Patent Application
20240293391
The invention relates to isoquinoline and pyridine based CXCR4 antagonists and their use in the treatment and prevention of viral infections such as HIV.
The CXC chemokine receptor 4 (CXCR4) is a seven transmembrane G protein-coupled receptor (GPCR), whose only endogenous ligand is the CXC chemokine ligand 12 (CXCL12), also known as stromal cell-derived factor 1 (SD33F-1). Binding of CXCL12 to CXCR4 activates different signaling pathways, leading to various biological responses such as chemotaxis, cell survival and proliferation, intracellular calcium flux and gene transcription. Under normal physiological conditions, binding of CXCL12 induces leukocytes to migrate along chemokine gradients towards sites with high concentrations of CXCL12 [Busillo & Benovic. Biochim. Biophys. Acta 2007, 1768, 952-963]. Aberrant CXCL12/CXCR4 signaling plays an important role in various pathological processes. Initially, CXCR4 was discovered as a coreceptor, in conjunction with the host CD4 receptor, for the entry of the T-tropic human immunodeficiency virus (HIV) into T-lymphocytes. CXCR4 is the main coreceptor of HIV-1 in the later stages of infection, leads to a decrease in CD4 cell count and is linked to a higher chance of advancing to the acquired immune deficiency syndrome (AIDS) [Alkhatib Curr Opin HIV AIDZ. 2009, 4, 96-103].
Mutations in the gene encoding for CXCR4 lead to truncation of its C-terminal tail and enhanced receptor activity. Clinically, it causes a rare combined immunodeficiency, characterized by warts, hypogammaglobulinemia, recurrent bacterial infection, and myelokathexis, which is known as the WHIM syndrome [Heusinkveld et al. J. Clin. Immunol. 2019, 1990, 532-556]. Overexpression of CXCR4 is reported for more than 20 types of solid and hematological cancers and is correlated with poor prognosis. CXCR4 was shown to be essential for various fundamental aspects of cancer, such as primary tumor growth, cancer cell migration, and the establishment of metastatic sites. This points towards CXCR4 as a general driver of human malignancies [Luker et al. J. Leukoc. Biol. 2021, 109, 969-989]. CXCR4 has also been implicated in various inflammatory disorders, such as rheumatoid arthritis, inflammatory bowel disease and asthma [García-cuesta et al. Front. Endocrinol. 2019, 10, 1-16].
The promise of CXCR4 as drug target spurred the search for small molecule CXCR4 antagonists [Debnath et al. Theranostics 2013, 3, 47-75; Tahirovic et al. Expert Opin. Ther. Pat 2020, 30, 87-101]. AMD3100 (Plerixafor, compound 1,
The invention is summarized in the following statements:
In 2016, a metal-free three-component reaction was published for the synthesis of 1,5-disubstituted [Thomas et al. Chem. Commun. 2016, 52, 2885-2888; Thomas et al. Chem. Commun. 2016, 52, 9236-9239; Parkash et al. Chem. Rec. 2021, 21, 376-385] 1,2,3-triazoles, known as the ‘triazolization reaction of ketones’. Acid-mediated denitrogenative ring opening of triazoloisoquinolines furnished various 1-methyleneisoquinolines [23]. Here, these synthetic methodologies were applied for the synthesis of a series of novel isoquinoline based CXCR4 antagonists, derived from lead compound 4. To probe the optimal substitution pattern for CXCR4 antagonism, various small substituents (R=F, Br, OCH3) were introduced on the isoquinoline moiety. In order to delineate if CXCR4 antagonism depends on the presence of a bicyclic isoquinoline moiety, the corresponding pyridine congeners were also prepared. As head groups, the classical 8-aminotetrahydroquinoline moiety, as well as the simplified 3-methylpyridinyl group, were selected (
The synthesis of the tetrahydroquinoline derivatives is shown in Scheme 1. A multi-component reaction between acetophenone derivatives 9a-d, 2,2-dimethoxyethan-1-amine 10 and 4-nitrophenyl azide (4-NPA) 11 in toluene at 100° C. yielded the dimethyl acetal-substituted triazoles 12a-d. Conversion into the triazolo[5,1-a]isoquinolines 13a-d was achieved using concentrated sulfuric acid in a modified Pomeranz-Fritsch reaction [Opsomer et al. Org. Lett. 2020, 22, 3596-3600]. This was followed by an acid-catalyzed ring opening of the triazole moiety [Jones et al. J. Chem. Soc., Perkin Trans. 1 1985, 19, 2719-2723] using water as the nucleophile to afford the 1-hydroxymethylisoquinolines 14a-d. Upon treatment with SOCl2 at room temperature, the desired 1-chloromethyl isoquinoline derivatives 15a-d were obtained.
The selective mono-protection [Gerber et al. Tetrahedron 2017, 73, 6347-6355; Manickam et al. Bioorg. Med. Chem. Lett. 2021, 31, 127685] of butane-1,4-diamine 16 with di-tert-butyl dicarbonate afforded intermediate 17, which was used for further reaction without any purification. Reductive amination with 6,7-dihydroquinolin-8(5H)-one in the presence of NaBH(OAc)3 at room temperature, yielded compound 18 in 73% yield. The subsequent alkylation of amine 18 by treatment with 1-chloromethylisoquinolines 15a-e in the presence of K2CO3 under reflux temperature was followed by acidic cleavage of the Boc protecting group, yielding final compounds 19a-e. A reductive amination between amine 18 and the commercially available picolinaldehyde in presence of NaBH(OAc)3, followed by Boc deprotection, furnished final compound 20 [Miller et al. cited above; Li et al. Eur. J. Med. Chem. 2018, 149, 30-44].
Reagents and conditions: a) toluene, 100° C., 17 h; b) H2SO4 (80-99 wt %), rt, 1-2 days; c) 2.5M H2SO4, 120° C., 1-2 days; d) SOCl2, DCM, rt, 3 h; e) Boc2O, DCM, rt, 17 h; f) 6,7-dihydroquinolin-8(5H)-one, NaBH(OAc)3, rt, 48 h; g) (i) K2CO3, ACN, reflux, 2 d; (ii) TFA, DCM, rt, 1 h; h) (i) picolinaldehyde, NaBH(OAc)3, DCE, rt, 2 d; (ii) TFA, DCM, rt, 1 h.
The synthesis of the 3-methylpyridine derivatives is depicted in Scheme 2. Formylation of the lithium salt of 2-bromo-3-methylpyridine 21 with DMF afforded 3-methylpicolinaldehyde 22 [28], which was then used in a reductive amination with amine 17, yielding key intermediate 23. Alkylation of 23 with 1-chloromethylisoquinolines 15a-e or a reductive amination with picolinaldehyde yielded target compounds 24a-e and 25, respectively [Eun et al. Arch. Pharmacal Res. 2009, 32, 1673-1679].
Reagents and conditions: a) nBuLi (1.6M in hexane), DMF, Et2O, 24 h; (b) 17, NaBH(OAc)3, DCE, rt, 48 h. c) 15a-d, K2CO3, ACN, reflux, 2 d; d) TFA, DCM, rt, 1 h; e) picolinaldehyde, NaBH(OAc)3, DCE, rt, 2 d.
The synthesis of the quinoline derivatives is shown in Scheme 3. The amino group of commercially available 4-aminobutan-1-ol 26 was reacted with phthalic anhydride yielding intermediate 27 in high yield. The next step involves a Swern oxidation to convert the primary alcohol to the corresponding aldehyde, affording compound 28. Reductive amination with 8-aminoquinoline, using sodium triacetoxyborohydride and acetic acid, furnished compound 29. Alkylation of the exocyclic amino group with various benzylhalides yielded compounds 30a-d. Finally, hydrazine mediated deprotection of the phthalic protecting group afforded the target compounds 31a-d.
Reagents and conditions: a) phthalic anhydride, toluene, reflux; b) (COCl)2, DMSO, CH2Cl2, −78° C. to 0° C.; c) 8-aminoquinoline, NaBH(OAc)3, CH3COOH, DCE, rt; d) benzylhalide, K2CO3, NaI, CH3CN, reflux; e) NH2NH2·H2O, EtOH, reflux.
Compounds 19a-e, 20, 24a-e and 25 were evaluated in a panel of in vitro cell-based assays (Table 1) [Hout et al. PLOS One 2017, 12, 1-18]. First, the ability of the compounds to compete with fluorescently labeled CXCL12 (CXCL12AF647) for binding at CXCR4 was determined. Secondly, since binding of CXCL12 to CXCR4 results into a transient increase of cytosolic calcium levels, the inhibition of this CXCL12-induced calcium mobilization by the various compounds was also investigated. Finally, given the role of CXCR4 as a major coreceptor for HIV entry, the antiviral activity of the compounds against the X4-tropic wild type HIV-1 NL4-3 strain and the HIV-2 ROD strain was determined in MT-4 cells. In parallel, the cytotoxicity of the compounds for uninfected MT-4 cells was assessed. In each of these assays, plerixafor was included as positive control and showed potent inhibitory activity in the various assays.
All newly synthesized isoquinoline based derivatives were able to compete with CXCL12 for binding to CXCR4, as evidenced by IC50 values of less than 40 nM for all analogues, the only exception being compound 24b that displayed an IC50 value of 0.89 μM in the binding assay. It is noteworthy that a structurally simplified analogue 25 carrying a 3-methylpyridinyl moiety (instead of the THQ group) and a pyridinyl ring (instead of the isoquinoline ring) is still endowed with very potent CXCR4 binding affinity (IC50=0.6 nM), making this a very ligand-efficient molecule. A similar trend was found in the CXCR4 calcium mobilization assay: all compounds were active (IC50 values of less than 1 μM), with compound 24b being the least active (IC50=3.85 μM). However, all compounds do show a diminished potency in the calcium flux assay, when compared to the binding assay, which is also observed for plerixafor. The HIV screening revealed a number of analogues (compounds 19a-c, 19e, 20, 24a and 24c) that show excellent antiviral activity against HIV-1 and HIV-2 (EC50<100 nM) and at the same time, lack cytotoxicity for the MT4 cell line. Compounds that were less active in the calcium mobilization assay (such as compounds 19b, 24b, 24 d-e and 25) also displayed a diminished activity as anti-HIV agent. Overall, the biological profile of compound 24c looks very promising, since it shows low nM activity in all assays (IC50 values between 0.6 and 6 nM) and lacks cytotoxicity (CC50=31 μM).
aConcentration needed to inhibit CXCL12 receptor binding by 50%
bConcentration needed to inhibit CXCL12-induced calcium signaling by 50%
cConcentration required to achieve 50% protection of MT-4 cells against HIV-induced cytopathicity
dConcentration required to reduce the viability of mock-infected MT4 cells by 50%
All chemicals were purchased from Acros Organics, Merck, Alfa Aesar, Fluorochem or TCI Europe and were used as received. For column chromatography, 70-230 mesh silica 60 (Acros) was used as the stationary phase. NMR spectra were recorded on commercial instruments (Bruker Avance 300 or Bruker Avance III HD 400 or Bruker Avance II+ 600) and chemical shifts (δ) are reported in parts per million (ppm) referenced to tetramethylsilane (1H), or the internal (NMR) solvent signal (13C). High-resolution mass spectra were acquired on a quadrupole orthogonal acceleration time-of-flight mass spectrometer (Synapt G2 HDMS, Waters, Milford, MA). Samples were infused at 3 μL/min and spectra were obtained in positive ionization mode with a resolution of 15000 (FWHM) using leucine enkephalin as lock mass. Melting points were determined using a Reichert Thermovar apparatus. Recombinant human CXCL12 was purchased from Peprotech and human CXCL12-Alexa 647® (CXCL12AF647) was obtained from Almac.
Commercially available substituted acetophenones 9a-d (1 eq., 2.5 mmol) and 2-aminoacetaldehyde dimethyl acetal 10 (1.1 eq., 2.75 mmol, 289 mg) were added to an oven-dried screw-capped reaction tube equipped with a magnetic stirring bar. The mixture was dissolved in dry toluene (2 ml) followed by addition of 4-nitrophenyl azide 11 (1.5 eq., 3.75 mmol, 615 mg) and stirred overnight at 100° C. in an aluminum heating block. The crude residue was purified via silica gel flash chromatography, yielding the title compounds. The following compounds were made according to this general procedure.
This compound was prepared from 3′,4′-dimethoxyacetophenone 9a (415.45 mg, 2.50 mmol). The crude residue was purified by silica gel flash chromatography using DCM/EtOAc (95:5) as mobile phase, yielding the title compound as a yellowish-orange solid (654 mg, 89%); mp 90-92° C. 1H NMR (400 MHZ, CDCl3) δ: 7.67 (s, 1H), 7.10 (d, J=2.0 Hz, 1H), 7.06 (dd, J=8.2, 2.0 Hz, 1H), 6.97 (d, J=8.2 Hz, 1H), 4.93 (t, J=5.6 Hz, 1H), 4.41 (d, J=5.7 Hz, 2H), 3.94 (s, 3H), 3.92 (s, 3H), 3.37 (s, 6H). 13C NMR (101 MHZ, CDCl3) δ: 150.0, 149.3, 139.0, 132.6, 122.1, 119.2, 112.4, 111.5, 103.7, 56.1, 56.1, 55.4, 49.7. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C14H19N3O4: 294.1448; found: 294.1455
This compound was prepared from 4′-methoxyacetophenone 9b (375.45 mg, 2.50 mmol). The crude residue was purified by silica gel flash chromatography using DCM/EtOAc (95:5) as mobile phase, yielding the title compound as a yellowish-orange viscous oil (488 mg, 74%). 1H NMR (400 MHZ, CDCl3) δ: 7.64 (s, 1H), 7.42 (d, J=8.7 Hz, 2H), 7.01 (d, J=8.7 Hz, 2H), 4.88 (t, J=5.6 Hz, 1H), 4.39 (d, J=5.7 Hz, 2H), 3.85 (s, 3H), 3.33 (s, 6H). 13C NMR (101 MHZ, CDCl3) δ: 160.4, 138.7, 132.5, 130.5, 118.9, 114.4, 103.3, 55.3, 55.1, 49.4. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C13H17N3O3: 264.1343; found: 264.1346.
This compound was prepared from 4′-fluoroacetophenone 9c (345.35 mg, 2.50 mmol). The crude residue was purified by silica gel flash chromatography using DCM/EtOAc (95:5) as mobile phase, yielding the title compound as a reddish viscous oil (447 mg, 71%). 1H NMR (400 MHZ, CDCl3) δ: 7.65 (s, 1H), 7.47 (dd, J=8.8, 5.2 Hz, 2H), 7.16 (t, J=8.6 Hz, 2H), 4.86 (t, J=5.6 Hz, 1H), 4.35 (d, J=5.6 Hz, 2H), 3.33 (s, 6H). 13C NMR (101 MHZ, CDCl3) δ: 164.6, 162.1, 138.0, 132.9, 131.2, 122.9, 116.2, 103.6, 55.5, 49.6. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C12H14FN3O2: 252.1143; found: 252.1139.
This compound was prepared from 4′-bromoacetophenone 9 d (497.6 mg, 2.50 mmol). The crude residue was purified by silica gel flash chromatography using DCM/EtOAc (95:5) as mobile phase, yielding the title compound as an orange viscous oil (485 mg, 62%). 1H NMR (400 MHZ, CDCl3) δ: 7.68 (s, 1H), 7.62 (d, J=8.5 Hz, 2H), 7.37 (d, J=8.5 Hz, 2H), 4.87 (t, J=5.6 Hz, 1H), 4.36 (d, J=5.6 Hz, 2H), 3.34 (s, 6H). 13C NMR (101 MHZ, CDCl3) δ: 138.1, 133, 132.4, 130.9, 125.9, 124.2, 103.7, 55.6, 49.8. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C12H14BrN3O2: 312.0343; found: 312.0338.
To an open round-bottom flask equipped with a magnetic stirring bar containing the triazole 12a-d (1.2 mmol) was added concentrated H2SO4 (80-95%, 2.5-4 mL), while stirring at 0° C. in an ice bath. After cooling for 30 minutes, the reaction mixture was stirred overnight at room temperature. The reaction mixture was poured over ice and neutralized by slowly adding a 3 M aqueous NaOH solution. The aqueous phase was subsequently extracted five times with dichloromethane. The combined organic layers were dried over MgSO4 and concentrated in vacuo. The crude residue was purified by flash chromatography, yielding the title compound. The following compounds were made according to this procedure.
This compound was prepared from triazole 12a (354 mg, 1.21 mmol). The crude residue was purified by silica gel flash chromatography using DCM/EtOAc (80:20) as mobile phase, yielding the title compound as an off-white solid (274 mg, 99%). mp 221-224° C. 1H NMR (400 MHZ, CDCl3) δ: 8.42 (d, J=7.3 Hz, 1H), 8.29 (s, 1H), 7.39 (s, 1H), 7.13 (s, 1H), 7.07 (d, J=7.3 Hz, 1H), 4.05 (s, 3H), 4.02 (s, 3H). 13C NMR (101 MHZ, CDCl3) δ: 150.7, 150.6, 132.3, 124.5, 123.9, 121.0, 117.3, 115.4, 107.9, 104.5, 56.3, 56.2. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C12H11N3O2: 230.0924; found: 230.0936.
This compound was prepared from triazole 12b (301 mg, 1.14 mmol). The crude residue was purified by silica gel flash chromatography using DCM/EtOAc (80:20) as mobile phase, yielding the title compound as an off-white solid (226 mg, 99%). mp 133-134° C. 1H NMR (400 MHZ, CDCl3) δ: 8.43 (d, J=7.4 Hz, 1H), 8.26 (s, 1H), 7.96 (d, J=8.8 Hz, 1H), 7.20 (dd, J=8.8, 2.6 Hz, 1H), 7.12 (d, J=2.5 Hz, 1H), 7.06 (d, J=7.4 Hz, 1H), 3.92 (s, 3H). 13C NMR (101 MHZ, CDCl3) δ: 160.0, 132.5, 130.7, 125.5, 124.6, 122.8, 118.3, 116.7, 115.6, 108.8, 55.5. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C11H9N3O: 200.0818; found: 200.0826
This compound was prepared from triazole 12c (301 mg, 1.2 mmol) and H2SO4 (80%, aq.). The crude residue was purified by silica gel flash chromatography using DCM/EtOAc (80:20) as mobile phase, yielding the title compound as an off-white solid (196 mg, 87%). mp 123-126° C. 1H NMR (400 MHZ, CDCl3) δ: 8.55 (d, J=7.4 Hz, 1H), 8.38 (s, 1H), 8.15 (dd, J=8.6, 5.3 Hz, 1H), 7.47 (d, J=9 Hz, 1H), 7.4 (d, J=8.4 Hz, 1H), 7.15 (d, J=7.4 Hz, 1H). 13C NMR (101 MHZ, CDCl3) δ 162.71 (d, J=250.3 Hz), 138.48, 133.55, 130.96 (d, J=8.5 Hz), 126.62 (d, J=9.1 Hz), 125.63, 123.90, 117.79 (d, J=23.9 Hz), 115.42 (d, J=3.4 Hz), 113.05 (d, J=22.2 Hz). HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C10H6FN3: 188.0618; found: 188.0617.
This compound was prepared from triazole 12 d (375 mg, 1.2 mmol). The crude residue was purified by silica gel flash chromatography using DCM/EtOAc (80:20) as mobile phase, yielding the title compound as an off-white solid (285 mg, 95%). mp 209-211° C. 1H NMR (400 MHZ, CDCl3) δ: 8.54 (d, J=7.4 Hz, 1H), 8.42 (s, 1H), 8.02 (d, J=8.5 Hz, 1H), 7.97 (d, J=1.9 Hz, 1H), 7.78 (dd, J=8.5, 1.9 Hz, 1H), 7.3 (d, J=7.4 Hz, 1H). 13C NMR (101 MHZ, CDCl3) δ: 131.8, 131.7, 130.3, 129.8, 125.7, 125.3, 123.5, 1229, 121.4, 114.6. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C10H6BrN3: 247.9818; found: 247.9812.
A solution of the triazoloisoquinoline 13a-d (1.2 mmol) in a 2.5M H2SO4 solution (8-10 ml) was heated at 120° C. until completion of the reaction (24-36 hours). Subsequently, the reaction mixture was poured over ice and neutralized by adding a saturated aqueous NaHCO3 solution. The aqueous phase was extracted three times with dichloromethane. The combined organic phases were dried over MgSO4 and concentrated in vacuo to afford the title compound, which was used for the next step without further purification. The following compounds were made according to this procedure.
This compound was prepared from triazoloisoquinoline 13a (275 mg, 1.2 mmol). The title compound was isolated as an off-white solid (242 mg, 92%). mp 131-133° C. 1H NMR (400 MHZ, CDCl3) δ: 8.33 (d, J=5.7, 1H), 7.46 (d, J=5.6, 1H), 7.11 (s, 1H), 7.05 (s, 1H), 5.13 (s, 2H), 4.04 (s, 3H), 4.03 (s, 3H). 13C NMR (101 MHZ, CDCl3) δ: 155.5, 153.7, 151.1, 140.1, 133.5, 119.9, 106.1, 101.9, 62.1, 56.8. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C12H13NO3: 220.0968; found: 220.0974.
This compound was prepared from triazoloisoquinoline 13b (240 mg, 1.2 mmol). The title compound was isolated as an off-white solid (173 mg, 76%); mp 63-65° C. 1H NMR (400 MHZ, CDCl3) δ: 8.30 (d, J=5.8 Hz, 1H), 7.76 (d, J=9.2 Hz, 1H), 7.43 (d, J=5.7 Hz, 1H), 7.19 (s, 1H), 7.04 (s, 1H), 5.11 (s, 2H), 3.88 (s, 3H). 13C NMR (101 MHz, CDCl3) δ: 160.7, 156.5, 140.5, 137.9, 124.8, 120.2, 120.1, 119.5, 104.8, 60.9, 55.2. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C11H1NO2: 190.0862; found: 190.0869.
This compound was prepared from triazoloisoquinoline 13c (225 mg, 1.2 mmol). The title compound was isolated as an off-white solid (194 mg, 91%). mp 72-74° C. 1H NMR (400 MHZ, CDCl3) δ: 8.45 (d, J=5.8 Hz, 1H), 7.96 (dd, J=9.1, 5.3 Hz, 1H), 7.57 (d, J=5.8 Hz, 1H), 7.48 (dd, J=9.2, 2.5 Hz, 1H), 7.3 (td, J=8.5, 2.5 Hz, 1H), 5.22 (s, 2H), 4.93 (s, 1H). 13C NMR (101 MHZ, CDCl3) δ: 163.9, 161.4, 156.9, 140.8, 137.1, 125.8, 121.6, 119.4, 117.4, 110.3. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C10H8FNO: 178.0663; found: 178.0659.
This compound was prepared from triazoloisoquinoline 13 d (298 mg, 1.2 mmol). The title compound was isolated as an off-white solid (238 mg, 83%). mp 143-145° C. 1H NMR (400 MHZ, CDCl3) δ: 8.47 (d, J=5.4 Hz, 1H), 8.03 (d, J=1.8 Hz, 1H), 7.79 (d, J=8.9 Hz, 1H), 7.69 (dd, J=8.9, 1.9 Hz, 1H), 7.51 (d, J=5.7 Hz, 1H), 5.20 (s, 2H), 4.89 (s, 1H). 13C NMR (101 MHZ, CDCl3) δ: 158.2, 137.5, 131.6, 130, 125.7, 125.4, 123.8, 119.7, 61.8. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C10H8BrNO: 237.9862; found: 237.9851
To a solution of 1-hydroxymethylisoquinoline 14a-d (1.2 mmol) in dry dichloromethane (1 ml) was added an excess of SOCl2. The resulting reaction mixture was stirred for 3 hours at room temperature. After completion of the reaction, the reaction mixture was neutralized with an aqueous saturated NaHCO3 solution. The aqueous phase was extracted several times with dichloromethane. The combined organic layers were dried over MgSO4 and concentrated under vacuum to afford the title compound, that was used for the next step without further purification. The following compounds were made according to this procedure:
This compound was prepared from 1-hydroxymethylisoquinoline 14a (263 mg, 1.2 mmol). The title compound was isolated as a red-orange solid; yield: 285 mg (99%); mp 148-150° C. 1H NMR (400 MHZ, CDCl3) δ: 8.34 (d, J=5.8 Hz, 1H), 7.50 (d, J=5.5 Hz, 1H), 7.42 (s, 1H), 7.09 (s, 1H), 5.09 (s, 2H), 4.07 (s, 3H), 4.03 (s, 3H). 13C NMR (101 MHZ, CDCl3) δ: 153.1, 153.0, 150.6, 141.0, 133.8, 122.7, 120.7, 105.4, 103.2, 56.2, 56.2, 45.7. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C12H12ClNO2: 238.0629; found: 238.0631.
This compound was prepared from 1-hydroxymethylisoquinoline 14b (227 mg, 1.2 mmol). The title compound was isolated as a dark red solid (225 mg, 90%). mp 85-87° C. 1H NMR (400 MHZ, CDCl3) δ: 8.40 (d, J=5.7 Hz, 1H), 8.15 (d, J=9.2 Hz, 1H), 7.55 (d, J=5.7 Hz, 1H), 7.29 (dd, J=9.2, 2.5 Hz, 1H), 7.10 (d, J=2.5 Hz, 1H), 5.09 (s, 2H), 3.96 (s, 3H). 13C NMR (101 MHZ, CDCl3) δ: 160.9, 154.7, 141.7, 138.9, 126.8, 121.0, 120.8, 104.8, 55.4, 44.3. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C11H10ClNO: 208.0524; found: 208.0516.
This compound was prepared from 1-hydroxymethylisoquinoline 14c (213 mg, 1.2 mmol). The title compound was isolated as a red solid (224 mg, 95%). mp 65-67° C. 1H NMR (600 MHZ, CDCl3) δ: 8.47 (d, J=5.7 Hz, 1H), 8.30 (dd, J=9.1, 5.3 Hz, 1H), 7.62 (d, J=5.7 Hz, 1H), 7.52-7.39 (m, 2H), 5.12 (s, 2H). 13C NMR (151 MHZ, CDCl3) δ: 164, 162.4, 155.9, 142.9, 138.6, 128.5, 123.8, 121.6, 118.5, 110.9, 45. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C10H7ClFN: 196.0324; found: 196.0316.
This compound was prepared from 1-hydroxymethylisoquinoline 14 d (286 mg, 1.2 mmol). The title compound was isolated as a redish solid (301 mg, 98%). mp 108-110° C. 1H NMR (400 MHZ, CDCl3) δ: 8.50 (d, J=5.7 Hz, 1H), 8.14 (d, J=9 Hz, 1H), 8.05 (d, J=1.9 Hz, 1H), 7.75 (dd, J=9, 1.9 Hz, 1H), 7.58 (d, J=7.6 Hz, 1H), 5.12 (s, 2H). 13C NMR (101 MHZ, CDCl3) δ: 156.1, 143.1, 137.9, 131.5, 129.8, 127, 125.4, 125, 120.8, 44.8. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C10H7BrClN: 255.9524; found: 255.9520
To a solution of 1,4-diaminobutane 16 (10 eq., 56.7 mmol, 5 g) in dichloromethane (62.5 mL) was added a solution of Boc2O (1 eq., 5.67 mmol, 1.238 g) in dichloromethane (62.5 mL) through a dropping funnel. The reaction mixture was stirred at room temperature for 16 h. After filtering the suspension, the filtrate was evaporated under vacuum. The oily residue was washed with brine and extracted with EtOAc to remove the excess amount of diamine. The organic layer was dried over anhydrous Na2SO4 and evaporated under vacuum, yielding the title compound as a colourless oil (6.9 g, 65%). 1H NMR (400 MHZ, CDCl3) δ: 4.77 (s, 1H), 3.07 (t, J=6.0 Hz, 2H), 2.66 (t, J=6.6 Hz, 2H), 1.52-1.42 (m, 4H), 1.39 (s, 9H), 1.09 (s, 2H). 13C NMR (75 MHZ, CDCl3) δ: 155.88, 78.41, 41.51, 40.09, 30.59, 28.17, 27.19. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C9H20N2O2: 189.1597; found: 189.1597.
6,7-Dihydroquinolin-8(5H)-one (1.03 eq., 3.40 mmol, 0.500 g) was added to a slurry of NaBH(OAc)3 (1.78 eq., 5.87 mmol, 1.244 g) in dichloroethane (3 ml), followed by the addition of tert-butyl (4-aminobutyl) carbamate 17 (1 eq., 3.30 mmol, 0.621 g). The reaction was stirred at room temperature for 48 hours. The reaction mixture was quenched with a 1N NaOH solution to obtain a PH˜8 of the aqueous layer. The reaction mixture was extracted three times with dichloromethane (3×) and the combined organic layers were concentrated to a volume of approximately 3 ml. Heptane (10 ml) was added and the volume was concentrated to 5 ml. Upon cooling the reaction mixture to room temperature, a precipitate was formed. After further cooling the suspension to 0° C., the precipitate was filtered off and dried under vacuum, yielding the title compound as a light brown solid (674 mg, 64%); mp 73-75° C. 1H NMR (400 MHZ, CDCl3) δ: 8.39 (d, J=4.1 Hz, 1H), 7.82 (s, 1H), 7.41 (d, J=7.8 Hz, 1H), 7.12 (t, J=7.4, 4.9 Hz, 1H), 4.01 (s, 1H), 3.09 (t, J=33.0 Hz, 2H), 2.90-2.70 (m, 4H), 2.32-2.00 (m, 2H), 1.95 (t, J=16.3 Hz, 2H), 1.73-1.62 (m, 2H), 1.62-1.51 (m, 2H), 1.42 (s, 9H). 13C NMR (101 MHZ, CDCl3) δ: 156.31, 147.03, 137.27, 132.95, 122.48, 57.79, 46.09, 28.64, 27.77, 27.40, 26.52, 22.02, 20.18. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C18H29N3O2: 320.2332; found: 320.2335.
To a solution of 1-(chloromethyl)isoquinoline 15a-d (1.5 eq) and tert-butyl (4-((5,6,7,8-tetrahydroquinolin-8-yl)amino)butyl)carbamate 18 (1 eq) in dry acetonitrile (5 ml) was added K2CO3 (8 eq). The reaction mixture was refluxed for 24-48 hours. The mixture was cooled to room temperature and filtered through a Celite® pad. The filtrate was evaporated in vacuo and the crude residue was purified via silica gel flash chromatography using a mixture of Et2O/MeOH as mobile phase (in a gradient gradually ranging from 100:0 to 90:10) yielding the title compounds. The following compounds were made according to this procedure.
This compound was prepared from 1-(chloromethyl)-6,7-dimethoxyisoquinoline 15a (86 mg, 0.36 mmol) and tert-butyl (4-((5,6,7,8-tetrahydroquinolin-8-yl) amino) butyl) carbamate 18 (100 mg, 0.32 mmol). The title compound was isolated as a brown viscous oil (114 mg, 69%). 1H NMR (300 MHZ, CDCl3) δ: 8.52 (s, 1H), 8.43 (d, J=4.2 Hz, 1H), 8.26 (d, J=5.6 Hz, 1H), 7.37 (d, J=5.6 Hz, 1H), 7.30 (d, J=7.5 Hz, 1H), 7.03-6.97 (m, 2H), 4.62 (s, 1H), 4.35 (dd, J=56.5, 12.4 Hz, 2H), 4.14 (s, 3H), 4.00 (s, 3H), 2.97-2.46 (m, 6H), 2.12-1.90 (m, J=22.3, 11.4 Hz, 3H), 1.70-1.54 (m, 1H), 1.39 (s, 9H), 1.34-1.16 (m, J=28.5, 11.3 Hz, 4H). 13C NMR (75 MHZ, CDCl3) δ: 156.09, 152.72, 149.61, 147.10, 140.57, 136.49, 134.38, 133.31, 124.39, 121.55, 119.29, 106.62, 104.75, 77.36, 60.90, 58.81, 56.51, 55.99, 50.53, 29.55, 28.54, 27.74, 25.31, 24.94, 21.80. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C30H40N4O4: 521.3122; found: 521.3124.
This compound was prepared from 1-(chloromethyl)-6-methoxyisoquinoline 15b (104 mg, 0.33 mmol) and tert-butyl (4-((5,6,7,8-tetrahydroquinolin-8-yl)amino)butyl) carbamate 18 (101.4 mg, 0.49 mmol). The title compound was isolated as a brown viscous oil (136 mg, 85%). 1H NMR (400 MHZ, CDCl3) δ: 8.66 (d, J=9.2 Hz, 1H), 8.54 (d, J=3.9 Hz, 1H), 8.32 (d, J=5.8 Hz, 1H), 7.43 (d, J=5.8 Hz, 1H), 7.36 (d, J=7.4 Hz, 1H), 7.18 (dd, J=9.3, 2.5 Hz, 1H), 7.08 (dd, J=7.6, 4.6 Hz, 1H), 7.01 (d, J=2.6 Hz, 1H), 4.89 (s, 1H), 4.38 (s, 1H) 4.24 (s, 2H), 3.94 (s, 3H), 2.94-2.56 (m, 6H), 2.42-2.12 (m, 4H), 2.15-1.99 (m, 4H), 1.39 (s, 9H). 13C NMR (101 MHz, CDCl3) δ: 161.5, 156.8, 147.8, 142.5, 139.1, 137.3, 135.2, 129.9, 122.5, 120.6, 120, 105, 56.1, 51.5, 40.5, 30.4, 30, 29.2, 28.2, 25.4, 22.2. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C29H38N4O3: 491.3016; found: 491.3017.
This compound was prepared from 1-(chloromethyl)-6-fluoroisoquinoline 15c (117 mg, 0.6 mmol) and tert-butyl (4-((5,6,7,8-tetrahydroquinolin-8-yl)amino) butyl) carbamate 18 (127.8 mg, 0.4 mmol). The title compound was isolated as a brown viscous oil (164 mg, 86%). 1H NMR (400 MHZ, CDCl3) δ: 8.95 (dd, J=8.9, 5.8 Hz, 1H), 8.51 (d, J=3.9 Hz, 1H), 8.36 (d, J=5.7 Hz, 1H), 7.47 (d, J=5.7 Hz, 1H), 7.38-7.29 (m, 3H), 7.05 (dd, J=7.6, 4.7 Hz, 1H), 4.80 (s, 1H), 4.36 (d, J=12.5 Hz, 1H), 4.20 (d, J=3.6 Hz, 2H), 2.95-2.52 (m, 6H), 2.14-1.97 (m, 2H), 1.88-1.63 (m, 4H), 1.39 (s, 9H), 1.30-1.22 (m, 2H). 13C NMR (101 MHZ, CDCl3) δ: 155.7, 146.8, 141.9, 137.7, 137.6, 136.3, 134.2, 130.8, 124.9, 121.4, 119.8, 109.6, 109.3, 78.5, 60.8, 57.2, 53.2, 50.6, 39.7, 33.3, 29.4, 29, 28.1, 27.2, 24.5, 24, 21.2. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C28H35FN4O2: 479.2817; found: 479.2808.
This compound was prepared from 1-(chloromethyl)-6-bromoisoquinoline 15 d (142 mg, 0.55 mmol) and tert-butyl (4-((5,6,7,8-tetrahydroquinolin-8-yl)amino) butyl) carbamate 18 (117.9 mg, 0.37 mmol). The title compound was isolated as a brown viscous oil (143 mg, 72%). 1H NMR (400 MHZ, CDCl3) δ: 8.78 (d, J=8.9 Hz, 1H), 8.53 (d, J=3.8 Hz, 1H), 8.40 (d, J=5.7 Hz, 1H), 7.92 (d, J=1.9 Hz, 1H), 7.64 (dd, J=9, 1.97 Hz, 1H), 7.43 (d, J=5.7 Hz, 1H), 7.35 (d, J=7.6 Hz, 1H), 7.06 (dd, J=7.6, 4.69 Hz, 1H), 4.80 (s, 1H), 4.36 (d, J=12, 1H), 4.20 (d, J=10 Hz, 2H), 2.95-2.54 (m, 6H), 2.11-2.03 (m, 2H), 1.75-1.58 (m, 4H), 1.41 (s, 9H), 1.21 (t, J=7 Hz, 2H). 13C NMR (101 MHZ, CDCl3) δ: 155.9, 146.8, 141.9, 137.6, 137.3, 136.6, 130.2, 128.9, 128.7, 124.8, 123.2, 121.8, 119.3, 61.2, 57.1, 50.8, 39.6, 30.7, 29.3, 28.9, 28.2, 27.2, 27.5, 26.9, 24.3, 24, 21.2, 19.9. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C28H35BrN4O2: 539.2016; found: 539.2021.
This compound was prepared from the commercially available 1-(bromomethyl)isoquinoline (200 mg, 0.9 mmol) and tert-butyl (4-((5,6,7,8-tetrahydroquinolin-8-yl)amino)butyl) carbamate 18 (191.78 mg, 0.6 mmol). The title compound was isolated as a brown viscous oil (169 mg, 61%). 1H NMR (400 MHZ, CDCl3) δ: 8.52 (d, J=3.8 Hz, 1H), 8.09 (d, J=8.5 Hz, 1H), 8.00 (d, J=8.4 Hz, 1H), 7.94 (d, J=8.5 Hz, 1H), 7.77 (d, J=7.7 Hz, 1H), 7.68-7.61 (m, 1H), 7.52-7.43 (m, 1H), 7.33 (d, J=7.5 Hz, 1H), 7.04 (dd, J=7.6, 4.7 Hz, 1H), 4.86 (s, 1H), 4.21 (dd, J=8.3, 6.6 Hz, 1H), 4.01-3.87 (m, 2H), 2.99 (s, 2H), 2.86-2.64 (m, 4H), 2.11-1.87 (m, 2H), 1.54-1.44 (m, 2H), 1.42 (s, 9H), 1.29-1.20 (m, 2H). 13C NMR (101 MHz, CDCl3) δ: 156.5, 147.7, 137.1, 136.6, 134.8, 129.6, 129.2, 128, 127.9, 126.3, 122.1, 121.9, 61.5, 58.9, 53.9, 52.8, 40.7, 30.2, 29.7, 28.9, 28.1, 26, 21.9. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C28H36N4O2: 461.2911; found: 461.2935.
To a flame-dried flask under an inert argon atmosphere was added 2-bromo-3-methyl-pyridine 21 (1 eq., 11.40 mmol, 2 g) and Et2O (1-2 ml) at −78° C., followed by addition of a 1.6M butyllithium solution in hexane (0.43 ml). After stirring for 3 hours at −78° C., DMF (2 ml) was added to this deep red-brown solution. The reaction temperature was gradually increased to room temperature and was stirred for an additional 22 hours. The reaction mixture was quenched through the dropwise addition of ice-water and extracted with Et2O. The organic layer was washed with a 5% aqueous NaHCO3 solution and brine. The combined organic layers were dried over MgSO4 and concentrated under vacuum. The crude residue was purified by silica gel flash chromatography using Et2O/petroleum ether (80:20) as the eluent, yielding the title compound as an orange-red oil (848 mg, 62%).
3-Methylpicolinaldehyde 22 (1 eq., 8.25 mmol, 1 g) was added to a slurry of NaBH(OAc)3 (1.9 eq., 15.68 mmol, 3.324 g) in dichloroethane (15 ml), followed by the addition of tert-butyl (4-aminobutyl) carbamate 17 (1.4 eq., 11.56 mmol, 2.176 g). The reaction was stirred at room temperature for 48 hours. The reaction mixture was quenched with a 1N NaOH solution to obtain a pH˜8 in the aqueous layer. After extracting the mixture with dichloromethane (3×), the combined organic phases were concentrated to a volume of approximately 3 ml. The crude residue was purified by silica gel column chromatography using Et2O/MeOH (96:4) as the eluent, affording the title compound as a yellowish viscous oil (1.8 g, 74%). 1H NMR (400 MHZ, CDCl3) δ: 8.37 (d, J=3.84 Hz, 1H), 7.41 (dd, J=7.6, 0.64 Hz, 1H), 7.06 (dd, J=7.5, 4.8 Hz, 1H), 4.79 (s, 1H), 3.86 (s, 2H), 3.17-3.06 (m, 2H), 2.94 (s, 1H), 2.72 (t, J=6.7 Hz, 2H), 2.29 (s, 3H), 1.64-1.49 (m, 4H), 1.42 (s, 9H). 13C NMR (101 MHz, CDCl3) δ: 157.1, 156.1, 146.5, 137.7, 130.9, 121.9, 79, 52, 49.5, 40.5, 28.5, 27.4, 18.1. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C16H27N3O2: 294.2176; found: 294.2169.
A similar procedure as mentioned earlier for the coupling with amine 18 was used. The following compounds were made according to this procedure.
This compound was prepared from 1-(chloromethyl)-6,7-dimethoxyisoquinoline 15a (50 mg, 0.21 mmol) and tert-butyl (4-(((3-methylpyridin-2-yl) methyl)amino) butyl) carbamate 23 (48 mg, 0.15 mmol). The title compound was isolated as a brownish viscous oil (76 mg, 73%). 1H NMR (400 MHZ, CDCl3) δ: 8.38 (d, J=4.1 Hz, 1H), 8.30 (d, J=5.6 Hz, 1H), 7.51 (s, 1H), 7.42 (d, J=5.6 Hz, 1H), 7.36 (d, J=7.4 Hz, 1H), 7.08 (dd, J=7.6, 4.8 Hz, 1H), 7.02 (s, 1H), 4.75 (s, 1H), 4.23 (s, 2H), 4.00 (s, 3H), 3.82 (s, 5H), 2.96-2.83 (m, 2H), 2.59 (s, 2H), 2.10 (s, 3H), 1.64-1.51 (m, 2H), 1.40 (s, 9H), 1.30 (t, J=6.9 Hz, 2H). 13C NMR (101 MHZ, CDCl3) δ: 162.5, 156.8, 155.9, 151.9, 151.3, 147.2, 141.2, 136.8, 133.1, 131.9, 123.9, 121.3, 117.7, 105.4, 79.5, 67.6, 59.1, 56.1, 54, 35.8, 28.4, 27.7, 25.8, 20.4. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C28H38N4O4: 495.2966; found: 495.2960.
This compound was prepared from 1-(chloromethyl)-6-methoxyisoquinoline 15b (100 mg, 0.48 mmol) and tert-butyl (4-(((3-methylpyridin-2-yl)methyl)amino) butyl) carbamate 23 (94.2 mg, 0.32 mmol). The title compound was isolated as a brownish viscous oil (118 mg, 57%). 1H NMR (400 MHZ, CDCl3) δ: 8.38 (d, J=3.8 Hz, 1H), 8.33 (d, J=5.8 Hz, 1H), 7.88 (d, J=9 Hz, 1H), 7.43 (d, J=5.8 Hz, 1H), 7.40 (d, J=7.1 Hz, 1H), 7.10 (dd, J=7.5, 4.8 Hz, 1H), 7.06-6.98 (m, 2H), 4.71 (s, 1H), 4.14 (s, 2H), 3.91 (s, 3H), 3.83 (s, 2H), 2.94-2.82 (m, 2H), 2.57 (t, J=7.2 Hz, 2H), 2.10 (s, 3H), 1.58-1.46 (m, 2H), 1.44 (s, 9H), 1.32-1.17 (m, 2H). 13C NMR (101 MHZ, CDCl3) δ: 160.2, 155.8, 146, 141.8, 138.2, 137.8, 133.2, 128.1, 123.2, 122.3, 119.7, 119.1, 104.1, 78.7, 59.1, 55.2, 53.7, 39.7, 28.2, 27.4, 22.7, 18. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C27H36N4O3: 465.2860; found: 465.2863.
This compound was prepared from 1-(chloromethyl)-6-fluoroisoquinoline 15c (120 mg, 0.61 mmol) and tert-butyl (4-(((3-methylpyridin-2-yl) methyl)amino)butyl) carbamate 23 (120 mg, 0.41 mmol). The title compound was isolated as a brown viscous oil (150 mg, 81%). 1H NMR (400 MHZ, CDCl3) δ: 8.41 (d, J=5.7 Hz, 2H), 8.06 (dd, J=9.2, 5.7 Hz, 1H), 7.49 (d, J=5.8 Hz, 1H), 7.42 (d, J=6.9 Hz, 1H), 7.35 (dd, J=9.3, 2.5 Hz, 1H), 7.17 (dd, J=8.8, 2.4 Hz, 1H), 7.12 (dd, J=7.6, 4.9 Hz, 1H), 4.66 (s, 1H), 4.16 (s, 2H), 3.83 (s, 2H), 2.97-2.86 (m, 2H), 2.56 (t, J=7.3 Hz, 2H), 2.13 (s, 3H), 1.59-1.48 (m, 2H), 1.42 (s, 9H), 1.31-1.19 (m, 2H). 13C NMR (101 MHZ, CDCl3) δ: 164.1, 161.6, 159, 156.9, 156.1, 146.4, 142.4, 138, 133.3, 129.9, 125, 122.6, 120.2, 116.9, 110, 79, 59.6, 59.4, 54.1, 40, 28.5, 27.8, 23, 18.3. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C26H33FN4O2: 453.2660; found: 453.2651.
This compound was prepared from 1-(chloromethyl)-6-bromoisoquinoline 15 d (100 mg, 0.39 mmol) and tert-butyl (4-(((3-methylpyridin-2-yl)methyl)amino)butyl) carbamate 23 (76.25 mg, 0.26 mmol). The title compound was isolated as a brown viscous oil (104 mg, 78%). 1H NMR (400 MHZ, CDCl3) δ: 8.43 (d, J=5.8 Hz, 1H), 8.40 (d, J=3.9 Hz, 1H), 7.93 (d, J=1.9 Hz, 1H), 7.85 (d, J=9 Hz, 1H), 7.50-7.41 (m, 3H), 7.12 (dd, J=7.6, 4.8 Hz, 1H), 4.65 (s, 1H), 4.15 (s, 2H), 3.83 (s, 2H), 2.96-2.87 (m, 2H), 2.56 (t, J=7.3 Hz, 2H), 2.16 (s, 3H), 1.75-1.60 (m, 2H), 1.58-1.47 (m, 2H), 1.42 (s, 9H). 13C NMR (101 MHZ, CDCl3) δ: 157.1, 156.4, 146.7, 142.8, 138.4, 137.9, 133.7, 130.4, 129.3, 128.7, 126.5, 125, 123, 119.9, 79.3, 59.7, 54.5, 40.3, 28.8, 28.1, 23.3, 18.7. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C26H33BrN4O2: 513.1860; found: 513.1854.
This compound was prepared from 1-(bromomethyl)isoquinoline (100 mg, 0.45 mmol) and tert-butyl (4-(((3-methylpyridin-2-yl)methyl)amino) butyl)carbamate 15 (88 mg, 0.3 mmol). The title compound was isolated as a brown viscous oil (110 mg, 84%). 1H NMR (400 MHZ, CDCl3) δ: 8.37 (dd, J=4.7, 1.1 Hz, 1H), 8.05 (d, J=8.4 Hz, 2H), 7.77 (dd, J=8.2, 1.3 Hz, 1H), 7.71-7.65 (m, 1H), 7.54-7.47 (m, 2H), 7.39 (dd, J=7.7, 0.7 Hz, 1H), 7.06 (dd, J=7.6, 4.8 Hz, 1H), 4.74 (s, 1H), 3.91 (s, 2H), 3.86 (s, 2H), 3.06-2.93 (m, 2H), 2.57 (t, J=7.2 Hz, 2H), 2.33 (s, 3H), 1.70-1.61 (m, 2H), 1.59-1.50 (m, 2H), 1.42 (s, 9H). 13C NMR (101 MHZ, CDCl3) δ: 161.2, 157.5, 156.4, 147.9, 146.6, 138.4, 136.4, 133.6, 129.7, 129.5, 127.9, 127.7, 126.5, 122.8, 121.9, 61.8, 60.1, 54.7, 40.6, 28.9, 28.2, 24.1, 19. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C26H34N4O2: 435.2754; found: 435.2757.
To a solution of the Boc-protected intermediates (0.1 mmol) in dry dichloromethane (1 ml) was added trifluoroacetic acid (0.3 ml). The reaction mixture was stirred for 1 hour at room temperature. The solvents were evaporated in vacuo. The residue was neutralized by adding a saturated aqueous NaHCO3 solution and the aqueous layer was extracted five times with dichloromethane. The combined organic layers were dried over MgSO4 and evaporated. The crude residue was purified via silica gel flash column chromatography using a mixture of DCM/MeOH (in a ratio gradually ranging from 95:5 to 90:10) as mobile phase. The following compounds were made according to this procedure.
This compound was prepared from its Boc-protected precursor (50 mg, 0.09 mmol). The title compound was isolated as a bright brown viscous oil (47 mg, 95%). 1H NMR (400 MHZ, DMSO) δ (ppm): 8.43 (d, J=4.5 Hz, 1H), 8.21 (d, J=5.6 Hz, 1H), 8.17 (s, 1H), 7.55 (d, J=5.4 Hz, 1H), 7.48 (d, J=7.9 Hz, 1H), 7.30 (s, 1H), 7.20-7.10 (m, 1H), 5.32 (s, 1H), 4.29 (dd, J=21.5, 12.0 Hz, 2H), 3.99 (s, 3H), 3.92 (s, 3H), 2.15-1.50 (m, 6H), 1.39 (s, 2H), 1.30-1.09 (m, 4H), 1.07-0.93 (m, J=8.3, 6.1 Hz, 4H). 13C NMR (101 MHZ, DMSO) δ: 158.51, 158.20, 157.89, 157.59, 152.18, 139.94, 132.61, 123.25, 121.82, 121.65, 119.06, 118.84, 116.52, 115.85, 112.78, 55.51, 52.06, 49.46, 48.60, 45.67, 43.69, 7.17. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C25H32N4O2: 421.2598; found: 421.2594.
This compound was prepared from its Boc-protected precursor (30 mg, 0.06 mmol). The title compound was isolated as a bright brown viscous oil (22 mg, 92%). 1H NMR (400 MHZ, CDCl3) δ: 9.99 (s, 2H), 8.54 (d, J=3.7 Hz, 1H), 8.51 (d, J=5.8 Hz, 1H), 8.03 (d, J=9.3 Hz, 1H), 7.51 (d, J=5.8 Hz, 1H), 7.41 (d, J=7.2 Hz, 1H), 7.22 (dd, J=9.3, 2.5 Hz, 1H), 7.12 (dd, J=7.7, 4.8 Hz, 1H), 7.08 (d, J=2.5 Hz, 1H), 4.47 (d, J=15 Hz, 1H), 4.05 (d, J=15 Hz, 2H), 3.07 (s, 3H), 3.01-2.46 (m, 6H), 2.12-1.91 (m, 4H), 1.81-1.54 (m, 4H). 13C NMR (101 MHZ, CDCl3) δ: 161.1, 156.6, 156, 146.9, 141.6, 139, 138, 135, 125.9, 123, 122.7, 120.9, 120.4, 105.3, 60.2, 55.7, 51.6, 51.3, 39.7, 31.1, 29.3, 27.7, 26.9, 21.7, 21.12. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C24H30N4O: 391.2492; found: 391.2487.
This compound was prepared from its Boc-protected precursor (27 mg, 0.06 mmol). The title compound was isolated as a bright brown viscous oil (21 mg, 99%). 1H NMR (400 MHZ, CDCl3) δ: 9.84 (s, 2H), 8.60 (d, J=5.8 Hz, 1H), 8.52 (d, J=3.5 Hz, 1H), 8.19 (dd, J=9.3, 5.3 Hz, 1H), 7.57 (d, J=5.8 Hz, 1H), 7.47-7.32 (m, 3H), 7.11 (dd, J=7.6, 4.8 Hz, 1H), 4.46 (d, J=15.1 Hz, 1H), 4.11 (d, J=14.9 Hz, 2H), 2.98-2.52 (m, 6H), 2.12-1.94 (m, 4H), 1.82-1.58 (m, 4H). 13C NMR (101 MHZ, CDCl3) δ: 164.5, 162, 157.3, 155.8, 147, 142.2, 138.2, 135, 127.4, 124.5, 122.8, 120.7, 118.3, 111.2, 60.3, 52.2, 51.5, 39.6, 29.8, 29.4, 27.7, 26.6, 21.7, 21.2. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C23H27FN4: 379.2292; found: 379.2286.
This compound was prepared from its Boc-protected precursor (22 mg, 0.04 mmol). The title compound was isolated as a bright brown viscous oil (18 mg, 99%). 1H NMR (600 MHZ, CDCl3) δ: 8.63 (d, J=5.8 Hz, 1H), 8.54 (d, J=3.7 Hz, 1H), 8.18 (d, J=8.9 Hz, 1H), 8.01 (d, J=1.8 Hz, 1H), 7.68 (dd, J=9, 1.9 Hz, 1H), 7.52 (d, J=5.8 Hz, 1H), 7.41 (d, J=7.2 Hz, 1H), 7.12 (dd, J=7.7, 4.7 Hz, 1H), 4.44 (d, J=14.8 Hz, 1H), 4.12 (d, J=14.9 Hz, 2H), 2.91-2.51 (m, 6H), 2.12-1.84 (m, 4H), 1.74-1.54 (m, 4H). 13C NMR (151 MHZ, CDCl3) δ: 162.4, 158, 147.3, 14306, 136.8, 134.5, 132, 129.7, 128.4, 127.9, 125.5, 123.3, 121.3, 118.1, 67.7, 59.2, 54, 41.8, 28.5, 26.3, 25.8, 20.5. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C23H27BrN4: 439.1492; found: 439.1495.
This compound was prepared from its Boc-protected precursor (21 mg, 0.05 mmol). The title compound was isolated as a bright brown viscous oil (16 mg, 99%). 1H NMR (400 MHZ, CDCl3) δ: 10.04 (s, 2H), 8.61 (d, J=3.7 Hz, 1H), 8.20 (d, J=8.7 Hz, 1H), 8.15 (d, J=8.4 Hz, 1H), 7.79 (d, J=7.7 Hz, 2H), 7.58-7.51 (m, 1H), 7.40 (d, J=7.2 Hz, 1H), 7.33 (d, J=8.4 Hz, 1H), 7.13 (dd, J=7.6, 4.8 Hz, 1H), 4.06 (d, J=14.2 Hz, 2H), 3.92 (d, J=14.2 Hz, 1H), 3.03-2.67 (m, 6H), 2.13-1.89 (m, 4H), 1.76-1.53 (m, 4H). 13C NMR (101 MHZ, CDCl3) δ: 162.4, 158.3, 147.3, 142.8, 136.8, 136.5, 132, 129.7, 127.5, 127.3, 126.8, 124.3, 121.3, 119.2, 67.7, 59.2, 54, 41.8, 28.5, 26.3, 25.8, 20.5. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C23H28N4: 361.2387; found: 361.2376.
This compound was prepared from its Boc-protected precursor (17 mg, 0.03 mmol). The title compound was isolated as a bright brown viscous oil (13 mg, 99%). 1H NMR (400 MHZ, CDCl3) δ: 8.52 (d, J=4 Hz, 1H), 8.43 (d, J=5.6 Hz, 1H), 7.47 (d, J=5.6 Hz, 1H), 7.44 (d, J=6.9 Hz, 1H), 7.27 (s, 1H), 7.12 (dd, J=7.7, 4.9 Hz, 1H), 7.07 (s, 1H), 4.20 (s, 2H), 4.02 (s, 3H), 4.00 (s, 3H), 3.83 (s, 2H), 3.15 (t, J=5.4 Hz, 2H), 2.69 (t, J=5.3 Hz, 2H), 2.26 (s, 3H), 1.89-1.84 (m, 2H), 1.82-1.77 (m, 2H). 13C NMR (101 MHZ, CDCl3) δ: 155.3, 153.9, 153, 150.4, 146.7, 140, 138.6, 1336, 131.8, 122.8, 122.6, 119.6, 105.4, 102.3, 57.1, 56.6, 561, 55.2, 39.5, 30.9, 29.6, 27.3, 26.2, 18.5. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C23H30N4O2: 395.2441; found: 395.2434.
This compound was prepared from its Boc-protected precursor (22 mg, 0.05 mmol). The title compound was isolated as a bright brown viscous oil (17 mg, 99%). 1H NMR (400 MHZ, CDCl3) δ: 8.46 (dd, J=4.8, 1.1 Hz, 1H), 8.41 (d, J=5.8 Hz, 1H), 7.96 (d, J=9.3 Hz, 1H), 7.45 (d, J=5.8 Hz, 1H), 7.41 (dd, J=7.7, 0.7 Hz, 1H), 7.15 (dd, J=9.3, 2.6 Hz, 1H), 7.09 (dd, J=7.6, 4.8 Hz, 1H), 7.03 (d, J=2.5 Hz, 1H), 4.13 (s, 2H), 3.92 (s, 3H), 3.78 (s, 2H), 2.99-2.89 (m, 2H), 2.59 (t, J=5.7 Hz, 2H), 2.20 (s, 3H), 1.76-1.63 (m, 2H). 13C NMR (101 MHZ, CDCl3) δ: 160.8, 156.9, 155.9, 146.7, 142, 138.7, 132.3, 126.6, 123, 122.7, 120.4, 120.2, 105, 57.7, 57.3, 55.6, 55.2, 39.8, 31.1, 28, 25.6, 18.6. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C22H28N4O: 365.2336; found: 365.2330.
This compound was prepared from its Boc-protected precursor (28 mg, 0.062 mmol). The title compound was isolated as a bright brown viscous oil (21.5 mg, 99%). 1H NMR (400 MHZ, CDCl3) δ: 8.51 (d, J=5.8 Hz, 1H), 8.45 (d, J=3.8 Hz, 1H), 8.12 (dd, J=9.3, 5.4 Hz, 1H), 7.52 (d, J=5.8 Hz, 1H), 7.44-7.36 (m, 2H), 7.32 (td, J=8.9, 2.5 Hz, 1H), 7.08 (dd, J=7.6, 4.8 Hz, 1H), 4.22 (s, 2H), 3.82 (s, 2H), 3.01 (t, J=5.3 Hz, 2H), 2.69 (t, J=5.2 Hz, 2H), 2.22 (s, 3H), 1.86-1.69 (m, 4H). 13C NMR (101 MHZ, CDCl3) δ: 163.9, 161.4, 158, 156, 146.3, 142.2, 137.9, 132.5, 128.7, 124.5, 122.4, 120.1, 117.1, 110.2, 58.3, 54.7, 40.5, 30.7, 29.5, 24.2, 18.2. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C21H25FN4: 353.2136; found: 353.2131.
This compound was prepared from its Boc-protected precursor (22 mg, 0.043 mmol). The title compound was isolated as a bright brown viscous oil (17.6 mg, 99%). 1H NMR (400 MHZ, CDCl3) δ: 8.57 (d, J=5.7 Hz, 1H), 8.46 (d, J=3.9 Hz, 1H), 7.97 (d, J=1.8 Hz, 1H), 7.94 (d, J=9.1 Hz, 1H), 7.63 (dd, J=9, 1.9 Hz, 1H), 7.47 (d, J=5.8 Hz, 1H), 7.41 (d, J=7.7 Hz, 1H), 7.09 (dd, J=7.6, 4.8 Hz, 1H), 4.19 (s, 2H), 3.82 (s, 2H), 3.04 (t, J=4.7 Hz, 2H), 2.68 (t, J=5.5 Hz, 2H), 2.22 (s, 3H), 1.87-1.70 (m, 4H). 13C NMR (101 MHZ, CDCl3) δ: 157.3, 155.2, 146.5, 142.3, 138.4, 137.3, 131.9, 130.8, 129.5, 126.1, 125.4, 125, 122.4, 119.5, 56.9, 55, 39.3, 30.8, 29.5, 27.1, 25.3, 18.4. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C21H25BrN4: 413.1336; found: 413.1339.
This compound was prepared from its Boc-protected precursor (22 mg, 0.05 mmol). The title compound was isolated as a bright brown viscous oil (17 mg, 99%). 1H NMR (400 MHZ, CDCl3) δ: 8.43 (dd, J=4.8, 1.1 Hz, 1H), 8.08 (dd, J=8.3, 4.7 Hz, 2H), 7.77 (dd, J=8.1, 1.1 Hz, 1H), 7.74-7.68 (m, 1H), 7.54-7.49 (m, 1H), 7.47 (d, J=8.5 Hz, 1H), 7.41 (dd, J=7.6, 0.9 Hz, 1H), 7.08 (dd, J=7.6, 4.8 Hz, 1H), 3.92 (s, 2H), 3.85 (s, 2H), 2.75 (t, J=6.5 Hz, 2H), 2.58 (t, J=6.8 Hz, 2H), 2.35 (s, 3H), 1.65-1.56 (m, 2H), 1.53-1.43 (m, 2H). 13C NMR (101 MHZ, CDCl3) δ: 158.1, 155.1, 147.2, 146.4, 138.4, 137.1, 131.7, 130.2, 128.2, 127.4, 127.1, 126.5, 122.4, 122.3, 61, 56.9, 55, 39.2, 30.7, 29.5, 26.9, 25.6, 18.3. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C21H26N4: 335.2230; found: 335.2228.
Picolinaldehyde (1 eq., 0.6887 mmol, 0.0655 ml) was added to a slurry of NaBH(OAc)3 (1.78 eq., 1.2259 mmol, 259.8 mg) in dichloromethane (3 ml), followed by the addition of tert-butyl (4-((5,6,7,8-tetrahydroquinolin-8-yl)amino) butyl)carbamate 18 (1 eq., 0.6887 mmol, 220 mg). The reaction mixture was stirred at room temperature for 48 hours. The, the reaction was quenched with a 1N NaOH solution to obtain pH˜8 of the aqueous layer. The aqueous layer was extracted three times with dichloromethane. The combined organic phases were concentrated and the crude residue was purified via silica gel flash chromatography using a mixture of DCM/MeOH (in a ratio of 96:4) as mobile phase, affording the title compound as a brown viscous oil (177 mg, 63%). 1H NMR (400 MHZ, CDCl3) δ: 8.49 (d, J=3.7 Hz, 1H), 8.45 (d, J=4.8 Hz, 1H), 7.73 (d, J=7.9 Hz, 1H), 7.63, (td, J=7.5, 1.7 Hz, 1H), 7.33 (d, J=7.5 Hz, 1H), 7.10 (dd, J=6.5, 5.6 Hz, 1H), 7.04 (dd, J=7.6, 4.7 Hz, 1H), 4.8 (s, 1H), 4.13 (d, J=7.4 Hz, 1H), 3.91 (d, J=14.6 Hz, 2H), 3.09-2.94 (m, 2H), 2.86-2.63 (m, 4H), 2.22-2.09 (m, 2H), 2.06-1.95 (m, 2H), 1.94-1.81 (m, 2H), 1.76-1.61 (m, 2H), 1.41 (s, 9H). 13C NMR (101 MHZ, CDCl3) δ: 162.4, 157.9, 155.9, 148.7, 147.3, 139.7, 136.7, 132, 124.2, 121.2, 121, 79.5, 67.7, 60.9, 54, 35.9, 28.4, 27.7, 25.8, 20.5. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C24H34N4O2: 411.2754; found: 411.2757.
Picolinaldehyde (1 eq., 1.02 mmol, 0.1 ml) was added to a slurry of NaBH(OAc)3 (1·78 eq., 1.82 mmol, 385.7 mg) in DCM (3 ml), followed by addition of tert-butyl (4-(((3-methylpyridin-2-yl)methyl)amino)butyl)carbamate 23 (1 eq., 1.02 mmol, 200 mg). The reaction was stirred at room temperature for 48 hours. After completion, the reaction mixture was quenched utilizing 1N NaOH solution to obtain pH˜8 in the aqueous layer. After extracting the mixture with DCM for 3 times, the combined organic phases were concentrated. Thereafter, the residue was purified through column chromatography using a DCM/MeOH (96:4) gradient elution to afford the corresponding compound. Brown viscous oil; yield: 206 mg (52%). 1H NMR (400 MHz, CDCl3) δ: 8.50 (d, J=4.1 Hz, 1H), 8.36 (dd, J=4.8, 1.2 Hz, 1H), 7.59 (td, J=7.7, 1.8 Hz, 1H), 7.39 (d, J=6.7 Hz, 1H), 7.34 (d, J=10.3 Hz, 1H), 7.12 (dddd, J=7.4, 5, 1.1 Hz, 1H), 7.07 (dd, J=7.6, 4.8 Hz, 1H), 4.81 (s, 1H), 3.80 (s, 2H), 3.75 (s, 2H), 3.06-2.92 (m, 2H), 2.53 (t, J=7.2 Hz, 2H), 2.30 (s, 3H), 1.57-1.47 (m, 2H), 1.43 (s, 9H), 1.41-1.31 (m, 2H). 13C NMR (101 MHZ, CDCl3) δ: 160.1, 157.3, 156.4, 149.1, 146.4, 138.3, 136.5, 133.5, 123.7, 122.7, 122.1, 60.7, 59.9, 54.4, 40.4, 28.8, 28.1, 24, 18.7. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C22H32N4O2: 385.2598; found: 385.2591.
This compound was synthesized starting from its Boc-protected precursor (23 mg, 0.06 mmol), according to the general deprotection procedure, yielding the title compound as a brownish viscous oil (17 mg, 99%). 1H NMR (600 MHZ, CDCl3) δ: 9.91 (s, 2H), 8.73 (dd, J=4.9, 0.86 Hz, 1H), 8.58 (d, J=3.9 Hz, 1H), 7.68 (td, J=7.7, 1.8 Hz, 1H), 7.41 (d, J=7 Hz, 1H), 7.29-7.21 (m, 2H), 7.15 (dd, J=7.7, 4.8 Hz, 1H), 3.92 (d, J=13.7 Hz, 2H), 3.73 (d, J=13.7 Hz, 1H), 2.99-2.64 (m, 4H), 2.39-2.20 (m, 2H), 2.11-1.92 (m, 2H), 1.75-1.56 (m, 6H). 13C NMR (151 MHZ, CDCl3) δ: 162.5, 157.9, 148.7, 17.2, 139.8, 136.7, 131.9, 124.2, 121.3, 121, 67.7, 60.9, 53.9, 41.8, 28.4, 26.3, 25.8, 20.5. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C19H26N4: 311.2230; found: 311.2226.
This compound was synthesized starting from its Boc-protected precursor (30 mg, 0.08 mmol), according to the general deprotection procedure, yielding the title compound as a brownish viscous oil (18 mg, 81%). 1H NMR (600 MHZ, CDCl3) δ: 9.36 (s, 2H), 8.66 (d, J=4.5 Hz, 1H), 8.52 (d, J=4.5 Hz, 1H), 7.65 (td, J=7.7, 1.6 Hz, 1H), 7.43 (d, J=7.5 Hz, 1H), 7.21 (d, J=7.4 Hz, 1H), 7.20 (d, J=5.1 Hz, 1H), 7.12 (dd, J=7.5, 4.9 Hz, 1H), 3.77 (s, 2H), 3.73 (s, 2H), 3.13 (t, J=5.4 Hz, 2H), 2.58 (t, J=5.2 Hz, 2H), 2.25 (s, 3H), 1.85-1.79 (m, 2H), 1.78-1.73 (m, 2H). 13C NMR (151 MHZ, CDCl3) δ: 157.4, 1552, 149.8, 146.8, 138.9, 137.4, 132, 124, 123.1, 122.8, 60.4, 57, 55.1, 39.6, 29.8, 27.3, 262, 18.6. HRMS (ESI-Q-TOF): m/z [M+H]+ calcd for C17H24N4: 285.2074; found: 285.2077.
To a reaction flask equipped with a stirring bar and a Dean-Stark receiver, containing 4-aminobutan-1-ol 26 (1.78 g, 20.0 mmol, 1.0 eq.) in toluene (60 ml) phthalic anhydride (2.96 g, 20.0 mmol, 1.0 eq.) was added. The mixture was refluxed for 3.5 h. The solvent was further evaporated in vacuo to give the title compound (4.25 g, 19.0 mmol, 97%) as a white solid. 1H NMR (400 MHZ, CDCl3) δ (ppm): 7.87-7.82 (m, 2H), 7.74-7.69 (m, 2H), 3.74 (t, J=7.1 Hz, 2H), 3.69 (t, J=6.4, 2H), 1.82-1.75 (m, 2H), 1.69 (s br, 1H), 1.65-1.59 (m, 2H).
In a flask equipped with a stirring bar, oxalyl chloride (0.34 mL, 4.0 mmol, 2.0 eq.) was dissolved in dichloromethane (15 mL) and cooled down to −78° C. To the mixture, a solution of DMSO (0.57 mL, 8.0 mmol, 4.0 eq.) in dichloromethane (4 mL) was added over 30 min. Upon addition, the mixture was stirred at −78° C. for 5 min, followed by the addition of a solution of the alcohol 27 (0.44 g, 2.0 mmol, 1.0 eq.) in dichloromethane (4 mL) over 30 min at −78° C. The resulting mixture was stirred for 40 min, after which Et3N (1.7 mL, 12.0 mmol, 6.0 eq.) was added dropwise over 10 min. The resulting mixture was allowed to warm up to 0° C. and stirred at 0° C. for 1 h. The reaction was quenched with water (10 mL). The organic layer was then separated and further washed twice with water (15 mL) and brine. The combined organic layers were dried over anhydrous magnesium sulfate and concentrated, yielding a pale yellow solid (2.370 g, 10.9 mmol, 99%). 1H NMR (400 MHZ, CDCl3) δ (ppm): 9.77 (t, J=1.1 Hz, 1H), 7.87-7.83 (m, 2H), 7.75-7.70 (m, 2H), 3.75 (t, J=6.8 Hz, 2H), 2.54 (dt, J=1.1, 7.3 Hz, 2H), 2.02 (quint, J=6.8 Hz, 2H).
To an oven-dried, Ar-flushed, screw-capped reaction tube equipped with a magnetic stirring bar, 4-(1,3-dioxoisoindolin-2-yl)butanal 27 (1.63 g, 7.5 mmol, 1.5 eq.), 8-quinolinamine (0.721 g, 5.000 mmol, 1.000 eq.) and sodium triacetoxyborohydride (2.86 g, 13.5 mmol, 2.7 eq.) were added. The reaction tube was evacuated and backfilled with argon three times. The mixture was dissolve in dry dichlorethane (17 mL) after which acetic acid (1.7 mL, 30.0 mmol, 6.0 eq.) was gradually added. The resulting mixture was stirred at room temperature under inert atmosphere for 20.5 h. The reaction was carefully quenched with aqueous NaHCO3 (15 mL) and extracted three times with dichloromethane (30 mL). The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. Silica gel column chromatography of the crude product using isohexanes/ethyl acetate (4/1) as the mobile phase provided the title compound as a yellow solid (1.27 g, 3.7 mmol, 74%). MP 93.8-95.0° C. 1H NMR (400 MHZ, CDCl3) δ (ppm): 8.69 (dd, J=1.7, 4.3 Hz, 1H), 8.03 (dd, J=1.7, 8.4 Hz, 1H), 7.86-7.81 (m, 2H), 7.72-7.67 (m, 2H), 7.37-7.33 (m, 2H), 7.02 (dd, J=0.8, 8.3 Hz, 1H), 6.65 (d, J=7.6 Hz, 1H), 6.12 (s br, 1H), 3.79-3.75 (m, 2H), 3.38-3.34 (m, 2H), 1.93-1.79 (m, 4H). 13C NMR (75 MHZ, CDCl3) δ (ppm): 168.4, 146.7, 144.7, 138.1, 135.9, 132.1, 128.6, 127.7, 121.3, 113.7, 104.5, 42.9, 37.7, 26.6, 26.4. HRMS (ESI-Q-TOF):m/z[M+H]+ calcd for C21H19N3O2: 346.15499; found: 346.1551
To an oven-dried, Ar-flushed, screw-capped reaction tube equipped with a magnetic stirring bar, 2-(4-(quinolin-8-ylamino)butyl)isoindoline-1,3-dione 28 (1.0 eq.), the benzyl halide derivative (1.5 eq.), potassium carbonate (8.0 eq.) and sodium iodide (0.250 eq.) were added. The reaction tube was evacuated and backfilled with argon three times. The mixture was dissolved in dry acetonitrile (67 mM) and refluxed up to 24 h. The mixture was cooled to room temperature and filtered through a Celite pad. The filtrate was evaporated in vacuo, basic alumina column chromatography of the crude product using isohexanes/ethyl acetate (varying between 4/1 and 3/2) as the mobile phase provided the corresponding protected/1-substituted-N1-(quinolin-8-yl)butane-1,4-diamines as semi-solids.
This compound was prepared according to general procedure using benzyl bromide (0.036 mL, 0.300 mmol), intermediate 29 (0.069 g, 0.20 mmol), potassium carbonate (0.221 g, 1.60 mmol) and dry acetonitrile (3 mL). Silica gel column chromatography of the crude product using isohexanes/ethyl acetate (3/2) as the mobile phase provided the title compound as a yellow viscous oil (0.062 g, 0.14 mmol, 71%). 1H NMR (400 MHZ, CDCl3) δ(ppm): 8.89 (dd, J=1.2, 3.9 Hz, 1H), 8.09 (dd, J=1.6, 8.2, 1H), 7.81-7.77 (m, 2H), 7.71-7.66 (m, 2H), 7.38-7.30 (m, 3H), 7.23-7.15 (m, 5H), 7.01-6.96 (m, 1H), 4.74 (s, 1H), 3.62-3.59 (m, 2H), 3.48-3.41 (m, 2H), 1.69-1.63 (m, 4H). 13C NMR (75 MHZ, CDCl3) δ(ppm): 168.3, 147.7, 147.1, 143.2, 138.7, 136.4, 133.8, 132.1, 129.8, 128.5, 128.0, 126.7, 126.3, 123.1, 120.8, 120.6, 118.6, 58.0, 51.7, 37.9, 26.3, 24.2. HRMS (ESI-Q-TOF): m/z[M+H]+ calcd for C28H25N3O2: 436.20194; found: 436.2003.
This compound was prepared according to the general procedure using 2-(chloromethyl)pyridine hydrochloride (0.049 g, 0.30 mmol), intermediate 29 (0.069 g, 0.20 mmol), potassium carbonate (0.221 g, 1.6 mmol), sodium iodide (0.007 g, 0.05 mmol) and dry acetonitrile (3 mL). Silica gel column chromatography of the crude product using isohexanes/ethyl acetate (4/1) as the mobile phase provided the title compound as a yellow viscous oil (0.044 g, 0.10 mmol, 50%). 1H NMR (400 MHz, CDCl3) ð (ppm): 8.86 (dd, J=1.6, 4.1 Hz, 1H), 8.51-8.50 (m, 1H), 8.08 (dd, J=1.6, 8.3 Hz, 1H), 7.81-7.77 (m, 2H), 7.71-7.66 (m, 2H), 7.51 (dt, J=1.4, 7.5 Hz, 1H), 7.46-7.41 (m, 1H), 7.38-7.30 (m, 3H), 7.11-7.03 (m, 2H), 4.84 (s, 2H), 3.64-3.57 (m, 4H), 1.74-1.61 (m, 4H). 13C NMR (75 MHZ, CDCl3) 0 (ppm): 168.3, 159.7, 148.7, 147.7, 146.8, 143.1, 136.4, 136.3, 133.7, 132.1, 129.8, 126.4, 123.1, 122.7, 121.8, 120.8, 120.6, 118.1, 59.6, 53.1, 37.9, 26.3, 24.3. HRMS (ESI-Q-TOF):m/z[M+H]+ calcd for C27H24N4O2: 437.19719; found: 437.1966
This compound was prepared according to general procedure using 1-(chloromethyl)isoquinoline hydrochloride (0.096 g, 0.45 mmol, intermediate 29 (0.104 g, 0.30 mmol), potassium carbonate (0.332 g, 2.4 mmol), sodium iodide (0.011 g, 0.075 mmol), dry acetonitrile (4.5 mL). Silica gel column chromatography of the crude product using isohexanes/ethyl acetate (3/2) as the mobile phase provided the title compound as a yellow viscous oil (0.093 g, 0.19 mmol, 64%). 1H NMR (400 MHZ, CDCl3) & (ppm): 8.88 (dd, J=2.3, 5.9 Hz, 1H), 8.54 (d, J=8.6 Hz, 1H), 8.42 (d, J=5.7 Hz, 1H), 8.09 (dd, J=1.8, 8.3 Hz, 1H), 7.78-7.76 (m, 2H), 7.73 (d, J=8.5 Hz, 1H), 7.68-7.65 (m, 2H), 7.57-7.53 (m, 1H), 7.51 (d, J=5.7 Hz, 1H), 7.39-7.33 (m, 4H), 7.21 (dd, J=2.6, 6.5 Hz, 1H), 5.30 (s, 2H), 3.58 (m, 4H), 1.64-1.52 (m, 4H)
This compound was prepared according to the general procedure using 1-(chloromethyl)-6-fluoroisoquinoline hydrochloride (0.139 g, 0.60 mmol), intermediate 29 (0.138 g, 0.40 mmol), potassium carbonate (0.442 g, 3.200 mmol), sodium iodide (0.015 g, 0.10 mmol), dry acetonitrile (6.0 mL). Silica gel column chromatography of the crude product using isohexanes/ethyl acetate (3/2) as the mobile phase provided the title compound as a yellow viscous oil (0.109 g, 0.22 mmol, 54%).
1H NMR (400 MHZ, CDCl3) δ (ppm): 8.88 (dd, J=1.8, 4.1 Hz, 1H), 8.85 (dd, J=5.7, 9.3 Hz, 1H), 8.41 (d, J=5.7 Hz, 1H), 8.10 (dd, J=1.8, 8.4 Hz, 1H), 7.77-7.74 (m, 2H), 7.69-7.65 (m, 2H), 7.48 (d, J=5.7 Hz, 1H), 7.39-7.31 (m, 4H), 7.22 (dd, J=2.0, 6.9 Hz, 1H), 7.13 (td, J=2.0, 13.5 Hz, 1H), 5.24 (s, 2H), 3.53-3.47 (m, 4H), 1.64-1.48 (m, 4H). 13C NMR (75 MHz, CDCl3) δ (ppm): 168.3, 162.8 (d, J=252.1 Hz), 158.6, 147.9, 147.3, 143.6, 142.4, 138.1 (d, J=10.3 Hz), 136.5, 133.8, 132.1, 130.4 (d, J=9.4 Hz), 129.8, 126.5, 125.2, 123.1, 121.5, 120.9, 120.1, 120.0, 117.0 (d, J=24.7 Hz), 109.8 (d, J=20.5 Hz), 59.5, 51.8, 37.8, 26.3, 23.7. HRMS (ESI-Q-TOF):m/z [M+H]+ calcd for C31H25FN4O2: 505.20341; found: 505.2029.
To a reaction tube equipped with a stirring bar, containing protected/1-substituted-N1-(quinolin-8-yl)butane-1,4-diamine (1.0 eq.) in ethanol (1.5 mL), hydrazine monohydrate (10 eq.) was added. The mixture was refluxed for 2 h. Upon completion and cooling, EtOH was evaporated in vacuo. Water (5 mL) was added to the remaining residue and further extracted 3 times with dichloromethane (10 mL). The combined organic layers were dried over anhydrous magnesium sulfate and concentrated to afford the corresponding N1-substituted-N1-(quinolin-8-yl)butane-1,4-diamines as semi-solids. No further purification was performed unless stated otherwise.
N1-benzyl-N1-(quinolin-8-yl)butane-1,4-diamine (31a)
This compound was prepared according to the general procedure using 2-(4-(Benzyl(quinolin-8-yl)amino)butyl)isoindoline-1,3-dione 30a (0.044 g, 0.10 mmol), hydrazine monohydrate (50 μL), ethanol (1.5 mL) affording the desired compound as a bright yellow semi-solid (0.030 g, 0.10 mmol, 98%). 1H NMR (400 MHZ, CDCl3) δ (ppm): 8.91 (dd, J=1.8, 4.1 Hz, 1H), 8.11 (dd, J=1.8, 8.2 Hz, 1H), 7.39-7.32 (m, 3H), 7.24-7.17 (m, 5H), 6.99 (dd, J=2.4, 6.7, 1H), 4.76 (s, 2H), 3.43-3.39 (m, 2H), 2.63-2.60 (m, 2H), 1.69-1.61 (m, 2H), 1.44-1.37 (m, 2H), 1.25 (s br, 1H). 13C NMR (75 MHZ, CDCl3) δ (ppm): 147.9, 147.7, 147.1, 143.3, 138.6, 136.5, 129.9, 128.7, 128.0, 126.8, 126.3, 120.9, 120.7, 118.8, 58.2, 51.9, 41.9, 31.2, 24.2. HRMS (ESI-Q-TOF):m/z[M+H]+ calcd for C20H23N3: 306.19646; found: 306.1961.
This compound was prepared according to the general procedure using 2-(4-((pyridin-2-ylmethyl)(quinolin-8-yl)amino)butyl)isoindoline-1,3-dione 30b (0.044 g, 0.10 mmol), hydrazine monohydrate (50 μL), ethanol (1.5 mL) affording the desired compound as a bright yellow semi-solid (0.030 g, 0.10 mmol, 97%). 1H NMR (400 MHz, CDCl3) δ(ppm): 8.89 (dd, J=1.8, 4.2 Hz, 1H), 8.55-8.52 (m, 1H), 8.11 (dd, J=1.8, 8.2 Hz, 1H), 7.52 (dt, J=1.8, 7.6 Hz, 1H), 7.42-7.34 (m, 4H), 7.13-7.06 (m, 2H), 4.84 (s, 2H), 3.57-3.51 (m, 2H), 2.67 (t, J=6.9 Hz, 2H), 1.74-1.66 (m, 2H), 1.50-1.41 (m, 2H), 1.25 (s, 2H). 13C NMR (75 MHZ, CDCl3) δ (ppm): 159.5, 148.8, 147.8, 146.9, 143.1, 136.5, 136.3, 129.9, 126.4, 122.8, 121.8, 120.9, 120.8, 118.2, 59.5, 53.4, 41.8, 29.7, 24.3. HRMS (ESI-Q-TOF):m/z[M+H]+ calcd for C19H22N4: 307.19171; found: 307.1912.
This compound was prepared according to the general procedure using 2-(4-((isoquinolin-1-ylmethyl) (quinolin-8-yl)amino) butyl)isoindoline-1,3-dione 30c (0.053 g, 0.11 mmol), hydrazine monohydrate (53 μL), ethanol (1.6 mL) affording the desired compound as a bright yellow semi-solid (0.037 g, 0.10 mmol, 96%). 1H NMR (400 MHZ, CDCl3) δ (ppm): 8.91 (dd, J=1.8, 4.1 Hz, 1H), 8.53 (d, J=8.5 Hz, 1H), 8.47 (d, J=5.8 Hz, 1H), 8.12 (dd, J=1.8, 8.3 Hz, 1H), 7.77 (d, J=8.3 Hz, 1H), 7.61-7.56 (m, 1H), 7.55 (d, J=5.8 Hz, 1H), 7.41-7.35 (m, 4H), 7.22 (dd, J=1.9, 6.9 Hz, 1H), 5.31 (s, 2H), 3.52 (t, J=7.6 Hz, 1H), 2.53 (t, J=6.9 Hz, 1H), 1.67-1.59 (m, 2H), 1.36-1.28 (m, 2H). 13C NMR (75 MHZ, CDCl3) δ (ppm): 158.1, 148.0, 147.6, 143.5, 141.4, 136.7, 136.4, 129.9, 127.7, 126.9, 126.8, 126.5, 126.2, 121.5, 120.9, 120.5, 119.6, 58.2, 52.4, 41.5, 30.7, 23.9.
This compound was prepared according to the general procedure using 2-(4-(((6-fluoroisoquinolin-1-yl) methyl)(quinolin-8-yl)amino) butyl)isoindoline-1,3-dione 30 d (0.053 g, 0.11 mmol), hydrazine monohydrate (37 μL), ethanol (1.1 mL). Reverse phase HPLC of the crude product using methanol/water (1/1) as the mobile phase provided the title compound as a bright yellow semi-solid (0.006 g, 0.02 mmol, 16.0%). 1H NMR (400 MHZ, CDCl3) δ (ppm): 8.91 (dd, J=1.6, 4.1 Hz, 1H), 8.49 (d, J=5.8 Hz, 1H), 8.20 (dd, J=5.8, 9.3 Hz, 1H), 8.13 (dd, J=1.5, 8.3 Hz, 1H), 7.51-7.47 (m, 2H), 7.42-7.38 (m, 1H), 7.38-7.34 (m, 2H), 7.27-7.25 (m, 1H), 7.20-7.15 (m, 1H), 5.04 (s, 2H), 3.48 (t, J=5.6 Hz, 2H), 2.85 (t, J=5.6 Hz, 2H), 1.83-1.81 (m, 4H).
The CXCL12AF647 binding assay with Jurkat cells has been described previously [Schoofs et al. JoVE 2018, 133, 1-8]. Briefly, Jurkat cells were resuspended in assay buffer [Hank's Balanced Salt Solution (HBSS, Thermo Fisher Scientific), 20 mM HEPES buffer, 0.2% bovine serum albumin (Sigma-Aldrich), pH 7.4] at 3×105 cells per sample and then treated with various concentrations of the compound at room temperature for 15 minutes. Afterwards, the cells were incubated with 2.9 nM CXCL12AF647 (in assay buffer) at room temperature for 30 minutes in the dark. Cells were fixed in 1% paraformaldehyde in DPBS and specific CXCL12AF647 binding [i.e. mean fluorescence intensity (MFI)] was quantified by flow cytometry (FACSArray™; Becton Dickinson). Data were analyzed with FlowJo® Software. The 50% inhibitory concentration (IC50) was calculated for each compound relative to the negative (i.e. autofluorescence of untreated and unlabeled cells) and positive (i.e. untreated cells exposed to CXCL12AF647 only) control.
The calcium mobilization assay has been described in detail previously [Claes et al. JoVE 2018, 132, 1-9]. U87.CD4.CXCR4 cells (2×104 cells per well in DMEM/10% FBS/0.01M HEPES) were seeded in gelatin-coated (Sigma-Aldrich; 0.1% gelatin in DPBS) black-walled 96-well plates and incubated overnight at 37° ° C. and 5% CO2. The next day, cells were loaded with the fluorescent calcium indicator Fluo-2 acetoxymethyl (AM) ester (4 μM; Abcam) and incubated at room temperature in the dark for 45 minutes. Then, cells were incubated with various concentrations of the compounds for 10 minutes prior to addition of 6.25 nM CXCL12 (in assay buffer). Fluctuations in intracellular calcium levels were measured in real time by the FLIPR Tetra® (Molecular Devices, Sunnyvale, CA, USA) in all 96 wells simultaneously. The response over baseline (after CXCL12 addition) was calculated with the ScreenWorks 4.0® software (Molecular Devices) by dividing the obtained relative light units (RLUs) through the base line measured just before CXCL12 addition. From this the IC50 value for each compound was determined taking into account the negative (i.e. untreated cells without CXCL12 stimulation) and positive (i.e. untreated cells with CXCL12 addition) control samples.
The anti-HIV assays in MT-4 cells been described in detail before [Hout et al. cited above; Vermeire et al. AIDS 2004, 18, 2115-2125]. Briefly, compound-treated MT-4 cells (5×104 cells per sample in cell culture medium; 30 min incubation at 37° C.) were infected with HIV-1 (NL4-3) and HIV-2 (ROD) virus stocks. After five days, the cytopathic effect was checked microscopically and cell viability (i.e. viral replication) was evaluated using the MTS/PES-based CellTiter 96 Aqueous One Solution Cell Proliferation assay (Promega, Fitchburg, WI, USA). Absorbance was recorded using the VersaMax ELISA™ microplate reader (Molecular Devices) and analyzed with the Softmax Pro® software (Molecular Devices).
| Number | Date | Country | Kind |
|---|---|---|---|
| 21181444.7 | Jun 2021 | EP | regional |
| 21204648.6 | Oct 2021 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2022/067318 | 6/24/2022 | WO |
