spiro atoms, spiro heterocyclyl is divided into mono-spiro heterocyclyl, di-spiro heterocyclyl, or poly-spiro heterocyclyl, and preferably refers to mono-spiro heterocyclyl or di-spiro heterocyclyl, more preferably 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered, or 5-membered/6-membered mono-spiro heterocyclyl. Representative examples of spiro heterocyclyl include, but are not limited to the following substituents:

“Fused Heterocyclyl” refers to a 5 to 20 membered polycyclic heterocyclyl group, wherein each ring in the system shares an adjacent pair of carbon atoms with the other ring, wherein one or more rings can contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system, and wherein said rings have one or more, sometimes preferably one to five, sometimes more preferably one to three, heteroatoms selected from the group consisting of N, O, and S (O) _p (wherein p is 0, 1, or 2) as ring atoms, the remaining ring atoms being C. Preferably a fused heterocyclyl is 6 to 14 membered, more preferably 7 to 10 membered, and most preferably 7 to 8 membered. According to the number of membered rings, fused heterocyclyl is divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclyl, preferably refers to bicyclic or tricyclic fused heterocyclyl, more preferably 5-membered/5-membered, or 5-membered/6-membered bicyclic fused heterocyclyl. Representative examples of fused heterocyclyl include, but are not limited to, the following substituents:

“Bridged Heterocyclyl” refers to a 5 to 14 membered polycyclic heterocyclic alkyl group, wherein every two rings in the system share two disconnected atoms, the rings can have one or more double bonds, but have no completely conjugated pi-electron system, and the rings have one or more heteroatoms selected from the group consisting of N, O, and S (O) _m (wherein m is 0, 1, or 2) as ring atoms, the remaining ring atoms being C. Preferably a bridged heterocyclyl is 6 to 14 membered, more preferably 7 to 10 membered, and most preferably 7 to 8 membered. According to the number of membered rings, bridged heterocyclyl is divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclyl, and preferably refers to bicyclic, tricyclic or tetracyclic bridged heterocyclyl, more preferably bicyclic or tricyclic bridged heterocyclyl. Representative examples of bridged heterocyclyl include, but are not limited to, the following substituents:

The ring of said heterocyclyl can be fused to the ring of an aryl, heteroaryl or cycloalkyl, wherein the ring bound to the parent structure is heterocyclyl. Representative examples include, but are not limited to the following substituents:

etc.

The heterocyclyl is optionally substituted or unsubstituted. When substituted, the substituent group (s) is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, group (s) independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfo, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic alkoxyl, cycloalkylthio, heterocylic alkylthio and -NR⁹R¹⁰. “Aryl” refers to a 6 to 14 membered all-carbon monocyclic ring or a polycyclic fused ring (a "fused" ring system means that each ring in the system shares an adjacent pair of carbon atoms with another ring in the system) group, and has a completely conjugated pi-electron system. Preferably aryl is 6 to 10 membered, such as phenyl and naphthyl, most preferably phenyl. The aryl can be fused to the ring of heteroaryl, heterocyclyl or cycloalkyl, wherein the ring bound to parent structure is aryl. Representative examples include, but are not limited to, the following substituents:

The aryl group can be substituted or unsubstituted. When substituted, the substituent group (s) is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfo, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic alkoxyl, cycloalkylthio, heterocylic and alkylthio.

“Heteroaryl” refers to an aryl system having 1 to 4 heteroatoms selected from the group consisting of O, S and N as ring atoms and having 5 to 14 annular atoms. Preferably a heteroaryl is 5-to 10-membered, more preferably 5-or 6-membered, for example, thiadiazolyl, pyrazolyl, oxazolyl, oxadiazolyl, imidazolyl, triazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrrolyl, N-alkyl pyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like. The heteroaryl can be fused with the ring of an aryl, heterocyclyl or cycloalkyl, wherein the ring bound to parent structure is heteroaryl. Representative examples include, but are not limited to, the following substituents:

The heteroaryl group can be substituted or unsubstituted. When substituted, the substituent group (s) is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfo, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic alkoxyl, cycloalkylthio, heterocylic alkylthio and -NR⁹R¹⁰. “Alkoxy” refers to both an -O- (alkyl) and an -O- (unsubstituted cycloalkyl) group, wherein the alkyl is defined as above. Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. The alkoxyl can be substituted or unsubstituted. When substituted, the substituent is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfo, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxyl, heterocylic alkoxyl, cycloalkylthio and heterocylic alkylthio.

“Bond” refers to a covalent bond using a sign of “-” .

"Hydroxyalkyl" refers to an alkyl group substituted by a hydroxy group, wherein alkyl is as defined above.

“Hydroxyl” refers to an -OH group.

“Halogen” refers to fluoro, chloro, bromo or iodo atoms.

“Amino” refers to a -NH₂ group.

“Cyano” refers to a -CN group.

“Nitro” refers to a -NO₂ group.

“Oxo group” refers to a =O group.

“Carboxyl” refers to a -C (O) OH group.

“Alkoxycarbonyl” refers to a -C (O) O (alkyl) or (cycloalkyl) group, wherein the alkyl and cycloalkyl are defined as above.

“Optional” or “optionally” means that the event or circumstance described subsequently can, but need not, occur, and the description includes the instances in whichthe event or circumstance may or may not occur. For example, “the heterocyclic group optionally substituted by an alkyl” means that an alkyl group can be, but need not be, present, and the description includes the case of the heterocyclic group being substituted with an alkyl and the heterocyclic group being not substituted with an alkyl. “Substituted” refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, independently substituted with a corresponding number of substituents. It goes without saying that the substituents exist in their only possible chemical position. The person skilled in the art is able to determine if the substitution is possible or impossible without paying excessive efforts by experiment or theory. For example, the combination of amino or hydroxyl group having free hydrogen and carbon atoms having unsaturated bonds (such as olefinic) may be unstable.

A “pharmaceutical composition” refers to a mixture of one or more of the compounds described in the present invention or physiologically/pharmaceutically acceptable salts or prodrugs thereof and other chemical components such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism, which is conducive to the absorption of the active ingredient and thus displaying biological activity.

“Pharmaceutically acceptable salts” refer to salts of the compounds of the invention, such salts being safe and effective when used in a mammal and have corresponding biological activity.

EXAMPLES

The following examples serve to illustrate the invention, but the examples should not be considered as limiting the scope of the invention. If specific conditions for the experimental method are not specified in the examples of the present invention, they are generally in accordance with conventional conditions or recommended conditions of the raw materials and the product manufacturer. The reagents without a specific source indicated are commercially available, conventional reagents.

The structure of each compound is identified by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS) . NMR chemical shifts (δ) were given in 10^-6 (ppm) . NMR is determined by Varian Mercury 300 MHz Bruker Avance III 400MHz machine. The solvents used were deuterated-dimethyl sulfoxide (DMSO-d₆) , deuterated-chloroform (CDCl₃) and deuterated-methanol (CD₃OD) .

High performance liquid chromatography (HPLC) is determined on an Agilent 1200DAD high pressure liquid chromatography spectrometer (Sunfire C18 150×4.6 mm chromatographic column) and a Waters 2695-2996 high pressure liquid chromatography spectrometer (Gimini C18 150×4.6 mm chromatographic column) . Liquid Chromatography Mass Spectrometry (LCMS) is determined on an Agilent 1200 high pressure liquid chromatography spectrometer &mass spectrometry (Sunfire C18 4.6*50mm 3.5 um chromatographic column) and an Agilent 19091S-433HP-5 high pressure liquid chromatography spectrometer &mass spectrometry (XBridge C18 4.6*50mm 3.5um chromatographic column) .

Chiral High performance liquid chromatography (HPLC) is determined on SFC Thar 80 &150 &200 (waters. )

The average rates of ATPase inhibition, and the IC₅₀ values are determined by Victor Nivo multimode plate reader (PerkinElmer, USA) .

The thin-layer silica gel plates used in thin-layer chromatography are Yantai Xinnuo silica gel plate. The dimension of the plates used in TLC was 0.15 mm to 0.2 mm, and the dimension of the plates used in thin-layer chromatography for product purification was 0.4 mm to 0.5 mm.

Column chromatography generally used Qingdao Haiyang 200 to 300 mesh silica gel as carrier.

The known starting material of the invention can be prepared by the conventional synthesis method in the prior art, or can be purchased from ABCR GmbH &Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc or Dari chemical Company, etc.

MS is mass spectroscopy with (+) referring to the positive mode which generally gives a M+1 (or M+H) absorption where M = the molecular mass.

Synthetic Procedures

Synthesis of 3-bromo-2- (1-methoxyethyl) pyridine (Int A) :

Step 1: Synthesis of 1- (3-bromopyridin-2-yl) ethan-1-ol (Int A-1)

To a solution of 3-Bromo-2-pyridinecarboxaldehyde (CAS: 405174-97-2, 1.80 g, 9.70 mmol) in THF (35 mL) is added a solution of MeMgBr in diethyl ether (6.50 mL, 19.4 mmol) . The solution is stirred for 30 minutes at -78℃ and then warmed to 25 ℃ over 1.5 h. The suspension was partitioned between aqueous NH₄Cl (50 mL) and EtOAc (50 mL*3) . The combined organic layers was washed with brine (50 mL) , dried over Na₂SO₄, filtered and concentrated. The crude product was purified by a silica gel column (petroleum ether/EtOAc = 10: 1to give 1- (3-bromopyridin-2-yl) ethan-1-ol (Int A-1) (1.80 g, yield: 87%) as a yellow oil. ESI-MS [M+H] ⁺: 201.9, 203.9

Step 2: Synthesis of 3-bromo-2- (1-methoxyethyl) pyridine (Int A)

To a solution of Int A-1 (1.80 g, 8.90 mmol) in DMF (30 mL) was added sodium hydride (0.71 g, 18 mmol) at 0 ℃. The resulting mixture was stirred for 45 min at 0 ℃. Methyl iodide (2.53 g, 17.8 mmol) was added to the above mixture dropwise. The resulting mixture was stirred at 25 ℃ for 2 h. The suspension was partitioned between H₂O (100 mL) and EtOAc (50 mL*3) . The combined organic layers was washed with brine (100 mL) , dried over Na₂SO₄, filtered and concentrated filtered and concentrated. The crude product was purified by a silica gel column (petroleum ether/EtOAc = 10: 1) to give 3- bromo-2- (1-methoxyethyl) pyridine (Int A) (2.1 g, yield: 95%) as a brown oil. ESI-MS [M+H] ⁺: 215.8, 217.8

Synthesis of di-tert-butyl 1- (5- (methoxycarbonyl) thiophen-2-yl) hydrazine-1, 2-dicarboxylate, Int B-S

A solution of isopropylmagnesium bromide (2.8 M, 19.39 mL) was added dropwise at -40 ℃ to a stirred solution of methyl 5-bromothiophene-2-carboxylate (10 g, 45.23 mmol) in THF (100 mL) under Ar atmosphere. The reaction mixture was stirred at -40 ℃ for 30min, and then a solution of Di-tert-Butyl azodicarboxylate (11.46 g, 49.76 mmol) in THF (50 mL) was added dropwise. The solution was allowed to warm to room temperature over 3 h. LCMS showed the reaction was complete. Quenched with a saturated aqueous ammonium chloride solution, extracted with EA, dried (Na2SO4) , filtered, and concentrated in vacuo. The crude product was purified by flash chromatography (20%EA in PE) to afford 37-2 (11g, 65%yield) .

Mass calc. C₁₆H₂₄N₂O₆S for 372.1, found 217.1 (M-Boc-tBu) ⁺.

¹H NMR (400 MHz, CDCl3) δ: 7.59 (d, J = 4.0 Hz, 1H) , 6.84 (s, 1H) , 6.79 (d, J = 4.0 Hz, 1H) , 3.85 (s, 3H) , 1.53 (m, 18H) .

Synthesis of 2- (2-bromo-5-hydrazineylphenyl) ethan-1-ol, Int B-T

Borane-THF complex (1M, 11.8 mL, 11.8 mmol) was added to a solution of (5-amino-2-bromo-phenyl) -acetic acid (CAS: 1261666-52-7 1.00 g, 4.37 mmol) in dry THF (10 mL) , under argon, over 20 min. Once the effervescence had ceased, the reaction mixture was heated to 50 ℃ for 2 h. The mixture was left to cool and then concentrated in vacuo. The residue was re-dissolved in DCM (25 mL) and treated with MeOH (2 mL) . The mixture was stirred vigorously and then concentrated in vacuo. The residue was dissolved in MeOH (25 mL) and re-concentrated in vacuo. The residue was purified by column chromatography, using 0-10% [2M NH₃ in MeOH] in DCM, to afford the title compound (0.54 g) . LCMS (ESI) : m/z 216 [MH⁺] .

Intermediate Int B-T’ (0.54 g, 2.51 mmol) was dissolved in concentrated aqueous HC1 (5 mL) and cooled to ^-5 ℃. To this was added a solution of sodium nitrite (0.18 g, 2.63 mmol) , in water (0.75 mL) , dropwise. Stirring continued for 5 min, before a solution of tin (II) chloride (1.01 g, 5.52 mmol) in concentrated aqueous HC1 (2.2 mL) was added. The reaction mixture was stirred just below 0 ℃ for 40 min and then basified using aqueous NaOH (4M, 30 mL) . The mixture was then stirred at room temperature for 1.5 h. This was extracted into EtOAc (3 x 50 mL) . The combined organics were washed with brine (2 x 50 mL) , dried (MgS0₄) , filtered and concentrated in vacuo to afford the title compound (294 mg, 51%) . LCMS (ESI) : m/z 231 [MH⁺]

The following compounds were synthesized with a similar route as Int B, using appropriate starting materials:

Synthesis of 2- (2-bromo-5-hydrazineylphenyl) ethan-1-ol, Int B-U

To a solution of (S) -3-bromo-5-iodo-2- (1-methoxyethyl) pyridine (15 g, 43.86 mmol) , and benzyl piperazine-1 -carboxylate (8.7 g, 39.48 mmol) in toluene (150 mL) at 0 ℃, were added cesium carbonate (71 . 46 g, 219.32 mmol) , BINAP (0.55 g, 0.88 mmol) and palladium acetate (0.49 g, 2.19 mmol) in portions. The reaction mixture was stirred at 90 ℃ for 12 h under an argon atmosphere. The resulting mixture was cooled down to room temperature, filtered and the filter cake was washed with EtOAc (150 mL x 3) . The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford benzyl (S) -4- (5-bromo-6- (1-methoxyethyl) pyridin-3-yl) piperazine-1 -carboxylate (16 g, 84%yield) as solid. LCMS (ESI) : m/z [M+H] calc'd for C^HbsNeOy 433.1 ; found 434.0.

Synthesis of 3- (1-ethyl-2- (2- (methoxymethyl) pyridine-3-yl) -5- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indol-3-yl) -2, 2-dimethylpropan-1-ol (Int B) :

Step 1: Synthesis of 5- (2- (methoxymethyl) pyridin-3-yl) -2, 2-dimethyl-5-oxopentanoic acid (Int B-1)

To a solution of 3-bromo-2- (methoxymethyl) pyridine (5.00 g, 24.9 mmol) in THF (15 mL) was added dropwise i-PrMgCl^. LiCl (1.30 M in THF, 21.4 mL, 27.8 mmol) at -10 ℃ under N₂ atmosphere. The resulting mixture was warmed to -5 ℃ and stirred for 1 h at -5 ℃. Then a solution of 3, 3-dimethyltetrahydropyran-2, 6-dione (cas: 2938-48-9) (3.70 g, 26.1 mmol) in THF (36 mL) was added dropwise at -5 ℃. The resulting mixture was warmed to 0 ℃ and stirred for 1.5 h at 0 ℃. Then the resulting mixture was warmed to room temperature and stirred at RT overnight. The reaction was quenched with HCl/Dioxane (4.0 M, 4.4 mL, 17 mmol) and H₂O (22 mL) . The mixture was stirred and separated. The aqueous layer was extracted with EtOAc (30 mL*3) . The organic layer was combined and dried over anhydrous Na₂SO₄. The organic layer was concentrated under vacuum. The residue was purified with column chromatography on silica gel (DCM/MeOH = 50: 1 to 20: 1) . 5- (2-(methoxymethyl) pyridin-3-yl) -2, 2-dimethyl-5-oxopentanoic acid (Int B-1) (3.70 g, 14.0 mmol, 55%yield) was obtained as light yellow solid. ESI-MS [M+H] ⁺: 265.98

Step 2: Synthesis of 3- (5-bromo-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropanoic acid (Int B-2) ; ethyl 3- (5-bromo-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropanoate (Int B-2’)

To a solution of 5- (2- (methoxymethyl) pyridin-3-yl) -2, 2-dimethyl-5-oxopentanoic acid (Int B-1) (3.70 g, 14.0 mmol) in EtOH (92 mL) was added (4-bromophenyl) hydrazine HCl salt (cas 622-88-8) (3.50 g, 15.7 mmol) . The resulting mixture was stirred at 85 ℃for 2 h. Then to the mixture was added HCl/Dioxane (4 M, 3.5 mL, 14.0 mmol) . The resulting mixture was stirred at 85 ℃ for another 3 h. The mixture was concentrated under vacuum. Trifluoroacetic acid (53.80 g, 471.9 mmol) was added to the residue. The mixture was stirred at 60 ℃ for another 1.5 h. The mixture was cooled to RT and concentrated under vacuum. The pH of the residue was adjusted to 5～6 with saturated NaHCO₃ aqueous (54 mL) , and EtOAc (128 mL) was added to the mixture. The mixture was stirred and separated. The organic layer was washed with brine (50 mL) and dried over anhydrous Na₂SO₄. The organic layer was concentrated under vacuum. A mixture of 3- (5-bromo-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropanoic acid (Int B-2) and ethyl 3- (5-bromo-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropanoate (Int B-2’) (8.70 g, 20.8 mmol) were obtained and used for next step directly without purification. ESI-MS [M+H] ⁺: 416.74, 418.77, 444.79, 446.81

Step 3: Synthesis of ethyl 3- (5-bromo-1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropanoate (Int B-3)

To a solution of mixture of 3- (5-bromo-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropanoic acid (Int B-2) and ethyl 3- (5-bromo-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropanoate (Int B-2’) (8.70 g, 20.8 mmol) in DMF (64 mL) was added Cs₂CO₃ (24.4 g, 74.1 mmol) . Then a solution of ethyl iodide (11.6 g, 75.0 mmol) in DMF (16 mL) was added. The resulting mixture was stirred at RT overnight. H₂O (300 mL) and EtOAc (300 mL) were added into the mixture. The mixture was stirred and separated. The organic layer was washed with brine (200 mL) and dried over anhydrous Na₂SO₄. The organic layer was concentrated under vacuum. The residue was purified with column chromatography on silica gel (petroleum ether/EtOAc = 3: 1 to 1: 1) . Ethyl 3- (5-bromo-1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropanoate (Int B-3) (3.8 g, 8.5 mmol, 57%yield over two steps) was obtained as brown liquid. ESI-MS [M+H] ⁺: 473.03, 474.94

Step 4: Synthesis of 3- (5-bromo-1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropan-1-ol (Int B-4)

To a solution of ethyl 3- (5-bromo-1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropanoate (Int B-3) (3.8 g, 8.0 mmol) in THF (159 mL) was added LiBH₄ (2.00 M in THF, 23.8 mL 47.6 mmol) . The resulting mixture was stirred at 60 ℃ overnight. The reaction mixture was quenched with saturated NH₄Cl aqueous solution (66 mL) at 0 ℃. H₂O (22 mL) and EtOAc (88 mL) were added into the mixture. The mixture was stirred and separated. The organic layer was washed with brine (40 mL) and dried over anhydrous Na₂SO₄. The organic layer was concentrated under vacuum. The residue was purified with column chromatography on silica gel (DCM/MeOH = 100: 1 to 50: 1) . 3- (5-bromo-1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropan-1-ol (Int B-4) (1.3 g, 3.1 mmol, 38%yield) was obtained as light yellow solid. ESI-MS [M+H] ⁺: 432.93

Step 5: Synthesis of 3- (1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -5- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indol-3-yl) -2, 2-dimethylpropan-1-ol (Int B)

To a solution of 3- (5-bromo-1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropan-1-ol (Int B-4) (1.3 g, 3.0 mmol) in toluene (35 mL) was added bis (pinacolato) diboron (1.1 g, 4.5 mmol) , potassium acetate (589 mg, 6.00 mmol) and Pd(dppf) Cl₂ (222 mg, 0.303 mmol) under N₂ atmosphere. The resulting mixture was stirred at 90 ℃ overnight. The mixture was concentrated to dryness under vacuum and the residue was purified with column chromatography on silica gel (DCM/MeOH =100: 1 to 50: 1) . 3- (1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -5- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indol-3-yl) -2, 2-dimethylpropan-1-ol (Int B) (600 mg, 1.25 mmol, 41%yield) was obtained as light yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ 8.74-8.68 (m, 1H) , 8.03 (s, 1H) , 7.90-7.84 (m, 1H) , 7.56-7.44 (m, 3H) , 4.45 (t, J = 5.4 Hz, 1H) , 4.22 (d, J = 10.8 Hz, 1H) , 4.15 (d, J = 10.8 Hz, 1H) , 4.04-3.94 (m, 1H) , 3.92-3.81 (m, 1H) , 3.14 (s, 3H) , 3.07-3.01 (m, 1H) , 2.98-2.92 (m, 1H) , 2.66 (d, J = 14.0 Hz, 1H) , 2.29 (d, J = 14.0 Hz, 1H) , 1.32 (s, 12H) , 1.02 (t, J = 7.2 Hz, 3H) , 0.60 (s, 3H) , 0.53 (s, 3H) . ESI-MS [M+H] ⁺: 479.11

The following intermediated were synthesized with a similar route as Int B, using appropriate starting materials:

Synthesis of (S) -2-bromo-4- (3- ( (tert-butyldiphenylsilyl) oxy) -2, 2-dimethylpropyl) -6-ethyl-5- (2- (1-methoxyethyl) pyridin-3-yl) -6H-thieno [2, 3-b] pyrrole, Int B-S4

To a solution of tert-butyl- [3- [6-ethyl-5- [2- [ (1S) -1-methoxyethyl] -3-pyridyl] thieno [2, 3-b] pyrrol-4-yl] -2, 2-dimethyl-propoxy] -diphenyl-silane, Int B-S4-H (350 mg, 572.90 μmol) , NBS (112.16 mg, 630.20 μmol) in DMF (5 mL) was added dropwise TEA (507.85 mg, 5.02 mmol, 700.00 μL) and stirred for 3h at RT. LCMS showed the reaction was complete. The reaction was extracted with EA and washed with water, the organic phases was concentrated under vacuum to give the crude product. The crude product was purified by flash chromatography (100%PE to 30%EA in PE) to afford Int B-S4 (350 mg, 88%yield) . Mass calc. C₃₇H₄₅BrN₂O₂SSi for 688.2, found 689.2, 691.2 (M+H) ⁺.

¹H NMR (400 MHz, DMSO-d6) δ: 8.69 (m, 1H) , 7.64 (m, 1H) , 7.45 (m, 11H) , 7.21 (m, 1H) , 4.18-4.02 (m, 1H) , 3.72-3.47 (m, 2H) , 3.16 (m, 1H) , 3.02 (s, 2H) , 2.89 (s, 2H) , 2.56-2.47 (m, 1H) , 2.34-2.18 (m, 1H) , 1.36-1.13 (m, 6H) , 0.92 (m, 9H) , 0.69 (m, 6H) .

Synthesis of 3- (5-bromo-1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropyl acetate (Int B-Ac)

To a solution of 3- (5-bromo-1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropan-1-ol (3.2 g, 7.4 mmol) (Int B-4) in DCM (15 mL) , was added at 0 ℃ AcCl (1.0 M in DCM, 8.9 mL, 8.9 mmol) . The reaction mixture was stirred at room temperature for overnight. LC-MS showed full conversion. The reaction mixture was washed by water, concentrated and purified by a silica gel column (0-5% MeOH/DCM) to afford 3- (5-bromo-1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropyl acetate (Int B-Ac) (3.3 g, 94%yield) as a yellow solid.

¹H NMR (400 MHz, CD3OD) δ 8.71 (dd, J = 4.9, 1.6 Hz, 1H) , 7.91 (dd, J = 7.8, 1.6 Hz, 1H) , 7.77 (d, J = 1.8 Hz, 1H) , 7.55 (dd, J = 7.8, 4.9 Hz, 1H) , 7.38 (d, J = 9.1 Hz, 1H) , 7.30 (dd, J = 8.7, 1.8 Hz, 1H) , 4.29-4.24 (m, 2H) , 4.07-3.85 (m, 2H) , 3.62-3.55 (m, 2H) , 3.25 (s, 3H) , 2.76-2.70 (m, 1H) , 2.50-2.46 (m, 1H) , 1.94 (s, 3H) , 1.10 (t, J =7.2 Hz, 1H) , 0.77 (s, 3H) , 0.75 (s, 3H) . ESI-MS [M+H] ⁺: 473.1, 475.1

The following compounds were synthesized with a similar route as Int B-Ac, using appropriate starting materials:

Synthesis of (S) -5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -1-ethyl-2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indole (Int Bi)

Step 1: ethyl 1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutane-1-carboxylate (Int Bi-1)

To a solution of ethyl 1- (hydroxymethyl) cyclobutane-1-carboxylate (CAS: 1195-81-9 10 g, 63 mmol) in DCM (80 mL) was added tert-butyl (chloro) diphenylsilane (21 mL, 83 mmol) and imidazole (8.61 g, 126 mmol) at 0 ℃, the reaction was warmed up to 25 ℃ and stirred for 16 h. The reaction was quenched with H₂O (80 mL) and extracted with DCM (100 mL*3) . The organic layer was combined, washed with brine (80 mL) , dried over Na₂SO₄ and concentrated under vacuum to give the crude, which was purified by combi-flash on silica gel (petroleum ether/EtOAc = 15: 1) to afford ethyl 1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutane-1-carboxylate (Int Bi-1) (19.5 g, 77%yield) as a colorless oil. ESI-MS [M+Na] ⁺: 419.0

Step 2: 1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutane-1-carboxylic acid (Int Bi-2)

To a solution of ethyl 1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutane-1-carboxylate (Int Bi-1) (19.5 g, 49.2 mmol) in MeOH (10 mL) stirred was added KOH (80 mL, 5.0 M, 0.4 mmol) dropwise. The reaction mixture was stirred at 100 ℃ for 19 h. The reaction was quenched with HCl (50 mL, 6.0 M) and extracted with EtOAc (150 mL*3) . The organic layer was combined, washed with brine (80 mL) , dried over Na₂SO₄ and concentrated under vacuum to give the crude, which was purified by combi-flash on silica gel (petroleum ether/EtOAc = 20: 1) to afford 1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutane-1-carboxylic acid (Int Bi-2) (9.10 g, 49%yield) as a colorless oil. ESI-MS [M+H] ⁺: 369.0

Step 3: 1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutane-1-carbonyl chloride (Int Bi-3)

A solution of 1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutane-1-carboxylic acid Int Bi-2 (9.10 g, 24.7 mmol) and sulfinyl dichloride (8.78 g, 73.5 mmol) in DCM (80 mL) was stirred at 25 ℃ for 3 h. The solvent was removed under vacuum to give the crude 1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutane-1-carbonyl chloride (Int Bi-3) (8.5 g, 74%yield) ESI-MS [M+H] ⁺: 387.1

Step 4: (5-bromo-1H-indol-3-yl) (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methanone (Int Bi-4)

To a solution of 1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutane-1-carbonyl chloride (Int Bi-3) (8.5 g, 21 mmol) in DCM (60 mL) stirred under nitrogen at -15 ℃was added a solution of SnCl₄ (6.27 g, 23.9 mmol) in DCM (15 mL) dropwise for 30 min. The reaction mixture was added a solution of 5-bromo-1H-indole (4.69 g, 23.9 mmol) in DCM (15 mL) dropwise. The reaction was quenched with H₂O (60 mL) and extracted with DCM (80 mL*3) . The organic layer was combined, washed with brine (60 mL) , dried over Na₂SO₄ to give (5-bromo-1H-indol-3-yl) (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methanone (Int Bi-4) (3.7 g, 31%yield) as a white solid. ESI-MS [M+H] ⁺: 546.1, 548, 0

Step 5: (5-bromo-1H-indol-3-yl) (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methanol (Int Bi-5)

To a solution of (5-bromo-1H-indol-3-yl) (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methanone (Int Bi-4) (3.70 g, 6.76 mmol) in THF (30 mL) was added LiBH₄ (0.89 g, 40.7 mmol) . The reaction mixture was warmed up to 60 ℃ and stirred for 20 h. After cooling, the reaction was quenched with ice water (50 mL) and extracted with EtOAc (80 mL*3) . The organic layer was combined, washed with brine (50 mL) , dried over Na₂SO₄ and concentrated under vacuum to give the crude, which was purified by combi-flash (petroleum ether/EtOAc = 2: 1) to afford (5-bromo-1H-indol-3-yl) (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methanol (Int Bi-5) (3.56 g, 90%purity, 86%yield) as a white solid. ESI-MS [M+Na] ⁺ = 570.1, 572.1

Step 6: 5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -1H-indole (Int Bi-6)

To a solution of (5-bromo-1H-indol-3-yl) (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methanol (Int Bi-5) (3.56 g, 6.49 mmol) in DCM (30 mL) were added diludine (0.66 g, 2.60 mmol) and p-toluenesulfonic acid monohydrate (0.06 g, 0.3 mmol) . The reaction mixture was stirred at 10 ℃ for 1 h. Filtered and the filtration was concentrated to give the crude, which was purified by combi-flash (petroleum ether/EtOAc = 3: 1) to afford 5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -1H-indole (Int Bi-6) (3.41 g, 90%purity, 89%yield) as a white solid. ESI-MS [M+Na] ⁺: 553.7, 555.7

Step7: 5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -2-iodo-1H-indole (Int Bi-7)

To a solution of 5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -1H-indole (Int Bi-6) (3.4 g, 6.4 mmol) and I₂ (1.6 g, 6.4 mmol) in THF (20 mL) was added AgOTf (1.97 g, 7.6 mmol) . The reaction mixture was stirred at 25 ℃ for 2 h. Filtered and the filtration was diluted with H₂O (50 mL) , extracted withEtOAc (80 mL*3) . The organic layer was combined, washed with brine (80 mL) , dried over Na₂SO₄ and concentrated under vacuum to give the crude, which was purified by combi-flash on silica gel (petroleum ether/EtOAc =1: 1) to afford 5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -2-iodo-1H-indole (Int Bi-7) (4.0 g, 90%purity, 86%yield) as a yellow solid. ESI-MS [M+Na] ⁺: 680.1, 682.1

Step 8: (S) - (2- (1-methoxyethyl) pyridin-3-yl) boronic acid

A solution of (S) -3-bromo-2- (1-methoxyethyl) pyridine (CAS: 2641451-44-5, 13.60 g, 62.96 mmol) , B₂Pin₂ (24.03 g, 94.44 mmol) , Pd (dppf) Cl₂ (4.61 g, 6.30 mmol) and KOAc (12.39 g, 125.9 mmol) in dioxane (100 mL) was stirred under nitrogen at 80 ℃for 16 h. The reaction was purified with reversion phase chromatography (30%water/70%acetonitrile) to give compound (S) - (2- (1-methoxyethyl) pyridin-3-yl)boronic acid (8.5 g, 74%yield) as a white solid. ESI-MS [M+H] ⁺: 182.0

Step 9: (S) -5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indole (Int Bi-8)

To a solution of 5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -2-iodo-1H-indole (Int Bi-7) (4.0 g, 6.1 mmol) in dioxane (10 mL) and H₂O (2.5 mL) were added (S) - (2- (1-methoxyethyl) pyridin-3-yl) boronic acid (1.32 g, 7.30 mmol) , [1, 1//'-Bis(diphenylphosphino) ferrocene] dichloropalladium (II) DCM (490 mg, 0.600 mmol) and K₂CO₃ (2.11 g, 15.2 mmol) . The reaction mixture was stirred at 70 ℃ for 12 h under N₂. Filtered and the filtration was diluted with H₂O (100 mL) , extracted with EtOAc (80 mL*3) . The organic layer was combined, washed with brine (80 mL) , dried over Na₂SO₄ and concentrated under vacuum to give the crude, which was purified by combi-flash on silica gel (petroleum ether/EtOAc = 3: 1) to afford (S) -5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indole (Int Bi-8) (1.1 g, 90%purity, 21%yield) as a yellow oil. ESI-MS [M+H] ⁺: 667.0, 669.0

Step 10: (S) -5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -1-ethyl-2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indole (Int Bi)

To a solution of (S) -5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -2- (2- (1-methoxyethyl) pyridin-3- yl) -1H-indole (Int Bi-8) (1.1 g, 1.6 mmol) in DMF (10 mL) were added iodoethane (500 mg, 3.20 mmol) and Cs₂CO₃ (1.04 g, 3.20 mmol) . The reaction mixture was stirred at 25 ℃ for 12 h. The reaction was diluted with H₂O (80 mL) and extracted with EtOAc (50 mL*3) . The organic layer was combined, washed with brine (80 mL) , dried over Na₂SO₄ and concentrated under vacuum to give the crude, which was purified by combi-flash on silica gel (petroleum ether/EtOAc = 2: 1) to afford (S) -5-bromo-3- ( (1- ( ( (tert-butyldiphenylsilyl) oxy) methyl) cyclobutyl) methyl) -1-ethyl-2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indole (Int Bi) (1.0 g, yield 81%) as a yellow oil. ESI-MS [M+H] ⁺: 695.1, 697.1Synthesis of methyl (2S) -3- (5- (3- (3-acetoxy-2, 2-dimethylpropyl) -1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-5-yl)hexahydropyrrolo [3, 4-b] pyrrol-1 (2H) -yl) -2- ( ( (benzyloxy) carbonyl) amino) propanoate (Int Bii)

Step 1: methyl N- ( (benzyloxy) carbonyl) -O- (methylsulfonyl) -L-serinate (Int Bii-1)

To a solution of methyl ( (benzyloxy) carbonyl) -L-serinate (50.0 g, 197 mmol) in DCM (300 mL) and TEA (29.97 g, 296.1 mmol) was added dropwise MsCl (33.92 g, 296.1 mmol) at 0 ℃ and stirred for 1 h at RT. The reaction solution was washed with water and brine the organic layers were dried over Na₂SO₄ and concentrated under reduced pressure to afford crude product. The crude product was purified by flash chromatography on silica gel (0-5%MeOH/DCM) to afford methyl N- ( (benzyloxy) carbonyl) -O- (methylsulfonyl) -L-serinate (Int Bii-1) (39.0 g, 60%yield) . ESI-MS [M+Na] ⁺: 354.0

Step 2: tert-butyl (3aR, 6aS) -1- ( (S) -2- ( ( (benzyloxy) carbonyl) amino) -3-methoxy-3-oxopropyl) hexahydropyrrolo [3, 4-b] pyrrole-5 (1H) -carboxylate (Int Bii-2)

To a solution of tert-butyl hexahydropyrrolo [3, 4-b] pyrrole-5 (1H) -carboxylate (5.5 g, 25 mmol) , N- ( (benzyloxy) carbonyl) -O- (methylsulfonyl) -L-serinate (Int Bii-1) (15 g, 45 mmol) in DMF (10 mL) and H₂O (1 mL) were added K₂CO₃ (2.2 g, 16 mmol) and stirred at 90 ℃ for 10 minutes, additional N- ( (benzyloxy) carbonyl) -O- (methylsulfonyl) -L-serinate (12 g, 36 mmol) and K₂CO₃ (2.5 g, 18 mmol) was added. The reaction diluted with H₂O (50 mL) extracted with EtOAc. The organic layer was washed with H₂O and brine then dried over Na₂SO₄, and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel 0-4%MeOH/DCM) to afford tert-butyl (3aR, 6aS) -1- ( (S) -2- ( ( (benzyloxy) carbonyl) amino) -3-methoxy-3-oxopropyl) hexahydropyrrolo [3, 4-b] pyrrole-5 (1H) -carboxylate (Int Bii-2) (7.0 g, 60%yield) as a brown oil. ESI-MS [M+H] ⁺: 448.3

Step 3: methyl (S) -2- ( ( (benzyloxy) carbonyl) amino) -3- ( (3aR, 6aS) -hexahydropyrrolo [3, 4-b] pyrrol-1 (2H) -yl) propanoate (Int Bii-3)

A solution of tert-butyl (3aR, 6aS) -1- ( (S) -2- ( ( (benzyloxy) carbonyl) amino) -3-methoxy-3-oxopropyl) hexahydropyrrolo [3, 4-b] pyrrole-5 (1H) -carboxylate (Int Bii-2) (7.0 g, 15 mmol) in TFA (20 mL) and DCM (20 mL) was stirred for 1 h at RT. The reaction was concentrated and adjusted to pH= 8 with NaHCO₃ (sat, aq) and extracted with EtOAc/MeOH (9: 1) , the organic phase was concentrated and purified by flash chromatography on silica gel (0-20%MeOH in DCM) to afford methyl (S) -2- ( ( (benzyloxy) carbonyl) amino) -3- ( (3aR, 6aS) -hexahydropyrrolo [3, 4-b] pyrrol-1 (2H) -yl)propanoate (Int Bii-3) (3.2g, 36%yield as a brown oil) .

¹H NMR (400 MHz, CDCl3) δ: 8.92 (br, 1H) , 7.61-7.58 (m, 1H) , 7.50-7.15 (m, 5H) , 5.10-4.99 (m, 2H) , 4.25-4.12 (m, 1H) , 3.66-3.56 (m, 3H) , 3.19-2.58 (m, 9H) , 2.21-2.14 (m, 1H) , 2.05-1.99 (m, 1H) , 1.57-1.46 (m, 1H) . ESI-MS [M+H] ⁺: 348.3.

Step 4: methyl (S) -3- ( (3aR, 6aS) -5- (3- (3-acetoxy-2, 2-dimethylpropyl) -1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-5-yl) hexahydropyrrolo [3, 4-b] pyrrol-1 (2H) -yl) -2- ( ( (benzyloxy) carbonyl) amino) propanoate (Int Bii)

A solution of (S) -2- ( ( (benzyloxy) carbonyl) amino) -3- ( (3aR, 6aS) -hexahydropyrrolo [3, 4-b] pyrrol-1 (2H) -yl) propanoate (3.0 g, 9.3mmol) , 3- (5-bromo-1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-3-yl) -2, 2-dimethylpropyl acetate (4.4 g, 8.6 mmol) , Ruphos (1.6 g, 3.4 mmol) , Pd2 (dba) 3 (0.79 g, 0.86 mmol) , cesium carbonate (8.4 g, 26 mmol) in Toluene (50 mL) was stirred for 15 h at 105 ℃ under argon. The reaction was concentrated under vacuum and purified by prep-HPLC (C18, 0.01 M NH4HCO3 in H2O, ACN) to afford methyl (S) -3- ( (3aR, 6aS) -5- (3- (3-acetoxy-2, 2-dimethylpropyl) -1-ethyl-2- (2- (methoxymethyl) pyridin-3-yl) -1H-indol-5-yl) hexahydropyrrolo [3, 4-b] pyrrol-1 (2H) -yl) -2- ( ( (benzyloxy) carbonyl) amino) propanoate (Int Bii) (2.2g, 34%yield) . ESI-MS [M+H] ⁺: 740.4

The compounds in the table below were synthesized with a similar route as Int Bii, as described above using appropriate starting materials:

Synthesis of tert-butyl (3aS, 6aR) -4- ( (S) -2- ( ( (benzyloxy) carbonyl) amino) -3-methoxy-3-oxopropyl) hexahydrocyclopenta [c] pyrrole-2 (1H) -carboxylate, Int Biii-3

Step 1: tert-butyl (3aR, 6aR, Z) -4- (methoxymethylene) hexahydrocyclopenta [c] pyrrole-2 (1H) -carboxylate (Int Biii-a)

To a solution of (Methoxymethyl) triphenylphosphonium chloride (2.74 g, 7.99 mmol) and Potassium tert-butoxide (996 mg, 8.88 mmol) in THF (30 mL) was added tert-butyl 4-oxohexahydrocyclopenta [c] pyrrole-2 (1H) -carboxylate (1.00 g, 4.44 mmol) at 0 ℃ under N₂, the solution was stirred at 25 ℃ for 16 h. TLC (petroleum ether/EtOAc = 5: 1) indicated the product was formed (Rf = 0.65, 0.7) . The residue after concentrated was purified by flash chromatography on silica gel (0-10%EtOAc/petroleum ether) to afford Int Biii-a (0.52 g, 42%yield) .

¹H NMR (400 MHz, DMSO-d6) δ: 5.95 (m, 1H) , 3.51 (m, 2H) , 3.47 (m, 1H) , 3.03 (m, 1H) , 3.00 (m, 1H) , 2.62 (m, 1H) , 1.93 (m, 2H) , 1.72 (m, 1H) , 1.46 (m, 1H) , 1.40 (s, 9H) , 1.30 (m, 2H) , 0.87 (m, 1H) . ESI-MS [M-tBu+MeCN] ⁺: 239.2

Step 2: tert-butyl (3aR, 6aR) -4-formylhexahydrocyclopenta [c] pyrrole-2 (1H) -carboxylate (Int Biii-b)

To a solution of tert-butyl (3aR, 4Z, 6aR) -4- (methoxymethylene) -1, 3, 3a, 5, 6, 6a- hexahydrocyclopenta [c] pyrrole-2-carboxylate Int Biii-a (1.00 g, 3.95 mmol) in MeCN (25 mL) was added 2, 2, 2-trichloroacetic acid (812 mg, 4.97 mmol) at 25 ℃. The solution was stirred for 16 h at 25℃. TLC indicated the reaction was complete and the reaction mixture was acidified to pH= 7-8 with NaHCO₃ aqueous solution and extracted with EtOAc. The organic layer was dried over Na₂SO₄, and concentrated under reduced pressure to afford product Int Biii-b. ESI-MS [M-tBu+MeCN] ⁺: 224.9

Step 3: tert-butyl (3aR, 6aR) -4- (hydroxymethyl) hexahydrocyclopenta [c] pyrrole-2 (1H) -carboxylate (Int Biii-c)

To a solution of tert-butyl (3aR, 6aR) -4-formyl-3, 3a, 4, 5, 6, 6a-hexahydro-1H-cyclopenta [c] pyrrole-2-carboxylate (2.50 g, 10.4 mmol) in MeOH (50 mL) was added Sodium borohydride (592 mg, 15.6 mmol) at 0 ℃. The solution was stirred for 2 h at 25 ℃. TLC showed the reaction was completed. The solution was quenched with NH₄Cl (aq. ) , solvent was evaporated under reduced pressure and the aqueous layer was extracted with EtOAc. The combined organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated. The resulting residue was purified by flash chromatography on silica gel (0-20%EtOAc/petroleum ether) to afford Int Biii-c (1.7g, 61%yield) . ¹H NMR (400 MHz, DMSO-d6) δ: 12.4 (s, 1H) , 4.51 (s, 1H) , 3.36 (m, 1H) , 3.30 (m, 3H) , 3.15 (dd, J₁ = 11.2 Hz, J₁ = 3.6 Hz, 1H) , 3.00 (m, 1H) , 2.26 (m, 1H) , 1.79 (m, 3H) , 1.38 (s, 9H) , 1.32 (m, 2H) . ESI-MS [M-tBu+MeCN] ⁺: 227.1

Step 4: tert-butyl (3aR, 6aR) -4- (iodomethyl) hexahydrocyclopenta [c] pyrrole-2 (1H) -carboxylate (Int Biii-d)

To a solution of tert-butyl (3aR, 6aR) -4- (iodomethyl) hexahydrocyclopenta [c] pyrrole-2 (1H) -carboxylate (500 mg, 1.86 mmol) , imidazole (279 mg, 4.10 mmol) , triphenylphosphine (782 mg, 2.98 mmol) in DCM (2.5 mL) and THF (0.3 mL) was added I₂ (757 mg, 2.98 mmol) at 15 ℃. The suspension was stirred for 1 h at 25℃. TLC (petroleum ether/EtOAc = 5: 1) indicated the reaction was complete (product Rf = 0.7) . The reaction was then quenched with Na₂SO₃, and the resulting product was extracted with EtOAc and washed with water and brine, the organic phase was concentrated under vacuum to give the crude product. The crude product was purified by combi-flash on silica gel (0-13%EtOAc/petroleum ether) to afford Int Biii-d (550 mg, 80%yield) .

¹H NMR (400 MHz, DMSO-d6) δ: 3.36 (m, 4H) , 3.20 (dd, J₁ = 12.0Hz, J₂ = 3.2 Hz, 1H) , 3.00 (m, 1H) , 2.67 (s, 1H) , 2.22 (s, 1H) , 1.91 (m, 3H) , 1.39 (s, 9H) , 1.34 (m, 2H) . ESI-MS [M-tBu+MeCN] ⁺: 377.1

Step 5: tert-butyl (3aS, 6aR) -4- ( ( (2S, 5R) -5-isopropyl-3, 6-dimethoxy-2, 5-dihydropyrazin-2-yl) methyl) hexahydrocyclopenta [c] pyrrole-2 (1H) -carboxylate (Int Biii-e)

To a solution of 3-isopropyl-2, 5-dimethoxy-1, 4-dihydropyrazine (1.26 g, 6.82 mmol) in THF (20 mL) was added drop-wise n-Butyllithium (2.50 M, 2.73 mL) at -78 ℃ and stirred for 15 min. Tert-butyl (3aR, 6aR) -4- (iodomethyl) -3, 3a, 4, 5, 6, 6a-hexahydro-1H-cyclopenta [c] pyrrole-2-carboxylate, Int Biii-d (2.10 g, 5.68 mmol) in THF (10 mL) was added and stirred for 2 h at 25 ℃. TLC (petroleum ether/EtOAc = 5: 1) indicated the reaction was complete. The reaction was then quenched with water, and the resulting mixture was extracted with EtOAc and washed with water. The organic phase was concentrated under vacuum to give the crude product. The crude product was purified by combi-flash on silica gel (0-13%petroleum ether/EtOAc) to afford Int Biii-e (2.0 g, 77%yield) .

¹H NMR (400 MHz, DMSO-d6) δ: 4.00 (m, 1H) , 3.91 (m, 1H) , 3.61 (m, 6H) , 3.25 (m, 2H) , 3.00 (m, 1H) , 2.57 (s, 1H) , 2.17 (m, 2H) , 1.85 (m, 3H) , 1.69 (m, 2H) , 1.37 (s, 9H) , 1.23 (m, 3H) , 1.00 (d, J = 4.0Hz, 3H) , 0.62 (d, J = 6.0Hz, 3H) . ESI-MS [M+H] ⁺: 408.4

Step 6: tert-butyl (3aS, 6aR) -4- ( (S) -2-amino-3-methoxy-3-oxopropyl) hexahydrocyclopenta [c] pyrrole-2 (1H) -carboxylate (Int Biii-f)

To a solution of tert-butyl (3aS, 6aR) -4- [ [ (2S, 5R) -5-isopropyl-3, 6-dimethoxy-2, 5-dihydropyrazin-2-yl] methyl] -3, 3a, 4, 5, 6, 6a-hexahydro-1H-cyclopenta [c] pyrrole-2-carboxylate (3.0 g, 7.3 mmol) in MeCN (36 mL) and water (36 mL) was added hydrochloric acid (1.0 M, 30 mL) at 25 ℃. The solution was stirred for 1 h at 25 ℃. LC-MS indicated the reaction was completed. The reaction solution was adjusted to pH= 9 with NaHCO₃ aqueous. The crude solution of Int Biii-f was used for the next step directly. ESI-MS [M+H] ⁺: 313.2

Step 7: tert-butyl (3aS, 6aR) -4- ( (S) -2- ( ( (benzyloxy) carbonyl) amino) -3-methoxy-3-oxopropyl) hexahydrocyclopenta [c] pyrrole-2 (1H) -carboxylate (Int Biii-3)

To a crude solution of Int Biii-f was added dropwise benzyl chloroformate (3.74 g, 21.9 mmol, 3.11 mL, 3 equiv) at 0 ℃. The solution was stirred for 2 h at 25 ℃. LC-MS indicated the reaction was completed. The reaction solution was extracted with EtOAc, washed with water and brine, the organic layers were dried over Na₂SO₄, and concentrated under reduced pressure to afford crude product. The crude product was purified by flash chromatography on silica gel (0-30%EtOAc/petroleum ether) to afford Int Biii-3 (1.8 g, 80%yield for two steps) as a colorless oil. ¹H NMR (400 MHz, DMSO-d6) δ: 7.82 (m, 1H) , 7.36 (m, 5H) , 5.05 (s, 2H) , 4.02 (m, 1H) , 3.64 (s, 3H) , 3.40 (m, 1H) , 3.20 (s, 1H) , 3.00 (m, 1H) , 2.60 (s, 1H) , 2.15 (s, 1H) , 1.86 (m, 2H) , 1.70 (m, 3H) , 1.38 (s, 9H) , 1.23 (m, 3H) . ESI-MS [M+Na] ⁺: 469.2

Synthesis of methyl (S) -1- ( (S) -2- ( (tert-butoxycarbonyl) amino) -3- (3- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) propanoyl) hexahydropyridazine-3-carboxylate Int C:

Step 1: 1, 2-di-tert-butyl 3-methyl (S) -tetrahydropyridazine-1, 2, 3-tricarboxylate Int C-1

To a solution of (S) -tetrahydropyridazine-1, 2, 3-tricarboxylic acid 1, 2-di-tert-butyl ester (CAS: 156699-39-7, 10.00 g, 30.28 mmol) in MeOH (150 mL) was added dropwise a solution of TMSCHN₂ (2 M solution in Et₂O) (66 mL, 150 mmol) at 0 ℃. The mixture was stirred at room temperature for 16 h. Then the reaction was quenched with aqueous 10%AcOH in water (100 mL) . The mixture was extracted with ethyl acetate (3 *300 mL) , and the combined extract was washed with NaHCO₃ (saturated, 300 mL) , dried over anhydrous Na₂SO₂₄, filtered and concentrated. The residue was purified by a silica-gel column (eluent: ethyl acetate /petroleum ether = 1: 3) to afford 1, 2-di-tert-butyl 3-methyl (S) -tetrahydropyridazine-1, 2, 3-tricarboxylate, Int C-1 (8.5 g, yield: 82%) as a colorless oil. MS: m/z = 345.2 (M+1, ESI+) .

Step 2: methyl (S) -hexahydropyridazine-3-carboxylate Int C-2

To a solution of 1, 2-di-tert-butyl 3-methyl (S) -tetrahydropyridazine-1, 2, 3-tricarboxylate, Int C-1 (8.50 g, 24.7 mmol) in dichloromethane (20 mL) was added TFA (20 mL) at 0 ℃. The mixture was allowed to room temperature for 3 h. The mixture was concentrated in vacuum to afford the crude methyl (S) -hexahydropyridazine-3-carboxylate, Int C-2 (9.20 g, yield: 100%) as a colorless oil. MS:m/z = 145.0 (M+1, ESI+) . This material was used directly in the next step.

Step 3: methyl (S) -1- ( (S) -3- (3-bromophenyl) -2- ( (tert-butoxycarbonyl) amino) propanoyl) hexahydropyridazine-3-carboxylate Int C-3

To a solution of (2S) -3- (3-bromophenyl) -2- [ (2-methylpropan-2-yl) oxycarbonylamino] propanoic acid (CAS: 82278-73-7, 3.01 g, 8.77 mmol) , HOBt (1.13 g, 12.45 mmol) and EDCI (1.60 g, 12.45 mmol) in dichloromethane (50 mL) were added methyl (S) -hexahydropyridazine-3-carboxylate, Int C-2 (3.26 g, 8.30 mmol) and NMM (7.64 g, 74.9 mmol) at 0 ℃. The resulting mixture was degassed and refilled with nitrogen for three times, then stirred for 16 h at 25℃. The suspension was partitioned between aqueous NH4Cl (50 mL) and DCM (3 *50 mL) . The combined organic layers were washed with brine (50 mL) , dried over Na2SO4, filtered and concentrated. The crude product was purified by a silica gel column eluting (eluent: ethyl acetate /petroleum ether = 1: 5) to give methyl (S) -1- ( (S) -3- (3-bromophenyl) -2- ( (tert-butoxycarbonyl) amino) propanoyl) hexahydropyridazine-3-carboxylate, Int C-3 (1.20 g, yield: 40%) as a yellow solid. MS: m/z = 469.7, 471.7 (M+ 1, ESI+) .

The compounds in the table below were synthesized with a similar route as Int C-3, as described above using appropriate starting materials:

Step 4: methyl (S) -1- ( (S) -2- ( (tert-butoxycarbonyl) amino) -3- (3- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) propanoyl) hexahydropyridazine-3-carboxylateInt C

To a mixture of methyl (S) -1- ( (S) -3- (3-bromophenyl) -2- ( (tert-butoxycarbonyl) amino) propanoyl) hexahydropyridazine-3-carboxylate, Int C-3 (1.20 g, 2.54 mmol) , potassium acetate (0.76 g, 7.63 mmol) and bis (pinacolato) diboron (1.30 g, 5.10 mmol) in dioxane (25 ml) was added Pd (dppf) Cl₂ (0.19 g, 0.25 mmol) . The resulting mixture was degassed and refilled with nitrogen for three times, then stirred for 4 h at 80 ℃ . The suspension was partitioned between aqueous NH₄Cl (50 mL) and ethyl acetate (3 *50 mL) . The combined organic layers were washed with brine (50 mL) , dried over Na₂SO₂, filtered and concentrated. The crude product was purified by a silica gel column (eluent: ethyl acetate /petroleum ether = 1: 10) to give methyl (S) -1- ( (S) -2- ( (tert-butoxycarbonyl) amino) -3- (3- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan- 2-yl) phenyl) propanoyl) hexahydropyridazine-3-carboxylate, Int C (0.75 g, yield: 55%) as a yellow solid. MS: m/z = 517.8 (M+ 1, ESI+) .

Synthesis of methyl N-methyl-N- ( (S) -1- (4-morpholinobut-2-ynoyl) pyrrolidine-3-carbonyl) -L-valine Int D:

Step 1: benzyl (S) -3- ( ( (S) -1- (tert-butoxy) -3-methyl-1-oxobutan-2 yl) (methyl) carbamoyl) pyrrolidine-1-carboxylate

To a stirred solution of tert-butyl methyl-L-valinate (CAS: 5616-87-5, 750 mg, 4.00 mmol) and DIPEA (1.03 g, 8.00 mmol) in dichloromethane (30 mL) was added (S) -1- ( (benzyloxy) carbonyl) pyrrolidine-3-carboxylic acid (CAS: 192214-00-9, 998 mg, 4.00 mmol) and CIP (1.67 g, 6.00 mmol) in portions at 0 ℃. The resulting mixture was stirred at 25 ℃ for 2 h. The mixture was washed with brine (30 mL) . The organic phase was dried over anhydrous Na₂SO₄. After filtration, the solvent was removed in vacuum and the residue was purified by flash column (eluent: petroleum ether /ethyl acetate =75:25) chromatography on silica gel to give benzyl (S) -3- ( ( (S) -1- (tert-butoxy) -3-methyl-1-oxobutan-2-yl) (methyl) carbamoyl) pyrrolidine-1-carboxylate Int D-1 (1.60 g, yield: 85%) as a white solid. MS: m/z = 418.9 (M+1, ESI+) .

Step 2: tert-butyl N-methyl-N- ( (S) -pyrrolidine-3-carbonyl) -L-valinate Int D-2

A solution of benzyl (S) -3- ( ( (S) -1- (tert-butoxy) -3-methyl-1-oxobutan-2-yl) (methyl) carbamoyl) pyrrolidine-1-carboxylate Int D-1 (1.00 g, 2.40 mmol) and Pd/C (0.50 g) in THF (20 mL) was stirred at 25 ℃ under H₂ for 4 h. The mixture was filtered, the filtrate was concentrated in vacuum to give tert-butyl N-methyl-N- ( (S) -pyrrolidine-3-carbonyl) -L-valinate Int D-2 (480 mg, 63%yield) as a brown oil, which was used for the next step without purification. MS: m/z = 284.9 (M+1, ESI+) .

Step 3: tert-butyl N-methyl-N- ( (S) -1- (4-morpholinobut-2-ynoyl) pyrrolidine-3-carbonyl) -L-valinate, Int D-3

A solution of tert-butyl N-methyl-N- ( (S) -pyrrolidine-3-carbonyl) -L-valinate Int D-2 (142 mg, 0.50 mmol) , 4-morpholinobut-2-ynoic acid (CAS: 38346-95-1, 169 mg, 1.00 mmol) , CIP (278 mg, 1.00 mmol) and DIPEA (258 mg, 4.00 mmol) in DMF (4 mL) was stirred at 0 ℃ for 2 h. The mixture was diluted with H₂O (30 mL) and extracted with ethyl acetate (2*20 mL) . The organic phase was dried over anhydrous Na₂SO₄. After filtration, the solvent was removed in vacuum and the residue was purified by flash column (eluent: dichloromethane/Methanol = 40: 1) chromatography on silica gel to give tert-butyl N-methyl-N- ( (S) -1- (4-morpholinobut-2-ynoyl) pyrrolidine-3-carbonyl) -L-valinate, Int D-3 (100 mg, yield: 41%) as a colorless oil. MS: m/z = 435.9 (M+1, ESI+) .

Step4: N-methyl-N- ( (S) -1- (4-morpholinobut-2-ynoyl) pyrrolidine-3-carbonyl) -L-valine, Int-D

A tert-butyl N-methyl-N- ( (S) -1- (4-morpholinobut-2-ynoyl) pyrrolidine-3-carbonyl) -L-valinate, Int D-3 (100 mg, 0.23 mmol) and TFA (1 mL) in dichloromethane (3 mL) was stirred at 25 ℃ for 2 h. The mixture was concentrated in vacuum to give N-methyl-N- ( (S) -1- (4-morpholinobut-2-ynoyl) pyrrolidine-3-carbonyl) -L-valine, Int-D (120 mg, crude) as a brown oil, which was used for the next step without purification. MS: m/z = 379.9 (M+1, ESI+) .

Synthesis of N-methyl-N- ( (S) -1- (2-oxopropanoyl) pyrrolidine-3-carbonyl) -L-valine, Int Di

To a solution of Int D-2 (202 mg, 0.71 mmol) and Et3N (86 mg, 0.85 mmol) in DCM (3 mL) was added 2-oxopropanoyl chloride (69 mg, 0.78 mmol) solution dropwise at 0 ℃. The reaction was stirred at RT for 3 h. The reaction was quenched with water (20 mL) and extracted with DCM (20 mL*3) . The organic layers were combined, washed with brine (30 mL) , dried over Na₂SO₄ and concentrated under vacuum to give the crude product, which was purified by combi-flash on silica gel (petroleum ether/EtOAc = 5: 1) to afford the ketoamide (200 mg, 79%yield) as a colorless oil. ) . ESI-MS [M+H] ⁺: 355.2To a solution of the ketoamide (200 mg, 0.58 mmol) in DCM (6 mL) was added TFA (100 mg, 0.87 mmol) , the reaction was stirred at 25 ℃ for 3 h. Evaporated to give N-methyl-N- ( (S) -1- (2-oxopropanoyl) pyrrolidine-3-carbonyl) -L-valine Int Di (135 mg, 77%yield) as a colorless oil. ESI-MS [M+H] ⁺: 299.10

Synthesis of 2-amino-2- (6-bromo-2, 3-dihydro-1H-inden-1-yl) acetic acid Int E:

Step 1: Synthesis of diethyl 2- ( (diphenylmethylene) amino) malonate Int E-1

A mixture of benzophenone imine (CAS: 1013-88-3, 25.0 g, 138 mmol) and dimethyl 2-aminopropanedioate hydrochloride (25.0 g, 136 mmol) in DCM (300 mL) was stirred at room temperature for 3 days. The solid material was filtered off and the filtrate was concentrated in vacuo. The residue was re-dissolved in TBME and washed with water, dried (MgS04) and concentrated in vacuo. The residue was purified by column chromatography (silica gel, eluting with 15-100%EtOAc in heptane) to give the title compound (27.0 g, 64%) as a colourless oil, which solidified on standing. 1H NMR (300 MHz, Chloroform-d) d 7.78 -7.63 (m, 2H) , 7.51 -7.28 (m, 6H) , 7.24 -7.11 (m, 2H) , 4.90 (s, 1H) , 3.78 (s, 6H) ; LCMS (ES) : m/z=312.2 [M + H] ⁺.

Step 2: Synthesis of dimethyl 2- (benzhydrylideneamino) -2- [ (lR) -6-bromoindan-l-yl] propanedioate, Int E-2

A solution of 6-bromoindan-l-ol (6.5 g, 31 mmol) , Int E-1 (14.0 g, 46 mmol) and trimethylphosphine (1M solution in THF, 46 mL, 46 mmol) in toluene (100 g) was cooled down to -75℃. Diethyl azodicarboxylate (40 wt%solution in toluene, 21 g, 48 mmol) was added dropwise over 30 minutes and the solution was stirred at -75℃ for 1.5 hours and at room temperature for 3 hours to give a dark brown solution. The solution was concentrated in vacuo and the residue was purified by column chromatography (silica gel, eluting with heptane/ethyl acetate 4: 1) to give the title compound (9.6 g, 62%) as a yellow oil. 1H NMR (300 MHz, Chloroform-d) d 7.84 (dd, J = 1.9, 0.9 Hz, 1H) , 7.64 -7.49 (m, 2H) , 7.47 -7.22 (m, 7H) , 7.22 -7.14 (m, 2H) , 7.05 (dd, J = 8.0, 1.1 Hz, 1H) , 4.26 -4.03 (m, 1H) , 3.44 (s, 3H) , 3.25 (s, 3H) , 2.94 (ddd, J = 15.6, 9.2, 6.0 Hz, 1H) , 2.76 (ddd, J = 15.6, 9.1, 5.9 Hz, 1H) , 2.55 -2.17 (m, 2H) ; LCMS (METHOD 3) (ES) : m/z =506.3, 508.3 [M + H] +.

Step 3: Synthesis of 2-Amino-2- [ (lR) -6-bromoindan-l-yl] acetic acid hydrobromide, Int E

To a solution of Int E-2 (9.6 g, 19 mmol) in THF (30 mL) at room temperature was added cone. HCI (10 mL) (exothermic) . The solution was stirred at room temperature for 30 min and then diluted with TBME (70 mL) and water (30 mL) . The phases were separated, the aqueous phase was extracted twice with TBME and then concentrated in vacuo, giving crude dimethyl 2-amino-2- [ (lR) -6-bromoindan-l-yl] propanedioate hydrochloride as a colourless oil, which was used without further purification.

To a solution of dimethyl 2-amino-2- [ (lR) -6-bromoindan-l-yl] propanedioate hydrochloride in water (20 mL) at room temperature was added cone. HBr (48%HBr, 20 mL) . The solution was heated at reflux for 3 hours and the product precipitated. The suspension was cooled to room temperature and filtered. The filter cake was washed with TBME and dried in vacuo, giving the title compound (5.2 g, 78%) as a white solid as a mixture of diastereomers. 1H NMR (300 MHz, Deuterium Oxide + 1 drop DCI) d 7.38 (s, 0.33H) , 7.34 -7.21 (m, 1.67H) , 7.15 -6.96 (m, 1H) , 4.41 (d, J =4.0 Hz, 0.33H) , 4.32 (d, J = 3.7 Hz, 0.67H) , 3.91 -3.81 (m, 0.33H) , 3.75 (dt, J = 9.3, 4.8 Hz, 0.67H) , 3.07 -2.54 (m, 2H) , 2.50 -2.01 (m, 1H) , 1.99 -1.64 (m, 1H) ; LCMS (METHOD 3) (ES) : m/z 270.2, 272.2 [M + H] ⁺.

Dioxane (20 mL) was added to the crude amino acid solution followed by Boc anhydride (2.0 g, 9.2 mmol) and the resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was washed with DCM (2 x 50 mL) , EtOAc (50 mL) was added and the mixture was acidified with 4N HCI to pH 2. The layers were separated and the aqueous phase was extracted with further EtOAc (30 mL) . The combined organic phases were dried (MgSO4) and concentrated in vacuo to give the title compound as a mixture of 2 diastereomers (1.22 g, 90%) . LCMS (ES) : m/z 382.4, 384.4 [M + H] +

Synthesis of methyl (S) -1- ( (S) -2- ( (tert-butoxycarbonyl) amino) -3- (1, 4-diazepan-1-yl) propanoyl) hexahydropyridazine-3-carboxylate, Int-F

Step 1: Synthesis of benzyl (S) -4- (2- ( (tert-butoxycarbonyl) amino) -3-methoxy-3-oxopropyl) -1, 4-diazepane-1-carboxylate Int F-1:

To a solution of benzyl 1, 4-diazepane-1-carboxylate (1.0 g, 4.3 mmol) and methyl (R) -2- ( (tert-butoxycarbonyl) amino) -3-iodopropanoate (4.20 g, 12.8 mmol) in acetonitrile (20 mL) was added triethylamine (2.75 g, 21.4 mmol) at 70 ℃ under nitrogen. Then the resulting solution was heated at the same temperature overnight. After 16 h, water (60 mL) was added to the reaction solution, extracted with dichloromethane (30 x 3 mL) . The organic layer was separated and evaporated to dryness. The residue was then purified by column chromatography (0-60%ethyl acetate in petroleum ether) to get benzyl (S) -4- (2- ( (tert-butoxycarbonyl) amino) -3-methoxy-3-oxopropyl) -1, 4-diazepane-1-carboxylate Int F-1 (631 mg, 1.45 mmol, 34%yield) as white solid. MS: m/z = 436.2 (M+1, ESI+) .

Step 2: Synthesis of (S) -3- (4- ( (benzyloxy) carbonyl) -1, 4-diazepan-1-yl) -2- ( (tert-butoxycarbonyl) amino) propanoic acid Int F-2

To a solution of benzyl (S) -4- (2- ( (tert-butoxycarbonyl) amino) -3-methoxy-3-oxopropyl) -1, 4-diazepane-1-carboxylate Int F-1 (630 mg, 1.45 mmol) in MeOH (9 mL) cooled to 0 ℃ was added aqueous solution of LiOH (120 mg, 5.0 mmol) in water (1 mL) slowly by syringe over 2 min. After addition, the solution was allowed to warm up to room temperature slowly and stirred for another 3 h. After complete hydrolysis, the solution was evaporated to dryness and diluted with water, neutralized with 0.1N HCl solution, extracted with DCM (20 x 2 mL) . The collected organic phase was combined and dried over sodium sulfate, then filtered and concentrated to get (S) -3- (4- ( (benzyloxy) carbonyl) -1, 4-diazepan-1-yl) -2- ( (tert-butoxycarbonyl) amino) propanoic acid Int F-2 (400 mg, 65%yield) as white solid which was pure according to LCMS. Used as such in the next step without further purification. MS: m/z = 422.3 (M+1, ESI+) .

Step 3: Synthesis of benzyl 4- ( (S) -2- ( (tert-butoxycarbonyl) amino) -3- ( (S) -3- (methoxycarbonyl) tetrahydropyridazin-1 (2H) -yl) -3-oxopropyl) -1, 4-diazepane-1-carboxylate Int F-3

To a solution of crude methyl (S) -hexahydropyridazine-3-carboxylate (CAS: 138323-07-6, 350 mg, 0.95 mmol) in dichloromethane (5 mL) at 0 ℃ was added N-methylmorpholine (1.0 g, 9.5 mmol) and (S) -3- (4- ( (benzyloxy) carbonyl) -1, 4-diazepan-1-yl) -2- ( (tert-butoxycarbonyl) amino) propanoic acid Int F-2 (400 mg, 0.95 mmol) , followed with addition of HOBt (27 mg, 0.2 mmol) and EDCI (570 mg, 1.5 mmol) . The resulting solution was stirred at room temperature for 16 h. Then the reaction solution was diluted with dichloromethane (30 mL) and washed with H2O (3 x 10 mL) . The organic layer was dried over anhydrous Na2SO4, filtered, the filtrate concentrated under reduced pressure. Then the mixture was purified by column chromatography (0-50%ethyl acetate in petroleum ether) to afford benzyl 4- ( (S) -2- ( (tert-butoxycarbonyl) amino) -3- ( (S) -3- (methoxycarbonyl) tetrahydropyridazin-1 (2H) -yl) -3-oxopropyl) -1, 4-diazepane-1-carboxylate Int F-3 (400 mg, 0.7 mmol, 76%yield) as white solid. MS: m/z = 548.3 (M+1, ESI+) .

Step 4: Synthesis of methyl (S) -1- ( (S) -2- ( (tert-butoxycarbonyl) amino) -3- (1, 4- diazepan-1-yl) propanoyl) hexahydropyridazine-3-carboxylate, Int-F

To a solution of benzyl 4- ( (S) -2- ( (tert-butoxycarbonyl) amino) -3- ( (S) -3- (methoxycarbonyl) tetrahydropyridazin-1 (2H) -yl) -3-oxopropyl) -1, 4-diazepane-1-carboxylate Int F-3 (150 mg, 0.27 mmol) in MeOH (5 mL) was added Pd/C (30 mg, 10%wt) at room temperature under nitrogen atmosphere, the mixture was then degassed and backfilled with hydrogen for three times, then stirred under the atmosphere of H₂ at room temperature for 2 h. After that, palladium catalyst was filtered off and washed the cake with more methanol, the filtrate was evaporated to dryness and purified by reverse-phase column chromatography (C18 column, eluting with 0-20%acetonitrile in water over 20 min) to get desired fractions, which was lyophilized to dryness to give methyl (S) -1- ( (S) -2- ( (tert-butoxycarbonyl) amino) -3- (1, 4-diazepan-1-yl) propanoyl) hexahydropyridazine-3-carboxylate, Int-F (24 mg, 0.060 mmol, 21%yield) as white solid. MS: m/z = 414.2 (M+1, ESI+) . ¹H NMR (400 MHz, CD₃OD) δ5.21 (ddd, J = 46.2, 8.4, 3.9 Hz, 1H) , 4.32 (d, J = 12.2 Hz, 1H) , 3.73 (t, J = 6.2 Hz, 3H) , 3.71 –3.57 (m, 1H) , 3.62 –3.34 (m, 2H) , 3.22 –3.12 (m, 3H) , 3.09 –2.95 (m, 1H) , 2.83 (ddt, J = 35.6, 26.3, 13.1 Hz, 5H) , 2.68 –2.55 (m, 1H) , 2.13 –1.57 (m, 7H) , 1.45 (s, 9H) ppm.

Synthesis of (3S) -N- ( (2S) -1- ( ( (63S, 4S) -11-ethyl-12- (2- (1-methoxyethyl) pyridin-3-yl) -10, 10-dimethyl-5, 7-dioxo-61, 62, 63, 64, 65, 66-hexahydro-11H-8-oxa-1 (5, 3) -indola-6 (1, 3) -pyridazina-2 (1, 3) -benzenacycloundecaphane-4-yl) amino) -3-methyl-1-oxobutan-2-yl) -N-methyl-1- (4-morpholinobut-2-ynoyl) pyrrolidine-3-carboxamide Example 1:

Step 1: Synthesis of methyl (3S) -1- ( (2S) -2- ( (tert-butoxycarbonyl) amino) -3- (3- (1-ethyl-3- (3-hydroxy-2, 2-dimethylpropyl) -2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indol-5-yl) phenyl) propanoyl) hexahydropyridazine-3-carboxylate 1

A solution of Int-C (290 mg, 0.56 mmol) , Int Bi (249 mg, 0.56 mmol) , Pd (DTBPF) Cl₂ (417 mg, 0.56 mmol) and Na₂CO₃ (83 mg, 1.40 mmol) in dioxane/H₂O (12 mL, 5: 1) was stirred at room temperature for 16 h. The mixture was filtered, and the filtrate was concentrated in vacuum and the residue was purified by silica-gel column chromatography (eluent: dichloromethane /Methanol = 98: 2 –95: 5) to give compound, methyl (3S) -1- ( (2S) -2- ( (tert-butoxycarbonyl) amino) -3- (3- (1-ethyl-3- (3-hydroxy-2, 2-dimethylpropyl) -2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indol-5-yl) phenyl) propanoyl) hexahydropyridazine-3-carboxylate 1 (110 mg, yield: 23%) as a brown solid. MS: m/z = 756.3 (M+1, ESI+) .

The compounds in the table below were synthesized with a similar route as Int 1, as described above using appropriate starting materials:

Step 2: Synthesis of (3S) -1- ( (2S) -2- ( (tert-butoxycarbonyl) amino) -3- (3- (1-ethyl-3- (3-hydroxy-2, 2-dimethylpropyl) -2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indol-5-yl) phenyl) propanoyl) hexahydropyridazine-3-carboxylic acid, 2

A solution of methyl (3S) -1- ( (2S) -2- ( (tert-butoxycarbonyl) amino) -3- (3- (1-ethyl-3- (3-hydroxy-2, 2-dimethylpropyl) -2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indol-5-yl) phenyl) propanoyl) hexahydropyridazine-3-carboxylate 1 (110 mg, 0.15 mmol) and trimethyltin hydroxide (132 mg, 0.73 mmol) in dichloroethane (2 mL) was stirred at 65 ℃ for 16 h. The mixture was diluted with dichloromethane (20 mL) and filtered, and the filtrate was washed with citric acid (10%in water) . The organic phase was dried over anhydrous Na₂SO₄. After filtration, the solvent was concentrated in vacuum to give (3S) -1- ( (2S) -2- ( (tert-butoxycarbonyl) amino) -3- (3- (1-ethyl-3- (3-hydroxy-2, 2-dimethylpropyl) -2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indol-5-yl) phenyl) propanoyl) hexahydropyridazine-3-carboxylic acid, 2 (130 mg, yield: 96%) as a brown solid, which was used for the next step without purification. MS: m/z =742.4 (M+1, ESI+) .

Synthesis of Example 16:

Step 1: methyl (S) -1- ( (S) -3- ( (3aR, 4R, 6aS) -2- (3- (3-acetoxy-2, 2-dimethylpropyl) -1- ethyl-2- (2- ( (S) -1-methoxyethyl) pyridin-3-yl) -1H-indol-5-yl) octahydrocyclopenta [c] pyrrol-4-yl) -2- ( ( (benzyloxy) carbonyl) amino) propanoyl) hexahydropyridazine-3-carboxylate To a solution Int B-vi (2.00 g, 4.10 mmol) in dioxane (10 mL) were added methyl Int C3-iii (2.25 g, 4.92 mmol) , Ruphos Pd G4 (697 mg, 0.82 mmol) , Ruphos (765 mg, 1.64 mmol) and Cs₂CO₃ (2.67 g, 8.21 mmol) . The reaction mixture was stirred at 100 ℃ for 12 h under N₂. After filtration, the solvent was removed in vacuum and the residue was purified by flash column (eluent: PE/EA = 80: 20 -10: 90) chromatography on silica gel to give compound 16A (1.31 g, yield: 29%) as a white solid. MS: m/z = 865.0 (M+H, ESI+) .

Step 2: (S) -1- ( (S) -2- ( ( (benzyloxy) carbonyl) amino) -3- ( (3aR, 4R, 6aS) -2- (1-ethyl-3- (3-hydroxy-2, 2-dimethylpropyl) -2- (2- ( (S) -1-methoxyethyl) pyridin-3-yl) -1H-indol-5-yl) octahydrocyclopenta [c] pyrrol-4-yl) propanoyl) hexahydropyridazine-3-carboxylic acid

To a solution of 16A (1.31 g, 1.51 mmol) in THF (13 mL) and water (6 mL) was added and LiOH·H₂O (252 mg, 6.01 mmol) , the mixture was stirred at rt for 16 h. The mixture was adjusted PH = 5-6 with HCl (1 mol/mL) and extracted with ethyl acetate (50 mL x 3) . The organic phase was washed with NaCl (saturated, 20 mL) and dried over anhydrous Na₂SO₄. After filtration, the solvent was removed and the crude 16B (1 g) was obtained as a light-yellow solid which was used for the next step without further purification. MS: m/z =809.0 (M+H, ESI+) .

Step 3: benzyl ( (23aR, 24R, 26aS, 63S, 4S) -11-ethyl-12- (2- ( (S) -1-methoxyethyl) pyridin-3-yl) -10, 10-dimethyl-5, 7-dioxo-21, 22, 23, 23a, 24, 25, 26, 26a, 61, 62, 63, 64, 65, 66-tetradecahydro-11H-8-oxa-1 (5, 3) -indola-2 (2, 4) -cyclopenta [c] pyrrola-6 (1, 3) -pyridazinacycloundecaphane-4-yl) carbamate

A mixture of 16B (500 mg, 0.62 mmol) , Py₂S₂ (1.91 g, 6.20 mmol) and PPh₃ (1.63 g, 6.20 mmol) in toluene (5 mL) was stirred at 85 ℃ for 3 h under N₂. The mixture was diluted with ethyl acetate (50 mL) and washed with NaCl (20 mL) , dried over anhydrous Na₂SO₄. After filtration, the solvent was removed in vacuum and the residue was purified by flash column (eluent: DCM/MeOH = 0 –5%) chromatography on silica gel to give compound 20C (385 mg, impure) as a yellow solid. Then the product was purified by Prep-HPLC (Column: Gemini-C18 150 x 21.2 mm, 5μm, ACN-H2O (0.05%NH3. H2O) , Flow rate: 20 ml/min; Wavelength: 214 nm /254 nm. 10 min gradient maintained at 25-40%CH3CN. ) to give P1: 20 mg, P2: 8 mg, P3: 10 mg, P4: 45 mg as a yellow solid.

MS: m/z = 791.0 (M+1, ESI+) .

Step 4: (4R, 5S, 8R, 10S, 16S) -10-amino-24-ethyl-23- {2- [ (1S) -1-methoxyethyl] pyridin-3-yl} -20, 20-dimethyl-18-oxa-2, 12, 24, 30-tetraazahexacyclo [20.5.2.1^ {2, 5} . 1^ {12, 16} . 0^ {4, 8} . 0^ {25, 29} ] hentriaconta-1 (27) , 22, 25, 28-tetraene-11, 17-dione

A solution of 16C (45 mg, 0.06 mmol) in ^tBuOH (4 mL) was added Pd/C (30 mg, 0.28 mmol) stirred at 40 ℃ for 2 h under H₂ at an atmosphere of balloon. The filtrate was collected by filtration through a cite washed with ^tBuOH (5 mL) and MeOH (5mL) . after evaporation to give 16D (23 mg, yield: 58%) as colorless oil. MS: m/z =657.1 (M+H, ESI+) .

Step 5: (1R, 2R) -N- [ (4R, 5S, 8R, 10S, 16S) -24-ethyl-23- {2- [ (1S) -1-methoxyethyl] pyridin-3-yl} -20, 20-dimethyl-11, 17-dioxo-18-oxa-2, 12, 24, 30-tetraazahexacyclo [20.5.2.1^ {2, 5} . 1^ {12, 16} . 0^ {4, 8} . 0^ {25, 29} ] hentriaconta-1 (28) , 22, 25 (29) , 26-tetraen-10-yl] -2-methylcyclopropane-1-carboxamide To a solution of compound 16D (23 mg, 0.03 mmol) , (1R, 2R) -2-methylcyclopropane-1-carboxylic acid (CAS: 6202-94-4; 7 mg, 0.07 mmol) and N, N-Diisopropylethylamine (23 mg, 0.17 mmol) in DMF (3 mL) stirred was added a solution of COMU (30 mg, 0.07 mmol) in DMF (0.5 mL) dropwise at -20℃. The reaction mixture was stirred at 25 ℃ for 2 h. Quenched with saturated NH₄Cl (100 mL) , extracted with DCM (20 mL x 3) , washed with brine (10 mL x 3) . The organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated in vacuum and the residue was purified by Prep-HPLC (Column: Gemini-C18 150 x 21.2 mm, 5μm, ACN--H2O (0.1%FA) , Flow rate: 20 ml/min; Wavelength: 214 nm. Sample injected in DMF, 12 min linear gradient from 45%to 95%CH3CN. ) to afford 5 isomers of Example 16.

Example 16 Isomer 1 (1.27 mg)

MS: m/z =739.0 (M+1, ESI+) . Retention Time (min) : 1.150 min.

1HNMR (400MHz, MeOD) δ 8.67 (s, 2H) , 8.06 (d, J = 8.1Hz, 1H) , 8.00 (d, J = 7.4Hz, 1H) , 7.84 (s, 1H) , 7.77 –7.69 (m, 2H) , 7.55 (dd, J = 13.0, 7.8Hz, 1H) , 7.47 (d, J = 7.6Hz, 1H) , 5.28 –5.21 (m, 3H) , 4.67 –4.59 (m, 2H) , 4.39 (s, 2H) , 4.34 –4.28 (m, 1H) , 3.77 (dd, J = 16.6, 11.8Hz, 2H) , 3.52 (d, J = 18.7Hz, 3H) , 3.38 (s, 1H) , 3.24 (d, J = 8.7Hz, 3H) , 3.03 (s, 2H) , 2.80 (s, 2H) , 2.56 (s, 3H) , 2.18 (s, 1H) , 2.14 –2.03 (m, 3H) , 1.93 (d, J = 4.4Hz, 3H) , 1.57 –1.46 (m, 3H) , 1.35 (dd, J = 12.2, 5.9Hz, 2H) , 1.19 (s, 12H) , 0.80 (t, J = 6.4Hz, 4H) , 0.50 (s, 1H) .

Example 16 Isomer 2 (0.86 mg)

MS: m/z =739.5 (M+1, ESI+) . Retention Time (min) : 1.281 min.

1HNMR (400MHz, MeOD) δ 8.62 (d, J = 5.1Hz, 1H) , 8.42 (s, 1H) , 7.81 (d, J = 7.8Hz, 1H) , 7.41 (dd, J = 7.5, 4.7Hz, 1H) , 7.23 –7.16 (m, 1H) , 7.00 (s, 1H) , 6.73 (d, J = 8.9Hz, 1H) , 5.27 –5.16 (m, 2H) , 4.51 (s, 1H) , 4.16 (dt, J = 14.7, 7.3Hz, 1H) , 3.86 (dd, J = 17.6, 9.5Hz, 1H) , 3.62 –3.46 (m, 3H) , 3.43 –3.30 (m, 2H) , 3.13 (dd, J = 14.9, 10.3Hz, 3H) , 2.98 (d, J = 12.6Hz, 1H) , 2.88 (d, J = 8.9Hz, 1H) , 2.74 (d, J = 12.7Hz, 1H) , 2.58 (d, J =14.0Hz, 2H) , 2.47 (s, 2H) , 2.17 (d, J = 15.0Hz, 2H) , 2.08 (dd, J = 17.1, 9.4Hz, 2H) , 1.93 (d, J = 5.7Hz, 2H) , 1.83 (d, J = 6.8Hz, 2H) , 1.67 (s, 2H) , 1.49 (d, J = 17.1Hz, 2H) , 1.27 –1.17 (m, 11H) , 1.16 –1.11 (m, 1H) , 1.08 (d, J = 6.4Hz, 1H) , 1.02 (d, J = 2.8Hz, 1H) , 0.81 (dd, J = 13.9, 7.4Hz, 3H) , 0.45 (d, J = 26.5Hz, 2H) .

Example 16 Isomer 3 (1.12 mg)

MS: m/z =739.4 (M+1, ESI+) . Retention Time (min) : 1.290 min.

1HNMR (400MHz, MeOD) δ 8.63 (d, J = 4.6Hz, 1H) , 8.05 (m, 1H) , 7.81 –7.75 (m, 1H) , 7.57 –7.52 (m, 1H) , 7.21 (d, J = 8.9Hz, 1H) , 6.88 (s, 1H) , 6.72 (d, J = 8.5Hz, 1H) , 5.48 (s, 1H) , 5.27 –5.22 (m, 1H) , 4.63 (dd, J = 13.8, 7.9Hz, 1H) , 4.42 (d, J = 12.4Hz, 1H) , 4.33 –4.26 (m, 1H) , 4.22 –4.14 (m, 1H) , 4.06 (dd, J = 12.5, 6.1Hz, 1H) , 3.90 (dd, J = 19.8, 9.3Hz, 2H) , 3.80 –3.68 (m, 2H) , 3.60 –3.52 (m, 2H) , 3.49 –3.43 (m, 2H) , 3.13 (s, 3H) , 3.00 (d, J = 14.6Hz, 2H) , 2.75 (s, 1H) , 2.57 (d, J = 11.1Hz, 2H) , 2.22 –2.16 (m, 1H) , 2.09 (t, J = 7.5Hz, 2H) , 1.95 –1.85 (m, 4H) , 1.71 (s, 1H) , 1.62 (dd, J =14.0, 5.6Hz, 1H) , 1.49 (d, J = 12.8Hz, 2H) , 1.34 (s, 1H) , 1.21 (d, J = 17.6Hz, 12H) , 1.12 (s, 3H) , 1.00 (t, J = 5.8Hz, 2H) , 0.84 (s, 2H) , 0.79 (d, J = 6.9Hz, 3H) , 0.49 (s, 2H) .

Example 16 Isomer 4 (2.35 mg)

MS: m/z =739.4 (M+1, ESI+) . Retention Time (min) : 1.291 min.

1HNMR (400MHz, MeOD) δ 8.60 (d, J = 3.5Hz, 1H) , 8.41 (s, 1H) , 7.69 (d, J = 7.7Hz, 1H) , 7.41 (dd, J = 7.7, 4.8Hz, 1H) , 7.22 (d, J = 8.8Hz, 1H) , 6.97 (s, 1H) , 6.72 (d, J =8.9Hz, 1H) , 5.23 (dd, J = 13.2, 8.5Hz, 2H) , 4.36 –4.27 (m, 1H) , 4.11 (dd, J = 14.6, 7.2Hz, 1H) , 3.97 (dd, J = 14.7, 7.2Hz, 1H) , 3.88 –3.74 (m, 2H) , 3.59 (d, J = 11.0Hz, 1H) , 3.38 (d, J = 7.9Hz, 1H) , 3.27 (s, 3H) , 3.13 (s, 1H) , 3.00 (d, J = 7.7Hz, 1H) , 2.92 –2.78 (m, 2H) , 2.64 (d, J = 7.4Hz, 2H) , 2.43 (d, J = 14.2 Hz, 1H) , 2.10 (dd, J = 16.3, 8.8Hz, 3H) , 1.93 (d, J = 5.8Hz, 1H) , 1.84 (dd, J = 15.2, 10.5Hz, 2H) , 1.65 (s, 1H) , 1.50 (d, J = 6.4Hz, 2H) , 1.33 (d, J = 6.1Hz, 3H) , 1.21 (d, J = 17.5Hz, 9H) , 1.00 (d, J = 4.4Hz, 2H) , 0.96 –0.87 (m, 1H) , 0.79 (d, J = 6.1Hz, 3H) , 0.74 (t, J = 6.9Hz, 1H) , 0.48 (d, J =7.6Hz, 1H) , 0.33 (s, 2H) .

Example 16 Isomer 5 (4.38 mg)

MS: m/z =739.4 (M+1, ESI+) . Retention Time (min) : 1.324 min.

1HNMR (400MHz, MeOD) δ 8.68 –8.58 (m, 1H) , 8.20 –8.02 (m, 1H) , 7.68 (dd, J =7.7, 1.4Hz, 1H) , 7.41 (dd, J = 7.7, 4.8Hz, 1H) , 7.22 (d, J = 8.3Hz, 1H) , 6.86 (s, 1H) , 6.71 (d, J = 7.9Hz, 1H) , 5.47 (d, J = 5.5Hz, 1H) , 5.35 –5.15 (m, 1H) , 4.42 (d, J = 12.9Hz, 1H) , 4.26 (dd, J = 12.2, 6.0Hz, 1H) , 4.09 (d, J = 6.0 Hz, 1H) , 3.95 (d, J = 6.6Hz, 1H) , 3.85 (d, J = 10.8Hz, 2H) , 3.69 (d, J = 10.9Hz, 1H) , 3.39 (s, 1H) , 3.24 (d, J = 17.2Hz, 3H) , 3.02 (dd, J = 32.1, 17.1Hz, 3H) , 2.77 (d, J = 18.6Hz, 1H) , 2.65 –2.52 (m, 1H) , 2.29 (d, J = 14.4Hz, 1H) , 2.22 –2.00 (m, 1H) , 1.99 –1.79 (m, 3H) , 1.78 –1.58 (m, 3H) , 1.55 –1.44 (m, 1H) , 1.33 (t, J = 14.0Hz, 3H) , 1.29 –1.09 (m, 5H) , 1.07 –0.86 (m, 3H) , 0.86 (s, 2H) , 0.75 (dd, J = 20.2, 13.2Hz, 3H) , 0.47 (s, 1H) , 0.32 (s, 2H) .

Step 3: Synthesis of tert-butyl ( (6³S, 4S) -1¹-ethyl-1²- (2- (1-methoxyethyl) pyridin-3-yl) -10, 10-dimethyl-5, 7-dioxo-6¹, 6², 6³, 6⁴, 6⁵, 6⁶-hexahydro-1¹H-8-oxa-1 (5, 3) -indola- 6(1, 3) -pyridazina-2 (1, 3) -benzenacycloundecaphane-4-yl) carbamate, 3

To a solution of (3S) -1- ( (2S) -2- ( (tert-butoxycarbonyl) amino) -3- (3- (1-ethyl-3- (3-hydroxy-2, 2-dimethylpropyl) -2- (2- (1-methoxyethyl) pyridin-3-yl) -1H-indol-5-yl) phenyl) propanoyl) hexahydropyridazine-3-carboxylic acid, 2 (100 mg, 0.14 mmol) , diisopropylethylamine (52 mg, 0.40 mmol) in dichloromethane (2 mL) was added HOBt (27 mg, 0.20 mmol) and EDCI (38 mg, 0.20 mmol) at 0 ℃. The mixture was stirred at 25 ℃ for 16 h. The mixture was diluted with dichloromethane (20 mL) washed with brine. The organic phase was dried over anhydrous Na₂SO₄. After filtration, the solvent was concentrated in vacuum and the residue was purified by a column chromatography on silica gel (eluent: dichloromethane /Methanol = 99: 1 –95: 5) to give tert-butyl ( (6³S, 4S) -1¹-ethyl-1²- (2- (1-methoxyethyl) pyridin-3-yl) -10, 10-dimethyl-5, 7-dioxo-6¹, 6², 6³, 6⁴, 6⁵, 6⁶-hexahydro-1¹H-8-oxa-1 (5, 3) -indola-6 (1, 3) -pyridazina-2 (1, 3) -benzenacycloundecaphane-4-yl) carbamate, 3 (20 mg, yield: 19%) as a light yellow solid. MS: m/z = 724.3 (M+1, ESI+) .

Step 4: Synthesis of (6³S, 4S) -4-amino-1¹-ethyl-1²- (2- (1-methoxyethyl) pyridin-3-yl) -10, 10-dimethyl-6¹, 6², 6³, 6⁴, 6⁵, 6⁶-hexahydro-1¹H-8-oxa-1 (5, 3) -indola-6 (1, 3) -pyridazina-2 (1, 3) -benzenacycloundecaphane-5, 7-dione, 4

A solution of tert-butyl ( (6³S, 4S) -1¹-ethyl-1²- (2- (1-methoxyethyl) pyridin-3-yl) -10, 10-dimethyl-5, 7-dioxo-6¹, 6², 6³, 6⁴, 6⁵, 6⁶-hexahydro-1¹H-8-oxa-1 (5, 3) -indola-6 (1, 3) -pyridazina-2 (1, 3) -benzenacycloundecaphane-4-yl) carbamate, 3 (20 mg, 0.03 mmol) and TFA (0.50 mL) in dichloromethane (1 mL) was stirred at 25 ℃ for 1 h. The mixture was concentrated in vacuum to give ( (63S, 4S) -4-amino-11-ethyl-12- (2- (1-methoxyethyl) pyridin-3-yl) -10, 10-dimethyl-61, 62, 63, 64, 65, 66-hexahydro-11H-8-oxa-1 (5, 3) -indola-6 (1, 3) -pyridazina-2 (1, 3) -benzenacycloundecaphane-5, 7-dione, 4 (25 mg, crude) as a brown solid, which was used for the next step without purification. MS: m/z = 624.3 (M+1, ESI+) .

Step 5: Synthesis of (3S) -N- ( (2S) -1- ( ( (63S, 4S) -11-ethyl-12- (2- (1-methoxyethyl) pyridin-3-yl) -10, 10-dimethyl-5, 7-dioxo-61, 62, 63, 64, 65, 66-hexahydro- 11H-8-oxa-1 (5, 3) -indola-6 (1, 3) -pyridazina-2 (1, 3) -benzenacycloundecaphane-4-yl) amino) -3-methyl-1-oxobutan-2-yl) -N-methyl-1- (4-morpholinobut-2-ynoyl) pyrrolidine-3-carboxamide Example 1

To a solution of N-methyl-N- ( (S) -1- (4-morpholinobut-2-ynoyl) pyrrolidine-3-carbonyl) -L-valine, Int D (6 mg, 0.016 mmol) , ( (63S, 4S) -4-amino-11-ethyl-12- (2- (1-methoxyethyl) pyridin-3-yl) -10, 10-dimethyl-61, 62, 63, 64, 65, 66-hexahydro-11H-8-oxa-1 (5, 3) -indola-6 (1, 3) -pyridazina-2 (1, 3) -benzenacycloundecaphane-5, 7-dione, 4 (10 mg, 0.016 mmol) and DIPEA (21 mg, 0.16 mmol) in DMF (1 mL) was added COMU (14 mg, 0.032 mmol) in a ice-salt bath. The mixture was stirred at room temperature for 4 h. The mixture was diluted with ethyl acetate (20 mL) and washed with brine (5 mL) . The organic phase was dried over anhydrous Na₂SO₄. After filtration, the solvent was removed in vacuum and the residue was purified by preparatory HPLC (Gemini 5u C₁₈ 150 x 21.2 mm, acteonitrile in water (0.1%formic acid) , flow rate: 20 mL/min; Wavelength: 214 nm. Sample injected in DMSO, 12.5 min linear gradient from 20%to 95%acetonitrile) to afford Example 1 (9 mg, yield: 56%) . MS: m/z = 1007.4 (M+Na, ESI+) . 1H NMR (400 MHz, DMSO) δ 8.80 –8.67 (m, 1.14H) , 8.29 (t, J = 8.7 Hz, 0.16H) , 8.16 –7.79 (m, 2.7H) , 7.76 –7.40 (m, 4.3H) , 7.33 –7.15 (m, 1H) , 7.12 –6.95 (m, 1H) , 5.70 –5.46 (m, 0.2H) , 5.45 –5.21 (m, 0.8H) , 5.15 –4.95 (m, 0.5H) , 4.75 –4.55 (m, 0.5H) , 4.40 –3.35 (m, 19H) , 3.29 –3.22 (m, 1H) , 3.19 –3.00 (m, 4H) , 2.99 –2.84 (m, 2H) , 2.82 –2.71 (m, 2H) , 2.71 –2.59 (m, 2H) , 2.59 –2.49 (m, 1H) , 2.45 –2.27 (m, 2H) , 2.26 –1.85 (m, 4H) , 1.85 –1.40 (m, 3H) , 1.34 –1.14 (m, 3H) , 1.12 –0.55 (m, 13.6H) , 0.45 (d, J = 3.3 Hz, 1.4H) .

Analytical conditions

Apparatus: SFC Thar X-5

Column: CHIRALPAK OJ-H 250 mm x 4.6 mm, 5 μm

Modifier : 40%EtOH (NH₄OH 0.2%)

Total Flow: 2.5 mL/min

Retention time = 8.44 min

The compounds in the table below were synthesized with a similar route as Example 1 using appropriate starting materials.

BIOLOGICAL ASSAYS

Biological Example 1: TR-FRET Assay for Disruption of Raf1 Ras-Binding Domain (RBD) Interaction with KRas by Compounds of the Invention (Raf1 disruption TR-FRET)

Material and Reagents

1. His-tagged recombinant human KRAS G12C/G12D (a.a. 1-164, N-terminal His-tagged, GppNHp-loaded, produced in-house)

2. FLAG-tagged recombinant human Raf1 Ras-binding domain (a.a. 54-131, C-terminal FLAG-tagged, produced in-house)

3. LANCE Detection Buffer (10x) (PerkinElmer, CAT#CR97-100)

4. Eu-W1024-labeled Anti-6xHis Antibody (PerkinElmer, CAT#AD0401)

5. Anti-FLAG IgG conjugated to SureLight-Allophycocyanin (PerkinElmer, CAT#AD0059F)

6. GppNHP (Non-hydrolyzable GTP analog) (Abcam, CAT#ab146659)

7. Bovine Serum Albumin heat shock fraction (Sigma-Aldrich, CAT#A9647-100G)

8. DMSO (Thermo Fisher Scientific)

9. Compounds -10 mM stock in DMSO

10. Victor Nivo multimode plate reader (PerkinElmer)

11. OptiPlate-384, white opaque 384-well microplate (PerkinElmer)

Experimental procedure

Disruption of Raf1-KRAS protein-protein binding of tested compound was determined using a time-resolved fluorescence resonance energy transfer (TR-FRET) techonology. The assay buffer contains 1X LANCE Detection buffer supplemented with 5 mM MgCl₂, 10 μM GppNHp and 2.5 μg/mL BSA. His-KRas (GppNHp-loaded) was pre-incubated with compound in an 8-point 4-fold dilution series starting at a final concentration of 10 μM. After incubation at RT for 20 hour, a mixture of FLAG-RAF1 RBD, anti-His Eu-W1024 and anti-FLAG allophycocyanin was then added to assay sample wells at final concentrations of 50 nM, 2nM and 100 nM, respectively. The reaction was incubated for an additional 1 hour or 20 hours (for covalent compounds) at RT. The TR-FRET signal was read on a plate reader with an excitation wavelength of 340 nm and detection wavelengths of 615 and 665 nm. The binding disruption curve was fitting using the sigmoidal dose-response curve with a variable slope model in GraphPad Prism. Compounds that facilitate disruption of a Raf1-KRas complex were identified as those eliciting a decrease in the TR-FRET ratio relative to DMSO control wells and IC₅₀ values (nM) are shown in the following Table 1.

Table 1. IC₅₀ values (nM) of the compounds in Raf1 disruption TR-FRET.

Biological Example 2: Mouse Pharmacokinetic Evaluation Experiment

Experiment Purpose:

To determine the drug concentrations in plasma at different times after intravenous and intragastric administration of the test compounds to the mice by using the LC/MS/MS method and using male C57BL/6 mice as the test animals; To study the pharmacokinetic behaviors of the test compounds in mice and to evaluate the pharmacokinetic characteristics thereof.

Experiment scheme: 36 healthy adult male C57BL/6 mice were used as experimental animals and divided into 4 groups according to the principle of similar body weight, with 9 in each group of the group IV (two groups) and 9 in each group of the group PO (two groups) . The animals were purchased from Jihui Laboratory Animal Co. LTD

Drug Preparation: Preparation of IV and PO dosing solutions in 10%DMSO+40%PEG300+5%Tween 80+45%Saline at 1 mg/mL

Administration: After fasting overnight, each rat in the group IV was administered intravenously with a volume of 1 mL/kg and a dose of 1 mg/kg; each mouse in the group PO was administered oral gavage with a volume of 5 mL/kg and a dose of 5 mg/kg.

Experiment Operation: After each of the male C57BL/6 mice in the group IV was administrated with the test compounds, 100 ul of blood samples were collected at 0.0833, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours, and were placed in a commercial anticoagulation tube pre-loaded with EDTA-K 2. After each rat in the group PO was administrated with the test compounds, 100 ul of blood samples were collected at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours respectively, and were placed in a commercial anticoagulation tube pre-loaded with EDTA-K 2. The test tube was centrifuged for 15 minutes to separate the plasma and stored at -70℃. 2 hours after administration, the animals can be fed. The content of the test compounds in plasma was determined by the LC/MS/MS method after intravenous and intragastric administration to the rats. The linear range of the method was from 2.00 to 6000 nM; plasma samples were analyzed after treatment of precipitating proteins by acetonitrile.

Experiment Results: Experiment results were shown in Table 2.

Experiment Conclusion: In the rat pharmacokinetic evaluation experiment, the compounds of the present invention show higher exposure and better oral availability than the reference compound RM-018.

Table 2: Evaluation of Experiment Results in Rat Pharmacokinetic

Note: Cl: clearance rate; AUC: exposure; T 1/2: half-life; C max: maximum compound concentration after oral administration; F: bioavailability

Groups RM-018 and Example A

Example A is a compound chosen from the novel Examples characterized above

Biological Example 3: CypA-Compound-KRAS Tri-complex Formation TR-FRET Assay by Compounds of the Invention (Tri-complex formation TR-FRET)

Material and Reagents

12. Avi-tagged, biotinylated recombinant human KRAS G12C (Biotin-KRAS^G12C, a.a. 1-164, C-terminal biotinylated, GppNHp-loaded, produced in-house)

13. His-Cyclophilin A recombinant human (His-CypA, a.a. 1-165, N-terminal His-tagged, produced in-house)

14. LANCE Detection Buffer (10x) (PerkinElmer, CAT#CR97-100)

15. LanthaScreen Eu-streptavidin (Invitrogen, CAT#PV5899)

16. SureLight Allophycocyanin-anti-6His antibody (PerkinElmer, CAT#AD0059H)

17. GppNHP (Non-hydrolyzable GTP analog) (Abcam, CAT#ab146659)

18. Bovine Serum Albumin heat shock fraction (Sigma-Aldrich, CAT#A9647-100G)

19. MgCl₂ (Sigma-Aldrich, CAT#M0250-500G)

20. DMSO (Thermo Fisher Scientific)

21. Compounds -10 mM stock in DMSO

22. Victor Nivo multimode plate reader (PerkinElmer)

23. OptiPlate-384, white opaque 384-well microplate (PerkinElmer)

Experimental procedure

The formation of CypA-compound-KRAS^G12C ternary complex was determined using a time-resolved fluorescence resonance energy transfer (TR-FRET) techonology. The assay buffer contains 1X LANCE Detection buffer supplemented with 5 mM MgCl₂, 10 μM GppNHp and 2.5 μg/mL BSA. Biotin-KRas^G12C (GppNHp-loaded, final concentration 30 nM) was incubated with his-CypA (final concentration of 300 nM) in the presence of tested compound in an 8-point 5-fold dilution series starting at a final concentration of 10 μM at RT for 1 hour. Then a mixture of Eu-streptavidin and allophycocyanin anti-His Ab was added to assay sample wells at final concentrations of 2nM and 100 nM, respectively. The reaction was incubated for an additional 1 hour or 19 hours (for covalent compounds) at RT. The TR-FRET signal was read on a plate reader with an excitation wavelength of 340 nm and detection wavelengths of 615 and 665 nm. The binding curve was fitting using the four-parameter sigmoidal dose-response curve with a variable slope model in GraphPad Prism. Compounds that facilitate ternary complex formation were identified as those eliciting a increase in the TR-FRET ratio relative to DMSO control wells and EC₅₀ values (nM) are shown in the following Table 3.

Table 3. EC₅₀ values (nM) of the compounds in tri-complex formation TR-FRET.

Conclusion: The compounds of the present invention disrupted the protein-protein binding between Raf1 RBD and KRAS.

Claims

A compound of formula (I) , formula (II) or formula (III) :

or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein,

ring A and ring B are each independently selected from the group consisting of cycloalkyl, heterocyclyl, aryl and heteroaryl;

R₁ is selected from the group consisting of cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein each of the cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, alkoxyalkyl, hydroxyl, hydroxyalkyl, haloalkyl, C_3-12 cycloalkyl, C_3-12 heterocyclyl, wherein the C_3-12 heterocyclyl is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl, heterocyclyl;

L and M are each independently selected from the group consisting of alkyl, alkoxy, alkyl-O-C (O) -, alkoxy-O-C (O) -, -alkyl-cycloalkyl-alkyl-O-C (O) -;

R₂, R₃, R₄, R₅, R₆, R₇ and R₈ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl, cycloalkyl and heterocyclyl;

or, R₄ and R₅ together with the carbon atom to which they are bound form cycloalkyl, heterocyclyl, wherein each of the cycloalkyl, heterocyclyl is optionally substituted with one or more substituents selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl;

n is 1, 2 or 3;

G iswherein, R_a and R_b are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl;

or, R_a and R_b together with the nitrogen atom to which they are bound form heterocyclyl and heteroaryl;

K is selected from the group consisting of vinyl ketone, vinyl sulfone, ynone, alkynyl sulfone, -S (O) ₂ and alkyl, -C (O) -C (O) -alkyl;

J is selected from the group consisting of cycloalkyl, heterocyclyl, aryl, heteroaryl and NR_cR_d,

or J is absent with the proviso that K is selected from alkyl, -C (O) -C (O) -alkyl;

R_c and R_d are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkenyl, alkynyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl.
The compound of claim 1, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, being a compound of formula (Ia) , formula (IIa) or formula (IIIa) :

wherein,

R₁₃ is selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl, heteroaryl;

m is 1, 2 or 3;

preferably, R₁₃ is selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-3 alkyl, C_1-3 alkoxy, C_1-3 hydroxyl, C_1-3 hydroxyalkyl, C_1-3 haloalkyl; m is 1.
The compound of anyone of claims 1 to 2, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, being a compound of formula (Ib) , formula (IIb) or formula (IIIb) :

wherein,

X₁ is bond, C or N;

X₂ is C, O, N;

R₉, R₁₀, R₁₁ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, alkyl, alkoxy, hydroxyl, hydroxyalkyl, haloalkyl, cycloalkyl and heterocyclyl.
The compound of anyone of claim 1 to 3, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein, ring A is selected from the group consisting of C_5-10 cycloalkyl, C_5-10 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-12 aryl and C_5-10 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S; and ring B are each independently selected from the group consisting of C_3-10 cycloalkyl, C_5-10 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-12 aryl and C_5-10 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S;

more preferably, ring B is selected from the group consisting of C_5-10 bridged cycloalkyl, C_5-10 bridged heterocyclyl comprising 1, 2 or 3 heteroatoms selected from N, O, S, C_5-10 fused heterocyclyl comprising 1, 2 or 3 heteroatoms selected from N, O, S;

more preferably, ring A and ring B are each independently selected from the group consisting of:

more preferably, ring A is selected from the group consisting of

ring B is selected from the group consisting of phenyl,

more preferably, ring B is selected from the group consisting of phenyl or

preferably, ring A is 7-12 membered fused heteraryl comprising 1, 2 or 3 heteroatoms selected from N, O, S; ring B is 7-12 membered fused heterocyclyl comprising 1, 2 or 3 heteroatoms selected from N, O, S;

more preferably, ring A isring B is selected from the group consisting of

preferably, ring B is selected from the group consisting of
The compound of anyone of claims 1 to 3, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein, R₁ is selected from the group consisting of C_3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-12 aryl and C_5-12 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S, wherein each of the C_3-12 cycloalkyl, C_5-12 heterocyclyl, C_3-12 aryl and C_5-12 heteroaryl is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkoxy, C_1-6 alkoxyalkyl, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-10 cycloalkyl, C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_7-10 bridged heterocyclyl containing 1 to 3 heteroatoms independently selected from N, O or S, wherein each of the C_3-6 heterocyclyl and the C_7-10 bridged heterocyclyl is optionally substituted with one or more substituents selected from C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_3-5 heterocyclyl.

R₁ is selected from the group consisting of C_5-8 cycloalkyl, C_5-8 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_5-8 aryl and C_5- ₈ heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S, wherein each of the C_5-8 cycloalkyl, C_5-8 heterocyclyl, C_5-8 aryl and C_5-8 heteroaryl is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, C_1-4 alkoxyalkyl, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl, C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, C_7-10 bridged heterocyclyl containing 1 to 3 heteroatoms independently selected from N or O, wherein each of the C_3-6 heterocyclyl and the C_7-10 bridged heterocyclyl is optionally substituted with one or more substituents selected from C_1-6 alkyl, C_3-5 heteroaryl containing a heteroatom selected from O.

R₁ is selected from the group consisting of phenyl, pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, wherein each of the phenyl, pyridinyl, pyrimidinyl, pyrazolyl and imidazolyl is optionally substituted with one or two substituents selected from hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, C_1-4 alkoxyalkyl, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl,
The compound of anyone of claims 1 to 3, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein, L and M are each independently selected from the group consisting of bond, C_1-8 alkyl, C_1-8 alkoxy, C_1-8 alkyl-O-C (O) -, C_1-8 alkoxy-O-C (O) -, -C_1-6 alkyl-C_3-6 cycloalkyl-C_1-6 alkyl-O-C (O) -;

preferably, L is independently selected from the group consisting of linear or branched C_3-6 alkyl, linear or branched C_3-6 alkoxy, linear or branched C_3-6 alkyl-O-C (O) -, linear or branched C_3-6 alkoxy-O-C (O) -, -C_1-3 alkyl-C_3-6 cycloalkyl-C_1-3 alkyl-O-C (O) -, linear or branched C_1-6 alkyl; M is independently selected from the group consisting of bond, C_1-3 alkyl, branched or linear C_1-3 alkoxy.
The compound of anyone of claims 1 to 3, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof,

wherein,

R₂, R₃, R₄, R₅, R₆, R₇, R₈ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-12 cycloalkyl and C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S;

or, R₄ and R₅ together with the carbon atom to which they are bound form C_3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, wherein each of the C_3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S is optionally substituted with one or more substituents selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl;

preferably, R₂, R₃, R₄, R₅, R₆, R₇, R₈ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl, C_3-6 cycloalkyl and C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S;

or, R₄ and R₅ together with the carbon atom to which they are bound form C_3-6 cycloalkyl, C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, wherein each of the C_3-6 cycloalkyl, C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S is optionally substituted with one or two substituents selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl.
The compound of claim 3, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein, R₉, R₁₀, R₁₁ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-12 cycloalkyl and C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S;

preferably, R₉, R₁₀, R₁₁ are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl, C_3-6 cycloalkyl and C_3-6 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S;

more preferably, R₁₁ is each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkenyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-8 cycloalkyl and C_3-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, wherein the C_3-8 cycloalkyl and C_3-8 heterocyclyl are optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, C_1-6 alkyl; preferably, the C_3-12 heterocyclyl is C₃-₇ mono-heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, or fused C_8-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S;
The compound of anyone of claims1 to 2, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein, n is 1.
The compound of claim 1, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof,

wherein,

G iswherein, R_a and R_b are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl;

or, R_a and R_b together with the carbon atom to which they are bound form C_3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-12 aryl and C_3-12 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S.

G iswherein, R_a and R_b are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl;

or, R_a and R_b together with the nitrogen atom to which they are bound form a C_3-8 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S and C_3-8 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S.

preferably, G is selected from the group consisting of pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, piperidyl, piperazinyl, pyrrolidyl, oxazolidinyl, thiazolidinyl.
The compound of claim 1, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein, K is selected from the group consisting of C_2-6 vinyl ketone, C_2-6 vinyl sulfone, C_3-6 ynone, C_2-6 alkynyl sulfone, C_1-3 alkyl, -C (O) -C (O) -C_1-3 alkyl.

preferably, K is selected from the group consisting of C_3-6 vinyl ketone, C_3-6 vinyl sulfone, C_4-6 ynone, C_3-6 alkynyl sulfone, methyl, ethyl, propyl, -C (O) -C (O) -methyl, -C (O) -C (O) -ethyl, -C (O) -C (O) -propyl.

more preferably, K iswherein, R₁₂ is C_1-6 alkyl;

more preferably, R₁₂ is C_1-3 alkyl.
The compound of claim 1, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein, J is selected from the group consisting of C_3-12 cycloalkyl, C_3-12 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-12 aryl, C_3-12 heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S and NR_cR_d;

R_c and R_d are each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-3 alkyl, C_1-3 alkenyl, C_1-3 alkynyl, C_1-3 alkoxy, hydroxyl, C_1-3 hydroxyalkyl, C_1-3 haloalkyl;

or J is absent, with the proviso that K is C_1-3 alkyl.

preferably, J is selected from the group consisting of C_3-8 cycloalkyl, C_3-8 heterocyclyl containing 1 to 2 heteroatoms independently selected from N, O or S, C_3-8 aryl and C_3- ₈ heteroaryl containing 1 to 2 heteroatoms independently selected from N, O or S.

more preferably, J is selected from the group consisting of pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, piperidyl, piperazinyl, pyrrolidyl, oxazolidinyl, thiazolidinyl, morpholinyl, thiomorpholinyl.
The compound of anyone of claims 1-12, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof

wherein,

X₂ is N, CH;

R₁₁ is each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkenyl, C_1-6 alkoxy, C_1-6 alkoxyalkyl, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-8 cycloalkyl and C_3-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, wherein the C_3-8 cycloalkyl and C_3-8 heterocyclyl are optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, C_1-6 alkyl;

preferably, the C_3-12 heterocyclyl is C₃-₇ mono-heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, or fused C_8-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S;

R₁₀ is ndependently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl;

R₃ is independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl;

or two of R₃ together with the atoms of ring B to which they are attached form a bridged C_3-8 cycloalkyl;

preferably, ring B is a C_5-10 fused heterocyclyl comprising 1, 2 or 3 heteroatoms selected from N, O, S; R₃ is independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl; r is 0, 1, 2 or 3;

s is 0, 1, 2 or 3; .
The compound of anyone of claims 1-13, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, being a compound of formula (IIIc) or (IIId) :

wherein,

X₂ is N, CH;

X₃ is N, CH and S;

R₁₁ is each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkenyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-8 cycloalkyl and C_3-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, wherein the C_3-8 cycloalkyl and C_3-8 heterocyclyl are optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, C_1-6 alkyl;

preferably, the C_3-12 heterocyclyl is C₃-₇ mono-heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, or fused C_8-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S;

R₁₀ is ndependently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl;

R₃ is independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl;

or two of R₃ together with ring B to which they are attached form a bridged C_3-8 cycloalkyl;

R₁₃ is selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-3 alkyl, C_1-3 alkoxy, C_1-3 hydroxyl, C_1-3 hydroxyalkyl, C_1-3 haloalkyl; m is 1.

r is 0, 1, 2 or 3;

s is 0, 1, 2 or 3.
The compound of anyone of claims 1-16, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, being a compound of formula (Id)

wherein,

X₂ is N, CH;

X₃ is N, CH and S;

R₁₁ is each independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-6 alkyl, C_1-6 alkenyl, C_1-6 alkoxy, hydroxyl, C_1-6 hydroxyalkyl, C_1-6 haloalkyl, C_3-8 cycloalkyl and C_3-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, wherein the C_3-8 cycloalkyl and C_3-8 heterocyclyl are optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, C_1-6 alkyl;

preferably, the C_3-12 heterocyclyl is C₃-₇ mono-heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S, or fused C_8-12 heterocyclyl containing 1, 2 or 3 heteroatoms independently selected from N, O or S;

R₁₀ is ndependently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl;

R₃ is independently selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, C_1-4 alkyl, C_1-4 alkoxy, hydroxyl, C_1-4 hydroxyalkyl, C_1-4 haloalkyl;

or two of R₃ together with ring B to which they are attached form a bridged C_3-8 cycloalkyl;

r is 0, 1, 2 or 3;

s is 0, 1, 2 or 3.
The compound of anyone of claims 1-17, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein,

R₁₁ is selected from the group consisting of linear or branched C_1-3 alkoxy, C_1-4 alkoxyalkyl, C_2-4 haloalkenyl, C_5-8 mono heterocyclyl containing 1, 2 or 3 heteroatoms selected from N, O, S, C_5-12 fused heterocyclyl containing 1, 2 or 3 heteroatoms selected from N, O, S, wherein the mono heterocyclyl and the fused heterocyclyl are optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, C_1-3 alkyl.

preferably, R₁₁ is selected from the group consisting of branched C_1-3 alkoxy, C_1-4 alkoxyalkyl, C_2-4 fluorine-alkenyl,

r is 1 or 2;
The compound of anyone of claims 1-18, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein, R₁₃ is selected from the group consisting of C_5-7 heteroaryl comprising 1, 2 or 3 heteroatoms selected from N, O or S;

preferably, R₁₃ is selected from the group consisting of pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, piperidyl, piperazinyl, pyrrolidyl, oxazolidinyl, thiazolidinyl, morpholinyl, thiomorpholinyl.
The compound of anyone of claims 1-17, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein the compound is:
A pharmaceutical composition comprising a therapeutically effective amount of the compound of claims 1-23, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt solvate, or prodrug thereof, and a pharmaceutically acceptable carrier.
A method for treating disease mediated by RAS mutation, comprising administering to a subject in need thereof an effective amount of a compound of any one of claims 1-23, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt solvate, or prodrug thereof, or the pharmaceutical composition of claim 19.
The method of claim 20, wherein the RAS mutation contain KRAS G12C, KRAS G12D, KRAS G12V, KRAS G13D, KRAS G12 R, HRAS or NRAS mutation.
A method for treating cancer comprising administering to a subject in need thereof an effective amount of a compound of any one of claims 1-18, or a tautomer, cis-or trans-isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt solvate, or prodrug thereof, or the pharmaceutical composition of claim 19, in which the disease or disorder is selected from the group consisting of Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma) , myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma) , alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma) , stomach (carcinoma, lymphoma, leiomyosarcoma) , pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma) , small bowel (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma) , large bowel (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma) ; Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor (nephroblastoma) , lymphoma, leukemia) , bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma) , prostate (adenocarcinoma, sarcoma) , testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma) ; Liver: hepatoma (hepatocellular carcinoma) , cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: gall bladder carcinoma, ampullary carcinoma, cholangiocarcinoma; Bone: osteogenic sarcoma (osteosarcoma) , fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma) , multiple myeloma, malignant giant cell tumor chordoma, osteochronfroma (osteocartilaginous exostoses) , benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans) , meninges (meningioma, meningiosarcoma, gliomatosis) , brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma) , glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors) , spinal cord neurofibroma, meningioma, glioma, sarcoma) ; Gynecological: uterus (endometrial 'carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma) , granulosa-thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma) , vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma) , vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma) , fallopian tubes (carcinoma) ; Hematologic: blood (myeloid leukemia (acute and chronic) , acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome) , Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma) ; Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma.