Methods for preparing KRAS G12C inhibitors imidazotriazine and pyrrolopyrimidine derivatives

Abstract

The present application discloses a method for preparing G12C inhibitors, imidazotriazine and pyrrolopyrimidine derivatives or stereoisomers thereof or pharmaceutically acceptable salts thereof, wherein the imidazotriazine and pyrrolopyrimidine derivatives or stereoisomers thereof or pharmaceutically acceptable salts thereof are used as G12C inhibitors for treating cancer.

Description

Technical Field

[0001] The present application discloses a method for preparing G12C inhibitors, imidazotriazine and pyrrolopyrimidine derivatives or stereoisomers thereof or pharmaceutically acceptable salts thereof, wherein the imidazotriazine and pyrrolopyrimidine derivatives or stereoisomers thereof or pharmaceutically acceptable salts thereof are used as G12C inhibitors for treating cancer. Background Art

[0002] RAS is one of the most well-known oncogenes. In humans, three RAS genes (HRAS, KRAS, and NRAS) encode four highly homologous RAS proteins (HRAS, KRAS-4A, KRAS-4B, and NRAS). RAS proteins are small GTPases that function as binary molecular switches and are involved in extracellular growth and differentiation signaling.

[0003] RAS normally cycles between a GDP-bound "off" state and a GTP-bound "on" state. This cycle is regulated by several factors. Guanine nucleotide exchange factors (GEFs), including SOS1 and SOS2, promote the exchange and formation of GTP-bound RAS. At the same time, GTPase activating proteins (GAPs), such as NF-1, promote GTP hydrolysis, thereby returning RAS to an inactive state bound to GDP (Kessler et al., PNAS, 2019, 116(32): 15823-15829). Once bound to GTP, RAS begins conformational changes in two specific regions, Switch 1 and Switch 2, allowing the binding and activation of downstream effector proteins to initiate a cascade of intracellular signaling pathways. These effectors include the RAF-MEK-ERK and PI3K-AKT-mTOR pathways, both of which play important roles in regulating cell proliferation, differentiation, and survival (Cox et al., Nature Reviews Drug Discovery, 2014, 13:828-851).

[0004] RAS mutations have been identified in approximately 30% of human tumors. These mutations often occur as single-base missense mutations in codons 12, 13, or 61, resulting in stabilization of the activated GTP-bound RAS form and constitutive activation of RAS downstream signaling pathways. KRAS is the most commonly mutated RAS in cancer, accounting for 85% of all RAS-driven cancers, followed by NRAS (12%) and HRAS (3%). KRAS mutations have been detected in approximately 95% of pancreatic ductal adenocarcinomas, 50% of colorectal adenocarcinomas, and 30% of lung adenocarcinomas. Most KRAS mutations occur at residue 12, and the mutation type varies in different cancers. In colon and pancreatic cancer, the predominant KRAS mutation is G12D (glycine to lysine), whereas in non-small cell lung cancer (NSCLC), almost half of KRAS mutations are G12C (glycine to cysteine) (Cox et al., Nature Reviews Drug Discovery, 2014, 13:828-851).

[0005] Based on the key role of RAS in cell proliferation and its high mutation rate in human cancer, RAS has long been regarded as a therapeutic target for many cancers. However, despite decades of research efforts, no anti-RAS small molecules have been clinically approved. The main reason is the lack of druggable pockets on the surface of RAS (Papke et al., Science, 2017, 355: 1158–1163). Recently, more and more studies have shown that RAS may be treated with small molecules. Several inhibitors directly targeting KRAS G12C are under study (Patricelli et al., Cancer Discovery, 2016, 6 (3); 316–29) (Fell et al., ACS Med. Chem. Lett. 2018, 9, 12, 1230-1234).

[0006] Small molecule selective inhibitors of KRAS are being developed to prevent or treat diseases, for example, WO2015 / 054572A1 provides compounds having activity as inhibitors of G12C mutant RAS proteins. WO2016 / 164675A1 and WO2017 / 015562A1 disclose substituted quinazoline compounds as KRAS G12C inhibitors. WO2014 / 152588A1, WO2016 / 049524A1, WO2016 / 168540A1, WO2017 / 058728A1, WO2017 / 058792A1, WO2017 / 058805A1, WO2017 / 058915A1, WO2017 / 087 528A1, WO2018 / 064510A1, WO2018 / 068017A1, WO2018 / 119183A2, WO2018 / 206539A1, WO2018 / 218069A1, WO2019 / 051291A1, WO2019 / 055540A1, WO201 9 / 137985A1, WO2019 / 141250A1, WO2019 / 150305A1, WO2019 / 155399A1, WO2019 / 213516A1, WO2019 / 213526A1, WO2019 / 215203A1, WO20192 / 17307A1, WO2019 / 217691A1, WO2019 / 232419A1, WO2020 / 028706A1, WO2020 / 027084A1, WO2020 / 027083A1, WO2020 / 035031A1, and WO2020 / 047192A1 further report compounds with KRAS G12C inhibitory activity.

[0007] Therefore, there is still a great need for new inhibitors that selectively target mutant KRAS with high efficacy and safety and methods for their preparation. Continued efforts to develop KRAS G12C inhibitors will suggest new treatment modalities for KRAS G12C-driven cancers. Summary of the invention

[0008] The present invention relates to the following aspects:

[0009] Item 1. A method for preparing a compound of formula (I) or (II),

[0010] The steps include:

[0011]

[0012] L1 and L2 are independently selected from a single bond, -CO-, -CO-NH-, -NH-CO-, -O-, -NR a -、-NR a (CH2) m -、-(CH2) m -、-O-(CH2) m -、-O-CH(R a )-、-CH(R a )-、-CH(R a )(CH2) m -, and -(CH2) m -O-;

[0013] R 1 is selected from cycloalkyl, heterocyclic, aryl, or heteroaryl, wherein the cycloalkyl, heterocyclic, aryl, or heteroaryl is optionally composed of at least one R 6 Substituent substitution (e.g. –(R 6 ) q1 , when q1 is greater than 1, each R 6 are the same or different substituents);

[0014] R 2 Selected from -NR b R c , cycloalkyl, heterocyclic, aryl, heteroaryl, the -NR b R c , cycloalkyl, heterocyclyl, aryl, heteroaryl, optionally with at least one R 6 Substituent substitution (e.g. –(R 6 ) q2 , when q2 is greater than 1, each R 6 are the same or different substituents);

[0015] R 6 Selected from -C 1-8 Alkyl, halogen, hydroxy, oxo, -C 1-8 Alkoxy, -NR b R c , cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein -C 1-8 Alkyl, -C 1-8 The alkoxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with at least one halogen, hydroxy, amino, CN, cycloalkyl, heterocyclyl, aryl, or heteroaryl;

[0016] Or when R 6 When connected to two adjacent atoms in the ring, the two R 6Together with the carbon atom to which it is attached, it forms a 5-8 membered ring, wherein the 5-8 membered ring contains 0, 1 or 2 heteroatoms selected from N, O or S which is optionally oxidized;

[0017] R 3 Selected from hydrogen, halogen, -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 Alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, the -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 Alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted by halogen, hydroxyl, -C 1-8 alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl;

[0018] R 3 ' is selected from hydrogen, -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 Alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, the -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 Alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted by halogen, hydroxyl, -C 1-8 Alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl; R 4 Selected from

[0019] R 5 Selected from hydrogen, halogen, -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 Alkynyl, oxo, -NR b R c 、-(CH2) m -C(O)-NR d R e , cycloalkyl, heterocyclyl, aryl, heteroaryl or -(CH2) m -CN;

[0020] Each R a , R b and R c are independently selected from hydrogen, deuterium, cyano, halogen, hydroxyl, -C 1-8 Alkoxy, -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 Alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NR d R e or -CO-NR dR e , the -C 1-8 Alkoxy, -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 Alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is optionally substituted by R f Replace, or

[0021] (R a and R b )、(R a and R c )、(R b and R c ) together with the atoms to which they are attached form a 4-6 membered ring, wherein the 4-6 membered ring is optionally substituted with at least one R g replace;

[0022] Each R f Selected from halogen, hydroxy, oxo, -C 1-8 Alkoxy, -NR d R e 、-CO-NR d R e 、-NR d -CO-R e , cycloalkyl, heterocyclyl, aryl or heteroaryl, the -C 1-8 The alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl groups are optionally substituted with halogen, hydroxyl or -C 1-4 Alkyl substitution;

[0023] R d , R e and R g Each independently selected from hydrogen, deuterium, halogen, oxo, or -C 1-8 Alkyl, the -C 1-8 The alkyl group is optionally substituted with at least one halogen, oxo, -CF3 or -COCH3;

[0024] PR is selected from Boc or Cbz;

[0025] p is independently selected from 0, 1, 2, 3 or 4;

[0026] q1 and q2 are independently selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8;

[0027] Each m and n is independently selected from 0, 1, 2, 3, 4, 5 or 6;

[0028] R 7 is selected from H or Br;

[0029] Selected from

[0030] Selected from

[0031] Item 2. The preparation method according to Item 1, characterized in that:

[0032] The reagents used in step 1 are phosphorus oxychloride, phosphorus trichloride, phosphorus pentachloride, Lucas reagent, a mixture of concentrated hydrochloric acid and anhydrous zinc chloride, Vilesmier-Haack reagent, thionyl chloride, sulfonyl chloride, chlorine, preferably POCl3, PCl5, SOCl2, more preferably POCl3.

[0033] Item 3. The preparation method according to Item 1, characterized in that:

[0034] The reagent used in step 2 is a nitrogen-containing organic base, preferably triethylamine, diethylamine, diisopropylethylamine, pyridine, methylimidazole, piperidine, morpholine, indoline, quinoline, isoquinoline, dimethylaminopyridine, more preferably triethylamine, diethylamine, diisopropylethylamine, dimethylaminopyridine, and further more preferably diisopropylethylamine.

[0035] Item 4. The preparation method according to Item 1, characterized in that:

[0036] In step 3, 1 The reagents for the CHO reaction are alkali metal salts of alcohols, alkyl lithium compounds, and lithium amide compounds, preferably sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide, potassium tert-butoxide, n-butyl lithium, sec-butyl lithium, tert-butyl lithium, lithium diisopropylamide, and lithium hexamethyldisilazide, and more preferably n-butyl lithium.

[0037] Item 5. The preparation method according to Item 1, characterized in that:

[0038] In step 3, 2 The reagents for the -L2-H reaction are alkali metal hydrides, alkali metal salts, alkyl lithium compounds, and lithium amide compounds, preferably sodium hydride, potassium hydride, lithium hydride, sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide, potassium tert-butoxide, n-butyl lithium, sec-butyl lithium, tert-butyl lithium, lithium diisopropylamide, and lithium hexamethyldisilazide, and more preferably sodium hydride.

[0039] Item 6. The preparation method according to Item 1, characterized in that:

[0040] The deprotection conditions in step 4 are catalytic hydrogenation or acidic deprotection, preferably a combination of trifluoroacetic acid and triethylsilane, a combination of BBr3, Pd / C and hydrogen, a combination of trifluoroacetic acid, hydrochloric acid, Pd / C and cyclohexadiene, and more preferably a combination of trifluoroacetic acid and triethylsilane, a combination of BBr3, Pd / C and hydrogen.

[0041] Item 7. The preparation method according to Item 1, characterized in that:

[0042] The reaction conditions with R4Cl in step 5 are an inorganic base, a nitrogen-containing organic base, an alkali metal hydride, preferably sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, triethylamine, diethylamine, diisopropylethylamine, pyridine, methylimidazole, piperidine, morpholine, indoline, quinoline, isoquinoline, dimethylaminopyridine, and more preferably sodium bicarbonate.

[0043] Item 8. The preparation method according to Item 1, characterized in that:

[0044] L1 is selected from a single bond, -CO-, -(CH2) m -、-CH(R a )-、-CH(R a )(CH2) m -、-(CH2) m -O-, further preferably selected from -(CH2) m -、-CH(R a )-; and / or

[0045] L2 is selected from a single bond, -O-, -O-(CH2) m -、-O-CH(R a )-、-O-CH(R a )-(CH2) m -; and / or

[0046] R 1 Selected from and / or

[0047] R 2 Selected from -NR b R c , and / or

[0048] R 3 Selected from hydrogen, halogen or -C 1-8 Alkyl; and / or

[0049] R 3 ' is selected from hydrogen, -C 1-8 Alkyl, -C 2-8 Alkenyl or -C 2-8 Alkynyl; and / or

[0050] R 4 Selected from and / or

[0051] R 5 Selected from hydrogen, -CH3, -C2H5, -C3H7, -C4H9, -C5H 11 、-(CH2) m -C(O)-NR d R e 、-(CH2) m -CN.

[0052] Item 9. The preparation method according to Item 1, characterized in that:

[0053] R 1 Selected from

[0054] and / or

[0055] R 2 Selected from (For example ), (For example ),

[0056] Item 10. The preparation method according to Item 1, characterized in that:

[0057] The compound of formula (I) or (II) is

[0058]

[0059] Example

[0060] The following examples are intended to be illustrative only and should not be construed as limiting in any way. Unless otherwise stated, the experimental methods in the following examples are conventional methods. Unless otherwise stated, reagents and materials are commercially available. All solvents and chemicals used are of analytical grade or chemical purity. Solvents are redistilled before use. Anhydrous solvents are prepared according to standard methods or reference methods. Silica gel (100-200 mesh) for column chromatography and silica gel (GF254) for thin layer chromatography (TLC) are commercially available from Qingdao Ocean Chemical Co., Ltd. or Yantai Chemical Co., Ltd. in China; unless otherwise stated, all silica gel is eluted with petroleum ether (60-90°C) / ethyl acetate (v / v) and developed with a solution of iodine or phosphomolybdic acid in ethanol. Unless otherwise stated, all extraction solvents are dried over anhydrous Na2SO4. Recorded on a Bruck-400 nuclear magnetic resonance spectrometer with TMS (tetramethylsilane) as an internal standard 1H NMR spectra. LC / MS data were recorded using an Agilent 1100 high performance liquid chromatograph-ion trap mass spectrometer (LC-MSD trap) equipped with a diode array detector (DAD) detecting at 214 nm and 254 nm and an ion trap (ESI source). All compound names except reagents are generate.

[0061] In the following examples, the following abbreviations are used:

[0062] Ac Acetyl

[0063] AcOH Acetic acid

[0064] Aq Water-based

[0065] Brine saturated sodium chloride aqueous solution

[0066] Bn Benzyl

[0067] Boc tert-Butyloxycarbonyl

[0068] Cbz benzyloxycarbonyl

[0069] DMF N,N-Dimethylformamide

[0070] Dppf 1,1"-Bis(diphenylphosphino)ferrocene

[0071] DBU 1,8-diazabicyclo[5.4.0]undec-7-ene

[0072] DCE 1,2-Dichloroethane

[0073] DCM Dichloromethane

[0074] DIEA or DIPEA N,N-diisopropylethylamine

[0075] DMAP 4-N,N-dimethylaminopyridine

[0076] DMF N,N-Dimethylformamide

[0077] DMSO Dimethyl sulfoxide

[0078] EA or EtOAc Ethyl acetate

[0079] eq equivalent

[0080] g

[0081] h or hr hour

[0082] HCl

[0083] HPLC High Performance Liquid Chromatography

[0084] IPA 2-Propanol

[0085] i-PrOH Isopropyl alcohol

[0086] mg milligram

[0087] mL milliliters

[0088] Mmol millimole

[0089] MeCN Acetonitrile

[0090] MeOH Methanol

[0091] Min Minutes

[0092] ms or MS mass spectrometry

[0093] NMR Nuclear Magnetic Resonance

[0094] Pd / C Palladium Carbon

[0095] PE Petroleum Ether

[0096] PMB 4-methoxybenzyl

[0097] PPA Polyphosphoric Acid

[0098] Rt., RT. or rt. Room temperature

[0099] Ru-Phos / Ru-PHOS 2-Dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl

[0100] SEM 2-Trimethylsilylethoxymethoxy

[0101] TBSCl tert-Butyldimethylsilyl chloride

[0102] TEA Triethanolamine

[0103] TFA Trifluoroacetic acid

[0104] THF Tetrahydrofuran

[0105] THP Tetrahydropyran

[0106] TLC Thin layer chromatography

[0107] synthesis

[0108] Example 1: Compound A1 (S) -1-(4-(2-((1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalen-1-ylmethyl)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazin-1-yl)prop-2-en-1-one

[0109] Step A: 2,4-Dichloroimidazo[2,1-f][1,2,4]triazine

[0110]

[0111] POCl3 (50 g, 328.9 mmol) was added to a solution of imidazo[2,1-f][1,2,4]triazine-2,4(1H,3H)-dione (10 g, 65.8 mmol) in toluene (60 ml) at room temperature. DIEA (25 g, 193 mmol) was then added dropwise at 60°C. The mixture was stirred at 90°C overnight. The solvent was removed under reduced pressure. The residue was used directly in the next step.

[0112] Step B: tert-Butyl 4-(2-chloroimidazo[2,1-f][1,2,4]triazin-4-yl)piperazine-1-carboxylate

[0113]

[0114] Tert-butyl piperazine-1-carboxylate (12 g, 63.8 mmol) was dissolved in THF (60 mL), and K2CO3 (26.4 g, 191.5 mmol) in water (50 ml) was then added. A mixture of 2,4-dichloroimidazo[2,1-f][1,2,4]triazine (12 g, 63.8 mmol) and DIEA (24.7 g, 191.5 mmol) in THF (60 ml) was slowly added. The reaction was then stirred at room temperature for 2 h. The solvent was evaporated and the residue was partitioned between EA (100 mL) and water (100 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The residue was slurried with MeOH overnight to give the target compound (9.3 g, 42%). 1 H NMR (400MHz, CD3Cl) δ7.71 (d, J = 0.9Hz, 1H), 7.54 (s, 1H), 4.87 (s, 2H), 4.04 (s, 2H), 3.61 (s, 4H), 1.50 (s, 9H) ppm. MS:M / e 339(M+1) + .

[0115] Step C: (S)-tert-butyl 4-(2-((1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazine-1-carboxylate

[0116]

[0117] To a solution of NaH (232 mg, 5.8 mmol, 60%) in THF (10 mL) at 0°C was added (S)-(1-methylpyrrolidin-2-yl)methanol (414 mg, 3.6 mmol). After 30 min, a solution of the product of step B (1 g, 2.9 mmol) in THF (5 mL) was added. The reaction was then stirred at 60°C overnight. The mixture was cooled to room temperature, diluted with water (30 mL) and extracted with EtOAc (60 mL x 2). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by combi flash to give the target compound (0.75 g, 62%). 1 H NMR(400MHz,DMSO-d6)δ7.96(d,J=0.9Hz,1H),7.54(d,J=0.9Hz,1H),4.77(s,2H) ,4.21(dd,J=10.5,5.0Hz,1H),4.05(dd,J=10.5,6.2Hz,1H),3.91(s,2H),3.50(s, 4H),2.95(dt,J=8.9,4.3Hz,1H),2.56(dd,J=13.2,6.3Hz,1H),2.35(s,3H),2.19( q,J=8.7Hz,1H),1.95(tt,J=12.0,8.4Hz,1H),1.74–1.54(m,3H),1.43(s,9H)ppm. MS:M / e 418(M+1) + .

[0118] Step D: tert-Butyl 4-(7-(hydroxy(naphthalen-1-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazine-1-carboxylate

[0119]

[0120] To a solution of the product of step C (500 mg, 1.2 mmol) in THF (10 mL) was added n-butyl lithium (1.12 mL, 2.8 mmol, 2.5 M in hexane), maintaining the temperature between -75 and -65 ° C. After 30 min, a mixture of 1-naphthaldehyde (224 mg, 1.44 mmol) in THF (2 mL) was added dropwise. The resulting mixture was stirred at -70 ° C for 1 h, then warmed to room temperature for 1 h. The reaction was quenched with saturated NH4Cl aqueous solution and extracted with EtOAc (60 mL x 2). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by combi flash to obtain the target compound (500 mg, 71%). NMR (400MHz, DMSO-d6) δ8.13–8.09(m,1H),7.97–7.92(m,1H),7.87(d,J=8.1Hz,1H),7.72–7.64(m,1H),7 .55–7.41(m,3H),7.30(d,J=7.0Hz,1H),6.77(d,J=5.1Hz,1H),6.27(d,J=5.2Hz,1H),5.76(s,1H),4.73( d,J=30.5Hz,2H),4.16(dd,J=10.7,4.8Hz,1H),4.03–3.78(m,3H),3.47(s,4H),2.96–2.86(m,1H),2.26( d,J=10.5Hz,3H),2.13(q,J=8.4Hz,1H),1.96–1.74(m,1H),1.71–1.46(m,3H),1.42(d,J=5.6Hz,9H)ppm. MS:M / e 574(M+1) + .

[0121] Step E: (S)-2-((1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalen-1-ylmethyl)-4-(piperazin-1-yl)imidazo[2,1-f][1,2,4]triazine

[0122]

[0123] To the product mixture of step D (100 mg) was added TFA (2 mL) and Et3SiH (2 mL). The reaction was heated at 25°C overnight. The mixture was concentrated to dryness and the residue was used directly in the next step without further purification. MS: M / e 458 (M+1) + .

[0124] Step F: (S)-1-(4-(2-((1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalen-1-ylmethyl)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazin-1-yl)prop-2-en-1-one

[0125]

[0126] To a mixture of the product of step E (50 mg, crude) in CH3CN (3 mL) was added saturated NaHCO3 (0.5 mL) at 0°C, followed by a solution of acryloyl chloride (6 mg, 0.07 mmol) in CH3CN (0.3 mL). The resulting mixture was stirred at room temperature for 30 min. The reaction was diluted with water, extracted with EA (60 mL), washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative TLC to give the target compound (9 mg, 8% over two steps). 1H NMR (400MHz, DMSO-d6) δ8.19(d,J=9.4Hz,1H),7.98–7.90(m,1H),7.83(d,J=8.1Hz,1H),7.52(dd,J =9.1,5.2Hz,2H),7.46–7.36(m,2H),7.31(s,1H),6.84(dd,J=16.7,10.5Hz,1H),6.16(dd,J=16.7,2 .0Hz,1H),5.73(dd,J=10.4,2.2Hz,1H),4.78(s,2H),4.63(s,2H),4.26(s,1H),4.13(s,1H),3.93(s ,2H),3.84–3.63(m,4H),3.02(s,1H),2.67(s,1H),2.37(s,4H),1.94(s,1H),1.74–1.67(m,3H)ppm. MS:M / e 512(M+1) + .

[0127] Example 2: Compound A2: (S)-1-(4-(7-((3-hydroxynaphthalen-1-yl)methyl)-2-((1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazin-1-yl)prop-2-en-1-one

[0128] Step A: tert-Butyl 4-(7-(hydroxy(3-hydroxynaphthalen-1-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazine-1-carboxylate

[0129]

[0130] To a solution of (S)-tert-butyl 4-(2-((1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazine-1-carboxylate (500 mg, 1.2 mmol) in THF (10 mL) was added n-butyl lithium (1.12 mL, 1.8 mmol, 2.5 M in hexanes) dropwise, maintaining the temperature between -75 and -65 °C. After 30 min, a mixture of 3-hydroxy-1-naphthaldehyde (246 mg, 1.44 mmol) in THF (5 mL) was added dropwise. The resulting mixture was stirred at -70 °C for 1 h and then warmed to room temperature for 1 h. The reaction was quenched with saturated NH4Cl solution, extracted with EtOAc, washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by combi flash to give the title compound (200 mg, 28%). MS:M / e 590(M+1) + .

[0131] Step B: (S)-4-((2-((1-methylpyrrolidin-2-yl)methoxy)-4-(piperazin-1-yl)imidazo[2,1-f][1,2,4]triazin-7-yl)methyl)naphthalen-2-ol

[0132]

[0133] To the product mixture of step A (200 mg) was added TFA (2 mL) and Et3SiH (2 mL). The reaction was stirred at 25 °C overnight. The mixture was concentrated to dryness. Water was added to dissolve the residue. The aqueous layer was adjusted to pH 12 with aqueous K2CO3 solution and extracted with EtOAc. The combined organic layers were dried over Na2SO4 and evaporated in vacuo to give a crude product (70 mg). MS: M / e 474 (M+1) + .

[0134] Step C: (S)-1-(4-(7-((3-hydroxynaphthalen-1-yl)methyl)-2-((1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazin-1-yl)prop-2-en-1-one

[0135]

[0136] To a mixture of the product of step B (70 mg, crude) in CH3CN (2 mL) was added saturated NaHCO3 (15 mg) in water (1 ml) at 0°C, followed by a solution of acryloyl chloride (15 mg, 0.16 mmol) in CH3CN (0.5 mL). The resulting mixture was stirred at room temperature for 30 min. The reaction was diluted with water and extracted with EA. The organic phase was washed with brine. The combined organic phases were dried over Na2SO4, filtered and concentrated. The residue was purified by preparative TLC to give the target compound (8 mg, 10% over two steps). 1 H NMR(400MHz,DMSO-d6)δ9.64(s,1H),8.04(d,J=8.4Hz,1H),7.70(d,J=8.1Hz,1H),7 .39(t,J=7.1Hz,2H),7.29(t,J=7.5Hz,1H),7.02(s,1H),6.93–6.76(m,2H),6.17(d, J=16.8Hz,1H),5.74(d,J=12.0Hz,1H),4.81(s,2H),4.57(s,2H),4.35(s,2H),3.95( s,2H),3.83–3.65(m,4H),2.99(s,2H),2.67(s,1H),2.43(s,1H),2.23–1.55(m,6H). MS:M / e 528(M+1) + .

[0137] Example 3: Compound A3: 2-((S)-1-acryloyl-4-(7-(3-methoxynaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazin-2-yl)acetonitrile

[0138] Step A: 7-Bromoimidazo[2,1-f][1,2,4]triazine-2,4(1H,3H)-dione

[0139]

[0140] To a solution of imidazo[2,1-f][1,2,4]triazine-2,4(1H,3H)-dione (30 g, 0.20 mol) in H2O (1.2 L) was added NBS (24.6 g, 0.14 mol) in several portions at below 25°C. The mixture was stirred at room temperature for 1 h. The mixture was filtered. The filtrate was concentrated to remove the solvent. The resulting residue and the previous filter cake were mixed and slurried in MeOH (20 V) and then MeOH:H2O (1:1, 20 V) to give the product (30.4 g, 94%). MS: M / e 231 (M+1) +.

[0141] Step B: 7-Bromo-2,4-dichloroimidazo[2,1-f][1,2,4]triazine

[0142]

[0143] A 350 ml sealed tube was charged with 7-bromoimidazo[2,1-f][1,2,4]triazine-2,4(1H,3H)-dione (10 g, 43 mmol), triethylamine hydrochloride (12 g, 88 mmol) and POCl3 (100 ml). The mixture was stirred at 120°C overnight. The mixture was concentrated to remove POCl3. The residue was diluted with EA (200 ml) and saturated NaHCO3 (aqueous solution) was added dropwise at below 20°C until the pH was above 7. The solution was separated. The organic layer was washed with H2O, dried over Na2SO4, filtered and concentrated. The resulting residue was purified by column chromatography using 0-20% EA in PE to give the product (8.5 g, 73%). MS: M / e 267 (M+1) + .

[0144] Step C: (S)-4-(7-bromo-2-chloroimidazo[2,1-f][1,2,4]triazin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester

[0145]

[0146] (S)-2-(Cyanomethyl)piperazine-1-carboxylic acid benzyl ester hydrochloride (974mg, 3.29mmol) was added to a solution of 7-bromo-2,4-dichloroimidazo[2,1-f][1,2,4]triazine (1g, 3.76mmol), DIEA (1.2g, 9.3mmol) in THF (10ml). The reaction was then stirred at room temperature for 2h. The reaction mixture was diluted in water and extracted with DCM. The combined organic extracts were washed with saturated aqueous NaCl solution, dried over Na2SO4 and concentrated. The crude product was purified by silica gel column chromatography (EA:PE=1:2) to give the target compound (1.4g, 86.8%). 1 H NMR (400MHz, CDCl3) δ7.55(s,1H),7.44–7.32(m,5H),6.03(s,1H),5.24–5.14(m ,2H),4.75(s,2H),4.27(s,1H),3.67–3.16(m,3H),2.83–2.58(m,2H)ppm,MS:M / e 490(M+1) + .

[0147] Step D: (S)-benzyl 4-(7-bromo-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0148]

[0149] To a solution of NaH (74 mg, 1.84 mmol, 60%) in THF (5 mL) at 0°C was added (S)-(1-methylpyrrolidin-2-yl)methanol (169 mg, 1.47 mmol). After 30 min, a solution of the product of step A (600 mg, 1.23 mmol) in THF (5 mL) was added. The reaction was then stirred at 60°C overnight. The mixture was cooled to room temperature, diluted with water and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by combi flash to give the target compound (180 mg, 26%). MS: M / e 569 (M+1) + .

[0150] Step E: (S)-benzyl 2-(cyanomethyl)-4-(7-(3-methoxynaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazine-1-carboxylate

[0151]

[0152] A flask was charged with Pd(dppf)Cl2 (12.4 mg, 0.017 mmol), the product of step B (100 mg, 0.17 mmol), (3-methoxynaphthalen-1-yl)boronic acid (38 mg, 0.19 mmol), Na2CO3 (22 mg, 0.2 mmol), dioxane (3 ml) and H2O (0.3 ml). The resulting mixture was stirred at 90°C for 4 h and then cooled to room temperature. The reaction was quenched with water, extracted with EtOAc, washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by combi flash to give the title compound (90 mg, 79%). MS: M / e 647 (M+1) + .

[0153] Step F: 2-((S)-4-(7-(3-methoxynaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazin-2-yl)acetonitrile

[0154]

[0155] BBr3 (1M in DCM, 0.61 ml) was added dropwise to a mixture of the product of step C (160 mg, 0.25 mmol) in DCM (2 mL) at -10°C. The reaction was stirred at -10°C for 1 hour. The mixture was filtered and the filter cake was dried in an oven. The filter cake (20 mg, crude) was used directly in the next step without further purification. MS: M / e 513 (M+1) + .

[0156] Step G: 2-((S)-1-acryloyl-4-(7-(3-methoxynaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazin-2-yl)acetonitrile

[0157]

[0158] To a mixture of the product of step D (20 mg, crude) in CH3CN (1 mL) was added saturated NaHCO3 (0.5 mL) at 0°C, followed by a solution of acryloyl chloride (4 mg) in CH3CN (0.3 mL). The resulting mixture was stirred at room temperature for 30 min. The reactant was diluted with water, extracted with EA, washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative TLC to give the target compound (1.7 mg, 10% over two steps). 1 H NMR (400MHz, DMSO-d6) δ7.93(d,J=8.2Hz,1H),7.83(s,1H),7.61–7.48(m,3H),7.34(s,2H),6.93–6.85(m,1H),6.23(d,J=16.6Hz ,1H),5.82(d,J=10.4Hz,1H),5.03(m,2H),4.17(s,2H),3.88(s,3H),3.67–3.44(m,5H),3.17–2.76(m,6H),2.07–1.38(m,6H)ppm. MS:M / e 567(M+1) + .

[0159] Example 17: Compound A17: 1-((S)-3-methyl-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalen-1-ylmethyl)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazin-1-yl)prop-2-en-1-one

[0160] Step A: (S)-tert-butyl 4-(7-bromo-2-chloroimidazo[2,1-f][1,2,4]triazin-4-yl)-3-methylpiperazine-1-carboxylate

[0161]

[0162] (S)-tert-Butyl 3-methylpiperazine-1-carboxylate (880 mg, 4.45 mmol) was dissolved in THF (5 mL). A mixture of 7-bromo-2,4-dichloroimidazo[2,1-f][1,2,4]triazine (1 g, 4 mmol) and DIEA (774 mg, 6 mmol) in THF (5 ml) was slowly added. The reaction was then stirred at room temperature for 2 h. The solvent was evaporated and the residue was partitioned between EA (100 mL) and water (100 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel column chromatography (eluted with ethyl acetate / PE, 0-30%) to give the target compound (1.3 g, 81%). MS: M / e 431 (M+1) + .

[0163] Step B: (S)-tert-butyl 4-(7-bromo-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)-3-methylpiperazine-1-carboxylate

[0164]

[0165] To a solution of NaH (60 mg, 1.5 mmol, 60%) in THF (5 mL) was added (S)-(1-methylpyrrolidin-2-yl)methanol (138 mg, 1.2 mmol) at 0°C. After 30 min, a solution of the product of step A (430 mg, 1 mmol) in THF (5 mL) was added. The reaction was then stirred at 60°C overnight. The mixture was cooled to room temperature, diluted with water (30 mL) and extracted with EtOAc (60 mL x 2). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by combi flash to give the target compound (320 mg, 63%). 1H NMR (400MHz, DMSO-d6) δ7.68(s,1H),4.28(s,1H),4.01(s,1H),3.85(d,J=12.2Hz,1H),3.60(s,1H),3.09(d,J=1 07.9Hz,4H),2.58(s,1H),2.24–2.11(m,1H),2.02–1.87(m,1H),1.74–1.54(m,4H),1.43(s,9H),1.24(s,3H)ppm. MS:M / e 510(M+1) +

[0166] Step C: tert-Butyl (3S)-4-(7-(hydroxy(naphthalen-1-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)imidazo[2,1-f][1,2,4]triazin-4-yl)-3-methylpiperazine-1-carboxylate

[0167]

[0168] To a solution of the product of step B (100 mg, 0.19 mmol) in THF (10 mL) was added n-butyl lithium (0.2 mL, 0.294 mmol, 1.6 M in hexane), maintaining the temperature between -75 and -65 ° C. After 30 min, a mixture of 1-naphthaldehyde (36 mg, 0.23 mmol) in THF (2 mL) was added dropwise. The resulting mixture was stirred at -70 ° C for 1 h and then warmed to room temperature for 1 h. The reaction was quenched with saturated NH4Cl aqueous solution and extracted with EtOAc (60 mL x 2). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by combi flash to give the target compound (90 mg, 78%). MS: M / e 588 (M+1) + .

[0169] Step D: 4-((S)-2-methylpiperazin-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalen-1-ylmethyl)imidazo[2,1-f][1,2,4]triazine

[0170]

[0171] To a mixture of the product of step C (90 mg) in TFA (1 mL) was added DCE (1 ml) and Et3SiH (1 mL). The reaction was heated at 25°C overnight. The mixture was concentrated to dryness and the residue was used directly in the next step without further purification. MS: M / e 472 (M+1) + .

[0172] Step E: 1-((S)-3-methyl-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-(naphthalen-1-ylmethyl)imidazo[2,1-f][1,2,4]triazin-4-yl)piperazin-1-yl)prop-2-en-1-one

[0173]

[0174] To a mixture of the product of step D (80 mg, crude) in CH3CN (3 mL) was added saturated NaHCO3 (1 mL) at 0°C, followed by a solution of acryloyl chloride (18 mg, 0.2 mmol) in CH3CN (0.3 mL). The resulting mixture was stirred at room temperature for 30 min. The reaction was diluted with water, extracted with EA (60 mL), washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by preparative TLC to give the target compound (18 mg, 20% over two steps). 1 H NMR (400MHz, DMSO-d6) δ8.19(d,J=6.2Hz,1H),7.94(d,J=7.7Hz,1H),7.83(d,J=7.9Hz,1H),7 .57–7.49(m,2H),7.47–7.25(m,3H),6.94–6.76(m,1H),6.18(d,J=15.7Hz,1H),5.74(d,J=10 .5Hz,1H),4.63(s,2H),4.22–4.05(m,4H),3.62-3.48(m,1H),3.18–2.87(m,4H),2.58(s,2H) ,2.30(s,3H),2.18(s,1H),1.94–1.85(m,1H),1.67–1.52(m,3H),1.21(d,J=14.4Hz,3H)ppm. MS:M / e 526(M+1) + .

[0175] Example 24: Compound B1: 2-((S)-1-acryloyl-4-(7-((3-hydroxynaphthalen-1-yl)methyl)-5-methyl-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0176] Step A: (S)-benzyl 4-(2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0177]

[0178] A mixture of 2,4-dichloro-5H-pyrrolo[3,2-d]pyrimidine (2.5 g, 0.013 mol), (S)-benzyl 2-(cyanomethyl)piperazine-1-carboxylate (3.44 g, 0.013 mol) and DIEA (5.15 g, 0.040 mol) in THF (50 ml) was stirred at 70 °C overnight. After completion, the solution was concentrated, diluted with EA (50 ml) and then washed with brine (20 ml). The organic layer was dried and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 50-100% EA in PE to give the product (1.7 g, 31%). MS: M / e 411 (M+1) + .

[0179] Step B: (2S)-benzyl 4-(7-((3-(benzyloxy)naphthalen-1-yl)(hydroxy)methyl)-2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0180]

[0181] A mixture of (S)-4-(2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (1 g, 2.43 mmol), 3-(benzyloxy)-1-naphthaldehyde (0.96 g, 3.66 mmol) and K2CO3 (0.5 g, 3.62 mmol) in MeCN (12 ml), H2O (10 ml) and MeOH (2 ml) was stirred at 40°C for 3 days. The mixture was poured into H2O (20 ml) and extracted with EA (20 ml x 2). The organic layer was washed with brine (10 ml), dried and concentrated. The resulting residue was purified by flash column chromatography using 10%-100% EA in PE to give the product (0.4 g, 24%). MS: M / e 673 (M+1) + .

[0182] Step C: (S)-benzyl 4-(7-((3-(benzyloxy)naphthalen-1-yl)methyl)-2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0183]

[0184] A solution of benzyl (2S)-4-(7-((3-(benzyloxy)naphthalen-1-yl)(hydroxy)methyl)-2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (500 mg, 0.74 mmol) in TFA (5 ml), triethylsilane (5 ml) and DCE (5 ml) was stirred at room temperature for 1 h. After completion, the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography using 0-60% EA in PE to give the product (0.42 g, 86%). MS: M / e 657 (M+1) + .

[0185] Step D: (S)-benzyl 4-(7-((3-(benzyloxy)naphthalen-1-yl)methyl)-2-chloro-5-methyl-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0186]

[0187] A mixture of (S)-4-(7-((3-(benzyloxy)naphthalen-1-yl)methyl)-2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (0.41 g, 0.62 mmol) and NaH (60%, 50 mg, 1.25 mmol) in DMF (10 ml) was stirred at room temperature for 10 min. MeI (0.13 g, 0.92 mmol) was added and stirred at room temperature for 1.5 h. After completion, the solution was quenched with H2O (10 ml) and then extracted with EA (20 ml x 2). The organic layer was dried and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-60% EA in PE to give the product (0.35 g, 84%). MS: M / e 671 (M+1) + .

[0188] Step E: (S)-benzyl 4-(7-((3-(benzyloxy)naphthalen-1-yl)methyl)-5-methyl-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0189]

[0190] A mixture of (S)-benzyl 4-(7-((3-(benzyloxy)naphthalen-1-yl)methyl)-2-chloro-5-methyl-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (0.32 g, 0.48 mmol), (S)-(1-methylpyrrolidin-2-yl)methanol (82.3 mg, 0.72 mmol), Pd2dba3 (131 mg, 0.14 mmol), RuPhos (67 mg, 0.14 mmol) and Cs2CO3 (311 mg, 0.95 mmol) in dioxane (10 ml) was stirred at 120° C. overnight. After completion, the mixture was poured into water (15 ml) and extracted with EA (20 ml×2). The organic layer was washed with brine (10 ml), dried and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography using 0-10% MeOH in DCM to give the product (170 mg, 48%). MS: M / e 750 (M+1) + .

[0191] Step F: 2-((S)-4-(7-((3-hydroxynaphthalen-1-yl)methyl)-5-methyl-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0192]

[0193] A mixture of (S)-4-(7-((3-(benzyloxy)naphthalen-1-yl)methyl)-5-methyl-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (170 mg, 0.23 mmol) and Pd / C (150 mg) in THF (10 ml) and MeOH (10 ml) was stirred at room temperature for 2 days under H2 atmosphere. After completion, the mixture was filtered through celite. The filtrate was concentrated under reduced pressure to give the product as a crude product (119 mg, 100%), which was used directly in the next step without further purification. MS: M / e 526 (M+1) + .

[0194] Step G: 2-((S)-1-acryloyl-4-(7-((3-hydroxynaphthalen-1-yl)methyl)-5-methyl-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0195]

[0196] To a mixture of 2-((S)-4-(7-((3-hydroxynaphthalen-1-yl)methyl)-5-methyl-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile (119 mg, 0.23 mmol) in MeCN (8 ml) was added NaHCO aqueous solution (saturated, 0.3 mL) at room temperature, followed by acryloyl chloride (30 mg, 0.33 mmol). The mixture was stirred at room temperature for 30 min. LiOH (20 mg, 0.83 mmol) in H2O (1 ml) was then added to the above solution and stirred at room temperature for 1 h. The mixture was poured into water and extracted with EA (15 ml x 3). The organic layer was dried and concentrated. The residue was purified by preparative TLC to give the product (4.07 mg). 1 H NMR (400MHz, CD3OD) δ8.00(d,J=8.3Hz,1H),7.64(d,J=8.0Hz,1H),7.48-7.30(m,2H),7.25(t,J=7.5Hz,1H),7. 21(s,1H),6.99(s,1H),6.88(s,1H),6.29(d,J=16.2Hz,1H),5.83(d,J=9.7Hz,1H),4.79(d,J=12.5Hz,1H),4.63 -4.54(m,1H),4.42(s,2H),4.08-3.94(m,2H),3.90(s,3H),3.87-3.83(m,1H),3.62–3.55(m,1H),3.50–3.37(m ,2H),3.13(s,2H),2.96(s,3H),2.41-2.29(m,1H),2.21-2.13(m,1H),2.08-2.01(m,2H),1.32-1.27(m,4H)ppm. MS:M / e 580(M+1) + .

[0197] Example 25: Compound B2: 2-((S)-1-acryloyl-4-(5-ethyl-7-((3-hydroxynaphthalen-1-yl)methyl)-2-(((S)-1-methylpyrrolidinone-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0198] Step A: (S)-benzyl 4-(2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0199]

[0200] A mixture of 2,4-dichloro-5H-pyrrolo[3,2-d]pyrimidine (5 g, 0.027 mol), (S)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (6.9 g, 0.027 mol) and DIEA (10 g, 0.078 mol) in THF (50 ml) was stirred at 70 ° C overnight. After completion, the solution was concentrated, diluted with EA (50 ml), and then washed with brine (20 ml). The organic layer was dried and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 50-100% EA in PE to give the product (11 g, 100%). 1 H NMR(400MHz,DMSO-d6)δ11.58(s,1H),7.65(s,1H),7.50-7.27(m,5H),6.43(s,1H),5.22-5.08(m,2H), 4.60(s,1H),4.29(d,J=11.6Hz,2H),3.99(d,J=10.9Hz,1H),3.60-3.35(m,3H),3.05–2.87(m,1H)ppm. MS:M / e 411(M+1) + .

[0201] Step B: (2S)-benzyl 4-(2-chloro-7-(hydroxy(3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0202]

[0203] A mixture of (S)-4-(2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (1.85 g, 4.5 mmol), 4-formylnaphthalene-2-yl pivalate (1.15 g, 4.5 mmol) and KCO (0.93 g, 6.7 mmol) in MeCN (20 ml) and HO (20 ml) was stirred at 45 ° C for 3 days. The mixture was poured into HO (20 ml) and extracted with EA (25 ml×2). The organic layer was washed with brine (10 ml), dried and concentrated. The resulting residue was purified by flash column chromatography with 10%-100% EA in PE to give the product (1.5 g, 50%). 1H NMR (400MHz, DMSO-d6) δ11.42(s,1H),7.98(d,J=8.3Hz,1H),7.91(d,J=8.2Hz, 1H),7.64(s,1H),7.58(s,1H),7.50-7.43(m,1H),7.42-7.31(m,5H),7.08(d,J =13.2Hz,1H),6.68(s,1H),5.22-5.06(m,2H),4.58(s,1H),4.34-4.17(m,2H), 3.96(d,J=11.4Hz,1H),3.63-3.35(m,5H),3.05-2.86(m,2H),1.38(s,9H)ppm. MS:M / e 667(M+1) + .

[0204] Step C: (S)-benzyl 4-(2-chloro-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0205]

[0206] A solution of (2S)-4-(2-chloro-7-(hydroxy(3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (500 mg, 0.75 mmol) in TFA (2 ml), triethylsilane (2 ml) and DCE (2 ml) was stirred at room temperature for 30 min. After completion, the mixture was poured into aqueous NaHCO3 solution (30 ml) and extracted with DCM (20 ml x 2). The organic layer was washed with brine (10 ml), dried and concentrated under reduced pressure to give the product (0.6 g, 100%, crude), which was used directly in the next step without further purification. MS: M / e 651 (M+1) + .

[0207] Step D: (S)-benzyl 4-(2-chloro-5-ethyl-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0208]

[0209] To a stirred mixture of (S)-4-(2-chloro-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (0.27 g, 0.41 mmol) and K2CO3 (86 mg, 0.62 mmol) in DMF (8 ml) was added iodoethane (0.13 g, 0.83 mmol), followed by stirring at room temperature for 4 h. Upon completion, the solution was poured into H2O (10 ml) and then extracted with EA (15 ml x 2). The organic layer was dried and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography using 10-60% EA in PE to give the product (0.2 g, 71%). MS: M / e 679 (M+1) + .

[0210] Step E: (S)-benzyl 2-(cyanomethyl)-4-(5-ethyl-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate

[0211]

[0212] A mixture of (S)-4-(2-chloro-5-ethyl-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (0.2 g, 0.29 mmol), (S)-(1-methylpyrrolidin-2-yl)methanol (68 mg, 0.59 mmol), Pd2dba3 (81 mg, 0.089 mmol), RuPhos (41 mg, 0.088 mmol) and Cs2CO3 (192 mg, 0.59 mmol) in dioxane (10 ml) was stirred at 120° C. overnight. After completion, the mixture was poured into water (15 ml) and extracted with EA (20 ml×2). The organic layer was washed with brine (10 ml), dried and concentrated under reduced pressure. The resulting residue was purified by preparative TLC using DCM:MeOH (10:1) to give the product (60 mg, 27%). MS: M / e 758 (M+1) + .

[0213] Step F: 4-((4-((S)-3-(cyanomethyl)piperazin-1-yl)-5-ethyl-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl)naphthalen-2-yl pivalate

[0214]

[0215] A mixture of (S)-2-(cyanomethyl)-4-(5-ethyl-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (60 mg, 0.079 mmol) and Pd / C (60 mg) in THF (5 ml) and MeOH (10 ml) was stirred at room temperature overnight under H2 atmosphere. After completion, the mixture was filtered through celite. The filtrate was concentrated under reduced pressure to give the product (45 mg, 91%), which was used directly in the next step without further purification. MS: M / e624 (M+1) + .

[0216] Step G: 2-((S)-1-acryloyl-4-(5-ethyl-7-((3-hydroxynaphthalen-1-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0217]

[0218] To a mixture of 4-((4-((S)-3-(cyanomethyl)piperazine-1-yl)-5-ethyl-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl)naphthalen-2-yl pivalate (45 mg, 0.072 mmol) in MeCN (8 ml) was added NaHCO aqueous solution (saturated, 0.3 mL) at room temperature, followed by acryloyl chloride (13 mg, 0.14 mmol). The mixture was stirred at room temperature for 30 min. LiOH (20 mg, 0.83 mmol) in H2O (1 ml) was then added to the above solution and stirred at room temperature for 1 h. The mixture was poured into water and extracted with EA (15 ml x 3). The organic layer was dried and concentrated. The residue was purified by preparative TLC to give the product (3.81 mg). 1H NMR (400MHz, DMSO-d6) δ9.86 (s, 1H), 9.60 (s, 1H), 8.10 (d, J = 9.1Hz, 1H), 7.67 (d, J = 8.2Hz, 1H), 7. 54(s,1H),7.37(t,J=7.4Hz,1H),7.26(t,J=7.5Hz,1H),6.97(d,J=7.2Hz,2H),6.19(d,J=16.4Hz,1 H),5.78(d,J=10.1Hz,1H),4.65-4.57(m,1H),4.55-4.48(m,1H),4.33(s,2H),4.28–4.02(m,3H), 3.89–3.49(m,7H),3.24–2.85(m,7H),2.29–2.10(m,1H),2.06–1.82(m,3H),1.23–1.19(t,3H)ppm. MS:M / e 594(M+1) + .

[0219] Example 26: Compound B3: 2-((S)-1-acryloyl-4-(2-(((2S,4R)-4-fluoro-1-methylpyrrolidin-2-yl)methoxy)-7-((3-hydroxynaphthalen-1-yl)methyl)-5-methyl-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0220] Step A: ((2S,4R)-4-Fluoro-1-methylpyrrolidin-2-yl)methanol

[0221]

[0222] To a stirred solution of 1-(tert-butyl)2-methyl(2S,4R)-4-fluoropyrrolidine-1,2-dicarboxylate (1 g, 4.0 mmol) in THF (15 ml) was added LAH (0.3 g, 7.9 mmol) in several portions. The mixture was then stirred at 50 °C overnight. Upon completion, the mixture was cooled to room temperature and quenched with Na2SO410H2O, which was then filtered through celite. The filtrate was concentrated under reduced pressure to give the product (0.54 g, 100%), which was used directly in the next step without further purification. MS: M / e 134 (M+1) + .

[0223] Step B: (S)-benzyl 4-(2-chloro-5-methyl-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0224]

[0225] To a stirred mixture of (S)-4-(2-chloro-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (0.98 g, 1.5 mmol, crude) and K2CO3 (410 mg, 3 mmol) in DMF (10 ml) was added iodomethane (0.57 g, 4 mmol), followed by stirring at room temperature for 2.5 h. Upon completion, the solution was poured into H2O (15 ml) and then extracted with EA (20 ml x 2). The organic layer was dried and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 0-60% EA in PE to give the product (0.93 g, 92%). MS: M / e 665 (M+1) + .

[0226] Step C: (S)-2-(cyanomethyl)-4-(2-(((2S.4R)-4-fluoro-1-methylpyrrolidin-2-yl)methoxy)-5-methyl-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester

[0227]

[0228] A mixture of (S)-4-(2-chloro-5-methyl-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (0.25 g, 0.38 mmol), ((2S,4R)-4-fluoro-1-methylpyrrolidin-2-yl)methanol (100 mg, 0.75 mmol), Pd2dba3 (103 mg, 0.11 mmol), RuPhos (53 mg, 0.11 mmol) and Cs2CO3 (245 mg, 0.75 mmol) in dioxane (10 ml) was stirred at 100° C. overnight. After completion, the mixture was poured into water (15 ml) and extracted with EA (20 ml×2). The organic layer was washed with brine (10 ml), dried and concentrated under reduced pressure. The resulting residue was purified by preparative TLC using DCM:MeOH (10:1) to give the crude product (50 mg), which was used directly in the next step without further purification. MS: M / e 762 (M+1) + .

[0229] Step D: 4-((4-((S)-3-(cyanomethyl)piperazin-1-yl)-2-(((2S,4R)-4-fluoro-1-methylpyrrolidin-2-yl)methoxy)-5-methyl-5H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl)naphthalen-2-yl pivalate

[0230]

[0231] A mixture of (S)-2-(cyanomethyl)-4-(2-(((2S,4R)-4-fluoro-1-methylpyrrolidin-2-yl)methoxy)-5-methyl-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (50 mg, crude) and Pd / C (50 mg) in THF (5 ml) and MeOH (5 ml) was stirred at room temperature overnight under H2 atmosphere. After completion, the mixture was filtered through celite. The filtrate was concentrated under reduced pressure to give the product (40 mg, crude), which was used directly in the next step without further purification. MS: M / e 628 (M+1) + .

[0232] Step E: 2-((S)-1-acryloyl-4-(2-(((2S,4R)-4-fluoro-1-methylpyrrolidin-2-yl)methoxy)-7-((3-hydroxynaphthalen-1-yl)methyl)-5-methyl-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0233]

[0234] To a mixture of 4-((4-((S)-3-(cyanomethyl)piperazine-1-yl)-2-(((2S,4R)-4-fluoro-1-methylpyrrolidin-2-yl)methoxy)-5-methyl-5H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl)naphthalen-2-ylpivalate (40 mg, crude) in MeCN (8 ml) was added NaHCO aqueous solution (saturated, 0.3 mL) at room temperature, followed by acryloyl chloride (12 mg, 0.13 mmol). The mixture was stirred at room temperature for 30 min. LiOH (20 mg, 0.83 mmol) in H2O (1 ml) was then added to the above solution and stirred at room temperature for 1 h. The mixture was poured into water and extracted with EA (15 ml x 3). The organic layer was dried and concentrated. The residue was purified by preparative TLC to give the product (1.14 mg). 1H NMR (400MHz, DMSO-d6) δ9.60 (s, 1H), 8.11 (t, J = 10.0Hz, 1H), 7.67 (d, J = 8.1Hz, 1H),7.35(d,J=7.0Hz,2H),7.26-7.19(m,2H),6.98(s,2H),6.18(d,J=16.8Hz,1 H),5.78(d,J=11.4Hz,1H),5.32(s,2H),4.31(s,2H),3.86(s,3H),3.80-3.61(m ,5H),3.51(s,1H),3.05-2.92(m,5H),2.01-1.96(m,5H),1.51-1.40(m,2H)ppm. MS:M / e 598(M+1) + .

[0235] Example 27: Compound B4: (S)-1-(4-(7-((3-hydroxynaphthalen-1-yl)methyl)-5-methyl-2-((1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one Step A: 4-(2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester

[0236]

[0237] A mixture of 2,4-dichloro-5H-pyrrolo[3,2-d]pyrimidine (5 g, 0.027 mol), benzyl piperazine-1-carboxylate (6.44 g, 0.029 mol) and DIEA (6.86 g, 0.053 mol) in THF (50 ml) was stirred at 70 ° C overnight. After completion, the solution was concentrated, diluted with EA (50 ml), and then washed with brine (20 ml). The organic layer was dried and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 30-100% EA in PE to give the product (8.45 g, 85%). 1 H NMR (400MHz, DMSO-d6) δ11.63(s,1H),7.64(s,1H),7.39(d,J=5.9Hz,4H),7.36–7.30(m,1H),6.42(s,1H),5.13(s,2H),3.79(s,4H),3.59(s,4H)ppm. MS:M / e 372(M+1) + .

[0238] Step B: Benzyl 4-(2-chloro-7-(hydroxy(3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate

[0239]

[0240] A mixture of benzyl 4-(2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate (1 g, 2.7 mmol), 4-formylnaphthalen-2-yl pivalate (0.83 g, 3.2 mmol) and K2CO3 (0.56 g, 4.1 mmol) in MeCN (20 ml) and H2O (10 ml) was stirred at 45°C overnight. The mixture was poured into H2O (20 ml) and extracted with EA (25 ml x 2). The organic layer was washed with brine (10 ml), dried and concentrated. The resulting residue was purified by flash column chromatography with 0-100% EA in PE to give the product (0.45 g, 27%). MS: M / e 628 (M+1) + .

[0241] Step C: Benzyl 4-(2-chloro-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate

[0242]

[0243] A solution of benzyl 4-(2-chloro-7-(hydroxy(3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate (450 mg, 0.72 mmol) in TFA (2 ml), triethylsilane (2 ml) and DCE (2 ml) was stirred at room temperature for 30 min. After completion, the mixture was poured into aqueous NaHCO3 solution (30 ml) and extracted with DCM (20 ml x 2). The organic layer was washed with brine (10 ml), dried and concentrated under reduced pressure to give the product (0.43 g, 98%), which was used directly in the next step without further purification. MS: M / e 612 (M+1) + .

[0244] Step D: Benzyl 4-(2-chloro-5-methyl-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate

[0245]

[0246] To a stirred mixture of 4-(2-chloro-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (0.43 g, 0.70 mmol) and K2CO3 (150 mg, 1.09 mmol) in DMF (5 ml) was added iodomethane (0.15 g, 1.06 mmol), followed by stirring at room temperature for 2 h. Upon completion, the solution was poured into H2O (10 ml) and then extracted with EA (15 ml x 2). The organic layer was dried and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography with 10-50% EA in PE to give the product (0.44 g, 100%). MS: M / e 626 (M+1) + .

[0247] Step E: (S)-benzyl 4-(5-methyl-2-((1-methylpyrrolidin-2-yl)methoxy)-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate

[0248]

[0249] A mixture of benzyl 4-(2-chloro-5-methyl-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate (0.44 g, 0.70 mmol), (S)-(1-methylpyrrolidin-2-yl)methanol (160 mg, 1.39 mmol), Pd2dba3 (190 mg, 0.21 mmol), RuPhos (100 mg, 0.21 mmol) and Cs2CO3 (460 mg, 1.41 mmol) in dioxane (10 ml) was stirred at 100° C. overnight. After completion, the mixture was poured into water (15 ml) and extracted with EA (20 ml×2). The organic layer was washed with brine (10 ml), dried and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography using 0-10% MeOH in DCM to give the crude product (250 mg) which was used directly in the next step without further purification. MS: M / e 705 (M+1) + .

[0250] Step F: (S)-4-((5-methyl-2-((1-methylpyrrolidin-2-yl)methoxy)-4-(piperazin-1-yl)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl)naphthalen-2-yl pivalate

[0251]

[0252] A mixture of (S)-4-(5-methyl-2-((1-methylpyrrolidin-2-yl)methoxy)-7-((3-(pivaloyloxy)naphthalen-1-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (250 mg, 0.35 mmol, crude) and Pd / C (50 mg) in THF (10 ml) and MeOH (10 ml) was stirred overnight at room temperature under H2 atmosphere. After completion, the mixture was filtered through celite. The filtrate was concentrated under reduced pressure to give the product (190 mg, crude), which was used directly in the next step without further purification. MS: M / e 571 (M+1) + .

[0253] Step G: (S)-1-(4-(7-((3-hydroxynaphthalen-1-yl)methyl)-5-methyl-2-((1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolidin[3,2-d]pyrimidin-4-yl)piperazin-1-yl)prop-2-en-1-one

[0254]

[0255] To a mixture of (S)-4-((5-methyl-2-((1-methylpyrrolidin-2-yl)methoxy)-4-(piperazine-1-yl)-5H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl)naphthalen-2-yl pivalate (190 mg, crude) in MeCN (2 ml) and THF (6 ml) was added NaHCO aqueous solution (saturated, 0.5 mL) at room temperature, followed by acryloyl chloride (61 mg, 0.67 mmol). The mixture was stirred at room temperature for 30 min. LiOH (60 mg, 2.5 mmol) in H2O (2 ml) was then added to the above solution and stirred at room temperature for 2 h. The mixture was poured into water and extracted with EA (15 ml x 3). The organic layer was dried and concentrated. The residue was purified by preparative TLC to give the product (1.83 mg). 1H NMR (400MHz, DMSO-d6) δ9.61 (s, 1H), 8.10 (d, J = 8.3Hz, 1H), 7.67 (d, J = 8.1Hz, 1H), 7.37 (t, J = 6. 0Hz,1H),7.34(s,1H),7.26(t,J=7.4Hz,1H),6.98(s,2H),6.83(dd,J=16.5,10.5Hz,1H),6.13( d,J=14.3Hz,1H),5.71(d,J=11.7Hz,1H),4.56(d,J=48.6Hz,2H),4.31(s,2H),3.83(s,3H),3.8 0-3.68(m,6H),3.09-2.77(m,6H),2.30-2.12(m,2H),2.06-1.96(m,2H),1.93-1.78(m,2H)ppm. MS:M / e 541(M+1) + .

[0256] Example 28: Compound B5: 2-((S)-1-acryloyl-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-((5-(trifluoromethyl)-1H-indazol-4-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0257] Step A: 4-Bromo-1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazole

[0258]

[0259] To a solution of 4-bromo-5-(trifluoromethyl)-1H-indazole (2 g, 7.6 mmol) in CH3CN (20 mL) cooled at 0°C was added DHP (1.9, 22.7 mmol) followed by PTSA (152 mg, 0.8 mmol). The resulting mixture was stirred at room temperature overnight. The solvent was evaporated, then water (8 mL) was added and extracted with EA (15 mL). The organic layer was dried, concentrated and purified by CombiFlash (PE:EA=9:1) to give the product (2.3 g, 88%). MS: M / e 349 (M+1) +

[0260] Step B: 1-(Tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazole-4-carbaldehyde

[0261]

[0262] n-Butyl lithium (1.6M in THF, 4.8mL) was added dropwise to a solution of 4-bromo-1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazole (2.2g, 6.3mmol) in THF (25mL) at -78°C. After addition, the solution was stirred for 30min, and then morpholine-4-carboxaldehyde (1.1g, 9.5mmol) in THF (5mL) was added. The solution was gradually stirred to room temperature for 2 hours. The reaction mixture was quenched with NH4Cl solution and extracted with EA. The organic layer was dried, concentrated and purified by CombiFlash (PE:EA=9:1) to give the product (1.3g, 70%). MS:M / e 299(M+1) + .

[0263] Step C: (S)-benzyl 4-(2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0264]

[0265] A solution of 2,4-dichloro-5H-pyrrolo[3,2-d]pyrimidine (20 g, 106 mmol), (S)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester (30.2 g, 117 mmol) and DIEA (20.5 g, 159 mmol) in THF (200 mL) was heated at 60 ° C overnight. The solution was cooled, water was added and extracted with EA. The organic layer was dried, concentrated and purified by gel column (PE:EA=1:2) to give the product (44 g, 100%). MS: M / e411 (M+1) + .

[0266] Step D: (S)-tert-butyl 4-(4-((benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2-chloro-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate

[0267]

[0268] DMAP (147 mg, 1.2 mmol) was added to a solution of (S)-benzyl 4-(2-chloro-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (5 g, 12 mmol) and (Boc)2O (3.1 g, 14 mmol) in DCM (50 mL). The solution was stirred at room temperature overnight, concentrated and purified by CombiFlash (PE:EA=50%) to give the product (6.2 g, 100%). MS: M / e 511 (M+1)+

[0269] Step E: tert-Butyl 4-((S)-4-((Benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate

[0270]

[0271] A solution of the product of step D (9.1 g, 17.8 mmol), (S)-(1-methylpyrrolidin-2-yl)methanol (4.1 g, 36 mmol), Pd2(dba)3 (4.9 g, 5.4 mmol), RuPhos (2.5 g, 5.4 mmol) and Cs2CO3 (11.6 g, 35.6 mmol) in dioxane (100 mL) was heated at 100° C. overnight under N2 atmosphere. The solution was cooled, evaporated, EA was added, slurried and filtered. The filtrate was washed with brine, dried, concentrated and purified by CombiFlash (DCM: NH3 (7M in MeOH) = 10:1) to give the product (5.1 g, 49%). MS: M / e 590 (M+1) +

[0272] Step F: (S)-benzyl 2-(cyanomethyl)-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate

[0273]

[0274] TFA (10 mL) was added to a solution of tert-butyl 4-((S)-4-((benzyloxy)carbonyl)-3-(cyanomethyl)piperazin-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidine-5-carboxylate (5.1 g, 8.7 mmol) in DCM (50 mL). The reaction mixture was stirred at room temperature overnight, then evaporated, water was added, and basified to pH = 7-8 with NaHCO3 solution. After extraction with DCM, the organic layer was dried, concentrated and purified by CombiFlash (DCM: NH3 (7M in MeOH) = 25:2) to give the product (2.1 g, 50%). MS: M / e 490 (M+1) + .

[0275] Step G: (2S)-benzyl 2-(cyanomethyl)-4-(7-(hydroxy(1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazol-4-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate

[0276]

[0277] A solution of (S)-benzyl 2-(cyanomethyl)-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate (500 mg, 1 mmol), 1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazole-4-carbaldehyde (298 mg, 1 mmol) and K2CO3 (276 mg, 2 mmol) in CH3CN / H2O (1:1, 6 mL) was heated at 50°C overnight. The mixture was extracted with EA, dried, concentrated and purified by CombiFlash (DCM:NH3 (7 M in methanol) = 25:2) to give the product (237 mg, 29%). 1 H NMR (400MHz, DMSO-d6) δ11.02(s,1H),8.40(s,1H),7.80(d,J=8.0Hz,1H),7.66(d,J=8.0Hz,1H ),7.39-7.35(m,5H),7.03-6.97(m,1H),6.57(s,1H),6.15(s,1H),5.90(d,J=12.0Hz,1H),5.17 -5.12(m,2H),4.57(s,1H),4.22(br.s,2H),4.07-3.87(m,4H),3.75(s,1H),3.35(s,1H),3.25 (s,1H),2.90(s,3H),2.40-2.31(m,4H),2.15(s,1H),2.01-1.91(m,3H),1.66-1.58(m,6H)ppm. MS:M / e 788(M+1) + .

[0278] Step H: (S)-benzyl 2-(cyanomethyl)-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-((5-(trifluoromethyl)-1H-indazol-4-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate

[0279]

[0280] TFA (0.5 mL) was added to a solution of (2S)-2-(cyanomethyl)-4-(7-(hydroxy(1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazol-4-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (50 mg, 0.06 mmol) in DCM (3 mL). It was stirred at 30 °C for 3 hours. The solvent was evaporated. The residue was dissolved in DCM (3 mL) and washed with NaHCO3 solution (3 mL). The organic layer was dried and concentrated to give the intermediate. It was dissolved in DCE (2 mL), then Et3SiH (0.5 mL) and TFA (0.5 mL) were added and heated at 30 °C overnight. The reaction mixture was evaporated, DCM was added, and washed with NaHCO3 solution. The crude product was purified by preparative TLC (DCM: NH3 (7M in methanol) = 25:2) to give the product (13 mg, 30%). MS: M / e 688 (M+1) + .

[0281] Step I: 2-((S)-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-((5-(trifluoromethyl)-1H-indazol-4-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0282]

[0283] Pd / C (10%, wet, 20 mg) was added to a solution of (S)-benzyl 2-(cyanomethyl)-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-((5-(trifluoromethyl)-1H-indazol-4-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylate (13 mg, 0.02 mmol) in EtOH / THF (1:1, 2 mL). It was stirred at room temperature under a balloon of H2 for 2 h. The catalyst was filtered off and the filtrate was concentrated to give the product (10 mg, crude). MS: M / e 554 (M+1) +

[0284] Step J: 2-((S)-1-acryloyl-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-((5-(trifluoromethyl)-1H-indazol-4-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0285]

[0286] NaHCO solution (saturated, 0.2 mL) was added to a solution of 2-((S)-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-7-((5-(trifluoromethyl)-1H-indazol-4-yl)methyl)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile (10 mg, crude) in CHCN (2 mL) followed by acryloyl chloride (1.3 mg, 0.02 mmol). The reaction mixture was stirred at room temperature for 10 min. The solution was evaporated, water (2 mL) was added, and extracted with EA (4 mL). The organic layer was dried, concentrated and purified by preparative TLC (DCM:NH (7 M in MeOH) = 8:1) to give the product (1 mg, 10%). 1 H NMR (400MHz, CD3OD) δ8.09(s,1H),7.72(d,J=12.0Hz,1H),7.59(d,J=8.0Hz ,1H),6.77(s,1H),6.30(d,J=16.0Hz,1H),5.85(s,1H),5.34(s,1H),4.62(s ,1H),4.54(s,2H),4.38(s,1H),4.12(s,1H),3.97(s,1H),3.75(s,2H),3.13 (s,3H),2.96-2.85(m,3H),2.44(s,1H),2.19-2.04(m,6H),1.61(s,1H)ppm. MS:M / e608(M+1) + .

[0287] Example 29: Compound B6: 2-((S)-4-(7-((5-chloro-6-fluoro-1H-indazol-4-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile

[0288] Step A: 4-Bromo-5-chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole

[0289]

[0290] To a solution of 4-bromo-5-chloro-6-fluoro-1H-indazole (2 g, 8.0 mmol) in CH3CN (20 mL) cooled at 0°C was added DHP (2 g, 24 mmol) followed by PTSA (138 mg, 0.80 mmol). The resulting mixture was stirred at room temperature overnight. The solvent was evaporated, then water (8 mL) was added and extracted with EA (15 mL). The organic layer was dried, concentrated and purified by CombiFlash (PE:EA=9:1) to give the product (2.4 g, 89.73%). MS: M / e 333 (M+1)+

[0291] Step B: 5-Chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-carboxylic acid methyl ester

[0292]

[0293] A solution of 4-bromo-5-chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (2.4 g, 7.2 mmol), Pd(dppf)Cl2 (0.53 g, 0.72 mmol) and TEA (2.18 g, 21.6 mmol) in MeOH (50 mL) was stirred overnight at 90 ° C under CO (6 atm) atmospheric pressure. The solution was cooled, water was added and extracted with EA. The organic layer was dried, concentrated and purified by gel column (PE:EA=1:2) to give the product (2.13 g, 94.42%). MS: M / e 313 (M+1) +

[0294] Step C: (5-Chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)methanol

[0295]

[0296] To a solution of 5-chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-carboxylic acid methyl ester (2.13 g, 6.81 mmol) in THF (20 mL) was added LAH (0.39 g, 10.21 mmol) in portions and stirred at room temperature for 2 h. The mixture was cooled, quenched with ice, and extracted with EA. The organic layer was dried, concentrated and purified by gel column (PE:EA=1:2) to give the product (1.72 g, 88.68%). MS: M / e 285 (M+1)+

[0297] Step D: 5-Chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-carbaldehyde

[0298]

[0299] To a solution of (5-chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)methanol (1.72 g, 6.03 mmol) in DCM (15 mL) was added Dess-Martin periodinane (3.07 g, 7.24 mmol) in portions and stirred at room temperature for 3 h. The mixture was quenched with NaHCO3 solution and extracted with DCM. The organic layer was dried, concentrated and purified by gel column (PE:EA=1:1) to give the product (1.65 g, 96.61%). MS: M / e 283 (M+1)+

[0300] Step E: (2S)-4-(7-((5-chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)(hydroxy)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylic acid benzyl ester

[0301]

[0302] A solution of (S)-2-(cyanomethyl)-4-(2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazine-1-carboxylic acid benzyl ester (500 mg, 1 mmol), 5-chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-4-carbaldehyde (289 mg, 1 mmol) and K2CO3 (282 mg, 2 mmol) in CH3CN / H2O (1:1, 6 mL) was heated at 50°C overnight. The mixture was extracted with EA, dried, concentrated, and purified by Combi-Flash (DCM:NH3 (7M in MeOH) = 25:2) to give the product (420 mg, 53.32%). MS: M / e 772 (M+1) + .

[0303] Step F: (S)-benzyl 4-(7-((5-chloro-6-fluoro-1H-indazol-4-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate

[0304]

[0305] TFA (0.5 mL) was added to a solution of (2S)-benzyl 4-(7-((5-chloro-6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)(hydroxy)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (50 mg, 0.06 mmol) in DCM (3 mL) and stirred at room temperature for 3 hours. The solvent was evaporated. The residue was dissolved in DCM (3 mL) and washed with NaHCO3 solution (3 mL). The organic layer was dried and concentrated to give the intermediate. It was dissolved in DCE (2 mL), then Et3SiH (0.5 mL) and TFA (0.5 mL) were added and heated at 30°C overnight. The reaction mixture was evaporated, DCM was added, and washed with NaHCO3 solution. The crude product was purified by preparative TLC (DCM: NH3 (7M in MeOH) = 25:2) to give the product (20 mg, 45.95%). MS: M / e 672 (M+1) + .

[0306] Step G: 2-((S)-4-(7-((5-chloro-6-fluoro-1H-indazol-4-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile

[0307]

[0308] Pd / C (10%, wet, 20 mg) was added to a solution of the product of step F (20 mg, 0.03 mmol) in EtOH / THF (1:1, 2 mL). It was stirred at room temperature for 2 hours under a balloon of H2. The catalyst was filtered and the filtrate was concentrated to give the product. (12 mg, 74.94) MS: M / e 538 (M+1) + .

[0309] Step H: 2-((S)-4-(7-((5-chloro-6-fluoro-1H-indazol-4-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile

[0310]

[0311] A mixture of 2-((S)-4-(7-((5-chloro-6-fluoro-1H-indazol-4-yl)methyl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5H-pyrrolo[3,2-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile (12 mg, 0.0372 mmol), 2-fluoroacrylic acid (4 mg, 0.0446 mmol), T3P (solution in DMF, 0.743 mmol) and TEA (8 mg, 0.0743) in THF was stirred at room temperature for 5 h, and the mixture was quenched with water (5 mL) and extracted with EA (20 mL). The organic layer was dried, concentrated and purified by preparative TLC (DCM:NH3 (7 M in MeOH) = 8:1) to give the product (3 mg, 13.25%). 1 H NMR(400MHz,CD3OD)δ13.30(s,1H),11.38(s,1H),10.44(s,1H),10.06(s,1H),8.29(s,1 H),7.44(d,J=9.1Hz,1H),7.26(s,1H),5.48–5.18(m,2H),4.66–4.56(m,1H),4.56–4.44 (m,2H),4.44–4.34(m,3H),3.83–3.73(m,1H),3.62–3.56(m,1H),3.44–3.32(m,4H),2.9 3(s,4H),2.38–2.18(m,2H),2.09–2.02(m,1H),1.98–1.91(m,1H),1.87–1.79(m,1H)ppm. MS:M / e 610(M+1) + .

[0312] Biochemical function assay

[0313] KRAS (aa 1-169) G12C, C51S, C80L, C118S with His tag were expressed, purified and loaded with GDP in-house. All protein and substrate solutions were prepared in assay buffer containing 25mM HEPES pH7.5, 10mM MgCl2 and 0.01% Triton X-100. Purified GDP-loaded KRAS (aa 1-169) G12C, C51S, C80L, C118S proteins were pre-incubated with serially diluted compounds at 24°C for 3 hours. Purified SOS1 (aa 564-1049) protein, GTP S (Sigma) and GST-cRaf RBD (aa 1-149) were then added to each well and incubated for another 3 hours at 24°C. This addition initiates the nucleotide exchange reaction and the conversion of inactive GDP-loaded KRASG12C to active GTP S KRASG12C bound to GST-cRaf RBD. After incubation, Mab anti-6HIS-Tb cryptate (Cisbio) and Mab anti-GST-XL665 (Cisbio) were added and further incubated at 24°C for 3 hours. The binding interaction between active GTP S KRASG12C and GST-cRaf RBD brings Tb and XL665 into close proximity, thereby increasing the FRET signal (Ex337nm, Em665nm / 620nm). The percentage inhibition of the nucleotide exchange reaction in the presence of increasing compound concentrations was calculated based on the fluorescence ratio at 665nm to 620nm detected on a BMG PHERAstar FSX instrument. The IC50 value for each compound was calculated by fitting the data to a four-parameter logistic model by Dotmatics.

[0314] Table 1: Compound A series

[0315] Compound No. IC50(nmol) Compound No. IC50(nmol) A1 3280 A2 427 A3 >10uM A17 2930

[0316] Table 2: Compound B series

[0317] Compound No. IC50(nmol) Compound No. IC50(nmol) B1 6.2 B2 13 B3 20.3 B4 329 B5 64.1 B6 639

Claims

1. A method for preparing a compound of formula (I) or (II), The steps include: L1 is selected from -(CH2) m -; L2 is selected from -O-(CH2) m -; R 1 Selected from R 2 Selected from R 3 is hydrogen; R 3 ' is selected from hydrogen or -C 1-8 alkyl; R 4 Selected from R 5 Selected from -CH3, -(CH2) m -CN; PR is selected from Boc or Cbz; p is independently selected from 0, or 1; m is selected from 1; R 7 is selected from H or Br; Selected from Selected from 2. The preparation method according to claim 1, characterized in that: The reagents used in step 1 are POCl3, PCl5, and SOCl2.

3. The preparation method according to claim 1, characterized in that: The reagents used in step 2 are triethylamine, diethylamine, diisopropylethylamine, and dimethylaminopyridine.

4. The preparation method according to claim 1, characterized in that: In step 3, 1 The reagents for the CHO reaction are sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide, potassium tert-butoxide, n-butyl lithium, sec-butyl lithium, tert-butyl lithium, lithium diisopropylamide, and lithium hexamethyldisilazide.

5. The preparation method according to claim 1, characterized in that: In step 3, 2 The reagents for the -L2-H reaction are sodium hydride, potassium hydride, lithium hydride, sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide, potassium tert-butoxide, n-butyl lithium, sec-butyl lithium, tert-butyl lithium, lithium diisopropylamide, and lithium hexamethyldisilazide.

6. The preparation method according to claim 1, characterized in that: The deprotection conditions in step 4 are a combination of trifluoroacetic acid and triethylsilane, a combination of BBr3, Pd / C and hydrogen, and a combination of trifluoroacetic acid, hydrochloric acid, Pd / C and cyclohexadiene.

7. The preparation method according to claim 1, characterized in that: The reaction conditions with R4Cl in step 5 are sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, triethylamine, diethylamine, diisopropylethylamine, pyridine, methylimidazole, piperidine, morpholine, indoline, quinoline, isoquinoline, and dimethylaminopyridine.

8. The preparation method according to claim 1, characterized in that: The compound of formula (I) or (II) is

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