Discovery of cyclic guanidine-linked sulfonamides as inhibitors of LMTK3 kinase

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The solid phase synthesis was performed using the “tea-bag” methodology [40]. 100 mg of p-methylbenzdrylamine (pMBHA) resin per compound (1.1mmol/g, 100-200 mesh) was sealed in a mesh “tea-bag,” neutralized with 5% diisopropylethylamine (DIEA) in dichloromethane (DCM), and subsequently swelled with additional DCM washes. The first diversity R1 was introduced by the coupling of Boc amino acid (6 eq) in dimethylformamide (0.1M DMF) for 60 minutes in the presence of diisopropylcarbodiimide (DIC, 6 equiv.) and 1-hydroxybenzotriazole hydrate (HOBt, 6 equiv.). The Boc protecting group was removed with 55% TFA/ DCM for 30 minutes and subsequently neutralized with 5% DIEA/DCM (3 x). The second diversity (R2) was introduced by the subsequent coupling of Boc-Amino Acid utilizing the same standard coupling procedures. The Boc protecting group was removed with 55% Trifluoroacetic Acid (TFA) in DCM for 30 minutes and subsequently neutralized with 5% DIEA/DCM (3 x) and the third diversity (R3) was introduced following the coupling of arylsulfonyl chloride (10 eq) in the presence of diisopropylamine (DIEA) (10 eq) in anhydrous DCM. All reactions were monitored for completion by Ninhydrin test. The reduction of the amide bonds was performed in a 1000 mL Wilmad LabGlass vessel under nitrogen in the presence of 1.0M Borane-Tetrahydrofuran (BH3-THF) complex solution (40-fold excess of BH3-THF per amide bond). The reaction vessel was heated to 65 °C and the temperature was maintained for 72 hours. The solution was then discarded and the bags were washed with THF and methanol. Once completely dry, the bags were treated overnight with piperidine at 65 °C and washed several times with methanol, DMF and DCM. Before proceeding, completion of the reduction was monitored by a control cleavage and analyzed by LCMS. The generated resin bound N-terminal sulfonated triamines were treated with cyanogen bromide (CNBr) in anhydrous DCM (5 eq) for 2 hours. The desired compounds were cleaved from the solid-support in the presence of HF and then extracted with acetic acid and lyophilized to obtain solid white powder. The obtained white powders were purified using preparative high-performance liquid chromatography and the desired cyclic guanidine-linked sulfonamides 1-6 were obtained in good yields and high purity (> 95%). N-((S)-1-((S)-5-(cyclohexylmethyl)-2-iminoimidazolidin-1-yl)-3-phenylpropan-2-yl)benzenesulfonamide (1): 1H NMR (400 MHz, DMSO-d6) δppm: 8.47 (s, 1H), 7-68-7.70 (m, 2H), 7.50-7.52 (m, 1H), 7.48-7.49 (m, 2H), 7.17-7.23 (m, 3H), 7.03 (d, J= 6.5 Hz, 2H), 3.54-3.6- (m, 2H), 3.34-3.43 (m, 2H), 2.95-3.03 (m, 2H), 2.55 (d, J= 6.1 Hz, 2H), 1.49-1.58 (m, 4H), 1.41-1.44 (m, 1H), 0.98-1.14 (m, 6H), 0.62-0.73 (m, 2H). 13C NMR (100 MHz, DMSO-d6) δppm: : 167.6, 159.5, 142.1, 138.0, 132.5, 129.5, 129.3, 128.9, 126.9, 126.6, 55.4, 53.2, 46.8, 44.2, 38.0, 33.9, 33.5, 32.0, 26.3, 25.9, 25.7. MS (ESI) m/z, calculated: 454.25, found [M+H]+: 455.0. N-((S)-1-((S)-2-imino-5-isobutylimidazolidin-1-yl)-3-phenylpropan-2-yl)benzenesulfonamide (2): 1H NMR (400 MHz, DMSO-d6) δppm: 8.46 (s, 1H), 7.69 (d, J= 8.56 Hz, 2H), 7-58-7.60 (m, 1H), 7-49-7.52 (m, 1H), 7.14-7.23 (m, 3H), 7.3 (d, J= 7.9 Hz, 2H), 3.57-3.63 (m, 2H), 3.34-3.43 (m, 3H), 2.96-3.02 (m, 2H), 2.56-2.67 (m, 2H), 1.34-1.38 (m, 1H), 0.94-1.04 (m, 2H), 0.72 (d, J= 6.6 Hz, 3H), 0.70 (d, J= 6.5 Hz, 3H) . 13C NMR (100 MHz, DMSO-d6) δppm: 159.9, 138.0, 132.6, 129.6, 129.3, 128.9, 126.9, 126.6, 55.9, 53.1, 46.6, 44.0, 24.3, 23.8, 21.8. MS (ESI) m/z, calculated: 414.21, found [M+H]+: 415.0. 2-chloro-N-((S)-1-((S)-5-(cyclohexylmethyl)-2-iminoimidazolidin-1-yl)-3-(4-hydroxyphenyl)propan-2-yl)benzenesulfonamide (3): 1H NMR (400 MHz, DMSO-d6) δppm: 8.00 (s, 1H), 7-87-7.89 (m, 1H), 7.75-7.86 (m, 3H), 7.58-7.60 (m, 1H), 7.51-7.56 (m, 1H), 7.46-7.50 (m, 1H), 6.76 (dd, J= 8.3 Hz, J= 1,4 Hz, 3H), 6.51 (dd, J= 8.4 Hz, J= 1.6 Hz, 3H).3.75-3.77 (m, 1H), 3.43-3.51 (m, 5H), 3.09—3.14 (m, 2H), 1.58-1.60 (m, 4H), 1.46-1.49 (m, 1H), 1.10-1.22 (m, 6H), 0.70-0.85 (m, 2H). 13C NMR (100 MHz, DMSO-d6) δppm: 158.6, 156.4, 138.7, 134.3, 132.3, 131.0, 130.3, 130.0, 127.9, 127.3, 115.6, 55.7, 53.6, 46.8, 44.3, 38.2, 38.0, 33.9, 33.5, 32.0, 26.3, 25.9, 25.7. MS (ESI) m/z, calculated: 505.05, found [M+H]+: 506.0. N-((S)-1-((S)-5-(cyclohexylmethyl)-2-iminoimidazolidin-1-yl)-3-(4-hydroxyphenyl)propan-2-yl)-4-methylbenzenesulfonamide (4): 1H NMR (400 MHz, DMSO-d6) δppm: 9.25 (s, 1H), 7.95 (s, 1H), 7.82 (brs, 1H), 7.67 (d, J= 8.5 Hz, 1H), 7.56 (d, J= 8.2 Hz, 2H), 7.31 (d, J= 7.9 Hz, 2H), 6.77 (d, J= 8.4 Hz, 2H), 6.57 (d, J= 8.4 Hz, 2H), 3-70-3.77 (m, 1H), 3.43-3.48 (m, 1H), 3.35-3.40 (m, 1H), 2.99-3.10 (m, 2H), 2.40-2.43 (m, 2H), 2.38 (s, 3H), 1.52-1.59 (m, 4H), 1.44-1.47 (m, 1H), 1.04-1.16 (m, 6H), 0.67-0.81 (m, 2H). 13C NMR (100 MHz, DMSO-d6) δppm: 158.6, 156.6, 143.1, 138.6, 130.1, 130.0, 127.5, 126.7, 115.6, 55.6, 53.0, 46.7, 44.6, 33.8, 33.5, 32.0, 26.3, 25.9, 25.7, 21.4. MS (ESI) m/z, calculated: 484.25.05, found [M+H]+: 485.0. N-((S)-1-(4-hydroxyphenyl)-3-((S)-2-imino-5-isobutylimidazolidin-1-yl)propan-2-yl)-4-methylbenzenesulfonamide (5): 1H NMR (400 MHz, DMSO-d6) δppm: 7.95 (brs, 1H), 7.83 (s, 1H), 7.68 (d, J= 8.4 Hz, 1H), 7.55 (d, J= 8.1 Hz, 2H), 7. 30 (d, J= 8.1 Hz, 2H), 6.77 (d, J= 8.4 Hz, 2H), 6.57 (d, J= 8.4 Hz, 2H), 3.69-3.73 (m, 1H), 3.37=3.48 (m, 2H), 3.01-3.09 (m, 2H), 2.33 (s, 3H), 1.38-1.43 (m, 1H), 1.03-1-16 (m, 2H), 0.77 (d, J= 6.8 Hz, 3H), 0.75 (d, J= 6.6 Hz, 3H). 13C NMR (100 MHz, DMSO-d6) δppm: 158.6, 156.6, 143.1, 138.6, 130.3, 130.1, 130.0, 127.6, 126.7, 115.7, 56.2, 53.0, 46.6, 44.6, 24.3, 23.8, 21.7, 21.4. MS (ESI) m/z, calculated: 444.22, found [M+H]+: 445.0. 2-chloro-N-((S)-1-(4-hydroxyphenyl)-3-((S)-2-imino-5-isobutylimidazolidin-1-yl)propan-2-yl)benzenesulfonamide (6): 1H NMR (400 MHz, DMSO-d6) δppm: 9.18 (s, 1H), 8.02 (s, 1H), 7.85-7.88 (m, 1H), 7.75-7.81 (m, 1H), 7.60-7.59 (m, 1H), 7.30-7.52 (m, 1H), 7.45-7.48 (m, 1H), 6.76 (d, J= 8.4 Hz, 2H), 6.50 (d, J= 8.4 Hz, 2H), 3.73 (m, 1H), 3-43-3-51 (m, 3H), 3.09-3.13 (m, 2H), 2-42-2.46 (m, 1H), 1.36-1-45 (m, 1H), 1.13-1.17 (m, 2H), 0.78 (d, J= 6.6 Hz, 3H), 0.77 (d, J= 6.6 Hz, 3H). 13C NMR (100 MHz, DMSO-d6) δppm: 158.7, 158.6, 156.4, 138.7, 134.2, 132.3, 131.0, 130.2, 130.0, 127.9, 127.3, 115.7, 115.6, 56.2, 53.5, 46.7, 44.4, 38.1, 24.3, 23.8, 21.7. MS (ESI) m/z, calculated: 465.01, found [M+H]+: 466.0.