Small molecule lanosterol synthase inhibitors

Abstract

A composition for treating a neurological disorder comprises a lanosterol synthase inhibitor.

Description

[0001] introduction

[0002] Glioblastoma or GBM is the most common primary malignant brain tumor in the United States (Ostrom et al., 2019). GBM has a poor prognosis with a median survival of 12 months, which has been extended to 14 months with current standard therapy (Stupp et al., 2005). Standard treatment, which requires surgical resection followed by concurrent radiation therapy and temozolomide, shows modest antitumor activity, but recurrence and tumor recurrence are inevitable. One of the key factors in the poor outcome of GBM patients is the lack of effective treatments for the disease. SUMMARY OF THE INVENTION

[0004] The present invention provides small molecule inhibitors of the cholesterol biosynthetic enzyme lanosterol synthase. By inhibiting lanosterol synthase, these compounds inhibit the biosynthesis of lanosterol and, therefore, the biosynthesis of cholesterol. They shift the biosynthetic flux to the synthesis of atypical sterols: 24,25-epoxycholesterol (EPC). This EPC upregulation using small molecule lanosterol synthase inhibitors results in potent specific killing of glioblastoma cell lines. The present invention provides a group of representative lanosterol synthase inhibitor analogs that kill glioblastoma cell lines, particularly in the low single-digit nanomolar range. In embodiments, the inhibitors can be orally administered and / or brain penetrating. The present invention provides methods and compositions for treating glioblastoma and developing new therapies for treating glioblastoma.

[0005] In one aspect, the present invention provides a compound of formula I or a salt, hydrate or stereoisomer thereof:

[0006]

[0007] in:

[0008] Q is selected from cyclopropane-1,2-diyl, CH2, (CH2)2, O;

[0009] X, Y, Z and W are each independently selected from CR and N; R = H, halogen (eg, F, Cl, Br); CH3, optionally fluorinated (eg, CF3), optionally deuterated (eg, CD3);

[0010] L is selected from: -OCHRC(O)- [R = H, Me, CF3, CHF2, CH2F], -NHCH2C(O)-, -NHCHMeC(O)-, -NMeCH2C(O)-, -NMeCHMeC(O)-, -SCH2C(O)-, -SCHMeC(O)-, -CH2OC(O)-, -CHMeOC(O)-, -CH2NHC(O)-, CHMeNHC(O)-, -OCH2NMeC(O)-, -CHMeNMeC(O)-, -CH2NHSO2-, -CHMeNHSO2-, -CH2NMeSO2-, -CMeNMeSO2-, -O(CH2)n- [n = 0 - 2], -OCHMe-, -OCHMeCH2-, -OCH2CHR- [R = Me, OH, OMe, OCF3, OCHF2, OCH2F], -OCHMeCHR- [R = OH, OMe, OCF3, OCHF2, OCH2F], -(CH2)nC(O)- [n = 0 - 2], -CHRCH2C(O)- [R = Me, OH, OMe, OCF3, OCHF2, OCH2F], -CH2CHRC(O)- [R = Me, OH, OMe, OCF3, OCHF2, OCH2F], -CHORCH2n- [n = 0 - 2; R = H, Me, CF3, CHF2, CH2F], -(CH2)nCHOR- [n = 1 - 2;R=H, Me, CF3, CHF2, CH2F], -CHORCH2CHOR'-[R, R' are independently selected from H, Me, CF3, CHF2, CH2F], -CHORCHMeCHOR'-[R, R' are independently selected from H, Me, CF3, CHF2, CH2F], -CHORCMeCH2-[R=H, Me, CF3, CHF2, CH2F], -CH(OR)CH=CH-[R=H, Me, CF3, CH HF2, CH2F], -(CH2)n-[n=0-3], -CH2NR-[R=H, Me]-, -CH=CHC(O)-, CH=CHCHR-[R=OH, OMe, OCF3, OCHF2, O CH2F], -CMe=CHC(O)-, CH=CMeC(O)-, CMe=CHCHOR-[R=H, Me, CF3, CHF2, CH2F], CH=CMeCHOR[R=H, Me, CF3 , CHF2, CH2F], -C□CC(O)-, -C□CCHR[R=H, Me, OH, OMe, OCF3, OCHF2, OCH2F], -(cyclopropane-1,2-diyl)C(O)-, -(cyclopropane-1,2-diyl)CHR-[R=H, Me, OH, OMe, OCF3, OCHF2, OCH2F], -(ethylene oxide-2,3-diyl)C(O)-, -(ethylene oxide-2,3-diyl)CHR-[R= H, Me, OH, OMe, OCF3, OCHF2, OCH2F], -C(O)(cyclopropane-1,2-diyl)-, -CHR(cyclopropane-1,2-diyl)-[R=H, Me, OH, OMe, OCF3, OCHF2, OCH2F], -C(O)(ethylene oxide-2,3-diyl)-, -CHR(ethylene oxide-2,3-diyl)-[R=H, Me, OH, OMe, OCF3, OCHF2, OCH2F]; and;

[0011] Ar is selected from 2- or 3- or 4-monosubstituted phenyl or disubstituted phenyl or trisubstituted phenyl (preferred substituents are selected from halogen (e.g. F, Cl, Br), Me or OMe, cycloPr, CN, -NHCHO, each optionally fluorinated and optionally deuterated (e.g. OCF3, CF3, CD3); and optionally substituted phenyl and heteroaryl), monocyclic and fused bicyclic heteroaryl, fused heteroaryl [e.g. pyridine, pyrimidine, pyrazine , pyridazine, oxazole, benzoxazole, pyrazole, quinoline, quinoxaline, isoquinoline], biaryl and fused aryl (naphthyl); each optionally fluorinated and optionally deuterated (e.g. OCF3, CF3, CD3); optional substituents are selected from halogen (e.g. F, Cl, Br), CF3, Me or OMe, cPr, CN, aryl and heteroaryl, each optionally fluorinated and optionally deuterated (e.g. OCF3, CF3, CD3).

[0012] In the implementation:

[0013] Q is cyclopropane-1,2-diyl;

[0014] X is N, and Y, Z and W are each CH; or, X, Y, Z and W are each CH;

[0015] L is selected from: -OCHC(O)-, -NHCH2C(O)-, -NHCHMeC(O)-, -NMeCH2C(O)-, -NMeCHMeC(O)-, -CH2OC(O)-, -CHMeOC(O)-, -CH2NHC(O)-, CHMeNHC(O)-, -OC H2NMeC(O)-, -CHMeNMeC(O)-, -OCHMe-, -OCHMeCH2-, -OCH2CHMe-, -OCHMeCHOH-, -(CH2)C(O)-, -CHMeCH2C(O)-, -CH2CHMeC(O)-, -CHOHCH2- , -CH2CHOH-, -CHOHCH2CHOH'-, -CHOHCHMeCHOH-, -CHOHCMeCH2-, -CH(OH)CH=CH2-, -CH2NH-, -(cyclopropane-1,2-diyl)C(O)-, -(cyclopropane-1,2-diyl)CH2-, -(ethylene oxide-2,3-diyl)C(O)-, -(ethylene oxide-2,3-diyl)CH2-, -C(O)(cyclopropane-1,2-diyl)-, -CH2(cyclopropane-1,2-diyl)-, -C(O)(ethylene oxide-2,3-diyl)-, -CH2(ethylene oxide-2,3-diyl)-; and / or

[0016] Ar is 2- or 3- or 4-monosubstituted phenyl or disubstituted phenyl or trisubstituted phenyl, the substituents being selected from halogen, Me or OMe, cycloPr, CN and -NHCHO, each optionally fluorinated and optionally deuterated.

[0017] In an embodiment, the compound:

[0018] Having the structure of Table 1, 2, 3, 4, 5, 6, 7, 8 or 9 herein;

[0019] Inhibits lanosterol synthase (LSS);

[0020] is orally bioavailable; and / or

[0021] Crosses the blood-brain barrier.

[0022] In one aspect, the present invention provides a pharmaceutical composition comprising a compound described herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0023] In one aspect, the invention provides a method of inhibiting lanosterol synthase comprising administering to a human in need thereof a small molecule lanosterol synthase inhibitor disclosed herein.

[0024] In one aspect, the present invention provides a method of upregulating 24,25-epoxycholesterol (EPC) comprising administering to a human in need thereof a small molecule lanosterol synthase inhibitor disclosed herein.

[0025] In one aspect, the invention provides a method of treating a neurological disease or disorder comprising administering to a human in need thereof a small molecule lanosterol synthase inhibitor disclosed herein.

[0026] In one aspect, the invention provides a method of screening for a candidate therapeutic for treating a neurological disease or disorder, the method comprising identifying an inhibitor of lanosterol synthase (p75).

[0027] In embodiments of the methods herein:

[0028] The neurological disease or disorder is glioblastoma (GBM) or a neurodegenerative disease such as amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, Alzheimer's disease, Huntington's disease; and / or

[0029] The method may also include a prior step of detecting or diagnosing the disease or condition and / or a subsequent step of detecting an amelioration or delay in progression of the resulting disease or condition.

[0030] The invention includes all combinations of specific embodiments described herein, as if each combination had been expressly described.

[0031] Description of Specific Embodiments of the Invention

[0032] Unless otherwise prohibited or indicated, in these descriptions and throughout the specification, the terms "a" and "an" refer to one or more, and the term "or" means and / or. It is understood that the examples and embodiments described herein are for illustrative purposes only, and that those skilled in the art will be advised of various modifications or changes to these examples and embodiments, which should be included in the spirit and scope of the present application and the scope of the appended claims. All publications, patents, and patent applications cited herein, including citations therein, are incorporated by reference in their entirety for all purposes.

[0033] We demonstrate that the tetracyclic dicarboximide MM0299 inhibits lanosterol synthase (denoted Lss in mice, or LSS in humans), the first enzyme in the post-squalene cholesterol biosynthetic pathway that synthesizes the tetracyclic sterol skeleton. Like other LSS inhibitors, MM0299 also blocks canonical cholesterol biosynthesis and diverts sterol synthesis flux toward the production of the bypass pathway product 24(S),25-epoxycholesterol (EPC), which is responsible for preventing the growth of GSCs by depleting cellular cholesterol. Compared with known LSS inhibitors, MM0299 exhibits superior selectivity for LSS over other sterol biosynthetic enzymes. We disclose MM0299 derivatives that are orally bioavailable, penetrate the blood-brain barrier, and stimulate EPC production in tumors but not in normal brain. The present invention provides novel lanosterol synthase inhibitors as therapeutic agents for glioblastoma or other neurological disorders.

[0034] Selective and brain-penetrating lanosterol synthase inhibitors target glioma stem-like cells by activating a bypass pathway that produces the toxic metabolite 24(S),25-epoxycholesterol

[0035] We conducted a medicinal chemistry campaign to generate pharmacologically optimized preclinical LSSi leads and subsequent studies to evaluate their target engagement, safety, and anti-GBM efficacy profiles. We exploited structure activity relationships (SARs) embedded in a large set of analogs generated through medicinal chemistry to correlate cellular phenotypic SAR data with cellular probe shift EC50s to distinguish on-target and off-target binding events. We used a high-throughput cell survival screen using Mut 6, a mouse GBM stem-like cell (GSC) line derived from a genetically engineered mouse GBM model, to identify chemicals that impair GBM cell fitness in which spontaneous tumor formation is driven by GFAP-Cre-mediated silencing of tumor suppressors Trp53, Pten, and Nf1 in mouse astrocytes.

[0036] We explored the mechanism of action of a tetracyclic dicarboximide (hereafter referred to as MM0299) that emerged from our screen. MM0299 inhibits lanosterol synthase (denoted Lss in mice, or LSS in humans), the first enzyme in the post-squalene cholesterol biosynthetic pathway that synthesizes the tetracyclic sterol backbone. Inhibition of LSS by MM0299 blocks canonical cholesterol biosynthesis and diverts sterol synthesis flux toward the production of the bypass pathway product 24(S),25-epoxycholesterol (EPC), which in turn blocks GSC growth.

[0037] Our invention provides a new approach to target tumor cell metabolism for brain tumor therapy. Metabolic reprogramming is a fundamental feature of the molecular pathogenesis of gliomas, and many strategies aimed at exploiting this process are undergoing preclinical and clinical testing (Zhou and Wahl, 2019). One such strategy involves targeting metabolic pathways that display higher flux in glioma cells relative to normal cells, as exemplified by studies evaluating the radiosensitizing effects of purine nucleotide synthesis inhibition in GBM (NCT04477200) (Zhou et al., 2020). Another strategy entails blocking the synthesis of the oncometabolite 2-hydroxyglutarate, which selectively accumulates and promotes the formation of isocitrate dehydrogenase (IDH)-mutated gliomas (NCT02481154) (Mellinghoff et al., 2021). In contrast, our approach reveals a potential new paradigm: activating rather than inhibiting metabolic pathways in GBM cells to trigger the production of tumor-specific cytotoxic metabolites EPCs.

[0038] In terms of clinical translation, we also report a derivative of MM0299 that is both orally bioavailable and penetrates the blood-brain barrier. These findings support the development of lanosterol synthase inhibitors as therapeutic agents for glioblastoma or other neurological disorders.

[0039] result

[0040] 1.MM0299 inhibits Mut6 cell growth by binding to lanosterol synthase

[0041] GFAP-Cre-mediated silencing of the tumor suppressors Trp53, Pten, and Nf1 in mouse astrocytes resulted in spontaneous tumors that were histologically comparable to human GBM (Kwon et al., 2008). A GBM tumor-derived glioma stem-like cell (GSC) line generated from this model (hereafter referred to as Mut6) was subsequently used in a high-throughput chemical screen to identify small molecules that impair the proliferation of these GSCs (Shi et al., 2019). MM0299 (1) was a tetracyclic dicarboximide that emerged from this screen and reduced the viability of Mut6 cells in a dose-dependent manner (IC 50=0.0182 μM).

[0042]

[0043] We sought to understand the mechanism of action (MoA) of MM0299 by identifying which proteins MM0299 binds to in Mut6 cells. To achieve this goal, we synthesized MM0299-probe (2), an analog of MM0299 that retains antiproliferative activity in Mut6 (IC 50 =1.18 μM) and contains both diazirine and alkyne functional groups. Upon ultraviolet (UV) light exposure, the photoreactive diazirine group forms a reactive carbenes, which have the potential to react with nearby amino acid residues, resulting in covalent addition of the compound to the bound protein. We incubated Mut6 cells with increasing concentrations of probe 2 with or without UV treatment (306 nm), and the resulting lysate underwent a copper (I)-catalyzed azide-alkyne cycloaddition reaction (commonly referred to as "click" chemistry) with fluorescent azide. Analysis of the click lysate by SDS-PAGE revealed multiple fluorescent protein bands representing proteins covalently bound to probe 2. Under UV conditions alone, the fluorescence intensity of several protein bands increased in a dose-dependent manner, with the most prominent band migrating at approximately 75 kDa (p75).

[0044] To determine whether binding to any of the observed bands was responsible for toxicity, we performed a series of probe displacement assays by incubating cells with a fixed concentration of probe 2 and increasing concentrations of competitor 1. We hypothesized that 1 would result in a dose-dependent decrease in the intensity of the fluorescence band of the putative target. Although probe 2 bound to a variety of proteins, we observed a dose-dependent decrease in fluorescence only for p75. We repeated the probe displacement assays for 12 structurally related MM0299 derivatives (analogs 3–14, Table 1 , see SI for synthesis and structure) and quantified the p75 binding EC 50 —the concentration of competitor required to displace 50% of the probe binding signal—to generate a structure-binding relationship. In parallel, we measured the antiproliferative activity of each derivative in a dose-dependent manner to determine its IC 50 , and the structure-activity relationship was obtained. 50 IC 50 After looking at the relationship between the two, we noticed that the slope of the best fit line was 1.06, indicating that the EC of the probe competition 50 IC 50 In addition, there was a significant correlation between p75 binding and cytotoxic activity over a 1000-fold potency range (R 2=0.864, p<0.0001), providing evidence that p75 is a functional target of MM0299.

[0045] To determine the identity of p75, we performed large-scale affinity purification of the probe-binding protein. Mut6 cells were incubated with 0.1 μM probe 2 and either DMSO or 1.0 μM of the active competitor analog 11 (IC 50 =0.000791 μM) and then UV cross-linked. The lysate was subjected to a click reaction using biotin-azide to biotinylate the probe-binding protein and then subjected to affinity purification using fixed streptavidin. The protein eluted from the streptavidin beads was digested with trypsin, and the resulting peptides were identified using liquid chromatography-mass spectrometry (LC-MS). Analysis of the LC-MS data from the above purification revealed that lanosterol synthase (Lss) was the most enriched protein in the pure probe purification, showing an enrichment of about 15.5 times compared to the probe + competitor purification. These data indicate that p75 is Lss (MW=83.1 kDa). Notably, no other protein enrichment exceeded 2 times, indicating that there are few alternative targets for MM0299.

[0046] We further tested whether p75 was indeed an LSS using a combination of pharmacology and genetics. First, we asked whether the structurally unique LSS inhibitor Ro 48-8071 could displace the MM0299 probe in a manner similar to the probe displacement assay described above for the MM0299 series (2). Ro 48-8071 displaced the MM0299 probe in a dose-dependent manner, as indicated by a decrease in p75 band intensity with increasing Ro 48-8071 concentrations. In addition, Ro 48-8071 bound to p75 in a manner similar to the observed p75 binding (EC 50 =0.00248 μM) at similar concentrations (IC 50 =0.0112 μM) impaired Mut6 cell proliferation. To further investigate whether p75 is LSS, we evaluated probe binding in HEK293T cells ectopically expressing FLAG epitope-tagged human LSS. In these cells, we observed that p75 band intensity increased after incubation and crosslinking with probe 2, while it was attenuated by the addition of excess competitor 1, demonstrating specificity. In summary, these results provide evidence that p75, which is considered a direct target of MM0299, is LSS.

[0047] 2. MM0299(1) inhibits Lss activity, leading to inhibition of the canonical pathway and induction of the alternative pathway.

[0048] Inhibitors of LSS block the canonical cholesterol biosynthetic pathway (known as the Bloch and Kandutsch-Russell pathways) while stimulating the production of 24(S),25-epoxycholesterol (EPC) via a "bypass" pathway (Mark et al., 1996; Morand et al., 1997). For sterol synthesis, squalene, a symmetrical polyene intermediate in the cholesterol biosynthetic pathway, is oxidized by squalene epoxidase (SQLE) in the presence of NADPH and oxygen to generate (S)-2,3-oxysqualene (OS), which is in turn converted to lanosterol by LSS. LSS inhibition bypasses OS back to SQLE, which catalyzes a second oxidation event to generate C2-symmetrical (S,S)-2,3:22,23-diepoxysqualene (DOS). DOS is a preferred substrate for LSS and is converted to 24(S),25-epoxylanosterol (EPL) by LSS even in the presence of LSS inhibitors (Boutaud et al., 1992). EPL is further converted to EPC by the same enzyme that converts lanosterol to cholesterol (Nelson et al., 1981a; Nelson et al., 1981b). The overall consequence of LSS inhibition is reduced levels of canonical pathway intermediates and increased levels of bypass pathway intermediates. Therefore, to test the hypothesis that 1 is an inhibitor of LSS, we quantified key intermediates in both the canonical pathway and bypass pathways after treatment with different doses of compound 1. Treatment of Mut6 cells with compound 1 resulted in a dose-dependent increase in the major LSS substrate OS (up to 6.48-fold) and a dose-dependent decrease in the product lanosterol (up to 24.5-fold) (IC 50 =0.0455 μM). Treatment with compound 1 also resulted in a dose-dependent increase in the bypass pathway intermediates DOS and EPL (up to 17.9-fold and 25.6-fold, respectively). Levels of the terminal product of the bypass pathway, EPC, peaked at 390 ng / mg protein (a 13.9-fold increase) when cells were treated with 0.1 μM compound 1, but then steadily declined at higher concentrations of compound 1. We believe that the decrease in EPC at higher concentrations may be a result of complete inhibition of LSS; however, at doses where EPC decreased, levels of DOS and EPL continued to rise. These data suggest that the decrease in EPC levels at higher doses is more likely a result of catabolism or efflux rather than a result of reduced synthesis. Related to this hypothesis, in astrocytes, EPC stimulates the expression of sterol transporters including ABCA1 and is itself prone to efflux (Wong et al., 2007). We hypothesize that the threshold level of EPC reached at higher concentrations of 1 activates expression of the transporter, leading to EPC efflux and the observed decrease in EPC levels. In summary, we conclude that inhibition of Lss by MM0299(1) results in a dose-dependent decrease in canonical intermediates and a dose-dependent increase in branch pathway intermediates.

[0049] We next evaluated whether MM0299(1) and related analogs could directly inhibit recombinant human LSS in a cell-free in vitro reaction. Recombinant LSS purified from E. coli converted OS to lanosterol in a time- and dose-dependent manner. Notably, lanosterol production saturated over time, which could be a result of substrate depletion or product inhibition. Regardless, we assessed enzyme activity over 30 min at 64 μM substrate, under which conditions the reaction remained linear. Under these conditions, compound 1 was highly potent at an IC 50 =2.22 μM inhibited the in vitro activity of LSS in a dose-dependent manner. Similar experiments using three different analogs (3, 6, 14) support the view that inhibition of Lss enzymatic activity is associated with the anti-GBM activity of Mut6 cells. It is noteworthy that the concentration of compound required to inhibit LSS activity in vitro is significantly higher than the concentration required to block cell growth. One possible explanation for this difference is that the conditions of the in vitro reaction, including non-ionic detergents in aqueous buffers, affect the solubility or binding affinity of the substrate or compound.

[0050] An unresolved question regarding the bypass pathway is how LSS is able to convert DOS to EPL in the presence of high concentrations of the inhibitor. In vitro kinetic experiments have shown that the conversion of DOS to EPL is faster than the conversion of OS to lanosterol, indicating that DOS is the preferred substrate (Boutaud et al., 1992). To address this question, we evaluated the efficacy of LSS in converting DOS to EPL in vitro. Consistent with previous results, the rate of conversion of DOS to EPL was 2.67 times that of the rate of conversion of OS to lanosterol. However, MM0299(1) inhibited the conversion of DOS to EPL with a potency comparable to that of OS to lanosterol (IC 50 =1.60 and 2.22 μM). Therefore, the conversion of DOS to EPL by LSS in the presence of MM0299 cannot be simply explained by an enhanced affinity of LSS for DOS. A more likely explanation comes from our observation that in cells treated with 1, DOS levels were 375-fold higher than OS. The effective concentration of DOS in the ER membrane is likely to be even higher, thus limiting the access of the inhibitor to the enzyme active site.

[0051] 3. Induction of the bypass pathway by MM0299 contributes to toxicity in Mut6 cells.

[0052] Next, we asked whether the increased EPC levels caused by LSS inhibition by MM0299 contribute to the antiproliferative activity observed in Mut6 cells. EPC is a potent ligand for LXR nuclear hormone transcription factors, which promote cholesterol efflux by upregulating cholesterol transporters such as ABCA1 and ABCG1 (Janowski et al., 1999; Janowski et al., 1996; Lehmann et al., 1997; Willy et al., 1995). In addition, EPC blocks the activation of SREBP, a master transcription factor that promotes the expression of enzymes involved in cholesterol biosynthesis and absorption (Horton et al., 2003; Radhakrishnan et al., 2007). Using mRNA sequencing, we found that treatment of Mut6 cells with 1 resulted in upregulation of LXR target genes including Abca1, Abcg1, and Srebf1 and concomitant downregulation of SREBP target genes, indicating that the levels of EPCs generated after 1 treatment are sufficient to affect these pathways ( Horton et al., 2003 ; Willy et al., 1995 ).

[0053] To better evaluate the role of EPCs in the anticancer activity of 1, we sought to determine whether the levels of EPCs achieved in cells treated with 1 were sufficient for toxicity. When cells were incubated with a polydeuterated derivative of 2,4(S),25-epoxylanosterol (d6-S-EPL), d6-EPCs were generated in a time-dependent manner, with d6-S-EPL having an IC of 0.483 μM. 50 Inhibition of growth by Mut6. Notably, the levels of d6-EPC produced when cells were incubated with an effective dose of d6-S-EPL (1 μM) were comparable to the levels of EPC detected after treatment with a lethal dose of 1 (0.1 μM) (390 ng d6-EPC / mg protein and 342 ng EPC / mg protein, respectively). These data suggest that EPCs synthesized after treatment with 1 are sufficient to block cell growth.

[0054] In summary, we conclude that 1 treatment results in sufficient levels of EPCs to downregulate SREBP target genes, upregulate LXR target genes, and block cell proliferation. Through their coordinated actions on LXRs and SREBPs, key transcriptional regulators of cholesterol homeostasis, EPCs may lead to a lethal reduction in cellular cholesterol levels. To test this hypothesis, we assessed cell proliferation following treatment with increasing doses of S-EPL or 1 in the presence of exogenous lanosterol or cholesterol conjugated to methyl-β-cyclodextrin (MCD), which facilitates solubilization and delivery of sterols. Supplementation of cells with lanosterol or cholesterol protected Mut6 cells from the toxic effects of S-EPL or 1. These observations provide evidence that the antiproliferative activity of 1 is due to the generation of EPCs and the resulting depletion of cellular cholesterol.

[0055] To assess whether the EPC is required for MM0299 activity, we considered pharmacological tools that could selectively block the bypass pathway. In the context of LSS inhibition, the generation of DOS requires two oxidation cycles by squalene epoxidase (SQLE), whereas OS requires only one. Therefore, we reasoned that co-administration of a SQLE inhibitor might have a greater effect on DOS levels and impede flux through the bypass pathway. Consistent with this prediction, we found that co-treatment with a sublethal dose of the SQLE inhibitor NB-598 completely blocked the accumulation of bypass pathway metabolites following 1 treatment, but did not reduce levels of typical sterols (Horie et al., 1990). In summary, we conclude that NB-598 is a pharmacological tool to selectively block the bypass pathway and assess the requirement for the EPC for MM0299 activity. We found that NB-598 rescued the toxicity of 1 in a dose-dependent manner, as measured by the IC 50 In contrast, NB-598 was unable to rescue the activity of bortezomib, a compound whose mechanism is unrelated to cholesterol synthesis, or TASIN-30, an inhibitor of emopamil binding protein (EBP), an enzyme downstream of LSS in cholesterol biosynthesis (Theodoropoulos et al., 2020). These observations are consistent with the hypothesis that LSS inhibition and the resulting increase in EPC are responsible for the low nanomolar range of toxicity. We note that 1 remained toxic to Mut6 cells at higher concentrations despite the addition of NB-598. Toxicity at these concentrations, which are unlikely to be achieved pharmacologically, is unlikely to be the result of EPC production and is more likely the result of sterol depletion through direct inhibition of cholesterol biosynthesis. Nonetheless, our combined observations conclude that specific induction of the branch metabolite EPC is necessary and sufficient for MM0299 to exert its most potent cytotoxic effects in Mut6 cells.

[0056] 4. Evaluation of LSS Selectivity of MM0299 and Known LSS Inhibitors

[0057] Enzyme selectivity is a major challenge in the development of inhibitors of the post-squalene cholesterol biosynthesis pathway (Korade et al., 2016; Moebius et al., 1998; Wages et al., 2018). This polypharmacology may be particularly problematic for the use of LSS inhibitors as anti-GBM agents, as inhibition of downstream biosynthetic enzymes blocks the production of the key metabolite EPC (Rabelo et al., 2017). Therefore, we systematically compared the off-target profiles of 1 with those of Ro 48-8071, an LSS inhibitor that has been extensively evaluated pharmacologically and biophysically, including structural determination of the complex with LSS (Morand et al., 1997; Thoma et al., 2004).

[0058] Similar to 1, Ro 48-8071 treatment resulted in a dose-dependent increase in intermediates in the bypass pathway, and the peak EPC levels induced by Ro 48-8071 treatment were associated with toxicity in Mut6 cells (IC 50 =0.0112 μM). Notably, the peak EPC level after Ro48-8071 treatment was only 62% of the peak EPC level after 1 treatment. This latter observation raises the hypothesis that Ro 48-8071 inhibits one or more enzymes downstream of LSS, thereby attenuating EPC synthesis. To evaluate whether Ro 48-8071 inhibits the synthesis of EPCs downstream of EPL, we cultured cells with isotope-labeled EPL (d6-EPL) and measured the production of d6-EPCs after compound treatment. Ro48-8071 inhibited the production of d6-EPCs, while 1 had no effect on d6-EPC production. These data suggest that Ro 48-8071 inhibits LSS, but may also have off-target effects in the sterol synthesis pathway downstream of LSS, thereby attenuating EPC synthesis. For small molecules that inhibit enzymes downstream of LSS, our prediction is that lanosterol will not be efficiently converted to cholesterol and therefore will not rescue toxicity as effectively as cholesterol. Consistent with our hypothesis, cholesterol is more effective than exogenous lanosterol (IC 50 Increased by 21.7 times) provides Ro-48-8071 (IC 50 In contrast, exogenous lanosterol and cholesterol provided complete and equivalent rescue in cells treated with 1. Together, these findings suggest that 1 does not inhibit enzymes downstream of LSS in the cholesterol synthesis pathway and therefore more efficiently produces the toxic metabolite EPC.

[0059] To enable unbiased identification of potential Ro 48-8071 off-targets, we synthesized Ro 48-8071 derivatives suitable for click chemistry. Ro 48-8071 contains a photoreactive benzophenone that, upon UV exposure, is predicted to form a reactive triple diradical capable of covalently modifying neighboring amino acids in binding proteins. We replaced the terminal olefinic group on Ro 48-8071 with a terminal alkynyl group (hereafter referred to as Ro-alkynyl), which serves as a functional handle for click chemistry reactions. After determining that Ro-alkyne retains antiproliferative activity against Mut6 (IC 50 =0.396 μM), we analyzed the cellular binding partners of Ro-alkynes. We identified multiple probe-dependent and UV-dependent bands that were displaced upon incubation with 10 μM Ro 48-8071; however, the band corresponding to LSS was only visible after ectopic expression of LSS in HEK293T cells.

[0060] To determine the identity of putative Ro 48-8071 off-targets, we performed affinity purification of Ro-alkyne binding proteins in the presence or absence of 10 μM Ro 48-8071. In stark contrast to MM0299, under Ro-alkyne-only conditions, 130 proteins showed greater enrichment than Lss (#131, 1.3-fold enrichment), suggesting many off-targets. Among the most enriched Ro-alkyne binding proteins were three enzymes in the cholesterol biosynthesis pathway: Ebp (#2, 8.4-fold enrichment), Lbr (#5, 3.1-fold enrichment), and Dhcr7 (#7, 2.5-fold enrichment). Together, these results suggest that, unlike M0299, Ro-48-8071 has many off-targets, including multiple enzymes involved in cholesterol biosynthesis.

[0061] Inhibition of these sterol biosynthetic enzymes may provide an explanation for the attenuated increase in EPCs observed by Ro-48-8071 after LSS inhibition. To assess how 1 and Ro 48-8071 affect these enzymes, we used a suite of small molecule tools that explore the cellular binding profiles of small molecules to several cholesterol biosynthetic enzymes (Theodoropoulos et al., 2020). We first tested the binding of 1 and Ro 48-8071 to EBP by incubating HCT116 cells with 1 μM TASIN-2 (EBP-specific photoaffinity probe) in the presence or absence of increasing doses of TASIN-30 (EBP-specific inhibitor), 1, or Ro 48-8071, followed by UV treatment and fluorescent dye binding. Consistent with the proteomics data, Ro 48-8071 and TASIN-30, but not 1, displaced TASIN-2 (band migrating at approximately 20 kDa). Additionally, Ro 48-8071 treatment resulted in the accumulation of 8,9-dehydrocholesterol (8,9-DHC), an atypical sterol that accumulates after genetic or pharmacological EBP inhibition (Braverman et al., 1999; Theodoropoulos et al., 2020), whereas treatment with 1 had no effect on cellular 8,9-DHC levels. Taken together, these data provide evidence that Ro 48-8071 binds to and inhibits Ebp in cells.

[0062] We next performed similar experiments using a probe specific for 7-dehydrocholesterol reductase (Dhcr7 in mice or DHCR7 in humans), called 4C12 (Theodoropoulos et al., 2020). The cross-linked Dhcr7 band migrated at 40 kDa and was displaced by 4C12 and Ro 48-8071; however, 1 did not displace the Dhcr7 probe at any dose tested. Consistent with inhibition, cellular levels of the Dhcr7 substrate 7-dehydrocholesterol accumulated after treatment with Ro 48-8071 but not 1. Compounds targeting EBP and DHCR7 can also inhibit DHCR24, which converts cholesterol to cholesterol (Theodoropoulos et al., 2020) (Wages et al., 2018). Although we did not have chemical tools to assess direct DHCR24 binding, we analyzed cholesterol levels in cells treated with 1 and Ro 48-8071. Our observation of accumulation of sterols in cells treated with high doses of Ro 48-8071, but not 1, supports the hypothesis that Ro 48-8071 also inhibits Dhcr24. Taken together, these data suggest that, although both 1 and Ro 48-8071 inhibit Lss, Ro 48-8071 also inhibits the activity of Ebp, Dhcr7, and Dhcr24 in an order of increasing potency.

[0063] Our observations suggest that Ro-48-8071 has significantly more off-targets than 1, some of which attenuate the production of the antiproliferative metabolite EPC. Therefore, although Ro-48-8071 inhibits LSS, its antiproliferative effects are more likely multifactorial. To assess the contribution of EPC to Ro 48-8071-induced toxicity, we co-incubated Ro 48-8071 with increasing concentrations of NB-598. NB-598 provided some protection against Ro 48-8071-induced toxicity (Ro 48-8071IC 50 However, the protective effect was not dose-dependent and disappeared at higher concentrations of NB-598. In contrast, NB-598 rescued the toxicity from 1 in a dose-dependent manner, resulting in an IC 50 The maximum increase is 11.8 times.

[0064] 5. MM0299 is effective in human GSC lines.

[0065] Given that our studies of MM0299 have primarily utilized Mut6 cells—a GSC line derived from a single genetically defined mouse model—we next sought to evaluate the antiproliferative activity and mechanism of action of MM0299 in human GSC lines with distinct genetic drivers. We generated two GSC lines from patients at UT Southwestern Medical Center, termed UTSW63 and UTSW71. UTSW63 was derived from a resected tumor from a 71-year-old male patient with temporal lobe malignancy. Genomic alterations that are bona fide GBM cancer drivers were identified in genomic analysis of this resected tumor. These included expected deletions of the tumor suppressors TP53 (deletion of 17p13.3-p11.1) and RB1 (deletion of 13q13.11-q14.3 and c.1215+1G>A, a typical splice donor site mutation predicted to cause a frameshift) and amplification of EGFR (93 copies of 7p11.2). UTSW71 was derived from a 52-year-old male patient with glioblastoma with alterations in different cancer drivers, including PTEN (deletion and P95S mutation at 10q21.1-q26.3) and amplification of a region containing the oncogenes CDK4 (34 copies at 12q13.3-q14.1), MDM2 (89 copies at 12q15), and c-MYC (51 copies at 8q24.13-q24.21).

[0066] Like Mut6 cells, both UTSW63 and UTSW71 grew as neurospheres in serum-free medium (data not shown). MM0299 inhibited the proliferation of both UTSW63 and UTSW71 cells. 50 0.0222 μM and 0.0212 μM, respectively. MM0299 toxicity in both human GSC lines was completely rescued by exogenous MCD-conjugated lanosterol or cholesterol. The naturally occurring mechanism of input of extracellular cholesterol is the uptake of circulating low-density lipoprotein (LDL). Therefore, we investigated whether human LDL could rescue MM0299 toxicity in human GBM cells. Addition of 20 μg / ml of human LDL to both UTSW63 and UTSW71 cells completely rescued the toxicity of 1. To investigate the role of the bypass pathway in MM0299 toxicity, we again turned to the tool compound NB-598. NB-598 was highly toxic in UTSW63 and UTSW71, with an IC 50 The lower doses of NB-598 that did not affect cell growth rescued the toxicity of 1, as shown by the IC 50In summary, the data suggest that 1 is effective in upregulating EPCs via a bypass pathway and thereby depleting cellular cholesterol pools in cultured human GSC lines.

[0067] 6. Analog 13 is bioavailable and able to cross the blood-brain barrier

[0068] Having established the mechanism and mode of action of MM0299, we sought to identify and evaluate analogs for bioavailability in the brain, a prerequisite for in vivo studies in glioblastoma. In vitro and in vivo pharmacological evaluation of a series of analogs (Table 1) led us to analog 13 as a candidate molecule for activating the bypass pathway in the mouse brain.

[0069]

[0070] Competing p75 / LSS (EC 50 =0.0287 μM) analog 13 has drug-like physicochemical properties (MW = 468 Da, CLogP = 4.3, LogP = 3.8, tPSA = 64), induced EPC upregulation, and in Mut6 cells (IC 50 =0.0443 μM) and human GSC line UTSW63 (IC 50 =0.0293 μM). Analog 13 was stable in plasma (>24 hours) and showed higher metabolic stability than other analogs when incubated with mouse S9 fraction (Table 1). After administration of 13 to mice at 5 mg / kg by intravenous injection (IV) or 20 mg / kg by oral administration (PO), pharmacokinetic (PK) analysis showed that AUC 口服 / AUC iv x dose iv / dose 口服 The defined oral bioavailability was 39% in plasma and 58% in brain. Importantly, 13 administered orally at 20 mg / kg also showed brain exposure, with total C 最大 5μM (2381ng / ml), free C 最大 The free AUC was 14 nM and the brain-blood ratio was 1.8 (defined as the free AUC 脑 / Free AUC 血浆 This is consistent with the free IC of the antiproliferative activity against the human GSC line calculated using the measured unbound fraction (fu) of 0.11 for the medium. 50Value (3nM) is better than that. Next, we used mass spectrometry to quantify the EPC levels after treatment with 13 in plasma and brain. After a single administration of 20mg / kg, the EPC level of plasma increased in a time-dependent manner. In the brain, we easily detected chain sterols, but no EPC was detected, even at the time point when plasma EPC was significantly increased. The cholesterol biosynthesis flux in the adult brain is expected to be low, which may explain the absence of detectable EPC. Alternatively, although we predict, the level of 13 in the brain may not be enough to inhibit LSS. In order to distinguish these possibilities, we asked whether we could detect EPC in orthotopic xenograft tumors derived from UTSW63 cells. 25 days after intracranial injection of UTSW63 cells, mice were orally administered with a dosing regimen of once or twice a day or 20mg / kg of 13 for 3 consecutive days. After treatment, we analyzed the EPC level and observed a dose-dependent increase in the EPC level in both serum and GBM tumors. These findings suggest that administration of 13 results in selective EPC induction in glioblastoma cells relative to normal brain and that EPCs may be used as a pharmacodynamic marker to guide the optimization of MM0299 for further preclinical evaluation.

[0071] discuss.

[0072] Our investigation of the mechanism of action of MM0299 reveals a novel approach for brain tumor therapy that targets tumor cell metabolism. Metabolic reprogramming is a fundamental feature of the molecular pathogenesis of gliomas, and many strategies aimed at exploiting this process are undergoing preclinical and clinical testing (Zhou and Wahl, 2019). One such strategy involves targeting metabolic pathways that display higher flux in glioma cells relative to normal cells, as exemplified by studies evaluating the radiosensitizing effects of purine nucleotide synthesis inhibition in GBM (NCT04477200) (Zhou et al., 2020). Another strategy entails blocking the synthesis of the oncometabolite 2-hydroxyglutarate, which selectively accumulates and drives the formation of isocitrate dehydrogenase (IDH)-mutated gliomas (NCT02481154) (Mellinghoff et al., 2021). In contrast, our approach relies on a new paradigm: activating rather than inhibiting metabolic pathways in GBM cells to trigger the production of tumor-specific cytotoxic metabolites, EPCs.

[0073] Like most cells, cancer cells regulate their non-esterified cholesterol pools through a balance of peripheral uptake, synthesis, and catabolism. Therefore, blocking cholesterol synthesis triggers a well-described negative feedback pathway that maintains homeostasis by increasing the uptake of low-density lipoprotein (LDL). However, low-density lipoprotein particles do not cross the blood-brain barrier, so the brain must synthesize its own cholesterol. Most of the synthesis of cholesterol in the brain occurs during embryonic development, and the synthesis rate of cholesterol in the adult brain drops sharply (Dietschy, 2009). However, relative to adult neurons, GBM cells that are expected to divide have an increased cholesterol demand and accordingly rely on both re-cholesterol synthesis and uptake of exogenous cholesterol (synthesized by astrocytes) to meet proliferation needs (Sassi et al., 2021). LSS inhibition has the potential to exploit the difference in cholesterol synthesis flux between normal brain and tumors. LSS inhibition shifts sterol flux from cholesterol synthesis to EPC synthesis, which ultimately blocks cell growth, predicting that tumors will synthesize more EPCs than normal brains. Consistent with this hypothesis, we found that in vivo administration of 13 did not result in the generation of detectable EPCs in normal brain but led to a dose-dependent increase in EPCs in orthotopic glioblastoma tumors. Thus, inhibition of cholesterol biosynthesis by LSS inhibition may selectively target rapidly dividing GBMs with elevated sterol synthesis.

[0074] Currently, there is multiple evidence that reducing the cellular cholesterol pool will hinder the progression of glioblastoma. The synthetic LXR agonists GW3965 and LXR-623 upregulate the expression of the cholesterol transporter ABCA1, which promotes cholesterol efflux and reduces the cellular cholesterol pool in GBM cells (Guo et al., 2011; Villa et al., 2016). As a result, LXR-623, which has brain penetrability, delays the progression of orthotopic glioblastoma tumors and prolongs the overall survival of mice bearing these tumors (Villa et al., 2016). However, there are mechanism-related adverse events that hinder the clinical development of LXR agonists for the treatment of glioblastoma or any other indications. For example, LXR agonists stimulate fatty acid synthesis, which leads to clinically unacceptable increases in circulating triglycerides and accelerates the development of fatty liver disease (Schultz et al., 2000; Joseph et al., 2002; Bradley et al., 2007).

[0075] Alternative strategies to reduce cholesterol levels in GBM include inhibitors of key enzymes in the sterol biosynthesis pathway. Cellular cholesterol levels are regulated within a narrow range of concentrations at the levels of synthesis, uptake of low-density lipoprotein, and catabolism (or efflux). Reduced cholesterol levels are sensed by the SREBP cleavage activator protein (SCAP), which promotes proteolysis and activation of SREBP-2 (Brown et al., 2018). Activated SREBP-2 translocates to the nucleus and promotes transcription of genes required for cholesterol synthesis and cholesterol uptake. Thus, these feedback pathways counteract the cholesterol-lowering effects of most enzyme inhibitors of cholesterol biosynthesis (Brown et al., 1978).

[0076] However, lanosterol synthase inhibition is unique in that it reduces cholesterol synthesis without stimulating these negative feedback pathways. LSS converts linear oxidized squalene to lanosterol, the first sterol ring-containing intermediate in the biosynthesis of cholesterol. Unlike other post-squalene enzymes, inhibition of LSS blocks cholesterol synthesis without leading to accumulation of sterol intermediates. Instead, partial inhibition of LSS diverts sterol flux to a "bypass" pathway that ultimately leads to the synthesis of atypical sterol EPCs. EPC accumulation triggers a feedback mechanism that blocks the activation of SREBPs, thereby preventing cells from upregulating genes for de novo cholesterol biosynthesis and cholesterol uptake (Radhakrishnan et al., 2007). In addition, EPCs are potent regulators of LXRs, nuclear hormone transcription factors whose activation induces cholesterol metabolism and efflux—(Janowski et al., 1996; Janowski et al., 1999). As an endogenous LXR ligand, EPC does not drive fatty acid synthesis (Rowe et al., 2003), and LSS inhibitors do not cause adverse events characteristic of synthetic LXR agonists in animals. Therefore, LSS inhibition has the potential to reduce cellular cholesterol pools in multiple ways, including direct inhibition of de novo cholesterol synthesis, inhibition of feedback pathways that promote cholesterol biosynthesis, and promotion of efflux through activation of LXRs. Here, we show that EPC synthesis via a bypass pathway is necessary and sufficient to inhibit the proliferation of glioblastoma cells by the LSS inhibitor MM0299.

[0077] Recent evidence also suggests that LSS is a potential glioblastoma target. Allis and colleagues reported that MI-2, a small molecule LSS inhibitor that increases EPC levels, exhibits antiproliferative activity in glioma. MI-2 has previously been reported to inhibit protein-protein interactions between Menin and MLL, important epigenetic regulators in leukemia (Grembecka et al., 2012; Shi et al., 2012). However, the activity of MI-2 in glioma does not involve Menin or MLL but is the result of direct inhibition of LSS. Their findings not only designate LSS as a potential target in GBM, but also reinforce the importance of continued target deconvolution for compounds evaluated in preclinical cancer studies (Phillips et al., 2019).

[0078] Most LSS inhibitors have been developed primarily to reduce LDL levels for the treatment of atherosclerosis, and data supporting their efficacy against cancer are sparse. One limitation to their development is the challenge of assessing LSS selectivity. In fact, most of the reported small molecule inhibitors of LSS are amphoteric amine drugs that bind to many other proteins, including multiple enzymes in the post-squalene cholesterol biosynthetic pathway (Rabelo et al., 2017). This multipharmacology is most prominent in LSS, EBP, and the sterol reductases DHCR7, DHCR14, and DHCR24 (Wages et al., 2018; Korade et al., 2016; Moebius et al., 1998). These enzymes are required to stabilize the carbocation charges generated during the conversion of highly lipophilic sterols and may explain why protonated amphoteric amines at physiological pH have the ability to bind to any or all of these enzymes. Given the similarity in substrate structure, as well as the physical proximity of these enzymes, photochemical probes whose binding and activity can be attributed to specific cholesterol biosynthetic enzymes have proven to be useful tools for studying these enzymes (Theodoropoulos et al., 2020). Using a combination of these photochemical probes and sterol mass spectrometry, we compared the selectivity of MM0299 with that of the well-characterized LSS inhibitor Ro 48-8071. Although Ro 48-8071 binds to and inhibits LSS, our studies show that it also binds to a number of off-targets and inhibits multiple enzymes in the cholesterol pathway, including EBP and DHCR7, which in turn attenuate EPC production. A similar off-target survey of MM0299 revealed no off-targets and no evidence of binding to or inhibition of other cholesterol biosynthetic enzymes.

[0079] Our studies reinforce the role of LSS as a target in glioblastoma and identify MM0299 as a highly selective LSS inhibitor. In addition to the cholesterol-lowering effects of EPCs described above, EPCs have been shown to stimulate the formation of myelinating oligodendrocytes through unknown mechanisms (Hubler et al., 2021) and promote midbrain dopaminergic neurogenesis through LXR activation (Theofilopoulos et al., 2013; Theofilopoulos et al., 2019). Neurodegenerative disorders including multiple sclerosis (MS) and Parkinson's disease (PD) are the result of neuronal demyelination and loss of midbrain dopaminergic neurons, respectively. These diseases require new therapies, and brain-penetrating MM0299 derivatives that induce EPC production also have clinical utility in these diseases.

[0080] A major challenge in developing therapies for GBM or other neurological conditions is to identify small molecule drugs that can cross the blood-brain barrier. Here, we describe derivatives of MM0299 that are orally bioavailable and have blood-brain barrier penetration. These derivatives lead to an increase in EPCs in orthotopic xenograft tumors, but not in normal brain.

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[0119] Thoma,R.,Schulz-Gasch,T.,D'Arcy,B.,Benz,J.,Aebi,J.,Dehmlow,H.,Hennig,M.,Stihle,M.,and Ruf,A.(2004).Insight into steroid scaffold formation fromthe structure of human oxidosqualene cyclase.Nature 432,118-122.10.1038 / nature02993.

[0120] Villa,G.R.,Hulce,J.J.,Zanca,C.,Bi,J.,Ikegami,S.,Cahill,G.L.,Gu,Y.,Lum,K.M.,Masui,K.,Yang,H.,et al.(2016).An LXR-Cholesterol Axis Creates aMetabolic Co-Dependency for Brain Cancers.Cancer Cell 30,683-693.10.1016 / j.ccell.2016.09.008.

[0121] Wages,P.A.,Kim,H.H.,Korade,Z.,and Porter,N.A.(2018).Identificationand characterization of prescription drugs that change levels of7-dehydrocholesterol and desmosterol.J Lipid Res 59,1916-1926.10.1194 / jlr.M086991.

[0122] Willy,P.J.,Umesono,K.,Ong,E.S.,Evans,R.M.,Heyman,R.A.,andMangelsdorf,D.J.(1995).LXR,a nuclear receptor that defines a distinctretinoid response pathway.Genes Dev 9,1033-1045.10.1101 / gad.9.9.1033.

[0123] Wong,J.,Quinn,C.M.,Guillemin,G.,and Brown,A.J.(2007).Primary humanastrocytes produce 24(S),25-epoxycholesterol with implications for braincholesterol homeostasis.J Neurochem 103,1764-1773.10.1111 / j.1471-4159.2007.04913.x.

[0124] Zhou,W.,and Wahl,D.R.(2019).Metabolic Abnormalities in Glioblastomaand Metabolic Strategies to Overcome Treatment Resistance.Cancers(Basel)11.10.3390 / cancers11091231.

[0125] Zhou, W., Yao, Y., Scott, AJ, Wilder-Romans, K., Dresser, JJ, Werner, CK, Sun, H., Pratt, D., Sajjakulnukit, P., Zhao, SG, et al.(2020).Purine metabolismregulates DNA repair and therapy resistance in glioblastoma.Nat Commun 11.3811.10.1038 / s41467-020-17512-x.

[0126] New snowflakes

[0127] Nat Chem Biol 12,218–225,doi:10.1038 / nchembio.2016(2016).

[0128] 2.McDonald,JG,Smith,DD,Stiles,AR&Russell,DWA comprehensive method for extraction and quantitative analysis of sterols and secosteroids from human plasma.J Lipid Res 53,1399-1409,doi:10.1194 / jlr.D022285(2012).

[0129] 3.Kurten,C.,Uhlen,M.&Syren,POOverexpression of functional humanoxidosqualene cyclase in Escherichia coli.Protein Expr Purif 115,46-53,doi:10.1016 / j.pep.2015.04.015(2015).

[0130] 4. McNaney, C. A. et al. An automated liquid chromatography - mass spectrometry process to determine metabolic stability half - life and intrinsic clearance of drug candidates by substrate depletion. Assay Drug Dev Technol 6, 121 - 129, doi:10.1089 / adt.2007.103(2008).

[0131] 5. Kalvass, J. C. & Maurer, T. S. Influence of nonspecific brain and plasma binding on CNS exposure: implications for rational drug discovery. Biopharm Drug Dispos 23, 327 - 338, doi:10.1002 / bdd.325(2002).

[0132] 6. Louis, D. N. et al. The 2021 WHO Classification of Tumors of the Central Nervous System: a summary. Neuro Oncol 23, 1231 - 1251, doi:10.1093 / neuonc / noab106(2021).

[0133] Synthesis

[0134] Unless otherwise stated, all reactions were carried out under nitrogen atmosphere using dry solvents under anhydrous conditions. Anhydrous solvents were obtained by passing them through commercially available alumina columns (Innovative technology, Inc., MA). All reagents were commercial compounds with the highest purity available. Analytical thin layer chromatography (TLC) was performed on aluminum plates with Merck Kieselgel 60 F254 and observed by UV irradiation (254 nm) or by staining with potassium permanganate solution. Flash column chromatography was performed under pressure using Merck Kieselgel 60 (230-400 mesh). Infrared spectra from thin films deposited on sodium chloride glass were obtained on a Perkin-Elmer I1000 FTIR series. The samples were analyzed at 20 ° C on a Rudolph Research Analytical Optical rotations were measured on an IV polarimeter. 1H NMR spectra were recorded in CDCl3, CD3OD, DMSOd6 and (CD3)2CO with residual protic solvent as internal reference on a Varian Inova-400 spectrometer at ambient temperature at 400 MHz (CDCl3, d H =7.26ppm; (CD3)2CO,d H =2.05ppm; CD3OD,d H =3.31ppm; DMSO-d6, d H = 2.50 ppm); chemical shifts (d) are given in parts per million (ppm), coupling constants (J) are given in Hertz (Hz). Proton spectra are reported as follows: d (multiplicity, coupling constant J, number of protons). The following abbreviations are used to explain multiplicity: app = apparent, b = broad, d = doublet, dd = doublet of doublet, ddd = doublet of doublet of doublet, dddd = doublet of doublet of doublet of doublet, m = multiplet, s = singlet, t = triplet. 13 C NMR spectra in CDCl3, CD3OD, DMSO-d6 and (CD3)2CO, CDCl3(d C =77.0ppm), CD3OD(d C =49.0ppm), DMSO-d6(d C =39.4ppm) or (CD3)2CO(d C =30.8 ppm) was used as an internal reference and recorded on the same spectrometer at 100 MHz at ambient temperature. Electrospray ionization mass spectra (ESI-MS) were recorded on a Shimadzu 2010-LCMS. HRMS was performed on a Shimadzu IT-TOF. Microwave reactions were performed on Initiator Classic. DMAP = 4-(dimethylamino)pyridine, DMF = N,N-dimethylformamide, DIPEA = N,N-diisopropylethylamine, DHP = tetrahydro-2H-pyran, EDC = N-ethylcarbodiimide hydrochloride, HATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, MW = microwave, NMP = 1-methyl-2-pyrrolidone, py = pyridine, TBDMS = tert-butyldimethylsilane, THF = tetrahydrofuran, THP = 3,4-dihydro-2H-pyran. Commercially available materials were used without further purification unless otherwise stated. All solvents were HPLC or ACS grade. Solvents used for moisture-sensitive operations were distilled from dry reagents under nitrogen: Et2O and THF were distilled from sodium benzophenonecarbonyl; benzene and toluene were distilled from sodium; CH2Cl2 was distilled from CaH2; pyridine was distilled from solid KOH; anhydrous N,N-dimethylformamide and CH3CN were purchased from commercial sources. Unless otherwise stated, reactions were performed under argon atmosphere with magnetic stirring. Flash chromatography (FC) was performed using E Merck silica gel 60 (240-400 mesh) according to the protocol of Still, Kahn and Mitra1.

[0135] General Procedure A for the Condensation of Anhydride 97 with (hetero)arylamines (Mechanisms 1-2). A mixture of anhydride 97 (1.0 eq.) and the corresponding (hetero)arylamine (1.0 eq.) in pyridine (0.1 M) was heated at 140° C. in a pressure sealed tube. The reaction was monitored by TLC analysis. The reaction was cooled to room temperature, diluted with CH2Cl2 (10 mL), and washed with excess 10% aqueous CuSO4. The organic phase was washed with H2O, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography to afford the corresponding dicarboximide.

[0136] General Procedure B for the Condensation of Anhydride 97 with (hetero)arylamines (Mechanisms 1-2). A mixture of anhydride 97 (1.0 eq.) and the corresponding (hetero)arylamine (1.0 eq.) in AcOH (0.5 M) was heated to 90 °C by conventional heating methods or to 100 °C for 45 minutes using a microwave reactor. The reaction was monitored by TLC analysis. The reaction was cooled to room temperature and crystallized. The crystals were collected by filtration and the residue was washed with Et2O / hexane (1:1) to remove excess AcOH without further purification. When no crystallization occurred, the mixture was diluted with EtOAc and neutralized with saturated aqueous NaHCO3. The organic phase was washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography as shown to give the corresponding dicarboximide.

[0137] General Procedure C (Mechanism 1) for the synthesis of compounds 92-95, 10, 11, 15, 16, 20, 29, 30a-45a, 47a-64a, 52b, 55b-58b, 60b-63b and 71-86 by alkylation. A mixture of (optionally substituted) nitrophenols or intermediates 98-106 (1.0 equiv), the corresponding bromide (1.5 equiv), K2CO3 (1.5 equiv) and NaI (0.5 equiv) in acetone (0.1 M) was heated to 80°C and stirred overnight. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography to give the alkylated product.

[0138] General procedure D for the one-pot reduction-cycloaddition synthesis of compound 7-9 from nitroarenes 93-95 (Mechanism 1). To a suspension of Fe (5 eq.) in AcOH (1.0 M) were added the corresponding nitroarenes 93-95 (1.0 eq.), maleic anhydride (1.2 eq.), and 1,3,5-cycloheptatriene (3.0 eq.). The mixture was heated to 120 °C and stirred for 24 h. The mixture was cooled to room temperature, filtered through a Celite pad, and the residue was washed with EtOAc (2×). The combined organic phases were washed with saturated aqueous NaHCO3, brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure, and the residue was purified by silica gel flash chromatography as shown to give the cycloadduct 7-9.

[0139] General procedure E for the synthesis of enones 110, 111a-f via Wittig olefination (Mechanism 2). A mixture of the corresponding benzaldehyde 107 or 108 (1 eq.) and the corresponding ylide (1.5 eq.) in CH2Cl2 (0.2 M) was refluxed at 70°C. The reaction was monitored by TLC. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography as shown to afford enones 110 or 111a-f.

[0140] General Procedure F for the Synthesis of Cyclopropane Analogs via Corey-Chaykovsky Cyclopropanation (Mechanism 2). A flame-dried flask was charged with trimethylsulfoxonium iodide (1.5 eq.) and NaH (1.5 eq., 60% dispersion in oil). The mixture was dissolved in anhydrous DMSO (0.5 M) and stirred at room temperature for 30 minutes. It was noted that the reaction mixture became homogeneous after 30 minutes. The corresponding enone (1.0 eq.) was added to the reaction mixture in one portion. The resulting mixture was heated to 85° C. and stirred overnight. After cooling to room temperature, the reaction was quenched with cold H2O. The mixture was extracted with EtOAc, and the combined organic layers were washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure, and the residue was purified by silica gel flash chromatography as shown to give the cyclopropyl derivative.

[0141] 1-(4-Methoxyphenyl)-2-(4-nitrophenoxy)ethan-1-one (92).

[0142]

[0143] General Procedure C. Purification by flash chromatography on silica gel (gradient elution, 0→30% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 92 (3.8 g, 13.2 mmol, 92%) as a light yellow solid. IR (cm -1 )2914,2842,1690,1600,1509,1341,1231,1172,1112,970,836; 1 H NMR (400MHz, CDCl3) δ8.19(d,J=9.2Hz,2H),7.97(d,J=8.8Hz,2H),6.99(d,J=9.2Hz,2H),6.98(d,J=9.2Hz,2H),5.36(s,2H),3.90(s,3H); 13 C NMR (100 MHz, CDCl3) δ 191.5, 164.6, 163.2, 130.6 (2C), 127.2, 126.1 (2C), 115.0 (2C), 114.5 (2C), 110.2, 70.7, 55.8; ES-API MS: m / z for C 15 H 14 NO5 calculated value 288.1, found value 288.1 [M+H] + .

[0144] 2-(2-Fluoro-4-nitrophenoxy)-1-(4-methoxyphenyl)ethan-1-one (93).

[0145]

[0146] General Procedure C. Purification by flash chromatography on silica gel (gradient elution, 0→25% EtOAc in hexanes) afforded 93 (45 mg, 0.14 mmol, 96%) as a yellow solid. IR (cm -1 )1686,1600,1508,1346,1293,1239,1171,970; 1 H NMR (400MHz, DMSO-d6) δ8.20 (dd, J=11.1, 2.6Hz, 1H), 8.06 (d, J=9.2Hz, 1H), 7.99 (d, J=8.8Hz,2H),7.34(t,J=8.9Hz,1H),7.11(d,J=8.8Hz,2H),5.88(s,2H),3.87(s,3H);13 C NMR (100 MHz, DMSO-d6) δ 191.3, 163.8, 152.0 (d, J = 10.0 Hz, 1C), 150.2 (d, J = 246.9 Hz, 1C), 140.3 (d, J = 7.0 Hz, 1C), 130.3 (2C), 126.8, 121.1 (d, J = 2.7 Hz, 1C), 114.7, 114.1 (2C), 112.1 (d, J = 21.3 Hz, 1C), 70.9, 55.7; ES-API MS: m / z for C 15 H 13 FNO5 calculated value 306.1, found value 306.1 [M+H] + .

[0147] 2-(3-Fluoro-4-nitrophenoxy)-1-(4-methoxyphenyl)ethan-1-one (94).

[0148]

[0149] General Procedure C. Purification by recrystallization in acetone / H2O gave 94 (376 mg, 1.23 mmol, 83%) as a brown solid. IR (cm -1 )1678,1600,1511,1332,1243,1171,1098,829; 1 H NMR (400MHz, DMSO-d6) δ8.15(dd,J=9.2,9.2Hz,1H),8.00(dd,J=8.8Hz,2H),7.29(d,J =14.0Hz,1H),7.11(d,J=8.4Hz,2H),7.02(d,J=9.2Hz,1H),5.78(s,2H),3.87(s,3H); 13 C NMR (100 MHz, DMSO-d6) δ 192.0, 164.9 (d, J = 11.3 Hz, 1C), 164.4, 157.3 (d, J = 260.0 Hz, 1C), 131.0 (2C), 128.6 (2C), 127.5, 114.8 (2C), 112.5 (d, J = 2.0 Hz, 1C), 104.8 (d, J = 24.6 Hz, 1C), 71.5, 56.3 (d, J = 2.3 Hz, 1C); ES-API MS: m / z for C 15 H 13 FNO5 calculated value 306.1, found value 306.1 [M+H] + .

[0150] 1-(4-Methoxyphenyl)-2-(4-nitro-3-(trifluoromethyl)phenoxy)ethan-1-one (95).

[0151]

[0152] General Procedure C. Purification by flash chromatography on silica gel (gradient elution, 0→30% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 95 (802 mg, 2.26 mmol, 94%) as an orange solid. IR (cm -1 )2940,1690,1601,1534,1314,1229,1174,1149,974,834; 1 H NMR (400MHz, CDCl3) δ7.98(d,J=8.4Hz,1H),7.96(d,J=8.4Hz,2H),7.36(d,J=2.8Hz ,1H),7.08(dd,J=9.2,2.4Hz,1H),7.00(d,J=8.4Hz,2H),5.41(s,2H),3.90(s,3H); 13 C NMR (100 MHz, CDCl3) δ 190.8, 164.8, 161.4, 130.6 (2C), 130.2, 128.3, 126.9, 126.6, 121.9 (q, J = 272.1 Hz, 1C), 117.0, 115.6 (q, J = 5.9 Hz, 1C), 114.6 (2C), 70.7, 55.9; ES-API MS: m / z for C 16 H 13 F3NO5 calculated value 356.1 , Measured value 356.1[M+H] + .

[0153] 2-(4-Aminophenoxy)-1-(4-methoxyphenyl)ethan-1-one (96).

[0154]

[0155] A mixture of nitroarene 92 (1.0 equiv.) and iron powder (5.0 equiv.) in saturated NH4Cl solution (1.0 M) and EtOH (1.0 M) was heated to 70 °C and stirred until TLC showed complete conversion. The mixture was cooled to room temperature, filtered through a Celite pad, and the residue was washed with EtOAc (2×). The combined organic layers were washed with saturated aqueous NaHCO3, brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→70% EtOAc in hexanes) to give aniline 96 (78 mg, 0.30 mmol, 90%) as a yellow solid. IR (cm -1 )3408,3334,1600,1506,1258,1213,1172,828; 1 H NMR (400MHz, CDCl3) δ7.96(d,J=8.8Hz,2H),6.92(d,J=8.8Hz,2H),6.76(d,J=8.8Hz,2H),6.59(d,J=8.8Hz,2H),5.10(s,2H),3.84(s,3H),3.42(br,2H); 13 C NMR (100 MHz, CDCl3) δ 193.8, 164.1, 151.4, 141.0, 130.7 (2C), 127.9, 116.4 (2C), 116.3 (2C), 114.1 (2C), 71.9, 55.7; ES-API MS: m / z for C 15 H 16 NO3 calculated value 258.1, found value 258.1 [M+H] + .

[0156] (3aR,4R,4aR,5aS,6S,6aS)-4,4a,5,5a,6,6a-Hexahydro-1H-4,6-vinylcyclopropane[f]isobenzofuran-1,3(3aH)-dione; (3aR,4S,4aS,5aR,6R,6aS)-4,4a,5,5a,6,6a-Hexahydro-1H-4,6-vinylcyclopropane[f]isobenzofuran-1,3(3aH)-dione (97).

[0157]

[0158] Endo-isomer Exo-isomer

[0159] A mixture of 1,3,5-cycloheptatriene (10 g, 0.108 mol) and maleic anhydride (12.0 g, 0.122 mol) in xylene (50 mL) was heated to 150 °C for 12 hours. The reaction was cooled to room temperature and concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel flash chromatography (gradient elution, 0→10% EtOAc in hexanes and 10→30% EtOAc in hexanes) to give exo-isomer 97-exo (1 g, 5.25 mmol, 5%) and endo-isomer 97-endo (14 g, 73.6 mmol, 70%), both as white solids. 97-endo: IR (cm -1 )3008,2945,1856,1823,1774,1376,1227,1093,1078,952,915; 1 H NMR (500MHz, CDCl3) δ5.87(m,2H),3.45(m,2H),3.23(dd,J=7.5,1.5Hz,2H),1.10(m,2H),0.35(m,1H),0.25(m,1H); 13 C NMR (100 MHz, CDCl3) δ 172.5 (2C), 128.6 (2C), 46.0 (2C), 33.8 (2C), 9.7 (2C), 5.3; ES-API MS: m / z for C 11 H 10 O3 calculated, found 191.1 [M+H] + .97-Appearance:IR(cm -1 )3016,2983,1848,1780,1305,1221,1086,1072,1042,960,915,851; 1 H NMR (500MHz, CDCl3) δ5.94(dd,J=4.0,4.0Hz,2H),3.41(m,2H),3.08(m,2H),1.12(m,2H),0.21(dd,J=13.5,7.0Hz,1H),0.13(m,1H); 13 C NMR (100 MHz, CDCl3) δ 172.5 (2C), 129.4 (2C), 46.7 (2C), 32.7 (2C), 5.6 (2C), 1.9; ES-API MS: m / z for C 11 H 11 O3 calculated value 191.1, found value 191.1 [M+H] + .

[0160] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (1).

[0161]

[0162] A mixture of anhydride 97-endo (1.0 equiv.) and the corresponding aniline (1.0 equiv.) in AcOH (0.2 M) was heated to 120 °C for 30 min. The mixture was cooled to room temperature, quenched with saturated aqueous NaHCO3, and extracted with EtOAc (3×). The combined organic layers were washed with brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by crystallization in EtOAc / hexanes to afford 1 (40 mg, 0.093 mmol, 35%) as a white solid. IR (cm -1 )1704,1601,1511,1389,1224,1172; 1 H NMR (400MHz, DMSO-d6) δ8.00(d,J=8.0Hz,2H),7.09(d,J=8.0Hz,2H),6.98(s,4H),5.82(m,2H ),5.54(s,2H),3.85(s,3H),3.27(m,2H),3.16(m,2H),1.17(m,2H),0.25(m,1H),0.07(m,1H); 13 C NMR (100 MHz, DMSO-d6) δ 192.8, 177.9 (2C), 163.8, 157.8, 130.4 (2C), 128.1 (2C), 127.7 (2C), 127.4, 125.2, 115.0 (2C), 114.3 (2C), 70.2, 55.8, 45.0 (2C), 33.4 (2C), 9.7 (2C), 4.5; ES-API MS: m / z for C 26 H 24 NO5 calculated value 430.2, found value 430.2 [M+H] + .

[0163] (3aR,4S,4aS,5aR,6R,6aS)-2-(4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (2).

[0164]

[0165] A mixture of anhydride 97-exo (1.0 equiv.) and the corresponding aniline (1.0 equiv.) in AcOH (0.2 M) was heated to 120 °C for 30 min. The mixture was cooled to room temperature, quenched with saturated aqueous NaHCO3, and extracted with EtOAc (3×). The combined organic layers were washed with brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by crystallization in EtOAc / hexanes to afford 2 (35 mg, 0.081 mmol, 33%) as a white solid. IR (cm -1 )1702,1599,1510,1387,1308,1228,1172; 1 H NMR (500MHz, DMSO-d6) δ8.00(d,J=9.0Hz,2H),7.12(d,J=9.5Hz,2H),7.09(d,J=9.5Hz,2H),7.02(d,J=8.5Hz, 2H),5.95(m,2H),5.56(s,2H),3.86(s,3H),3.24(m,2H),2.89(m,2H),0.95(m,2H),0.12(m,1H),-0.02(m,1H); 13 C NMR (100 MHz, DMSO-d6) δ 192.3, 177.3 (2C), 163.2, 157.3, 129.9 (2C), 128.8 (2C), 127.7 (2C), 126.8, 124.8, 114.5 (2C), 113.7 (2C), 69.5, 55.2, 44.7 (2C), 31.5 (2C), 4.7 (2C), 0.86; ES-API MS: m / z for C 26 H 24 NO5 calculated value 430.2, found value 430.2 [M+H] + .

[0166] 1-(4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-1H-pyrrole-2,5-dione (5).

[0167]

[0168] A mixture of aniline 96 (219 mg, 0.851 mmol) and maleic anhydride (100 mg, 1.02 mmol) in AcOH (8.5 mL) was heated to 120 ° C and stirred for 12 hours. The mixture was cooled to room temperature, quenched with cold saturated aqueous NaHCO3, and extracted with EtOAc (3×). The combined organic layers were washed with brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→40% EtOAc in hexanes) to give compound 5 (148 mg, 0.44 mmol, 52%) as a yellow solid. IR (cm -1 )1714,1601,1511,1397,1224,1172; 1 H NMR (400MHz, CDCl3) δ8.00(d,J=9.2Hz,2H),7.24(d,J=8.8Hz,2H),7.02(d,J=8.8Hz,2H),6.97(d,J=8.8Hz,2H),6.83(s,2H),5.23(s,2H),3.89(s,3H); 13 C NMR (100 MHz, CDCl3) δ 192.8, 169.9 (2C), 164.4, 157.8, 134.4 (2C), 130.8 (2C), 127.8 (2C), 127.7, 124.9, 115.6 (2C), 114.3 (2C), 71.1, 55.8; ES-API MS: m / z for C 19 H 16 NO5 calculated value 338.1, found value 338.1 [M+H] + .

[0169] (3aR,4S,7R,7aS)-2-(4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-3a,4,7,7a-tetrahydro-1H-4,7-ethanoisoindole-1,3(2H)-dione (3).

[0170]

[0171] To a solution of maleimide 5 (1 eq.) in CH2Cl2 (0.1 M) was added cyclohexadiene (5.0 eq.). The resulting mixture was heated to 60 °C until TLC showed complete consumption of the starting material. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→40% EtOAc in hexanes) to give the cycloadduct 3 (9.0 mg, 0.021 mmol, 36%) as a white solid. IR (cm -1)2954,1707,1601,1512,1389,1299,1233,1172; 1 H NMR (500MHz, CDCl3) δ7.98(d,J=8.8Hz,2H),7.09(d,J=9.0Hz,2H),6.98(d,J=9.0Hz,2H),6.96(d,J=9.0Hz,2H),6.27(d d,J=4.0,3.5Hz,2H),5.19(s,2H),3.89(s,3H),3.24(m,2H),2.99(m,2H),1.65(d,J=7.5Hz,2H),1.43(d,J=8.0Hz,2H); 13 C NMR (100 MHz, CDCl3) δ 192.8, 178.4 (2C), 164.3, 158.1, 132.6 (2C), 130.8 (2C), 128.0 (2C), 125.6, 115.5 (2C), 114.3 (2C), 105.0, 71.2, 55.8, 44.4 (2C), 32.2 (2C), 23.9 (2C); ES-API MS: m / z for C 25 H 24 NO5 calculated value 418.2, found value 418.2 [M+H] + .

[0172] (3aR,4S,7R,7aS)-2-(4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-3a,4,7,7a-tetrahydro-1H-4,7-methanoisoindole-1,3(2H)-dione (4).

[0173]

[0174] To a solution of maleimide 5 (1 eq.) in CH2Cl2 (0.1 M) was added cyclopentadiene (5.0 eq.). The resulting mixture was heated to 60 °C until TLC showed complete consumption of the starting material. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→70% EtOAc in hexanes) to give the cycloadduct 4 (30 mg, 0.074 mmol, 63%) as a light yellow solid. IR (cm -1 )2843,1706,1601,1512,1386,1263,1227,1173; 1H NMR (500MHz, CDCl3) δ7.98(d,J=8.5Hz,2H),7.05(d,J=9.0Hz,2H),6.97(d,J=9.0Hz,2H),6.96(d,J=8.5Hz,2H) ,6.24(s,2H),5.18(s,2H),3.89(s,3H),3.49(m,2H),3.41(m,2H),1.78(d,J=9.0Hz,1H),1.60(d,J=9.0Hz,1H); 13 C NMR (100 MHz, CDCl3) δ 192.8, 177.2 (2C), 164.3, 158.1, 134.8 (2C), 130.8 (2C), 128.1 (2C), 127.6, 125.5, 115.5 (2C), 114.2 (2C), 71.1, 55.7, 52.4, 45.8 (2C), 45.6 (2C); ES-API MS: m / z for C 24 H 22 NO5 calculated value, found 404.1[M+H] + .

[0175] (3aR,4R,4aR,5aS,6S,6aS)-2-(2-fluoro-4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (7).

[0176]

[0177] Step D. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) afforded 7 (30 mg, 0.067 mmol, 14%) as a white solid. IR (cm -1 )1707,1693,1600,1518,1385,1291,1243,1166; 1 H NMR (400MHz, CDCl3) δ7.97(d,J=8.8Hz,2H),7.06(m,1H),6.97(d,J=8.8Hz,2H),6.78(m,1H),6.75(m,1H),5.8 5(dd,J=4.4,3.6Hz,2H),5.19(s,2H),3.89(s,3H),3.48(m,2H),3.15(m,2H),1.14(m,2H),0.35-0.26(m,2H); 13C NMR (100 MHz, DMSO-d6) δ 192.6, 177.5 (2C), 164.1, 160.0, 159.1, 156.6, 130.7 (2C), 128.0, 127.8, 127.6 (2C), 114.5 (2C), 110.0, 103.4, 70.7, 56.1, 45.7, 45.2, 33.7, 33.6, 9.9 (2C), 4.9; ES-API MS: m / z for C 26 H 23 FNO5 calculated value 448.2, found value 448.2 [M+H] + .

[0178] (3aR,4R,4aR,5aS,6S,6aS)-2-(3-fluoro-4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (8).

[0179]

[0180] Step D. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) afforded 8 (183 mg, 0.40 mmol, 59%) as a white solid. IR (cm -1 )1774,1709,1600,1515,1441,1388,1268,1240,1174; 1 H NMR(400MHz,DMSO-d6)δ7.99(d,J=8.8Hz,2H),7.15(dd,J=9.2,9.2Hz,1H),7.1 0(d,J=8.8Hz,2H),7.04(dd,J=11.6,2.0Hz,1H),6.84(ddd,J=8.8,1.6,1.6Hz, 1H),5.82(dd,J=4.4,3.6Hz,2H),5.68(s,2H),3.86(s,3H),3.28(m,2H),3.18( m,2H),1.18(m,2H),0.26(dd,J=12.8,7.3Hz,1H),0.08(dd,J=8.8,3.7Hz,1H); 13C NMR (100 MHz, DMSO-d6) δ 192.2, 177.4 (2C), 163.7, 150.7 (d, J = 243.5 Hz, 1C), 145.8 (d, J = 10.1 Hz, 1C), 130.2 (2C), 127.5 (2C), 127.1, 124.8 (d, J = 9.1 Hz, 1C), 123.1 (d, J = 3.3 Hz, 1C), 115.0, 114.8, 114.1 (2C), 70.5, 55.7, 44.8 (2C), 33.3 (2C), 9.5 (2C), 4.4; ES-API MS: m / z for C 26 H 23 FNO5 calculated value 448.2, found value 448.1 [M+H] + .

[0181] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-methoxyphenyl)-2-oxoethoxy)-2-(trifluoromethyl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (9).

[0182]

[0183] Step D. Purification by flash chromatography on silica gel (gradient elution, 0→30% EtOAc in hexanes) afforded 9 (432 mg, 0.87 mmol, 72%) as a white solid. IR (cm -1 )3062,3016,1715,1694,1600,1506,1316,1174; 1 H NMR (500MHz, CDCl3) δ7.97(d,J=9.2Hz,2H),7.31(d,J=2.8Hz,1H),7.12(dd,J=8.4,2.8Hz,1H),7.03(d,J=8.4Hz,1H),6 .98(d,J=8.8Hz,2H),5.86(m,2H),5.25(s,2H),3.89(s,3H),3.47(m,2H),3.16(m,2H),1.12(m,2H),0.35-0.26(m,2H); 13C NMR (100 MHz, CDCl3) δ 191.8, 177.8 (2C), 164.5, 158.9, 132.5, 132.1, 130.8 (2C), 128.2, 127.8, 127.4, 118.8, 118.6, 114.7 (q, J = 5.3 Hz, 1C), 114.4 (2C), 71.0, 55.8, 46.3, 45.6, 33.8, 33.6, 10.3, 10.1, 5.05, 4.96; ES-API MS: m / z for C 27 H 23 F3NO5 calculated value 498.2, found value 498.1 [M+H] + .

[0184] (3aR,4R,4aR,5aS,6S,6aS)-2-(3-Chloro-4-hydroxyphenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (98).

[0185]

[0186] Step B (microwave). Compound 98 was obtained by crystallization as a white solid (466 mg, 1.48 mmol, 94%). IR (cm -1 )3396,1694,1496,1291,1195,1166,736; 1 H NMR (400MHz, CDCl3) δ7.08(d,J=2.4Hz,1H),6.93(d,J=8.8Hz,1H),6.87(dd,J=8.8,2.4Hz,1H),5.81(dd,J =4.8,3.2Hz,2H),3.43(m,2H),3.08(dd,J=1.6,1.6Hz,2H),2.66(br,1H),1.11(m,2H),0.31-0.21(m,2H); 13 CNMR (100 MHz, CDCl3) δ 178.2 (2C), 152.8, 128.0, 127.9 (2C), 126.4, 124.1, 120.7, 116.7, 45.4 (2C), 33.9 (2C), 10.0 (2C), 4.8; ES-API MS: m / z for C 17 H 15 ClNO3 calculated value 316.1, found value 316.1 [M+H] + .

[0187] (3aR,4R,4aR,5aS,6S,6aS)-2-(3,5-Dichloro-4-hydroxyphenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (99).

[0188]

[0189] Step B (microwave). Compound 99 was obtained by crystallization as a light brown solid (423 mg, 1.21 mmol, 77%). IR (cm -1 )3234,1699,1489,1416,1184,1162,732; 1 H NMR (400MHz, CDCl3) δ7.13(s,2H),6.08(br,1H),5.86(dd,J=4.4,3.2Hz,2H),3.49(m,2H),3.13(dd,J=1.6,1.6Hz,2H),1.15(m,2H),0.36-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.5 (2C), 148.3, 128.1 (2C), 126.8 (2C), 124.7, 121.4 (2C), 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 17 H 14 Calculated value of Cl2NO3: 350.0 Found: 350.2 [M+H] + .

[0190] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-Mercaptophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (100).

[0191]

[0192] Step B (microwave). Compound 100 was obtained by crystallization as yellow crystals (80%). IR (cm -1 )3003,2942,1704,1496,1389,1193,1172,812,733; 1H NMR (400MHz, CDCl3) δ7.31(d,J=8.4Hz,2H),7.04(d,J=8.8Hz,2H),5.84(dd,J=4.8,3.2Hz, 2H),3.50(s,1H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.7 (2C), 132.1, 129.9 (2C), 129.6, 128.0 (2C), 127.2 (2C), 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 17 H 16 NO2S calculated value 298.1, found value 298.1 [M+H] + .

[0193] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-Aminophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (101).

[0194]

[0195] Step A. Purification by flash chromatography on silica gel (gradient elution, 0→75% EtOAc in hexanes) afforded 101 (97 mg, 0.35 mmol, 66%) as a yellow solid. IR (cm -1 )3336,3060,3029,2948,1699,1517,1404,1199,1165,916,734; 1 H NMR (400MHz, CDCl3) δ6.90(d,J=8.8Hz,2H),6.67(d,J=8.8Hz,2H),5.83(dd,J=4.8,3.2H z,2H),3.76(br,2H),3.47(m,2H),3.09(dd,J=2.0,1.6Hz,2H),1.13(m,1H),0.29(m,2H); 13 C NMR (100 MHz, CDCl3) δ 178.1 (2C), 146.7, 127.8 (2C), 127.5 (2C), 122.2, 115.1 (2C), 45.2 (2C), 33.8 (2C), 9.9 (2C), 4.6; ES-API MS: m / z for C 17 H 17N2O2 calculated value 281.1, found value 281.1 [M+H] + .

[0196] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-Hydroxyphenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (102a).

[0197]

[0198] Step B (microwave). Compound 102a was obtained by crystallization as a white solid (145 mg, 0.52 mmol, 98%). IR (cm -1 )3398,1772,1692,1517,1188; 1 H NMR (500MHz, DMSO-d6) δ9.71 (s, 1H), 6.87 (d, J = 8.5Hz, 2H), 6.79 (d, J = 9.0Hz, 2H), 5.81 (dd, J = 4. 0,4.0Hz,2H),3.27(m,2H),3.15(m,2H),1.17(m,2H),0.26(dd,J=13.0,7.0Hz,1H),0.08(m,1H); 13 C NMR (100 MHz, CDCl3) δ 177.8 (2C), 157.2, 128.0 (2C), 127.5 (2C), 123.3, 115.3 (2C), 44.7 (2C), 33.2 (2C), 9.5 (2C), 4.3; ES-API MS: m / z for C 17 H 16 NO3 calculated value 282.1, found value 282.1 [M+H] + .

[0199] (3aR,4R,4aR,5aS,6S,6aS)-2-(2-Fluoro-4-hydroxyphenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (102b).

[0200]

[0201] Step B. Purification by flash chromatography on silica gel (gradient elution, 0→55% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 102b (95%) as a brown solid. IR (cm -1 )3372,1698,1518,1401,1311,1186;1 H NMR (500MHz, CDCl3) δ6.86 (ddd, J=67.5, 8.0, 7.5Hz, 1H), 6.51 (m, 2H), 5.94 (br, 1H) ,5.86(m,2H),3.48(m,2H),3.17(d,J=18.0Hz,1H),1.15(m,2H),0.35-0.27(m,2H); 13 C NMR (100 MHz, DMSO-d6) δ 177.6 (2C), 159.5 (d, J = 63.0 Hz, 1C), 156.7, 135.5, 130.8, 128.0, 127.7, 112.1 (d, J = 20.8 Hz, 1C), 103.5 (d, J = 9.7 Hz, 1C), 45.7, 45.1, 33.7, 33.5, 9.8 (2C), 4.8; ES-API MS: m / z for C 17 H 15 FNO3 calculated value 300.1, found value 300.1 [M+H] + .

[0202] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-Hydroxypyridin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (103).

[0203]

[0204] Step B (microwave). Purification by flash chromatography on silica gel (gradient elution, 0→70% EtOAc in CH2Cl2) followed by recrystallization from CH2Cl2 / hexanes afforded 103 (1.64 g, 5.81 mmol, 72%) as an orange solid. IR (cm -1 )3290,3010,2360,1710,1580,1488,1288,1181; 1 H NMR (400MHz, CDCl3) δ7.93 (d, J=2.8Hz, 1H), 7.08 (dd, J=8.4, 2.8Hz, 1H), 6.81 (d, J=8.8Hz, 1H),5.68(dd,J=4.0,4.0Hz,2H),3.26(m,2H),2.96(m,2H),0.96(m,2H),0.15-0.06(m,2H); 13C NMR (100 MHz, CDCl3) δ 177.8 (2C), 154.1, 137.1, 136.7, 127.5 (2C), 124.8, 122.8, 45.2 (2C), 33.4 (2C), 9.5 (2C), 4.4; ES-API MS: m / z for C 16 H 15 N2O3 calculated value 283.1, found value 283.1 [M+H] + .

[0205] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-Hydroxypyrimidin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (104).

[0206]

[0207] Step A. Purification by flash chromatography on silica gel (gradient elution, 0→5% MeOH in CH2Cl2) followed by recrystallization from MeOH / CH2Cl2 / hexanes afforded 104 (1.08 g, 3.81 mmol, 48%) as an orange solid. IR (cm -1 )3008,1710,1567,1422,1291,1183,731; 1 H NMR (400MHz, CD3OD) δ8.38(s,1H),5.86(dd,J=4.8,3.6Hz,2H),3.39(m,2H),3.26(dd,J=1.6,1.6Hz,2H),1.21(m,2H),0.36-0.26(m,2H); 13 CNMR (100 MHz, CD3OD) δ 177.5 (2C), 152.2, 145.7 (2C), 144.3, 127.4 (2C), 45.6 (2C), 33.5 (2C), 9.3 (2C), 3.7; ES-API MS: m / z for C 15 H 14 N3O3 calculated value 284.1, found value 284.1 [M+H] + .

[0208] (3aR,4R,4aR,5aS,6S,6aS)-2-(3-chloro-4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (10).

[0209]

[0210] Method C. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 10 (112 mg, 0.24 mmol, 77%) as a white solid. IR (cm -1 )2957,1711,1601,1505,1233,1174,972,733; 1 H NMR (400MHz, CDCl3) δ8.01(d,J=8.8Hz,2H),7.23(d,J=2.4Hz,1H),7.00(dd,J=8.8,2.4Hz,1H),6.96(d,J=9.2Hz,2H),6.88(d,J=8.8Hz ,1H),5.84(dd,J=4.8,3.2Hz,2H),5.26(s,2H),3.88(s,3H),3.47(m,2H),3.11(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.4, 177.6 (2C), 164.4, 153.9, 131.0 (2C), 128.8, 128.0 (2C), 127.5, 126.1, 125.9, 123.7, 114.3 (2C), 114.0, 72.1, 55.8, 45.4 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 23 ClNO5 calculated value 264.1, found value 464.1 [M+H] + .

[0211] (3aR,4R,4aR,5aS,6S,6aS)-2-(3,5-dichloro-4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (11).

[0212]

[0213] Method C. Purification by flash chromatography on silica gel (gradient elution, 0→35% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 11 (121 mg, 0.24 mmol, 85%) as a white solid. IR (cm -1)3077,3010,2954,1714,1601,1471,1172,971,733; 1 H NMR (400MHz, CDCl3) δ7.98(d,J=9.2Hz,2H),7.22(s,2H),6.96(d,J=8.8Hz,2H),5.86(dd,J=4.4,3.6Hz, 2H),5.20(s,2H),3.88(s,3H),3.50(m,2H),3.14(dd,J=1.6,1.6Hz,2H),1.16(m,2H),0.37-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.2, 177.1 (2C), 164.2, 151.1, 130.7 (2C), 129.8 (2C), 128.9, 128.1 (2C), 127.6, 127.3 (2C), 114.2 (2C), 74.7, 55.8, 45.5 (2C), 34.1 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 22 Cl2NO5 calculated value 498.1, found value 498.1 [M+H] + .

[0214] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((2-(4-methoxyphenyl)-2-oxoethyl)thio)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (15).

[0215]

[0216] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→55% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 15 (88 mg, 0.20 mmol, 59%) as a light yellow solid. IR (cm -1 )2955,2358,1706,1669,1599,1497,1379,1261,1175,733; 1H NMR (400MHz, CDCl3) δ7.92(d,J=9.2Hz,2H),7.43(d,J=8.8Hz,2H),7.10(d,J=8.8Hz,2H),6.93(d,J=8.8Hz,2H),5.84(dd ,J=4.8,3.6Hz,2H),4.26(s,2H),3.87(s,3H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.6, 177.6 (2C), 164.1, 136.3, 131.2 (2C), 130.5, 130.3 (2C), 128.5, 128.0 (2C), 127.1 (2C), 114.1 (2C), 55.7, 45.5 (2C), 41.0, 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 23 NO4SNa calculated value 468.1, measured value 468.1 [M+Na] + .

[0217] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((2-(4-methoxyphenyl)-2-oxoethyl)amino)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (16).

[0218]

[0219] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 16 (32 mg, 0.07 mmol, 71%) as a white solid. IR (cm -1 )3394,2959,2924,2852,1702,1602,1524,1260,1172,818,735; 1H NMR (400MHz, CDCl3) δ7.99(d,J=8.8Hz,2H),6.98(d,J=8.8Hz,4H),6.70(d,J=8.8Hz,2H),5.85(dd,J=4.8,3.6Hz,2H ),5.12(br,1H),4.52(s,2H),3.89(s,3H),3.48(m,2H),3.10(dd,J=1.6,1.6Hz,2H),1.13(m,2H),0.33-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.0, 178.2 (2C), 164.1, 147.2, 130.1 (2C), 127.7 (3C), 127.5 (2C), 121.4, 114.1 (2C), 113.0 (2C), 55.6, 49.6, 45.2 (2C), 33.8 (2C), 9.9 (2C), 4.6; ES-API MS: m / z for C 26 H 25 N2O4 calculated value 429.2, found value 429.1 [M+H] + .

[0220] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-oxo-2-phenylethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (20).

[0221]

[0222] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 20 (32 mg, 0.08 mmol, 45%) as a light yellow solid. IR (cm -1 )1704,1513,1218,1184,734; 1 H NMR (400MHz, CDCl3) δ7.99(d,J=7.2Hz,2H),7.62(t,J=7.6Hz,1H),7.50(dd,J=8.0,7.6Hz,2H),7.09(d,J=8.8Hz,2H),6. 98(d,J=8.8Hz,2H),5.84(dd,J=4.0,4.0Hz,2H),5.25(s,2H),3.48(m,2H),3.12(m,2H),1.14(m,2H),0.34-0.25(m,2H); 13CNMR (100 MHz, CDCl3) δ 194.3, 178.0 (2C), 158.0, 134.6, 134.2, 129.1 (2C), 128.4 (2C), 128.00 (2C), 127.98 (2C), 125.6, 115.5 (2C), 71.2, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 21 NO4Na calculated value 422.1, measured value 422.1 [M+Na] + .

[0223] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(4-methoxyphenethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (29).

[0224]

[0225] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→35% EtOAc in hexanes) afforded 29 (77 mg, 0.19 mmol, 52%) as a white solid. IR (cm -1 )3007,2953,1705,1513,1245,1180,1031,826; 1 H NMR (400MHz, CDCl3) δ7.19(d,J=8.8Hz,2H),7.05(d,J=8.8Hz,2H),6.92(d,J=9.2Hz,2H),6.85(d,J=8.8Hz,2H),5.85(dd,J=4.8,3.6Hz, 2H),4.12(t,J=7.2Hz,2H),3.79(s,3H),3.48(m,2H),3.11(dd,J=1.6,1.6Hz,2H),3.03(t,J=7.2Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 178.1 (2C), 158.9, 158.5, 130.2, 130.1 (2C), 127.94 (2C), 127.86 (2C), 124.6, 115.2 (2C), 114.1 (2C), 69.3, 55.5, 45.4 (2C), 34.9, 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 26NO4 calculated value 416.2, found value 416.2 [M+H] + .

[0226] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (30a).

[0227]

[0228] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 30a (140 mg, 0.33 mmol, 92%) as a light yellow foam. IR (cm -1 )2954,2359,1704,1512,1392,1236,1185,1041,735; 1 H NMR (400MHz, CDCl3) δ7.52(d,J=8.0Hz,1H),7.47(dd,J=2.4,1.2Hz,1H),7.36(dd,J=8.4,7.6Hz,1H),7.13(dd,J=8.0,2.4Hz,1H),7.06(d,J=8.8Hz,2H) ,6.94(d,J=8.8Hz,2H),5.81(dd,J=4.8,3.6Hz,2H),5.21(s,2H),3.82(s,3H ),3.43(m,2H),3.08(dd,J=1.6,1.6Hz,2H),1.11(m,2H),0.31-0.22(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.9, 177.9 (2C), 160.0, 157.9, 135.7, 129.9, 127.9 (2C), 127.8 (2C), 125.5, 120.6, 120.5, 115.3 (2C), 112.4, 71.0, 55.6, 45.3 (2C), 33.9 (2C), 10.0 (2C), 4.7; ES-API MS: m / z for C 26 H 23 NO5Na calculated value 452.1, measured value 452.1 [M+Na] + .

[0229] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(2-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (31a).

[0230]

[0231] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→60% EtOAc in hexanes) gave a mixture of starting material phenol and product, which was further repurified by HPLC (reverse phase, MeCN / H2O / 0.1% TFA) to give product 31a as a light yellow foam (33 mg, 0.077 mmol, 43%). IR (cm -1 )3053,3008,2952,1706,1597,1512,1184,973,735; 1 H NMR (400MHz, CDCl3) δ7.93(dd,J=8.0,1.6Hz,1H),7.54(ddd,J=8.0,8.0,1.6Hz,1H),7.06(d,J=8.8Hz,2H),7.06(m,1H),7.01(d,J=8.4Hz,2 H),6.94(d,J=9.2Hz,2H),5.84(dd,J=4.8,3.6Hz,2H),5.22(s,2H),3.95(s,3H),3.47(m,2H),3.12(m,2H),1.13(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 195.1, 178.1 (2C), 159.3, 158.2, 135.0, 131.1, 127.7 (2C), 127.6 (2C), 124.9, 124.6, 121.2, 115.4 (2C), 111.5, 74.4, 55.7, 45.2 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 26 H 23 NO5Na calculated value 452.1, measured value 452.2 [M+Na] + .

[0232] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-ethoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (32a).

[0233]

[0234] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 32a (93 mg, 0.21 mmol, 73%) as a yellow solid. IR (cm -1 )1705,1600,1511,1393,1226,1172; 1 H NMR (400MHz, CDCl3) δ7.96(d,J=8.8Hz,2H),7.08(d,J=8.8Hz,2H),6.97(d,J=9.2Hz,2H),6.94(d,J=8.8Hz,2H),5.84(dd,J=4.4, 3.2Hz,2H),5.18(s,2H),4.11(q,J=6.8Hz,2H),3.48(m,2H),3.11(m,2H),1.45(t,J=7.2Hz,3H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.8, 178.0 (2C), 163.8, 158.1, 130.8 (2C), 128.1, 128.0 (4C), 127.5, 125.5, 116.1, 115.5 (2C), 114.7 (2C), 71.1, 64.1, 45.5 (2C), 34.0 (2C), 14.9, 10.1 (2C), 4.9; ES-API MS: m / z for C 27 H 25 NO5Na calculated value 466.2, measured value 466.1 [M+Na] + .

[0235] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-oxo-2-(4-(prop-2-yn-1-yloxy)phenyl)ethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (33a).

[0236]

[0237] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 33a (32 mg, 0.07 mmol, 86%) as a colorless solid. IR (cm -1)3298,3052,3011,2956,1772,1704,1600,1511,1223,1173,734; 1 H NMR (400MHz, CDCl3) δ7.98(d,J=8.8Hz,2H),7.06(d,J=8.8Hz,2H),7.03(d,J=9.2Hz,2H),6.96(d,J=9.2Hz,2H),5.82(dd,J=4.8,3.2Hz, 2H),5.17(s,2H),4.76(d,J=2.4Hz,2H),3.46(m,2H),3.10(dd,J=2.0,2.0Hz,2H),2.55(t,J=2.4Hz,1H),1.12(m,2H),0.32-0.23(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.9, 178.0 (2C), 162.1, 158.1, 130.8 (2C), 128.4, 128.1, 127.99 (2C), 127.98, 125.6, 115.5 (2C), 115.1 (2C), 76.6, 71.2, 56.1, 45.5 (2C), 34.0 (2C), 10.1 (2C); ES-APIMS: m / z for C 28 H 24 NO5 calculated value 454.2, found value 454.1 [M+H] + .

[0238] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-butoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (34a).

[0239]

[0240] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 34a (110 mg, 0.23 mmol, 56%) as colorless needles. IR (cm -1 )2957,2874,1705,1601,1512,1224,1173,965,830,734; 1H NMR (400MHz, CDCl3) δ7.96(d,J=8.8Hz,2H),7.08(d,J=9.2Hz,2H),6.97(d,J=9.2 Hz,2H),6.94(d,J=8.8Hz,2H),5.84(dd,J=4.4,3.2Hz,2H),5.18(s,2H),4.04(t, J=6.8Hz,2H),3.48(m,2H),3.11(dd,J=2.0,2.0Hz,2H),1.79(pent,J=7.2Hz,2H) ,1.50(hex,J=7.6Hz,2H),1.14(m,2H),0.98(t,J=7.2Hz,3H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.8, 178.0 (2C), 164.0, 158.1, 130.8 (2C), 128.0 (4C), 127.4, 125.5, 115.5 (2C), 114.7 (2C), 71.1, 68.2, 45.5 (2C), 34.0 (2C), 31.3, 19.4, 14.0, 10.1 (2C), 4.9; ES-API MS: m / z for C 29 H 29 NO5Na calculated value 494.2, measured value 494.1 [M+Na] + .

[0241] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-fluorophenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (35a).

[0242]

[0243] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 35a (150 mg, 0.36 mmol, 44%) as a colorless solid. IR (cm -1 )1704,1509,1225,1186,971,835,736; 1H NMR (400MHz, CDCl3) δ8.03(dd,J=8.8,5.2Hz,2H),7.17(dd,J=8.4,8.8Hz,2H),7.09(d,J=8.8Hz,2H),6.97(d,J=9.2Hz, 2H),5.84(dd,J=4.8,3.6Hz,2H),5.19(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.0, 178.0 (2C), 166.4 (d, J = 255 Hz, 1C), 157.9, 131.3 (d, J = 9.5 Hz, 2C), 131.1 (d, J = 3.0 Hz, 1C), 128.04 (2C), 127.97 (2C), 125.7, 116.3 (d, J = 21.8 Hz, 2C), 115.5 (2C), 71.3, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 20 FNO4Na calculated value 440.1, measured value 440.1 [M+Na] + .

[0244] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-chlorophenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (36a).

[0245]

[0246] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 36a (223 mg, 0.51 mmol, 51%) as a colorless solid. IR (cm -1 )1704,1515,1397,1219,1174,1087,966,736; 1H NMR (400MHz, CDCl3) δ7.94(d,J=8.4Hz,2H),7.47(d,J=8.4Hz,2H),7.09(d,J=8.8Hz,2H),6.96(d,J=9.2Hz,2H),5 .84(dd,J=4.8,3.6Hz,2H),5.18(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 CNMR (100 MHz, CDCl3) δ 193.2, 177.8 (2C), 157.6, 140.5, 132.7, 129.7 (2C), 129.2 (2C), 127.84 (2C), 127.76 (2C), 125.6, 115.2 (2C), 71.1, 45.2 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 25 H 21 ClNO4 calculated value 434.1, found value 434.1 [M+H] + .

[0247] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-bromophenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (37a).

[0248]

[0249] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 37a (35 mg, 0.073 mmol, 69%) as a light yellow solid. IR (cm -1 )1703,1585,1514,1399,1229,1194,984,822; 1 H NMR (400MHz, CDCl3) δ7.85(d,J=8.4Hz,2H),7.63(d,J=8.4Hz,2H),7.09(d,J=8.8Hz,2H),6.96(d,J=9.2Hz ,2H),5.84(dd,J=4.0,3.6Hz,2H),5.18(s,2H),3.47(m,2H),3.11(m,2H),1.14(m,2H),0.34-0.25(m,2H); 13C NMR (100 MHz, CDCl3) δ 193.6, 178.0 (2C), 157.8, 133.3, 132.4 (2C), 130.0 (2C), 129.5, 128.04 (2C), 127.96 (2C), 125.8, 115.4 (2C), 71.2, 45.4 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-APIMS: m / z for C 25 H 20 BrNO4Na calculated value 500.0, found value 500.0 [M+Na] + .

[0250] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-oxo-2-(p-tolyl)ethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (38a).

[0251]

[0252] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 38a (100 mg, 0.24 mmol, 65%) as a white solid. IR (cm -1 )3055,3008,2952,1705,1609,1514,1386,1310,1228,1183,964,818,734; 1 H NMR (400MHz, CDCl3) δ7.89(d,J=8.0Hz,2H),7.29(d,J=8.0Hz,2H),7.08(d,J=8.8Hz,2H),6.97(d,J=8.8Hz,2H),5.84(dd ,J=4.8,3.6Hz,2H),5.22(s,2H),3.48(m,2H),3.11(dd,J=1.6,1.2Hz,2H),2.43(s,3H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.9, 178.0 (2C), 158.1, 145.2, 132.2, 129.7 (2C), 128.5 (2C), 128.0 (4C), 125.6, 115.5 (2C), 71.2, 45.5 (2C), 34.0 (2C), 22.0, 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H24 NO4 calculated value 414.2, found value 414.1 [M+H] + .

[0253] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-oxo-2-(4-(trifluoromethyl)phenyl)ethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (39a).

[0254]

[0255] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 39a (25 mg, 0.053 mmol, 30%) as a colorless solid. IR (cm -1 )1704,1516,1326,1176,1123,1067,718; 1 H NMR (400MHz, CDCl3) δ8.10(d,J=8.4Hz,2H),7.77(d,J=8.0Hz,2H),7.10(d,J=8.8Hz,2H),6.97(d,J=9.2Hz,2H),5 .84(d,J=4.4,3.2Hz,2H),5.23(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 CNMR (100 MHz, CDCl3) δ 193.9, 178.0 (2C), 157.7, 129.0 (2C), 128.1 (2C), 128.0 (2C), 126.19, 126.15, 126.11, 126.07, 125.9, 115.5 (2C), 71.5, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 20 F3NO4Na calculated value 490.1, measured value 490.1 [M+Na] + .

[0256] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-oxo-2-(4-(trifluoromethoxy)phenyl)ethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (40a).

[0257]

[0258] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 40a (35 mg, 0.072 mmol, 51%) as a colorless solid. IR (cm -1 )1704,1515,1398,1260,1230,1194,1174,989,734; 1 H NMR (400MHz, CDCl3) δ8.06(d,J=8.8Hz,2H),7.32(d,J=8.4Hz,2H),7.09(d,J=8.8Hz,2H),6.97(d,J=8.8Hz,2H),5 .84(dd,J=4.8,3.6Hz,2H),5.19(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.2, 178.0 (2C), 157.8, 153.4, 132.8, 130.7 (2C), 128.08 (2C), 127.98 (2C), 125.8, 121.7, 120.8 (2C), 115.5 (2C), 71.3, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 20 F3NO5Na calculated value 506.1, found value 506.1 [M+Na] + .

[0259] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-hydroxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (41a).

[0260]

[0261] Acetate 42a (530 mg, 1.16 mmol) was dissolved in MeOH (4.0 mL) and KCO (320 mg, 2.31 mmol) was added. The resulting mixture was stirred at room temperature for 2 h. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→70% EtOAc in hexanes) to give phenol 41a (375 mg, 0.90 mmol, 78%) as a white solid. IR (cm -1 )3334,1704,1700,1602,1512,1392,1186,1173,735; 1 H NMR (400MHz, CDCl3) δ7.92(d,J=8.0Hz,2H),7.07(d,J=9.2Hz,2H),6.97(d,J=8.8Hz,2H),6.86(d,J=8.8Hz,2H),5 .84(dd,J=4.8,3.6Hz,2H),5.18(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.15(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.8, 178.2 (2C), 161.0, 158.2, 131.1 (2C), 128.03 (2C), 127.99 (2C), 127.7, 125.5, 115.9 (2C), 115.5 (2C), 71.0, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 22 NO5 calculated value 416.1, found value 416.1 [M+H] + .

[0262] 4-(2-(4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcycloprop[f]isoindol-2(1H)-yl)phenoxy)acetyl)phenyl acetate (42a).

[0263]

[0264] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 42a (45 mg, 0.098 mmol, 55%) as a white solid. IR (cm -1 )1760,1705,1600,1513,1193,1166;1 H NMR (400MHz, CDCl3) δ8.03(d,J=8.8Hz,2H),7.24(d,J=8.8Hz,2H),7.09(d,J=9.2Hz,2H),6.97(d,J=8.8Hz,2H),5.84(dd ,J=4.4,3.6Hz,2H),5.21(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.2Hz,2H),2.33(s,3H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.1, 178.0 (2C), 168.9, 157.9, 155.2, 132.2, 130.2 (2C), 128.03 (2C), 127.98 (2C), 125.7, 122.3 (2C), 115.5 (2C), 71.2, 45.5 (2C), 34.0 (2C), 21.4, 10.1 (2C), 4.9; ES-API MS: m / z for C 27 H 23 NO6Na calculated value 480.1, measured value 480.1 [M+Na] + .

[0265] 4-(2-(4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcycloprop[f]isoindol-2(1H)-yl)phenoxy)acetyl)phenyl methyl carbonate (43a).

[0266]

[0267] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→55% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 43a (47 mg, 0.099 mmol, 56%) as a white solid. IR (cm -1 )1770,1704,1508,1262,1216,734; 1H NMR (400MHz, CDCl3) δ8.04(d,J=8.8Hz,2H),7.33(d,J=8.8Hz,2H),7.09(d,J=9.2Hz,2H),6.97(d,J=8.8Hz,2H),5.84(dd ,J=4.8,3.6Hz,2H),5.21(s,2H),3.93(s,3H),3.48(m,2H),3.12(dd,J=1.6,2.0Hz,2H),1.15(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.1, 178.0 (2C), 157.9, 155.3, 153.6, 132.3, 130.3 (2C), 128.04 (2C), 127.98 (2C), 125.7, 121.7 (2C), 115.5 (2C), 71.2, 55.9, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 27 H 23 NO7Na calculated value 496.1, measured value 496.1 [M+Na] + .

[0268] Methyl (4-(2-(4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcyclopropyl[f]isoindol-2(1H)-yl)phenoxy)acetyl)phenyl)carbamate (44a).

[0269]

[0270] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→60% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 44a (55 mg, 0.11 mmol, 68%) as a light yellow solid. IR (cm -1 )3315,2954,1706,1683,1516,1393,1224,1190,1084,972; 1H NMR (400MHz, CDCl3 / CD3OD) δ7.83(d,J=8.8Hz,2H),7.46(d,J=8.4Hz,2H),6.95(d,J=9.2Hz,2H),6.88(d,J=9.2Hz,2 H),5.73(dd,J=4.4,3.6Hz,2H),5.15(s,2H),3.68(s,3H),3.35(m,2H),3.03(m,2H),1.05(m,2H),0.24-0.14(m,2H); 13 C NMR (100 MHz, CDCl3 / CD3OD) δ 193.1, 178.3 (2C), 157.9, 154.3, 144.2, 129.5 (2C), 128.6, 127.8 (2C), 127.7 (2C), 125.1, 117.7 (2C), 115.2 (2C), 70.5, 52.3, 45.2 (2C), 33.7 (2C), 9.8 (2C), 4.5; ES-API MS: m / z for C 27 H 25 N2O6 Calculated value 473.2, Found 473.2 [M+H] + .

[0271] Methyl 4-(2-(4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcycloprop[f]isoindol-2(1H)-yl)phenoxy)acetyl)benzoate (45a).

[0272]

[0273] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 45a (27 mg, 0.060 mmol, 33%) as a yellow solid. IR (cm -1 )1705,1513,1282,1218,1186,1110,734; 1 H NMR (400MHz, CDCl3) δ8.15(d,J=8.4Hz,2H),8.04(d,J=8.4Hz,2H),7.09(d,J=8.8Hz,2H),6.98(d,J=8.8Hz,2H), 5.84(dd,J=4.8,3.6Hz,2H),5.24(s,2H),3.96(s,3H),3.48(m,2H),3.12(m,2H),1.15(m,2H),0.34-0.25(m,2H);13 C NMR (100 MHz, CDCl3) δ 194.1, 178.0 (2C), 166.2, 157.8, 137.8, 134.8, 130.2 (2C), 128.4 (2C), 128.1 (2C), 128.0 (2C), 125.8, 115.5 (2C), 71.4, 52.8, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 27 H 23 NO6Na calculated value 480.1, measured value 480.1 [M+Na] + .

[0274] (4R,4aR,5aS,6S,6aS)-2-(5-(2-(6-methoxypyridin-3-yl)-2-oxoethoxy)pyridin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (46a).

[0275] Step C. Purification by silica gel flash chromatography (0-45% EtOAc / hexanes) and recrystallization from CH2Cl2 / hexanes afforded 46a (63 mg, 0.15 mmol, 42%) as a white solid. 1 H NMR (400MHz, CDCl3) δ8.10 (d, J = 8.4Hz, 2H), 7.89 (d, J = 8.4Hz, 2H), 7.81 (dd, J = 8. 4,1.2Hz,2H),7.63(dddd,J=7.2,7.6,1.2,1.2Hz,1H),7.51(dd,J=7.6,7.6Hz,2H ),7.10(d,J=9.2Hz,2H),6.99(d,J=8.8Hz,2H),5.84(dd,J=4.8,3.6Hz,2H),5.27 (s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.15(m,2H),0.35-0.25(m,2H); 13 C NMR (100MHz, CDCl3) δ196.0,194.2,178.0(2C),157.8,142.3,137.1,136.9,133.3,130.4(2C),130.3(2C),1 28.7(2C),128.4(2C),128.1(2C),128.0(2C),125.8,115.5(2C),71.5,45.5(2C),34.0(2C),10.1(2C),4.9.

[0276] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(naphthalen-2-yl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (47a).

[0277]

[0278] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 47a (33 mg, 0.076 mmol, 43%) as a white solid. IR (cm -1 )3410,2964,1710,1261,1029,801; 1 H NMR (400MHz, CDCl3) δ7.97(dd,J=2.0,1.6Hz,1H),7.86(ddd,J=7.6,1.6,1.2Hz,1H),7.59(ddd,J=8.0,1.2,1.2Hz,1H),7.44(dd,J=8.0,8.0Hz,1H),7.09( d,J=9.2Hz,2H),6.97(d,J=9.2Hz,2H),5.84(dd,J=4.8,3.2Hz,2H),5.20(s,2 H),3.48(m,2H),3.12(dd,J=1.6,1.2Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.3, 178.0 (2C), 157.8, 136.1, 135.4, 134.1, 130.4, 128.6, 128.1 (2C), 128.0 (2C), 126.5, 125.8, 115.5 (2C), 71.3, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 21 ClNO4 calculated value 434.1, found value 434.1 [M+H] + .

[0279] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(2,3-dichlorophenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (48a).

[0280]

[0281] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 48a (140 mg, 0.30 mmol, 42%) as a white solid. IR (cm -1 )3056,3010,2955,1705,1512,1187,735; 1 H NMR (400MHz, CDCl3) δ7.60(dd,J=8.0,2.0Hz,1H),7.37(dd,J=7.6,1.6Hz,1H),7.30(dd,J=8.0,7.6Hz,1H),7.09(d,J=9.2Hz,2H),6.9 2(d,J=8.8Hz,2H),5.84(dd,J=4.8,3.6Hz,2H),5.10(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 197.8, 178.0 (2C), 157.7, 138.9, 134.4, 133.3, 128.06, 128.04 (2C), 127.99 (2C), 127.7, 125.9, 115.5 (2C), 73.0, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 20 Cl2NO4 calculated value 468.1, found value 468.1 [M+H] + .

[0282] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(2,5-dichlorophenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (49a).

[0283]

[0284] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 49a (37 mg, 0.080 mmol, 17%) as a white solid. IR (cm -1 )2962,1710,1261,1048,803; 1H NMR (400MHz, CDCl3) δ7.55(d,J=2.4Hz,1H),7.42(d,J=8.8,2.4Hz,1H),7.38(d,J=8.8Hz,1H),7.10(d,J=8.8Hz,2H),6.93(d,J= 8.8Hz,2H),5.84(dd,J=4.8,3.6Hz,2H),5.13(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 196.6, 178.0 (2C), 157.7, 137.6, 133.6, 132.9, 132.9, 131.9, 130.1, 128.1 (2C), 128.0 (2C), 125.9, 115.5 (2C), 73.1, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 20 Cl2NO4 calculated value 468.1, found value 468.1 [M+H] + .

[0285] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(2,6-dichlorophenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (50a).

[0286]

[0287] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 50a (155 mg, 0.33 mmol, 47%) as a white solid. IR (cm -1 )2961,1709,1513,1184,1045,785; 1 H NMR (400MHz, CDCl3) δ7.35(m,3H),7.10(d,J=8.8Hz,2H),6.99(d,J=9.2Hz,2H),5.85(dd,J=4.8,3 .6Hz,2H),5.02(s,2H),3.48(m,2H),3.12(dd,J=1.6,2.0Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13C NMR (100 MHz, CDCl3) δ 197.2, 178.0 (2C), 157.8, 136.7, 131.7, 131.6, 128.4 (2C), 128.0 (5C), 125.9, 115.7 (2C), 73.1, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 20 Cl2NO4 calculated value 468.1, found value 468.1 [M+H] + .

[0288] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-oxo-2-(2,3,4-trichlorophenyl)ethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (51a).

[0289]

[0290] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 51a (133 mg, 0.26 mmol, 41%) as a white solid. IR (cm -1 )2964,1709,1512,1261,1026,801; 1 H NMR (400MHz, CDCl3) δ7.48(d,J=8.4Hz,1H),7.33(d,J=8.4Hz,1H),7.09(d,J=9.2Hz,2H),6.90(d,J=9.2Hz,2H),5 .84(dd,J=4.8,3.6Hz,2H),5.08(s,2H),3.48(m,2H),3.12(dd,J=1.6,2.0Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 197.1, 178.0 (2C), 157.6, 137.8, 136.9, 133.3, 131.8, 129.1, 128.08 (2C), 127.99 (2C), 127.7, 126.0, 115.4 (2C), 73.0, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 19 Cl3NO4 calculated value 502.0, found value 502.1 [M+H]+ .

[0291] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3,5-dichlorophenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (52a).

[0292]

[0293] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 52a (142 mg, 0.30 mmol, 43%) as a white solid. IR (cm -1 )2960,1708,1513,1214,801; 1 H NMR (400MHz, CDCl3) δ7.85(d,J=2.0Hz,2H),7.60(t,J=2.0Hz,1H),7.10(d,J=9.2Hz,2H),6.97(d,J=9.2Hz,2H),5 .84(dd,J=4.8,3.2Hz,2H),5.17(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.4, 178.0 (2C), 157.6, 136.9, 136.1, 133.9, 128.1 (2C), 128.0 (3C), 127.0 (2C), 126.0, 115.5 (2C), 71.3, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 20 Cl2NO4 calculated value 468.1, found value 468.1 [M+H] + .

[0294] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3,5-dichlorophenyl)-2-oxoethoxy)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (52b).

[0295]

[0296] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→30% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 52b (133 mg, 0.27 mmol, 32%) as a light yellow solid. IR (cm -1 )3085,3005,1716,1516,1221,1171,734; 1 H NMR(400MHz, CDCl3)δ7.83(d,J=1.6Hz,2H),7.61(m,1H),7.02(br,1H),6.76(s,1H),6.74(s,1H),5 .85(dd,J=4.0,3.6Hz,2H),5.16(s,2H),3.47(m,2H),3.15(m,2H),1.14(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.7 (2C), 177.2, 159.4 (d, J = 4.8 Hz, 1C), 158.1 (d, J = 237.1 Hz, 1C), 136.7, 136.2 (2C), 134.0 (2C), 130.1 (d, J = 11.3 Hz, 1C), 127.9 (d, J = 15.4 Hz, 1C), 126.9 (2C), 113.7 (d, J = 13.9 Hz, 1C), 111.0 (d, J = 17.3 Hz, 1C), 104.0 (d, J = 26.0 Hz, 1C), 71.3, 45.9, 45.6, 33.9 (2C), 10.0 (2C), 4.9; ES-API MS: m / z for C 25 H 19 Cl2FNO4 calculated value 486.1, found value 486.1 [M+H] + .

[0297] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3-chloro-5-fluorophenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (53a).

[0298]

[0299] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→35% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 53a (37 mg, 0.082 mmol, 22%) as a white solid. IR (cm -1)2964,1711,1261,1029,802; 1 H NMR (400MHz, CDCl3) δ7.77(dd,J=1.2,1.2Hz,1H),7.59(ddd,J=8.4,1.2,1.2Hz,1H),7.34(ddd,J=8.0,2.0,2.0Hz,1H),7.10(d,J=9.2Hz,2H ),6.97(d,J=8.8Hz,2H),5.84(dd,J=4.8,3.2Hz,2H),5.16(s,2H),3.48(m,2H),3.12(dd,J=1.6,2.0Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.4, 178.0 (2C), 162.9 (d, J = 251.2 Hz, 1C), 157.6, 137.2 (d, J = 7.1 Hz, 1C), 136.3 (d, J = 9.5 Hz, 1C), 128.1 (2C), 128.0 (2C), 126.0, 124.7 (d, J = 3.3 Hz, 1C), 121.7 (d, J = 24.7 Hz, 1C), 115.5 (2C), 114.0 (d, J = 22.6 Hz, 1C), 71.4, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 20 FClNO4 calculated value 452.1, found value 452.1 [M+H] + .

[0300] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3-chloro-5-(trifluoromethyl)phenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (54a).

[0301]

[0302] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→35% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 54a (133 mg, 0.26 mmol, 28%) as a white solid. IR (cm -1 )3079,2963,2857,1710,1513,1326,1175,803; 1H NMR (400MHz, CDCl3) δ8.14(m,2H),7.84(m,1H),7.10(d,J=8.8Hz,2H),6.96(d,J=8.8Hz,2H),5.84(dd,J =4.8,3.6Hz,2H),5.19(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.6, 178.0 (2C), 157.5, 136.4 (d, J = 37.6 Hz, 1C), 133.2 (d, J = 33.6 Hz, 1C), 132.0, 130.6 (d, J = 3.6 Hz, 1C), 128.2 (2C), 128.0 (2C), 126.1, 123.7 (d, J = 4.6 Hz, 1C), 122.9 (d, J = 271.7 Hz, 1C), 115.4 (2C), 71.5, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-APIMS: m / z for C 26 H 20 F3ClNO4 calculated value 502.1, found value 502.1 [M+H] + .

[0303] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3,4-dimethoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (55a).

[0304]

[0305] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→60% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 55a (100 mg, 0.22 mmol, 61%) as a white solid. IR (cm -1 )3056,3008,2957,1705,1513,1267,1169,1021,734; 1H NMR (400MHz, CDCl3) δ7.63(dd,J=8.4,2.0Hz,1H),7.55(d,J=2.0Hz,1H),7.08(d,J=8.8Hz,2H),6.97(d,J=8.8Hz,2H),6.90(d,J=8.4Hz,1H), 5.84(dd,J=4.8,3.2Hz,2H),5.20(s,2H),3.96(s,3H),3.94(s,3H),3.48(m,2H),3.11(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.9, 178.0 (2C), 158.1, 154.2, 149.5, 127.99 (2C), 127.97 (2C), 127.8, 125.6, 123.1, 115.5 (2C), 110.5, 110.3, 71.1, 56.4, 56.3, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-APIMS: m / z for C 27 H 26 NO6 calculated value 460.2, found value 460.1 [M+H] + .

[0306] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3,4-dimethoxyphenyl)-2-oxoethoxy)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (55b).

[0307]

[0308] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→65% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 55b (130 mg, 0.27 mmol, 82%) as an orange solid. IR (cm -1 )3009,2957,1715,1595,1515,1265,1165,733; 1H NMR (400MHz, CDCl3) δ7.61(dd,J=8.4,2.0Hz,1H),7.53(d,J=2.0Hz,1H),7.00(m,1H),6.91(d,J=8.4Hz,1H),6.77(m,1H),6.75(m, 1H),5.84(dd,J=4.0,4.0Hz,2H),5.20(s,2H),3.97(s,3H),3.94(s,3H),3.47(m,2H),3.15(m,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100MHz, CDCl3) δ192.2, 177.3 (2C), 159.9 (d, J=10.4Hz, 1C), 158.1 (d, J= 251.8Hz,1C),154.3,149.6,129.9(d,J=19.1Hz,1C),127.9(d,J=19.5Hz,1C), 127.6,123.1,113.2(d,J=13.0Hz,1C),111.1(d,J=10.3Hz,1C),110.5,110.4 ,110.2,104.0,71.1,56.4,56.3,45.9,45.6,33.9(2C),10.1(2C),4.9; ES-API MS: m / z for C 27 H 25 FNO6 calculated value 478.2, found value 478.1 [M+H] + .

[0309] 3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(Benzo[d][1,3]dioxol-5-yl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (56a).

[0310]

[0311] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 56a (95 mg, 0.21 mmol, 61%) as a white solid. IR (cm -1 )3009,2955,2909,1704,1512,1252,1037,734; 1H NMR (400MHz, CDCl3) δ7.60(dd,J=8.4,1.6Hz,1H),7.46(d,J=1.6Hz,1H),7.08(d,J=8.8Hz,2H),6.97(d,J=8.8Hz,2H),6.87(d,J=8.4Hz ,1H),6.06(s,2H),5.84(dd,J=4.8,3.6Hz,2H),5.15(s,2H),3.48(m,2H),3.11(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.4, 178.0 (2C), 158.0, 152.7, 148.6, 129.4, 127.99 (2C), 127.98 (2C), 125.6, 124.9, 115.5 (2C), 108.4, 108.2, 102.2, 71.1, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 22 NO6 calculated value 444.1, found value 444.1 [M+H] + .

[0312] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(Benzo[d][1,3]dioxol-5-yl)-2-oxoethoxy)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (56b).

[0313]

[0314] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 56b (88 mg, 0.19 mmol, 58%) as a cream solid. IR (cm -1 )3010,2955,2913,1715,1516,1254,1170,1038,733; 1H NMR (400MHz, CDCl3) δ7.58(dd,J=8.4,1.6Hz,1H),7.45(d,J=1.6Hz,1H),7.01(m,1H),6.88(d,J=8.0Hz,1H),6.76(m,1H),6. 74(m,1H),6.07(s,2H),5.84(dd,J=4.0,4.0Hz,2H),5.15(s,2H),3.47(m,2H),3.14(m,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.7, 177.3 (2C), 159.8 (d, J = 10.3 Hz, 1C), 158.1 (d, J = 251.7 Hz, 1C), 152.8, 148.7, 129.9 (d, J = 18.3 Hz, 1C), 129.1, 127.9 (d, J = 12.3 Hz, 1C), 124.8, 113.3, 113.2, 111.1 (d, J = 18.2 Hz, 1C), 108.4, 108.2, 104.0 (d, J = 28.3 Hz, 1C), 102.3, 71.2, 45.9, 45.6, 33.9 (2C), 10.0 (2C), 4.9; ES-API MS: m / z for C 26 H 21 FNO6 calculated value 462.1, found value 462.1 [M+H] + .

[0315] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3,4-dimethylphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (57a).

[0316]

[0317] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 57a (124 mg, 0.29 mmol, 82%) as a white solid. IR (cm -1 )3052,3008,2953,2921,1705,1513,1236,1185,735; 1H NMR (400MHz, CDCl3) δ7.75(d,J=1.6Hz,1H),7.71(dd,J=8.0,1.6Hz,1H),7.24(d,J=8.0Hz,1H),7.08(d,J=9.2Hz,2H),6.97(d,J=9.2Hz,2H), 5.84(dd,J=4.8,3.2Hz,2H),5.22(s,2H),3.48(m,2H),3.11(dd,J=1.6,1.6Hz,2H),2.33(s,3H),2.32(s,3H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 194.0, 178.0 (2C), 158.1, 144.0, 137.6, 132.5, 130.2, 129.4, 127.97 (2C), 127.96 (2C), 126.1, 125.5, 115.5 (2C), 71.1, 45.5 (2C), 34.0 (2C), 20.4, 20.0, 10.1 (2C), 4.9; ES-API MS: m / z for C 27 H 26 NO4 calculated value 428.2, found value 428.1 [M+H] + .

[0318] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3,4-dimethylphenyl)-2-oxoethoxy)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (57b).

[0319]

[0320] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 57b (105 mg, 0.24 mmol, 71%) as a light pink solid. IR (cm -1 )3014,2952,1715,1516,1166,1124,733; 1H NMR (400MHz, CDCl3) δ7.74(m,1H),7.70(dd,J=7.6,1.2Hz,1H),7.25(d,J=8.0Hz,1H),7.00(m,1H),6.77(m,1H),6.74(m,1H), 5.84(dd,J=4.0,3.6Hz,2H),5.22(s,2H),3.47(m,2H),3.14(m,2H),2.33(s,3H),2.32(s,3H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100MHz, CDCl3) δ193.3, 177.3 (2C), 159.9 (d, J=10.3Hz, 1C), 158.1 (d, J= 251.5Hz,1C),144.1,137.6,132.3,130.3,129.9(d,J=14.0Hz,1C),129.3,127 .9(d,J=16.3Hz,1C),126.0,113.2,113.1,111.2(d,J=17.2Hz,1C),103.9(d, J=33.2Hz,1C),71.1,45.9,45.6,33.9(2C),20.4,20.0,10.0(2C),4.9; ES-API MS: m / z for C 27 H 25 FNO4 calculated value 446.2, found value 446.1 [M+H] + .

[0321] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3,4-dichlorophenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (58a).

[0322]

[0323] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 58a (33 mg, 0.070 mmol, 40%) as a white solid. IR (cm -1 )2955,1704,1512,1393,1212,1185,995,734; 1H NMR (400MHz, CDCl3) δ8.08(d,J=2.0Hz,1H),7.82(dd,J=8.4,2.0Hz,1H),7.58(d,J=8.4Hz,1H),7.10(d,J=8.8Hz,2H),6.96(d,J =9.2Hz,2H),5.84(dd,J=4.8,3.6Hz,2H),5.16(s,2H),3.48(m,2H),3.12(dd,J=2.0,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.7, 178.0 (2C), 157.7, 138.9, 134.1, 133.9, 131.2, 130.6, 128.1 (2C), 128.0 (2C), 127.6, 125.9, 115.5 (2C), 71.4, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 19 Cl2NO4Na Calculated value 490.1 Measured value 490.5 [M+Na] + .

[0324] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3,4-dichlorophenyl)-2-oxoethoxy)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (58b).

[0325]

[0326] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→35% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 58b (122 mg, 0.25 mmol, 75%) as a light brown solid. IR (cm -1 )2956,1716,1517,1396,1170,735; 1H NMR (400MHz, CDCl3) δ8.07(d,J=2.0Hz,1H),7.81(dd,J=8.4,2.0Hz,1H),7.59(d,J=8.4Hz,1H),7.02(br,1H),6.76(m, 1H),6.74(m,1H),5.85(dd,J=4.0,3.6Hz,2H),5.16(s,2H),3.47(m,2H),3.15(m,2H),1.13(m,2H),0.35-0.26(m,2H); 13 C NMR (100MHz, CDCl3) δ192.0 (2C), 177.3, 159.4 (d, J = 2.4Hz, 1C), 158.1 (d, J = 24 4.4Hz, 1C), 139.1, 134.0, 133.9, 131.3, 130.5, 130.1, (d, J = 15.8Hz, 1C), 127. 9(d,J=15.1Hz,1C),127.5(2C),113.6(d,J=13.8Hz,1C),111.0(d,J=11.6Hz,1 C),104.0(d,J=25.8Hz,1C),71.3,45.9,45.6,33.9(2C),10.0(2C),4.9; ES-API MS: m / z for C 25 H 19 Calculated value of Cl2FNO4: 486.1 Found: 486.1 [M+H] + .

[0327] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3-fluoro-4-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (59a).

[0328]

[0329] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→65% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 59a (32 mg, 0.072 mmol, 62%) as a light yellow solid. IR (cm -1 )3010,2956,1705,1613,1513,1434,1392,1286,1227,1186,734; 1H NMR (400MHz, CDCl3) δ7.80(ddd,J=8.4,2.0,0.8Hz,1H),7.75(dd,J=11.6,2.0Hz,1H),7.08(d,J=9.2Hz,2H),7.01(dd,J=8.4,8.4Hz,1H),6.97(d ,J=9.2Hz,2H),5.84(dd,J=4.8,3.6Hz,2H),5.15(s,2H),3.97(s,3H),3 .48(m,2H),3.11(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 CNMR (100 MHz, CDCl3) δ 192.3 (d, J = 1.8, 1C), 178.0 (2C), 157.9, 153.1 (d, J = 75.5, 1C), 152.0 (d, J = 182.6, 1C), 128.03 (2C), 127.98 (2C), 127.8 (d, J = 5.2, 1C), 126.0 (d, J = 3.3, 1C), 125.7, 116.2 (d, J = 19.1, 1C), 115.5 (2C), 112.7 (d, J = 1.9, 1C), 71.2, 56.6, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 22 FNO5Na calculated value 470.1, found value 470.1 [M+Na] + .

[0330] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3-chloro-4-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (60a).

[0331]

[0332] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→65% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 60a (57 mg, 0.12 mmol, 57%) as a light yellow solid. IR (cm -1 )1705,1594,1508,1207,1061,734; 1H NMR (400MHz, CDCl3) δ8.04(d,J=2.0Hz,1H),7.92(dd,J=8.8,2.4Hz,1H),7.08(d,J=9.2Hz,2H),6.99(d,J=8.8Hz,1H),6.97(d,J=8.8Hz ,2H),5.84(dd,J=4.4,3.6Hz,2H),5.15(s,2H),3.98(s,3H),3.48(m,2H),3.11(dd,J=1.2,1.6Hz,2H),1.13(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.2, 178.0 (2C), 159.6, 157.9, 130.8, 129.1, 128.1, 128.03 (2C), 127.97 (2C), 125.7, 123.4, 115.5 (2C), 111.7, 71.2, 56.7, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 22 ClNO5Na calculated value 486.1, found value 486.1 [M+Na] + .

[0333] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3-chloro-4-methoxyphenyl)-2-oxoethoxy)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (60b).

[0334]

[0335] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→55% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 60b (94 mg, 0.20 mmol, 58%) as a white solid. IR (cm -1 )3057,3011,2954,1716,1595,1516,1260,1169,1061,1008,735; 1H NMR(400MHz, CDCl3)δ8.02(d,J=2.4Hz,1H),7.91(dd,J=8.8,2.0Hz,1H),7.01(br,1H),6.99(d,J=8.8Hz,1H),6.77(m,1H),6 .74(m,1H),5.84(dd,J=4.0,4.0Hz,2H),5.15(s,2H),3.99(s,3H),3.47(m,2H),3.14(m,2H),1.13(m,2H),0.34-0.26(m,2H); 13 C NMR (100MHz, CDCl3) δ191.5 (2C), 177.3, 159.7, 159.6 (d, J = 10.3Hz, 1C), 158.1 ( d,J=251.7Hz,1C),130.7,130.0(d,J=12.8Hz,1C),129.1,127.9(d,J=14.3Hz,1C ),127.9,123.5,113.4(d,J=13.8Hz,1C),111.7(2C),111.1(d,J=14.3Hz,1C),1 03.9(d,J=23.2Hz,1C),71.2,56.7,45.9,45.6,33.9(2C),10.0(2C),4.9; ES-API MS: m / z for C 26 H 22 ClFNO5 calculated value 482.1, found value 482.1 [M+H] + .

[0336] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-chloro-3-methylphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (61a).

[0337]

[0338] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 61a (42 mg, 0.094 mmol, 53%) as a white solid. IR (cm -1 )2964,1710,1261,1046,801; 1H NMR (400MHz, CDCl3) δ7.86(d,J=1.6Hz,1H),7.75(dd,J=8.4,2.0Hz,1H),7.45(d,J=8.4Hz,1H),7.09(d,J=8.8Hz,2H),6.97(d,J=9.2Hz ,2H),5.84(dd,J=4.8,3.6Hz,2H),5.19(s,2H),3.48(m,2H),3.12(dd,J=2.0,1.6Hz,2H),2.44(s,3H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.6, 178.0 (2C), 157.9, 140.9, 137.3, 133.0, 130.8, 129.8, 128.04 (2C), 127.98 (2C), 127.2, 125.7, 115.5 (2C), 71.2, 45.5 (2C), 34.0 (2C), 20.4, 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 23 ClNO4 calculated value 482.1, found value 448.1 [M+H] + .

[0339] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-chloro-3-methylphenyl)-2-oxoethoxy)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (61b).

[0340]

[0341] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→35% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 61b (80 mg, 0.17 mmol, 52%) as a light pink solid. IR (cm -1 )1716,1519,1508,1168,1049,734; 1H NMR (400MHz, CDCl3) δ7.84(d,J=1.6Hz,1H),7.75(ddd,J=8.4,8.4,1.6Hz,1H),7.46(d,J=8.4Hz,1H),7.02(br,1H),6.77 (m,1H),6.74(m,1H),5.85(dd,J=4.0,3.6Hz,2H),5.19(s,2H),3.47(m,2H),3.15(m,2H),1.13(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.9 (2C), 177.3, 159.6 (d, J = 10.3 Hz, 1C), 158.1 (d, J = 251.9 Hz, 1C), 143.3, 141.1, 137.4, 132.8, 131.6, 130.8, 129.9 (2C), 129.1, 127.9 (d, J = 13.8 Hz, 1C), 127.1, 126.5, 71.2, 45.9, 45.6, 33.9 (2C), 20.4, 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 22 ClFNO4 calculated value 466.1, found value 466.1 [M+H] + .

[0342] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3-chloro-4-fluorophenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (62a).

[0343]

[0344] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 62a (25 mg, 0.055 mmol, 31%) as a white solid. IR (cm -1 )2964,1710,1261,1045,802; 1H NMR (400MHz, CDCl3) δ8.09(dd,J=7.2,2.0Hz,1H),7.92(ddd,J=8.4,4.4,2.0Hz,1H),7.26(dd,J=8.4,8.4Hz,1H),7.09(d,J=9.2Hz,2H),6 .96(d,J=9.2Hz,2H),5.84(dd,J=4.8,3.6Hz,2H),5.15(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.3, 178.0 (2C), 161.7 (d, J = 257.1 Hz, 1C), 157.7, 131.8 (d, J = 3.7 Hz, 1C), 131.6 (d, J = 1.2 Hz, 1C), 129.1 (d, J = 8.6 Hz, 1C), 128.1 (2C), 128.0 (2C), 125.9, 122.5 (d, J = 18.3 Hz, 1C), 117.4 (d, J = 21.8 Hz, 1C), 115.5 (2C), 71.3, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 20 FClNO4 calculated value 452.1, found value 452.1 [M+H] + .

[0345] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3-chloro-4-fluorophenyl)-2-oxoethoxy)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (62b).

[0346]

[0347] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 62b (86 mg, 0.18 mmol, 55%) as a light yellow solid. IR (cm -1 )2956,1716,1517,1253,1170,734; 1H NMR (400MHz, CDCl3) δ8.07(d,J=8.8Hz,1H),7.90(m,1H),7.27(dd,J=8.4,8.4Hz,1H),7.02(br,1H),6.76(m,1H), 6.74(m,1H),5.84(dd,J=4.0,3.6Hz,2H),5.16(s,2H),3.47(m,2H),3.15(m,2H),1.14(m,2H),0.34-0.27(m,2H); 13 C NMR (100MHz, CDCl3) δ191.3 (2C), 177.0, 161.6 (d, J = 257.6Hz, 1C), 159.2 (d, J = 11.0Hz, 1C), 157 .9(d,J=252.0Hz,1C),131.4(2C),131.4(d,J=1.2Hz,1C),129.9(d,J=20.6Hz,1C),128.8(d,J=8 .6Hz,1C),127.7(d,J=12.9Hz,1C),122.5,117.2(d,J=21.7Hz,1C),113.4(d,J=13.8Hz,1C),110.8(d,J=12.6Hz,1C),103.7(d,J=24.7Hz,1C),71.1,45.7,45.4,33.7(2C),9.8(2C),4.7; ES-API MS: m / z for C 25 H 19 ClF2NO4 calculated value 470.1, found value 470.1 [M+H] + .

[0348] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-chloro-3-(trifluoromethyl)phenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (63a).

[0349]

[0350] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→35% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 63a (97 mg, 0.19 mmol, 34%) as a white solid. IR (cm -1 )2961,1710,1604,1513,1261,1180,1036,803,735; 1H NMR (400MHz, CDCl3) δ8.33(d,J=2.0Hz,1H),8.10(d,J=8.4,2.0Hz,1H),7.64(d,J=8.4Hz,1H),7.09(d,J=8.8Hz,2H),6.95(d,J= 8.8Hz,2H),5.83(dd,J=4.8,3.6Hz,2H),5.17(s,2H),3.47(m,2H),3.11(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.9, 178.0 (2C), 157.5, 138.5 (d, J = 1.6 Hz, 1C), 133.0, 132.8, 123.4, 129.4 (d, J = 3.2 Hz, 1C), 128.1 (2C), 128.0 (2C), 127.9, 127.8 (d, J = 3.0 Hz, 1C), 126.0, 122.5 (d, J = 272.4 Hz, 1C), 115.4 (2C), 71.5, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 20 F3ClNO4 calculated value 502.1, found value 502.1 [M+H] + .

[0351] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-chloro-3-(trifluoromethyl)phenyl)-2-oxoethoxy)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (63b).

[0352]

[0353] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 63b (31 mg, 0.06 mmol, 18%) as a light yellow solid. IR (cm -1 )2957,1716,1519,1319,1172,1141,1114,1036,735; 1H NMR (400MHz, CDCl3) δ8.32(d,J=1.6Hz,1H),8.09(dd,J=8.4,2.0Hz,1H),7.66(d,J=8.4Hz,1H),7.03(br,1H),6.77(m, 1H),6.74(m,1H),5.85(dd,J=4.0,3.6Hz,2H),5.18(s,2H),3.47(m,2H),3.15(m,2H),1.14(m,2H),0.35-0.26(m,2H); 13 C NMR (100MHz, CDCl3) δ191.9 (2C), 177.0, 157.9 (d, J = 252.3Hz, 1C), 159.0 (d, J = 10.3H z,1C),138.5(d,J=1.7Hz,1C),132.6,132.5,132.3,129.9(d,J=12.1Hz,1C),129.4( d, J = 32.0 Hz, 1C), 127.8, 127.6 (q, J = 5.2 Hz, 1C), 123.6, 120.9, 113.6 (d, J = 13.8 Hz, 1C), 110.7, 103.7 (d, J = 23.0 Hz, 1C), 71.3, 45.7, 45.4, 33.7 (2C), 9.8 (2C), 4.7; ES-API MS: m / z for C 26 H 19 Calculated value of ClF4NO4: 520.1, found: 520.1[M+H] + .

[0354] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(naphthalen-2-yl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (64a).

[0355]

[0356] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 64a (116 mg, 0.25 mmol, 73%) as a light yellow solid. IR (cm -1 )3055,3010,2954,1704,1512,1188,734; 1H NMR (400MHz, CDCl3) δ8.53(s,1H),8.03(dd,J=8.8,1.6Hz,1H),7.98(d,J=8.0Hz,1H ),7.93(d,J=8.8Hz,1H),7.90(d,J=8.0Hz,1H),7.64(dd,J=6.8,7.2Hz,1H),7.58(d d,J=7.2,7.2Hz,1H),7.10(d,J=8.8Hz,2H),7.02(d,J=8.8Hz,2H),5.84(dd,J=4.4, 3.6Hz,1H),5.38(s,2H),3.48(m,2H),3.12(m,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 194.2, 178.0 (2C), 158.1, 136.2, 132.6, 132.0, 130.3, 129.9, 129.2, 129.0, 128.1, 128.03 (2C), 127.98 (2C), 127.3, 125.6, 123.8, 115.6 (2C), 71.3, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 29 H 34 NO4 calculated value 460.2, found value 460.1 [M+H] + .

[0357] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(4-methoxyphenyl)-2-oxoethoxy)pyridin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (71).

[0358]

[0359] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→70% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 71 (86 mg, 0.20 mmol, 57%) as a light yellow solid. IR (cm -1 )3063,2940,2840,1710,1601,1401,1232,1173,972; 1H NMR (400MHz, CDCl3) δ8.31(d,J=2.8Hz,1H),7.96(d,J=8.8Hz,2H),7.32(dd,J=8.8,3.2Hz,1H),7.06(d,J=8.8Hz,1H),6.97(d,J=8.8Hz ,2H),5.88(dd,J=4.8,3.2Hz,2H),5.27(s,2H),3.89(s,3H),3.49(m,2H),3.14(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.8, 177.5 (2C), 164.5, 154.6, 139.6, 137.9, 130.7 (2C), 128.0 (2C), 127.3, 123.8, 122.7, 114.4 (2C), 71.1, 55.8, 45.8 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 25 H 23 N2O5 calculated value 431.2, found value 431.2 [M+H] + .

[0360] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(6-methoxypyridin-3-yl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (72).

[0361]

[0362] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 72 (40 mg, 0.093 mmol, 26%) as a white solid. IR (cm -1 )3053,3009,2953,1705,1602,1513,1377,1298,1226,1186,735; 1H NMR (400MHz, CDCl3) δ8.86(d,J=2.4Hz,1H),8.16(dd,J=8.4,2.4Hz,1H),7.08(d,J=8.8Hz,2H),6.96(d,J=9.2Hz,2H),6.81(d, J=8.8Hz,1H),5.83(dd,J=3.6,4.4Hz,2H),5.13(s,2H),4.01(s,3H),3.47(m,2H),3.11(m,2H),1.13(m,2H),0.33-0.24(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.8, 178.0 (2C), 167.3, 157.8, 149.6, 138.6, 128.0 (2C), 127.9 (2C), 125.7, 124.7, 115.4 (2C), 111.8, 71.3, 54.4, 45.4 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 25 H 23 N2O5 calculated value 431.2, found value 431.2 [M+H] + .

[0363] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(6-methoxypyridin-3-yl)-2-oxoethoxy)pyridin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (73).

[0364]

[0365] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→70% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 73 (63 mg, 0.15 mmol, 42%) as a light yellow solid. IR (cm -1 )3054,3011,2953,1710,1602,1490,1376,1298,1231,969,730; 1H NMR (400MHz, CDCl3) δ8.84(d,J=2.4Hz,1H),8.32(d,J=2.8Hz,1H),8.15(dd,J=8.8,2.4Hz,1H),7.33(dd,J=8.8,2.8Hz,1H),7.07(d,J=8.8Hz,1H),6. 83(d,J=8.8Hz,1H),5.88(dd,J=4.4,3.6Hz,2H),5.22(s,2H),4.02(s,3H) ,3.49(m,2H),3.14(dd,J=1.6,1.6Hz,2H),1.13(m,2H),0.35-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.6, 177.5 (2C), 167.5, 154.4, 149.5, 139.8, 138.5, 137.8, 128.1 (2C), 124.4, 123.9, 122.8, 112.0, 71.2, 54.5, 45.8 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 24 H 22 N3O5 calculated value 432.2, found value 432.2 [M+H] + .

[0366] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(4-methoxyphenyl)-2-oxoethoxy)pyrimidin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (74).

[0367]

[0368] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→25% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 74 (58 mg, 0.13 mmol, 79%) as a white solid. IR (cm -1 )2975,1714,1601,1428,1237,1177,973,730; 1H NMR (400MHz, CDCl3) δ8.45(s,2H),7.93(d,J=8.8Hz,2H),6.98(d,J=8.8Hz,2H),5.91(dd,J=4.4,3.6Hz, 2H),5.36(s,2H),3.90(s,3H),3.50(m,2H),3.17(dd,J=1.6,1.6Hz,2H),1.13(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 190.7, 176.8 (2C), 164.7, 152.1, 146.7, 146.1 (2C), 130.6 (2C), 128.0 (2C), 126.9, 114.5 (2C), 71.0, 55.8, 46.0 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 24 H 22 N3O5 calculated value 432.2, found value 432.2 [M+H] + .

[0369] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(3-chloro-4-methoxyphenyl)-2-oxoethoxy)pyridin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (75).

[0370]

[0371] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in CH2Cl2) followed by recrystallization from CH2Cl2 / hexanes afforded 75 (86 mg, 0.18 mmol, 52%) as a yellow solid. IR (cm -1 )3055,3009,2954,2846,1775,1711,1595,1478,1400,1263,1209,1061,730; 1H NMR (400MHz, CDCl3) δ8.31(d,J=3.2Hz,1H),8.02(d,J=2.4Hz,1H),7.90(dd,J=8.4,2.4Hz,1H),7.32(dd,J=8.8,2.8Hz,1H),7.07(d,J=8.8Hz,1H),7. 00(d,J=8.8Hz,1H),5.88(dd,J=4.8,3.6Hz,2H),5.24(s,2H),3.99(s,3H) ,3.49(m,2H),3.14(dd,J=2.0,2.0Hz,2H),1.14(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.1, 177.5 (2C), 159.8, 154.4, 139.7, 137.8, 130.7, 129.0, 128.1 (2C), 127.8, 123.8, 123.6, 122.8, 111.8, 71.1, 56.7, 45.8 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 25 H 22 Calculated value of ClN2O5: 465.1, found: 465.1 [M+H] + .

[0372] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(4-chloro-3-methylphenyl)-2-oxoethoxy)pyridin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (76).

[0373]

[0374] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→65% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 76 (94 mg, 0.21 mmol, 59%) as a yellow solid. IR (cm -1 )2956,1710,1580,1478,1400,1291,1232,1186,1050,912,730; 1H NMR (400MHz, CDCl3) δ8.31(d,J=2.8Hz,1H),7.83(d,J=1.6Hz,1H),7.72(dd,J=8.4,2.4Hz,1H),7.47(d,J=8.4Hz,1H),7.31(dd,J=8.8,3.2Hz,1H),7. 07(d,J=8.8Hz,1H),5.88(dd,J=4.8,3.6Hz,2H),5.27(s,2H),3.49(m,2H) ,3.14(dd,J=2.0,2.0Hz,2H),2.45(s,3H),1.14(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 192.6, 177.5 (2C), 154.4, 141.2, 139.8, 137.8, 132.7, 131.7, 130.7, 129.9, 128.1 (2C), 127.0, 123.9, 122.8, 71.2, 45.8 (2C), 33.9 (2C), 20.4, 10.1 (2C), 5.0; ES-API MS: m / z for C 25 H 22 Calculated value of ClN2O4: 449.1, found: 449.1 [M+H] + .

[0375] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(3-chloro-4-fluorophenyl)-2-oxoethoxy)pyridin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (77).

[0376]

[0377] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→65% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 77 (71 mg, 0.16 mmol, 71%) as a yellow solid. IR (cm -1 )3055,3010,2957,1776,1714,1581,1488,1402,1292,1253,1206,1059,831,731; 1H NMR (400MHz, CDCl3) δ8.31(d,J=2.8Hz,1H),8.07(dd,J=6.8,2.4Hz,1H),7.90(ddd,J=8.4,4.4,2.0Hz,1H),7.32(dd,J=8.8,3.2Hz,1H),7.28(d,J=9 .2Hz,1H),7.08(d,J=8.8Hz,1H),5.88(dd,J=4.8,3.6Hz,2H),5.25(s,2H) ,3.49(m,2H),3.15(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.2, 177.5 (2C), 161.9 (d, J = 257.9 Hz, 1C), 154.2, 140.0, 137.7, 131.5, 131.4 (d, J = 3.8 Hz, 1C), 128.9 (d, J = 8.6 Hz, 1C), 128.1 (2C), 123.4 (d, J = 110.2 Hz, 1C), 122.7 (d, J = 18.7 Hz, 1C), 117.5 (d, J = 21.7 Hz, 1C), 71.2, 45.8 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 24 H 19 Calculated value of ClFN2O4: 453.1, found: 453.1 [M+H] + .

[0378] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(4-chloro-3-(trifluoromethyl)phenyl)-2-oxoethoxy)pyridin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (78).

[0379]

[0380] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→65% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 78 (37 mg, 0.074 mmol, 44%) as a white solid. IR (cm -1 )3055,3012,2957,1774,1710,1604,1478,1401,1320,1209,1181,1141,730; 1H NMR (400MHz, CDCl3) δ8.32(d,J=3.6Hz,1H),8.31(d,J=2.4Hz,1H),8.09(dd,J=8.4,2.0Hz,1H),7.68(d,J=8.4Hz,1H),7.33(dd,J=8.8,2.8Hz, 1H),7.10(d,J=8.8Hz,1H),5.88(dd,J=4.8,3.6Hz,2H),5.28(s,2H),3.50(m,2H),3.15(dd,J=2.0,2.0Hz,2H),1.14(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.7, 177.5 (2C), 154.1, 140.1, 138.9 (q, J = 2.0 Hz, 1C), 137.7, 132.7, 132.6, 129.9, 129.5, 128.1 (2C), 127.8 (q, J = 5.0 Hz, 1C), 124.0, 122.9, 122.4 (q, J = 273.0 Hz, 1C), 71.5, 45.8 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 25 H 19 Calculated value of ClF3N2O4: 503.1, found: 503.1 [M+H] + .

[0381] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(3,4-dichlorophenyl)-2-oxoethoxy)pyridin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (79).

[0382]

[0383] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→60% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 79 (133 mg, 0.28 mmol, 80%) as a yellow solid. IR (cm -1 )3060,3038,3010,2956,1776,1709,1582,1479,1399,1211,1187,1032,730; 1H NMR (400MHz, CDCl3) δ8.31(d,J=2.8Hz,1H),8.06(d,J=2.0Hz,1H),7.80(dd,J=8.4,2.0Hz,1H),7.60(d,J=8.4Hz,1H),7.32(dd,J=8.8,3.2Hz, 1H),7.08(d,J=8.8Hz,1H),5.88(dd,J=4.8,3.6Hz,2H),5.25(s,2H),3.49(m,2H),3.15(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.6, 177.5 (2C), 154.2, 140.0, 139.2, 137.7, 134.1, 133.7, 131.4, 130.4, 128.1 (2C), 127.4, 123.9, 122.8, 71.3, 45.8 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 24 H 19 Cl2N2O4 calculated value 469.1, found value 469.1 [M+H] + .

[0384] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(3,5-dichlorophenyl)-2-oxoethoxy)pyridin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (80).

[0385]

[0386] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→55% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 80 (44 mg, 0.094 mmol, 27%) as a white solid. IR (cm -1 )3076,2955,1710,1568,1488,1399,1291,1216,1187,730; 1H NMR (400MHz, CDCl3) δ8.31(d,J=2.8Hz,1H),7.83(d,J=2.0Hz,2H),7.62(t,J=2.0Hz,1H),7.32(dd,J=8.4,3.2Hz,1H),7.09(d,J =8.8Hz,1H),5.88(dd,J=4.4,3.6Hz,2H),5.25(s,2H),3.49(m,2H),3.15(dd,J=1.5,1.5Hz,2H),1.14(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.3, 177.5 (2C), 154.1, 140.0, 137.8, 136.5, 136.3, 134.1 (2C), 128.1 (2C), 126.8 (2C), 123.9, 122.8, 71.3, 45.8 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 24 H 19 Cl2N2O4 calculated value 469.1, found value 469.1 [M+H] + .

[0387] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(5,6-dichloropyridin-3-yl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (81).

[0388]

[0389] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 81 (50 mg, 0.10 mmol, 30%) as a light yellow solid. IR (cm -1 )3009,1704,1512,1187,734; 1 H NMR (400MHz, CDCl3) δ8.91(d,J=2.0Hz,1H),8.35(d,J=2.0Hz,1H),7.11(d,J=9.2Hz,2H),6.96(d,J=9.2Hz,2H),5 .84(dd,J=4.8,3.6Hz,2H),5.13(s,2H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.25(m,2H);13 C NMR (100 MHz, CDCl3) δ 192.6, 178.0 (2C), 157.3, 154.2, 147.4, 138.5, 137.7, 131.8, 130.1, 128.3 (2C), 128.0 (2C), 126.2, 115.4 (2C), 71.8, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 24 H 19 Cl2N2O4 calculated value 469.1, found value 469.1 [M+H] + .

[0390] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(3-chloro-4-methoxyphenyl)-2-oxoethoxy)pyrimidin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (82).

[0391]

[0392] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→30% EtOAc in CH2Cl2) followed by recrystallization from CH2Cl2 / hexanes afforded 82 (106 mg, 0.23 mmol, 65%) as a light yellow solid. IR (cm -1 )2952,1714,1595,1428,1263,1212,731; 1 H NMR (400MHz, CDCl3) δ8.49(s,2H),7.99(d,J=2.0Hz,1H),7.88(dd,J=8.8,2.0Hz,1H),7.02(d,J=8.8Hz,1H),5.91(dd, J=4.4,3.6Hz,2H),5.33(s,2H),4.08(s,3H),3.50(m,2H),3.17(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.36-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 190.0, 176.8 (2C), 160.0, 152.0, 146.9, 146.1 (2C), 130.5, 128.9, 128.0 (2C), 127.4, 123.8, 111.9, 110.2, 71.0, 56.8, 46.0 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 24H 21 Calculated value of ClN3O5: 466.1, found: 466.1[M+H] + .

[0393] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(4-chloro-3-methylphenyl)-2-oxoethoxy)pyrimidin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (83).

[0394]

[0395] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→15% EtOAc in CH2Cl2) followed by recrystallization from CH2Cl2 / hexanes afforded 83 (67 mg, 0.15 mmol, 43%) as a white solid. IR (cm -1 )2953,1714,1428,1290,1236,1050,731; 1 H NMR (400MHz, CDCl3) δ8.48(s,2H),7.82(d,J=2.0Hz,1H),7.70(dd,J=8.4,2.4Hz,1H),7.49(d,J=8.4Hz,1H),5.91(dd, J=4.8,3.6Hz,2H),5.36(s,2H),3.50(m,2H),3.17(dd,J=1.6,1.6Hz,2H),2.46(s,3H),1.14(m,2H),0.36-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.5, 176.8 (2C), 152.0, 146.9, 146.1 (2C), 141.5, 137.7, 132.3, 130.6, 130.1, 128.0 (2C), 126.8, 71.1, 46.0 (2C), 33.9 (2C), 20.4, 10.1 (2C), 5.0; ES-API MS: m / z for C 24 H 21 Calculated value of ClN3O4: 450.1, found: 450.1 [M+H] + .

[0396] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(3-chloro-4-fluorophenyl)-2-oxoethoxy)pyrimidin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (84).

[0397]

[0398] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→25% EtOAc in CH2Cl2) followed by recrystallization from CH2Cl2 / hexanes afforded 84 (35 mg, 0.077 mmol, 45%) as a light yellow solid. IR (cm -1 )3010,1713,1428,1251,1187,731; 1 H NMR (400MHz, CDCl3) δ8.49(s,2H),8.05(dd,J=6.8,2.0Hz,1H),7.89(ddd,J=8.8,4.4,2.0Hz,1H),7.30(dd,J=8.4,8.4Hz ,1H),5.91(dd,J=4.8,3.6Hz,2H),5.35(s,2H),3.50(m,2H),3.17(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.35-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 190.1, 176.8 (2C), 162.0 (d, J = 258.3 Hz, 1C), 151.8, 147.1, 146.1 (2C), 131.4 (d, J = 1.3 Hz, 1C), 131.1 (d, J = 3.7 Hz, 1C), 128.8 (d, J = 8.6 Hz, 1C), 128.1 (2C), 117.7 (d, J = 21.7 Hz, 1C), 110.2, 71.1, 46.0 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 23 H 17 ClFN3O4 calculated value 453.1 measured value 453.1 [M] + .

[0399] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(3,4-dichlorophenyl)-2-oxoethoxy)pyrimidin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (85).

[0400]

[0401] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→35% EtOAc in CH2Cl2) followed by recrystallization from CH2Cl2 / hexanes afforded 85 (65 mg, 0.14 mmol, 40%) as a white solid. IR (cm -1 )3009,1712,1429,1397,1216,1184,732; 1 H NMR (400MHz, CDCl3) δ8.49(s,2H),8.04(d,J=2.0Hz,1H),7.78(dd,J=8.4,2.0Hz,1H),7.62(d,J=8.4Hz,1H),5.9 2(dd,J=4.8,3.2Hz,2H),5.35(s,2H),3.50(m,2H),3.18(dd,J=1.6,2.0Hz,2H),1.14(m,2H),0.35-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 190.5, 176.8 (2C), 151.8, 147.1, 146.1 (2C), 139.6, 134.2, 133.4, 131.5, 130.3, 128.1 (2C), 127.2, 71.1, 46.0 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 23 H 18 Cl2N3O4 calculated value 470.1, found value 470.1 [M+H] + .

[0402] (3aR,4R,4aR,5aS,6S,6aS)-2-(5-(2-(3,5-dichlorophenyl)-2-oxoethoxy)pyrimidin-2-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (86).

[0403]

[0404] Step C. Purification by flash chromatography on silica gel (gradient elution, 0→20% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 86 (33 mg, 0.07 mmol, 42%) as a light yellow solid. IR (cm -1 )3008,1712,1428,1220,731; 1H NMR (400MHz, CDCl3) δ8.50 (s, 2H), 7.81 (d, J = 2.0Hz, 2H), 7.65 (t, J = 2.0Hz, 1H), 5.92 (dd, J = 4.8, 3 .6Hz,2H),5.34(s,2H),3.51(m,2H),3.18(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.36-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 190.3, 176.8 (2C), 151.8, 147.1, 146.1 (2C), 136.5, 136.2, 134.4 (2C), 128.1 (2C), 126.7 (2C), 71.1, 46.0 (2C), 33.9 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 23 H 18 Cl2N3O4 calculated value 470.1, found value 470.1 [M+H] + .

[0405] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(Hydroxymethyl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (105).

[0406]

[0407] Step A. Purification by flash chromatography on silica gel (gradient elution, 0→80% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 105 (80 mg, 0.27 mmol, 52%) as a light yellow solid. IR (cm -1 )3504,3052,3033,2963,2886,1696,1508,1395,1191,1175,1006; 1 H NMR (400MHz, CDCl3) δ7.42(d,J=8.4Hz,2H),7.15(d,J=8.4Hz,2H),5.86(dd,J=4.8,3.6Hz,2H), 4.69(s,2H),3.49(m,2H),3.13(dd,J=2.0,1.6Hz,2H),1.79(br,1H),1.15(m,2H),0.30(m,2H); 13C NMR (100 MHz, CDCl3) δ 177.7 (2C), 141.4, 131.1, 127.8 (2C), 127.4 (2C), 126.6 (2C), 64.8, 45.3 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 18 H 18 NO3 calculated value 296.1, found value 296.1 [M+H] + .

[0408] 4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-Dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcycloprop[f]isoindol-2(1H)-yl)-3-fluorobenzonitrile (106).

[0409]

[0410] Step B (microwave). Crystallization from CH2Cl2 / Et2O / hexanes gave benzonitrile 106 as yellow crystals (2.55 g, 8.27 mmol, 80%). IR (cm -1 )3067,2989,2360,1722,1421,1266,1254,1169,1045,899,728; 1 H NMR (400MHz, CDCl3) δ7.52(d,J=8.4Hz,1H),7.49(d,J=8.8Hz,1H),7.30(br,1H),5. 87(dd,J=4.4,4.0Hz,2H),3.50(m,2H),3.21(m,2H),1.16(m,2H),0.38-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.9 (2C), 176.2 (2C), 157.2 (d, J = 256.6 Hz, 1C), 130.7, 128.7 (d, J = 4.2 Hz, 1C), 128.0 (2C), 124.7 (d, J = 13.3 Hz, 1C), 120.8 (d, J = 23.1 Hz, 1C), 117.0 (d, J = 2.6 Hz, 1C), 114.5 (d, J = 9.1 Hz, 1C), 46.00, 45.96, 34.0 (2C), 10.0 (2C), 5.0; ES-API MS: m / z for C 18 H 14 FN2O2 calculated value 309.1, found value 309.1 [M+H] + .

[0411] 4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-Dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcycloprop[f]isoindol-2(1H)-yl)benzaldehyde (107).

[0412]

[0413] To a solution of benzyl alcohol 105 (100 mg, 0.34 mmol) in anhydrous toluene was added MnO2 (176 mg, 2.03 mmol) and the mixture was heated to 70 °C and stirred for 5 hours. The reaction was cooled to room temperature, filtered through a Celite pad, and the residue was washed with CH2Cl2. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→45% EtOAc in hexanes) to give benzaldehyde 105 (98 mg, 0.33 mmol, 98%) as an amorphous yellow solid. IR (cm -1 )2999,2954,2824,1773,1704,1700,1601,1386,1186,734,720; 1 H NMR (400MHz, CDCl3) δ10.01(s,1H),7.93(d,J=8.0Hz,2H),7.40(d,J=8.0Hz,2H),5.86(br,2H),3.50(br,2H),3.16(s,2H),1.16(br,2H),0.32(m,2H); 13 C NMR (100 MHz, CDCl3) δ 191.1, 177.0 (2C), 137.0, 135.7, 130.2 (2C), 127.9 (2C), 126.9 (2C), 45.4 (2C), 33.9 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 18 H 16 NO3 calculated value 294.1, found value 294.1 [M+H] + .

[0414] 4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-Dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcycloprop[f]isoindol-2(1H)-yl)-3-fluorobenzaldehyde (108).

[0415]

[0416] In a pressure sealed tube, benzonitrile 106 (2.46 g, 7.98 mmol) was dissolved in formic acid (16 mL, 80% in deionized water). Raney nickel (Raney-Ni) (1.2 g) was added to the reaction in one go. The resulting mixture was sealed and refluxed at 110 ° C for 4 hours, cooled to room temperature, filtered through a Celite pad and washed with EtOAc. The eluent was neutralized with saturated NaHCO3, washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→45% EtOAc in hexane) and then recrystallized in CH2Cl2 / hexane to obtain the desired benzaldehyde 108 as a colorless solid (1.95 g, 6.26 mmol, 79%). IR (cm -1 )3069,3045,3015,1780,1698,1588,1512,1391,1253,1181,816,736,718; 1 H NMR (400MHz, CDCl3) δ9.99(s,1H),7.74(d,J=8.8Hz,1H),7.69(d,J=9.6Hz,1H),7.35(br,1 H),5.88(dd,J=4.4,3.6Hz,2H),3.50(m,2H),3.21(m,2H),1.17(m,2H),0.37-0.28(m,2H); 13 C NMR (100 MHz, CDCl3) δ 190.1 (d, J = 1.8 Hz, 1C), 176.4 (2C), 157.9 (d, J = 255.7 Hz, 1C), 138.4 (d, J = 6.0 Hz, 1C), 130.4, 128.0 (2C), 126.2 (d, J = 3.5 Hz, 1C), 125.4 (d, J = 14.0 Hz, 1C), 116.9 (d, J = 20.3 Hz, 1C), 46.04, 45.99, 34.0 (2C), 10.1 (2C), 5.0; ES-API MS: m / z for C 18 H 15 FNO3 calculated value 312.1, found value 312.1 [M+H] + .

[0417] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(Aminomethyl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (109).

[0418]

[0419] Step 1. (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(azidomethyl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcycloprop[f]isoindole-1,3(2H,3aH)-dione. To a solution of benzyl alcohol 105 (50 mg, 0.17 mmol) and PPh3 (67 mg, 0.25 mmol) in anhydrous DMF (0.85 mL) at 0°C was added tetrabromomethane (84 mg, 0.25 mmol). The resulting yellow mixture was stirred at 0°C until TLC showed complete conversion of the alcohol, then NaN3 (33 mg, 0.51 mmol) was added in one portion and stirring was continued for another 2 h at room temperature. The reaction was diluted with H2O (5 mL) and extracted with EtOAc (3×5 mL). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→35% EtOAc in hexanes) to give the azide (49 mg, 0.15 mmol, 91%) as a white solid. IR (cm -1 )3078,3052,3009,2979,2949,2099,1706,1516,1387,1190,734,714; 1 H NMR (400MHz, CDCl3) δ7.36(d,J=8.0Hz,2H),7.19(d,J=8.4Hz,2H),5.84(dd,J=4.4,3.2Hz,2H) ,4.33(s,2H),3.47(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.15-1.11(m,2H),0.33-0.24(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.5 (2C), 135.9, 131.7, 128.6 (2C), 127.8 (2C), 126.8 (2C), 54.2, 45.3 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 18 H 18 N2O2K calculated value 333.1, measured value 333.1 [M+K-N2] + .

[0420] Step 2: To a solution of the above azide (step 1) (1.31 g, 4.09 mmol) in THF / H2O (vol:vol / 10:1, 40 mL) was added PMe3 solution (8.2 mL, 1.0 M in THF). The resulting mixture was stirred at room temperature for 1 hour. The reaction was then diluted with H2O (20 mL) and extracted with EtOAc (3×20 mL). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was recrystallized in CH2Cl2 / hexanes to afford amine 109 (1.15 g, 0.15 mmol, 91%) as a yellow solid. IR (cm -1 )3461,3372,3008,2954,2867,1772,1713,1704,1514,1385,1186,734; 1 H NMR(400MHz, CDCl3) δ7.36(d,J=8.4Hz,2H),7.10(d,J=8.4Hz,2H),5.83(dd,J=4 .8,3.2Hz,2H),3.85(m,2H),1.76(m,2H),1.14-1.10(m,2H),0.32-0.23(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.7 (2C), 130.4, 128.8, 127.8 (2C), 127.7 (2C), 126.6 (2C), 126.4, 45.3 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 18 H 19 N2O2Na calculated value 318.1, found value 318.1 [M+Na+H] + .

[0421] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((E)-3-(4-methoxyphenyl)-3-oxoprop-1-en-1-yl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (110).

[0422]

[0423] Step E. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) afforded enone 110 (70 mg, 0.16 mmol, 85%) as an amorphous yellow solid. IR (cm -1)3465,3010,2957,2841,1707,1660,1604,1512,1378,1335,1261,1222,1172,1022,819,733; 1 H NMR (400MHz, CDCl3) δ8.03(d,J=9.2Hz,2H),7.77(d,J=15.6Hz,1H),7.69(d,J=8.4Hz,2H),7.52(d,J=15.6Hz,1H),7.26(d,J=8.4Hz,2H), 6.99(d,J=8.8Hz,2H),5.87(dd,J=4.8,3.2Hz,2H),3.89(s,3H),3.51(m,2H),3.16(dd,J=1.6,1.6Hz,2H),1.16(m,2H),0.36-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 188.4, 177.4 (2C), 163.5, 142.6, 135.2, 133.3, 130.9, 130.8 (2C), 128.8 (2C), 127.8 (2C), 126.8 (2C), 122.9, 113.9 (2C), 55.5, 45.4 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 27 H 24 NO4 calculated value 426.2, found value 426.0 [M+H] + .

[0424] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((E)-3-(3-chloro-4-methoxyphenyl)-3-oxoprop-1-en-1-yl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (111a).

[0425]

[0426] Step E. Purification by silica gel flash chromatography (gradient elution, 0→50% EtOAc in hexanes) afforded enone 111a (263 mg, 0.55 mmol, 86%) as a light yellow solid. IR (cm -1 )3056,3010,2954,1722,1662,1607,1593,1392,1276,734; 1H NMR (400MHz, CDCl3) δ8.04(d,J=1.6Hz,1H),7.91(dd,J=8.4,2.0Hz,1H),7.66(d,J=15.6Hz,1H),7.44(d,J=15.6Hz,1H),7.40(m,1H),7.38(m, 1H),7.12(br,1H),6.96(d,J=8.8Hz,1H),5.84(dd,J=4.0,3.6Hz,2H),3.93(s,3H),3.45(m,2H),3.16(m,2H),1.12(m,2H),0.32-0.24(m,2H); 13 C NMR (100 MHz, CDCl3) δ 186.9, 176.6 (2C), 158.8, 157.4 (d, J = 252.9 Hz, 1C), 142.0 (d, J = 2.0 Hz, 1C), 137.7 (d, J = 7.6 Hz, 1C), 131.1, 130.8, 129.6, 129.1, 127.7, 124.7 (d, J = 2.1 Hz, 1C), 123.3, 123.0, 121.2 (d, J = 13.9 Hz, 1C), 115.6 (d, J = 20.2 Hz, 1C), 111.4, 56.4, 45.7 (2C), 33.8 (2C), 9.9 (2C), 4.8; ES-API MS: m / z for C 27 H 22 ClFNO4 calculated value 478.1, found value 478.1 [M+H] + .

[0427] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((E)-3-(4-chloro-3-methylphenyl)-3-oxoprop-1-en-1-yl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (111b).

[0428]

[0429] Step E. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) afforded enone 111b (228 mg, 0.49 mmol, 77%) as a light yellow solid. IR (cm -1 )3055,2947,3007,1716,1666,1609,1517,1391,1181,1050,734; 1H NMR (400MHz, CDCl3) δ7.88(d,J=1.2Hz,1H),7.77(dd,J=8.4,2.4Hz,1H),7.72(d,J=15.6Hz,1H),7.48(d,J=2.4Hz,1H),7.46-7 .43(m,3H),7.18(br,1H),5.88(dd,J=4.0,4.0Hz,2H),3.50(m,2H),3.20(m,2H),2.46(s,3H),1.16(m,2H),0.37-0.28(m,2H); 13 C NMR (100 MHz, CDCl3) δ 189.0, 176.8 (2C), 157.7 (d, J = 253.2 Hz, 1C), 142.6 (d, J = 2.3 Hz, 1C), 140.0, 137.9 (d, J = 7.5 Hz, 1C), 137.0, 136.3, 131.2, 129.9, 129.7, 128.0, 127.5, 124.9 (d, J = 2.9 Hz, 1C), 124.0, 123.8, 121.5 (d, J = 13.9 Hz, 1C), 115.9 (d, J = 20.0 Hz, 1C), 46.0, 45.9, 34.0 (2C), 20.4, 10.1 (2C), 5.0; ES-API MS: m / z for C 27 H 22 Calculated value of ClFNO3: 462.1, found: 462.1[M+H] + .

[0430] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((E)-3-(3-chloro-4-fluorophenyl)-3-oxoprop-1-en-1-yl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (111c).

[0431]

[0432] Step E. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) afforded enone 111c (189 mg, 0.41 mmol, 63%) as a light yellow solid. IR (cm -1 )3008,1713,1609,1516,1391,1250,1205,814,740; 1H NMR (400MHz, CDCl3) δ8.08(dd,J=7.2,2.0Hz,1H),7.92(m,1H),7.73(d,J=15.6Hz,1H),7.44(m,1H),7.42(m,1H),7.42(d,J=15.6Hz,1 H),7.26(dd,J=8.8,8.4Hz,1H),7.17(br,1H),5.87(dd,J=3.6,4.0Hz,2H),3.49(m,2H),3.19(m,2H),1.15(m,2H),0.36-0.27(m,2H); 13 C NMR (100MHz, CDCl3) δ187.2,176.7(2C),161.2(d,J=255.9Hz,1C),157.6(d,J=253.2Hz,1C),143.3(d, J=2.4Hz,1C),137.6(d,J=7.5Hz,1C),135.0(d,J=3.6Hz,1C),131.6(d,J=0.8Hz,1C),130.0,129.1(d, ES-API MS: m / z for C 26 H 19 Calculated value of ClF2NO3: 466.1, found: 466.1[M+H] + .

[0433] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((E)-3-(4-chloro-3-(trifluoromethyl)phenyl)-3-oxoprop-1-en-1-yl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (111d).

[0434]

[0435] Step E. Purification by silica gel flash chromatography (gradient elution, 0→40% EtOAc in hexanes) afforded enone 111d (192 mg, 0.37 mmol, 58%) as a light yellow solid. IR (cm -1)3009,2997,1713,1610,1515,1311,1206,1175,737; 1 H NMR (400MHz, CDCl3) δ8.31(s,1H),8.10(dd,J=8.4,1.2Hz,1H),7.75(d,J=15.6Hz,1H),7.64(d,J=8.4Hz,1H),7.44(d,J=15.6Hz,1 H),7.45(m,1H),7.43(m,1H),7.18(br,1H),5.87(dd,J=4.0,3.6Hz,2H),3.49(m,2H),3.19(m,2H),1.15(m,2H),0.37-0.27(m,2H); 13 C NMR (100MHz, CDCl3) δ187.4, 176.7 (2C), 157.6 (d, J = 253.4Hz, 1C), 143.8 (d, J = 2.2Hz, 1C), 137. 5(q,J=1.5Hz,1C),137.4(d,J=7.5Hz,1C),136.3,132.7,132.2,130.0,129.2(q,J=31.8Hz,1C) ,127.9,127.8(q,J=5.3Hz,1C),125.1(d,J=2.0Hz,1C),122.9(2C),122.6(q,J=272.3Hz,1C),121.9(d,J=13.8Hz,1C),116.0(d,J=20.3Hz,1C),45.93,45.87,34.0(2C),10.0(2C),5.0; ES-API MS: m / z for C 27 H 19 ClF4NO3 calculated value 516.1, found value 516.1 [M+H] + .

[0436] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((E)-3-(3,4-dichlorophenyl)-3-oxoprop-1-en-1-yl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (111e).

[0437]

[0438] Step E. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) afforded enone 111e (175 mg, 0.36 mmol, 57%) as a light yellow solid. IR (cm -1)2957,1714,1607,1516,1391,1210,1031,739; 1 H NMR (400MHz, CDCl3) δ8.08(d,J=1.6Hz,1H),7.82(dd,J=8.4,1.6Hz,1H),7.73(d,J=16.0Hz,1H),7.57(d,J=8.4Hz,1H),7.44(m,1H),7. 42(m,1H),7.41(d,J=16.0Hz,1H),7.17(br,1H),5.87(dd,J=4.0,4.0Hz,2H),3.49(m,2H),3.19(m,2H),1.15(m,2H),0.36-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 187.6, 176.7 (2C), 157.6 (d, J = 253.3 Hz, 1C), 143.5 (d, J = 2.2 Hz, 1C), 137.9, 137.5 (d, J = 7.5 Hz, 1C), 137.4, 133.6, 131.0, 130.7, 129.9, 128.0, 127.7, 125.0 (d, J = 2.8 Hz, 1C), 123.2 (2C), 121.8 (d, J = 13.8 Hz, 1C), 115.9 (d, J = 20.3 Hz, 1C), 45.94, 45.90, 34.0 (2C), 10.0 (2C), 5.0; ES-API MS: m / z for C 26 H 19 Cl2FNO3 calculated value 482.1, found value 482.1 [M+H] + .

[0439] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((E)-3-(3,5-dichlorophenyl)-3-oxoprop-1-en-1-yl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (111f).

[0440]

[0441] Step E. Purification by silica gel flash chromatography (gradient elution, 0→40% EtOAc in hexanes) afforded enone 111f (271 mg, 0.56 mmol, 88%) as a light yellow solid. IR (cm -1 )3077,3009,2957,1714,1564,1516,1391,1214,1178,738,700;1 H NMR (400MHz, CDCl3) δ7.81 (s, 2H), 7.66 (d, J = 15.6Hz, 1H), 7.51 (m, 1H), 7.41 (m, 1H), 7.38 (m, 1H), 7.38 ( d,J=15.6Hz,1H),7.13(br,1H),5.83(m,2H),3.45(m,2H),3.16(m,2H),1.12(m,2H),0.32-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 187.1, 176.6 (2C), 157.4 (d, J = 253.2 Hz, 1C), 143.6 (d, J = 1.8 Hz, 1C), 140.2, 137.2 (d, J = 7.5 Hz, 1C), 135.7 (2C), 132.7, 129.8, 127.8, 127.0 (2C), 125.1, 122.9 (2C), 121.7 (d, J = 13.9 Hz, 1C), 115.8 (d, J = 20.4 Hz, 1C), 45.8 (2C), 33.8 (2C), 9.9 (2C), 4.9; ES-API MS: m / z for C 26 H 19 Calculated value of Cl2FNO3: 482.1, found: 482.0 [M+H] + .

[0442] 4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcyclopropyl[f]isoindol-2(1H)-yl)benzyl 4-methoxybenzoate (12).

[0443]

[0444] To a mixture of benzyl alcohol 105 (50 mg, 0.17 mmol), p-methoxybenzoic acid (39 mg, 0.25 mmol) and PPh3 (67 mg, 0.25 mmol) in anhydrous THF (1.7 mL) at 0°C was added DIAD (51 mg, 0.25 mmol) dropwise. The resulting mixture was stirred and slowly warmed to room temperature over 3 hours. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→45% EtOAc in hexanes) and then recrystallized from CH2Cl2 / hexanes to afford ester 12 (52 mg, 0.12 mmol, 71%) as a yellow solid. IR (cm -1)3450,3056,3008,1707,1606,1512,1377,1274,1257,1169,736; 1 H NMR (400MHz, CDCl3) δ8.01(d,J=8.4Hz,2H),7.51(d,J=8.0Hz,2H),7.20(d,J=8.4Hz,2H),6.91(d,J=8.8Hz,2H),5.86(dd ,J=4.8,3.6Hz,2H),5.33(s,2H),3.86(s,3H),3.50(m,2H),3.14(dd,J=1.2,1.2Hz,2H),1.15(m,2H),0.35-0.26(m,2H); 13 CNMR (100 MHz, CDCl3) δ 177.6 (2C), 166.0, 163.5, 136.7, 131.7 (2C), 131.6, 128.82 (2C), 127.79 (2C), 126.6 (2C), 122.3, 113.6 (2C), 65.7, 55.4, 45.3 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 26 H 24 NO5 calculated value 430.2, found value 430.1 [M+H] + .

[0445] N-(4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcyclopropyl[f]isoindol-2(1H)-yl)benzyl)-4-methoxybenzamide (17).

[0446]

[0447] To a solution of benzylamine 109 (45 mg, 0.15 mmol) in anhydrous CH2Cl2 (0.5 mL) was added Et3N (18.6 mg, 0.18 mmol) and p-methoxybenzoyl chloride (27.5 mg, 0.16 mmol). The resulting mixture was stirred at room temperature for 2 hours. The solvent was removed under reduced pressure and the residue was purified by flash chromatography on silica gel (gradient elution, 0→90% EtOAc in hexanes) and then recrystallized from CH2Cl2 / hexanes to afford amide 17 as a light yellow solid (54 mg, 0.13 mmol, 83%). IR (cm -1 )3338,3010,2956,1704,1607,1505,1387,1255,1181,732; 1H NMR (400MHz, CDCl3) δ7.74(d,J=8.8Hz,2H),7.41(d,J=8.4Hz,2H),7.14(d,J=8.4Hz,2H),6.91(d,J=8.8Hz,2H),6.35(t,J=5.6Hz,1H),5 .85(dd,J=4.8,3.2Hz,2H),4.62(d,J=5.6Hz,2H),3.84(s,3H),3.49(m,2H),3.13(dd,J=2.0,2.0Hz,2H),1.14(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.6 (2C), 166.8, 162.3, 138.9, 131.1, 128.77 (2C), 128.76 (2C), 127.8 (2C), 126.8 (2C), 126.4, 113.8 (2C), 55.4, 45.3 (2C), 43.6, 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 26 H 25 N2O4 calculated value 429.2, found value 429.2 [M+H] + .

[0448] N-(4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcycloprop[f]isoindol-2(1H)-yl)benzyl)-4-methoxybenzenesulfonamide (18).

[0449]

[0450] To a solution of p-methoxybenzenesulfonyl chloride (1.0 equiv) in anhydrous CH2Cl2 (0.2 M) at 0°C was added benzylamine 109 (1.15 equiv) followed by Et3N (2.0 equiv). The resulting yellow mixture was warmed to room temperature and stirring was continued at this temperature for 2 h. The reaction was quenched with aqueous hydrochloric acid (1 N) and extracted with EtOAc. The combined organic layers were washed with saturated NaHCO3 solution, brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes to afford sulfonamide 18 (156 mg, 0.34 mmol, 58%) as a colorless solid. IR (cm -1 )3278,3007,2952,1703,1597,1389,1260,1184,1157,734;1 H NMR (400MHz, CDCl3) δ7.76(d,J=9.2Hz,2H),7.24(d,J=8.4Hz,2H),7.06(d,J=8.4Hz,2H),6.95(d,J=8.8Hz,2H),5.83(dd,J=4.4,3.2Hz,2H) ,4.64(t,J=6.4Hz,1H),4.11(d,J=6.0Hz,2H),3.86(s,3H),3.46(m,2H),3.11(dd,J=2.0,1.6Hz,2H),1.15-1.11(m,2H),0.33-0.24(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.5 (2C), 163.0, 136.7, 131.4, 131.3, 129.3 (2C), 128.5 (2C), 127.8 (2C), 126.8 (2C), 114.3 (2C), 55.6, 46.7, 45.3 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 25 H 24 N2O5SNa calculated value 487.1, found value 487.1 [M+Na] + .

[0451] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(3-(4-methoxyphenyl)-3-oxopropyl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (19).

[0452]

[0453] To a solution of enone 110 (53 mg, 0.12 mmol) and diethyl 1,4-dihydro-2,6-dimethyl-3,5-pyridinedicarboxylate (also known as Hantzsch ester) (63 mg, 0.25 mmol) in anhydrous THF (1.2 mL) was added dropwise TiCl4 solution (0.18 mL, 0.18 mmol, 1 M in CH2Cl2). The resulting heterogeneous mixture slowly turned into a brown homogeneous solution. After 20 minutes, the reaction mixture was poured into a separating funnel containing a biphasic layer of EtOAc (10 mL) and saturated NaHCO3 solution (10 mL). The aqueous phase was extracted with EtOAc (2×10 mL), and the combined organic layers were washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) and then recrystallized from CH2Cl2 / hexanes to give the desired compound 19 as a white solid (44 mg, 0.10 mmol, 86%). IR (cm -1 )3010,2956,1706,1674,1601,1515,1384,1259,1176; 1 H NMR (400MHz, CDCl3) δ7.92(d,J=8.8Hz,2H),7.31(d,J=8.4Hz,2H),7.09(d,J=8.4Hz,2H),6.92(d,J=9.2Hz,2H),5.85(dd,J=4.8,3.6Hz,2H),3 .86(s,3H),3.49(m,2H),3.23(dd,J=7.6,8.4Hz,2H),3.13(dd,J=2.0,1.6Hz,2H),3.06(dd,J=8.0,7.2Hz,2H),1.14(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 197.4, 177.7 (2C), 163.5, 142.1, 130.2 (2C), 129.82, 129.79, 129.2 (2C), 127.8 (2C), 126.5 (2C), 113.7 (2C), 55.5, 45.3 (2C), 39.9, 33.8 (2C), 29.9, 9.9 (2C), 4.7; ES-APIMS: m / z for C 27 H 26 NO4 calculated value 428.2, found value 428.1 [M+H] + .

[0454] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-methoxybenzoyl)cyclopropyl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (24).

[0455]

[0456] Step F. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 24 (70 mg, 0.16 mmol, 85%) as a light yellow solid. IR (cm -1 )3007,2956,1707,1599,1393,1232,1178,1029,734; 1 H NMR (400MHz, CDCl3) δ7.97(d,J=8.8Hz,2H),7.23(d,J=8.4Hz,2H),7.10(d,J=8.4Hz,2H),6.94(d,J=8.8Hz,2H),5.86(dd,J=4.8,3.6Hz,2H) ,3.87(s,3H),3.50(m,2H),3.14(dd,J=1.6,1.6Hz,2H),2.83(m,1H),2 .65(m,1H),1.90(m,1H),1.49(m,1H),1.15(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 196.5, 177.7 (2C), 163.5, 141.3, 130.6, 130.4 (2C), 130.1, 127.78, 127.76, 126.9 (2C), 126.6 (2C), 113.7 (2C), 55.5, 45.3 (2C), 33.8 (2C), 29.0, 18.7, 14.1, 9.9 (2C), 4.7; ES-APIMS: m / z for C 28 H 26 NO4 calculated value 440.2, measured value 440.2 [M+H] + .

[0457] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(3-(4-methoxybenzoyl)oxiran-2-yl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (25).

[0458]

[0459] To a solution of enone 110 (220 mg, 0.52 mmol) in MeOH / CH2Cl2 (5 mL, 1:1 / vol:vol) at 0°C were added H2O2 (234 mg, 2.07 mmol, 30 wt% solution) and NaOH (114 mg, 2.84 mmol, 0.5 M in H2O). The resulting heterogeneous yellow mixture slowly turned into a colorless mixture. The reaction was slowly warmed to room temperature and stirred for 2 days and poured into a separatory funnel containing a biphasic layer of EtOAc (15 mL) and saturated aqueous NaHCO3 (15 mL). The aqueous phase was extracted with EtOAc (2×10 mL), dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by flash chromatography on silica gel (gradient elution, 0→60% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes to afford the desired epoxide 25 as an amorphous white solid (200 mg, 0.45 mmol, 89%). IR (cm -1 )3464,3010,2957,1708,1600,1517,1382,1242,1174,734; 1 H NMR (400MHz, CDCl3) δ7.98(d,J=8.8Hz,2H),7.44(d,J=8.8Hz,2H),7.22(d,J=8.4Hz,2H),6.95(d,J=8.8Hz,2H),5.87(dd,J=4.0,4.0Hz, 2H),4.20(d,J=2.0Hz,1H),4.08(d,J=1.6Hz,1H),3.88(s,3H),3.50(m,2H),3.16(dd,J=1.6,1.6Hz,2H),1.16(m,2H),0.36-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 190.8, 177.5 (2C), 164.3, 136.2, 132.3, 130.7 (2C), 128.5, 127.83, 127.79, 126.8 (2C), 126.4 (2C), 114.1 (2C), 60.8, 58.6, 55.6, 45.4 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 27 H 24 NO5 calculated value 442.2, found value 442.1 [M+H] + .

[0460] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3-chloro-4-methoxybenzoyl)cyclopropyl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (65).

[0461]

[0462] Step F. Purification by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 65 (43 mg, 0.087 mmol, 84%) as a light yellow solid. IR (cm -1 )3055,3009,2954,1714,1663,1594,1395,1259,1211,1183,1063,1014,736,698; 1 H NMR (400MHz, CDCl3) δ8.03 (d, J=2.4Hz, 1H), 7.89 (dd, J=8.4, 2.4Hz, 1H), 7.1 1(br,1H),7.02(d,J=8.0Hz,1H),6.98(d,J=8.8Hz,1H),6.96(d,J=10.8Hz,1H ),5.86(dd,J=4.0,4.0Hz,2H),3.98(s,3H),3.49(m,2H),3.17(m,2H),2.79(m ,1H),2.67(m,1H),1.90(m,1H),1.51(m,1H),1.14(m,2H),0.35-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 195.5, 177.1 (2C), 159.1, 157.6 (d, J = 251.9 Hz, 1C), 144.5 (d, J = 7.5 Hz, 1C), 131.1, 130.6, 129.4, 128.8, 128.0, 123.3, 122.8 (d, J = 2.3 Hz, 1C), 118.1 (d, J = 14.3 Hz, 1C), 114.4 (d, J = 20.2 Hz, 1C), 111.5 (2C), 110.2, 56.6, 53.7, 46.0, 34.0 (2C), 29.1 (2C), 19.4, 10.1 (2C), 5.0; ES-API MS: m / z for C 28 H 24 ClFNO4 calculated value 492.1, found value 492.2 [M+H] + .

[0463] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-chloro-3-methylbenzoyl)cyclopropyl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (66).

[0464]

[0465] Step F. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 66 (40 mg, 0.084 mmol, 78%) as a yellow solid. IR (cm -1 )3054,3009,2956,1716,1666,1521,1395,1180,1049,912,735; 1 H NMR (400MHz, CDCl3) δ7.84 (d, J=1.6Hz, 1H), 7.73 (dd, J=8.4, 2.0Hz, 1H), 7.4 3(d,J=8.4Hz,1H),7.11(br,1H),7.02(d,J=8.0Hz,1H),6.96(d,J=10.4Hz,1H ),5.86(dd,J=4.0,4.0Hz,2H),3.49(m,2H),3.17(m,2H),2.82(m,1H),2.68(m ,1H),2.44(s,3H),1.92(m,1H),1.53(m,1H),1.15(m,2H),0.36-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 197.1, 177.1 (2C), 157.6 (d, J = 251.9 Hz, 1C), 144.4, 140.0, 136.4 (d, J = 96.8 Hz, 1C), 130.7, 129.6, 127.9, 127.1, 122.8, 118.1 (d, J = 13.6 Hz, 1C), 114.4 (d, J = 19.3 Hz, 1C), 46.0, 45.7, 34.0 (2C), 31.8, 29.5, 29.4, 22.9, 20.3, 19.6, 14.4, 10.1 (2C), 5.0; ES-API MS: m / z for C 28 H 24 Calculated value of ClFNO3: 476.1, found: 476.2 [M+H] + .

[0466] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3-chloro-4-fluorobenzoyl)cyclopropyl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (67).

[0467]

[0468] Step F. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 67 (18 mg, 0.038 mmol, 35%) as a white solid. IR (cm -1 )3056,3010,2957,1722,1668,1520,1394,1250,1182,1065,736; 1 H NMR (400MHz, CDCl3) δ8.05(dd,J=6.8,2.4Hz,1H),7.88(ddd,J=8.4,4.4,2.0Hz,1H),7.24(dd,J=8.8,8.8Hz,1H),7.11(d,J=8.8Hz,1H),7.02(d,J=7.6Hz ,1H),6.96(d,J=10.8Hz,1H),5.86(m,2H),3.49(m,2H),3.17(m,2H),2.79(m ,1H),2.70(m,1H),1.93(m,1H),1.56(m,1H),1.15(m,2H),0.36-0.27(m,2H); 13 C NMR (100MHz, CDCl3) δ195.5,177.1(2C),161.3(d,J=256.0Hz,1C),157.6(d,J=252.3Hz,1C),144.0(d,J=7.5 Hz,1C),134.7(d,J=3.6Hz,1C),131.3(d,J=0.5Hz,1C),129.5(d,J=15.3Hz,1C),128.7(d,J=8.4Hz,1C),128 .2,127.9(d,J=14.9Hz,1C),126.2(d,J=7.3Hz,1C),122.5(d,J=39.7Hz,1C),118.3(d,J=13.7Hz,1C),117.1 ES-API MS: m / z for C27 H 21 Calculated value of ClF2NO3: 480.1, found: 480.1[M+H] + .

[0469] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(4-chloro-3-(trifluoromethyl)benzoyl)cyclopropyl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (68).

[0470]

[0471] Step F. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 68 (48 mg, 0.091 mmol, 78%) as a white solid. IR (cm -1 )3055,3010,2957,1715,1674,1603,1520,1394,1315,1036,737; 1 H NMR (400MHz, CDCl3) δ8.30(d,J=1.6Hz,1H),8.06(dd,J=8.4,2.0Hz,1H),7.63(d,J=8.0Hz,1H),7.12(m,1H),7.02(d,J=7.6Hz,1H),6.96(d,J=10.4 Hz,1H),5.86(dd,J=3.6,3.6Hz,2H),3.49(m,2H),3.17(m,2H),2.81(m,1H ),2.74(m,1H),1.95(m,1H),1.61(m,1H),1.15(m,2H),0.36-0.29(m,2H); 13 CNMR (100 MHz, CDCl3) δ 195.8, 177.1 (2C), 157.6 (d, J = 252.3 Hz, 1C), 145.7, 143.8, 137.7, 135.9, 132.4, 132.2, 131.8, 129.4, 128.0, 127.5, 122.8, 118.5, 114.6, 60.6, 45.9, 45.8, 34.0 (2C), 29.8, 29.4, 20.0, 10.1 (2C), 5.0; ES-API MS: m / z for C 28 H 21 Calculated value of ClF4NO3: 530.1, found: 530.1[M+H] + .

[0472] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3,4-dichlorobenzoyl)cyclopropyl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (69).

[0473]

[0474] Step F. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 69 (44 mg, 0.088 mmol, 86%) as a white solid. IR (cm -1 )3009,2919,1715,1668,1520,1393,1181,736; 1 H NMR (400MHz, CDCl3) δ8.05(d,J=2.0Hz,1H),7.79(dd,J=8.4,2.0Hz,1H),7.56(d,J=8.4Hz,1H),7.12(m,1H),7.02(d,J=7.6Hz,1H),6.96(d,J=10.8 Hz,1H),5.86(dd,J=3.6,3.6Hz,2H),3.49(m,2H),3.17(m,2H),2.79(m,1H ),2.71(m,1H),1.93(m,1H),1.57(m,1H),1.15(m,2H),0.35-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 195.9, 177.1 (2C), 158.9, 143.9, 138.0, 137.0, 133.7, 131.0, 130.7 (d, J = 4.0 Hz, 1C), 130.3, 127.9, 127.4, 125.7, 122.7, 122.3, 46.0, 45.7, 34.0, 31.8, 29.8, 29.5, 22.9, 19.9, 14.4, 10.1, 5.0; ES-API MS: m / z for C 27 H 21 Calculated value of Cl2FNO3: 496.1, found: 496.1[M+H] + .

[0475] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-(3,5-dichlorobenzoyl)cyclopropyl)-2-fluorophenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (70).

[0476]

[0477] Step F. Purification by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded 70 (44 mg, 0.088 mmol, 86%) as a yellow solid. IR (cm -1 )3077,3009,2956,1715,1566,1520,1397,1217,1182,736; 1 H NMR (400MHz, CDCl3) δ7.80 (s, 2H), 7.53 (s, 1H), 7.09 (m, 1H), 6.99 (d, J = 6.8Hz, 1H), 6.94 (d, J = 10.4Hz, 1H), 5.84 (dd, J = 3.6 ,3.6Hz,2H),3.46(m,2H),3.15(m,2H),2.77(m,1H),2.70(m,1H),1.89(m,1H),1.55(m,1H),1.13(m,2H),0.33-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 195.6, 177.0 (2C), 157.5 (d, J = 252.3 Hz, 1C), 143.7 (d, J = 7.4 Hz, 1C), 139.8, 135.8 (2C), 132.8 (2C), 129.4 (d, J = 17.0 Hz, 1C), 127.8 (d, J = 7.5 Hz, 1C), 126.7 (2C), 122.6, 118.2 (d, J = 13.7 Hz, 1C), 114.3 (d, J = 20.8 Hz, 1C), 45.9, 45.6, 33.8 (2C), 29.8, 29.4, 20.2, 10.0 (2C), 4.9; ES-API MS: m / z for C 27 H 21 Calculated value of Cl2FNO3: 496.1, found: 496.1[M+H] + .

[0478] 4-((3aR,4R,4aR,5aS,6S,6aS)-1,3-dioxo-3,3a,4,4a,5,5a,6,6a-octahydro-4,6-vinylcyclopropyl[f]isoindol-2(1H)-yl)phenyl 2-(4-methoxyphenyl)acetate (13).

[0479]

[0480] To a solution of phenol 102 (50 mg, 0.18 mmol) in anhydrous THF (1.8 mL) was added NaH (17 mg, 0.71 mmol, 60% dispersion in mineral oil) and heated to 60 °C for 20 min. The mixture was cooled to room temperature and neat 2-(4'-methoxyphenyl)-acetyl chloride (66 mg, 0.36 mmol) was added. The turbid mixture became transparent and the reaction was complete within 30 min. The reaction was quenched with H2O (5 mL) and extracted with EtOAc (2×10 mL). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→50% EtOAc in hexanes) to give ester 13 as a white solid (65 mg, 0.15 mmol, 86%). IR (cm -1 )3006,2959,1748,1713,1506,1393,1204,1124,1038,812,735; 1 H NMR (400MHz, CDCl3) δ7.28(d,J=8.4Hz,2H),7.18(d,J=8.8Hz,2H),7.13(d,J=9.2Hz,2H),6.89(d,J=8.8Hz,2H),5.84(dd ,J=4.8,3.2Hz,2H),3.80(s,3H),3.78(s,2H),3.47(m,2H),3.11(dd,J=1.6,1.6Hz,2H),1.13(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.6 (2C), 170.0, 159.0, 150.5, 130.5 (2C), 129.4, 127.9 (2C), 127.6 (2C), 125.4, 122.2 (2C), 114.3 (2C), 55.4, 45.4 (2C), 40.6, 34.0 (2C), 10.0 (2C), 4.8; ES-API MS: m / z for C 26 H 23 NO5Na calculated value 452.1, measured value 452.1 [M+Na] + .

[0481] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((E)-2-(methoxyimino)-2-(4-methoxyphenyl)ethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (26).

[0482]

[0483] A mixture of ketone 1 (50 mg, 0.12 mmol), methoxylamine hydrochloride (26.3 mg, 0.31 mmol) and sodium acetate (42 mg, 0.51 mmol) was dissolved in EtOH (0.4 mL) and H2O (1.2 mmol). The resulting suspension was heated to 70 °C and stirred for 2 h. The reaction was cooled to room temperature and extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→40% EtOAc in hexanes) and then recrystallized from CH2Cl2 / hexanes to give 26 as a white solid (27 mg, 0.059 mmol, 49%). IR (cm -1 )2938,1707,1608,1512,1250,1180,1048,1030,830,734; 1 H NMR (400MHz, CDCl3) δ7.61(d,J=8.8Hz,2H),7.05(d,J=8.8Hz,2H),6.95(d,J=9.2Hz,2H),6.86(d,J=8.8Hz,2H),5.84(d d,J=4.4,3.6Hz,2H),5.16(s,2H),4.02(s,3H),3.81(s,3H),3.48(m,2H),3.11(m,2H),1.13(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 178.0 (2C), 160.8, 158.2, 153.7, 128.6 (2C), 128.0 (2C), 127.9 (2C), 126.1, 125.2, 115.3 (2C), 114.0 (2C), 62.6, 60.3, 55.5, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 27 H 27 N2O5 calculated value 459.2, found value 459.2 [M+H] + .

[0484] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((E)-2-(hydroxyimino)-2-(4-methoxyphenyl)ethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (27).

[0485]

[0486] A mixture of ketone 1 (48 mg, 0.11 mmol), aminohydroxide hydrochloride (31 mg, 0.44 mmol) and sodium acetate (55 mg, 0.67 mmol) was dissolved in EtOH (0.55 mL) and H2O (0.55 mmol). The resulting suspension was heated to 70°C and stirred for 2 hours. The reaction was cooled to room temperature and extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→40% EtOAc in hexanes) and then recrystallized in CH2Cl2 / hexanes to give 27 as a white solid (38 mg, 0.085 mmol, 78%). IR (cm -1 )3370,2838,1704,1513,1250,1181,1029,830,734; 1 H NMR (400MHz, CDCl3) δ8.77(br,1H),7.59(d,J=8.8Hz,2H),7.05(d,J=8.8Hz,2H),6.98(d,J=8.8Hz,2H),6.88(d,J=8.8H z,2H),5.84(dd,J=4.4,3.6Hz,2H),5.24(s,2H),3.81(s,3H),3.48(m,2H),3.11(m,2H),1.13(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 178.2 (2C), 160.9, 158.2, 155.0, 128.6 (2C), 128.0 (2C), 127.9 (2C), 125.9, 125.2, 115.3 (2C), 114.1 (2C), 59.8, 55.5, 45.4 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 24NO5 calculated value 430.2, found value 430.1 [M+H2O-NHOH] + .

[0487] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(2-hydroxy-2-(4-methoxyphenyl)ethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (28).

[0488]

[0489] To a solution of ketone 1 (50 mg, 0.12 mmol) in MeOH / CH2Cl2 (vol:vol / 1:1, 2.30 mL) was added NaBH4 (5.7 mg, 0.15 mmol) at 0°C. The resulting mixture was stirred at this temperature for 2 hours. The reaction was quenched with cold H2O (1 mL) and extracted with EtOAc (2×5 mL). The combined organic layers were washed with brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→65% EtOAc in hexanes) to give benzyl alcohol 28 as a white solid (33 mg, 0.076 mmol, 66%). IR (cm -1 )3462,2955,1704,1513,1247,1182,734; 1 H NMR (400MHz, CDCl3) δ7.36(d,J=8.8Hz,2H),7.07(d,J=9.2Hz,2H),6.95(d,J=8.8Hz, 2H), 6.92 (d, J=8.8Hz, 2H), 5.85 (dd, J=4.4, 3.6Hz, 2H), 5.05 (ddd, J=8.4, 2.8, 2.8Hz, 1H),4.05(dd,J=9.6,3.2Hz,1H),3.99(dd,J=8.8,9.2Hz,1H),3.81(s,3H),3.48(m,2H ),3.11(dd,J=2.0,2.0Hz,2H),2.71(d,J=2.8Hz,1H),1.14(m,2H),0.34-0.25(m,2H); 13C NMR (100 MHz, CDCl3) δ 178.1 (2C), 159.8, 158.5, 131.8, 128.00 (2C), 127.97 (2C), 127.8 (2C), 125.2, 115.3 (2C), 114.2 (2C), 73.7, 72.2, 55.5, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 24 NO4 calculated value 414.2, measured value 414.2 [M-OH] + .

[0490] (1R,2R,4S,5S,6S,7R)-Tricyclo[3.2.2.02,4]non-8-ene-6,7-dicarboxylic acid (112).

[0491]

[0492] Anhydride 97 (4 g, 21.0 mmol) was dissolved in a 40% MeOH in H2O solution (105 mL) containing NaOH (3.36 g, 84.1 mmol). The resulting mixture was heated to reflux at 90°C and stirred overnight. After stirring for 1 hour, a white precipitate was formed. The mixture was cooled to 0°C, neutralized with cold aqueous hydrochloric acid (6N), and extracted with a mixture of CHCl3 and i-PrOH (volume:volume / 3:1). The remaining aqueous phase was concentrated under reduced pressure and the precipitated NaCl salt was removed by filtration. The precipitated salt was washed several times with CHCl3. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→15% MeOH in CH2Cl2) and then recrystallized from MeOH / CH2Cl2 / hexanes to give acid 112 (2.7 g, 13.0 mmol, 61%) as a white solid. IR (cm -1 )2998,2737,1705,1416,1230,943,702; 1 H NMR (400MHz, DMSO-d6) δ11.8(s,2H),5.72(dd,J=4.8,3.6Hz,2H),2.99(m,2H),2.94(m,2H),0.96(m,2H),0.08--0.02(m,2H); 13 CNMR (100 MHz, DMSO-d6) δ 173.9 (2C), 127.2 (2C), 47.8 (2C), 33.7 (2C), 9.2 (2C), 2.6 (2C); ES-API MS: m / z for C 11 H 12O4 calculated value 208.1, measured value 208.1[M] + .

[0493] (1R,3aS,4S,5S,5aS,6aR,6bR,7R)-4-Bromo-2-oxooctahydro-2H-1,5-methanocycloprop[e]benzofuran-7-carboxylic acid (113).

[0494]

[0495] To a mixture of acid 112 (500 mg, 2.40 mmol) and NaHCO (504 mg, 6.00 mmol) was added H2O (24 mL). It was noted that bubbling occurred immediately. The mixture was stirred and heated to 50 ° C for 10 minutes to ensure complete formation of the dicarboxylate. The mixture was cooled to 0 ° C and bromine solution (575 mg, 3.6 mmol) was added dropwise until no further decolorization occurred. The resulting mixture was warmed to room temperature and stirred for another 6 hours. The mixture was extracted with CHCl (3×). The combined organic layers were washed with 15% Na2S2O5 aqueous solution, brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→5% MeOH in CH2Cl2) and then recrystallized in EtOAc / hexane to obtain bromolactone 113 (380 mg, 1.32 mmol, 55%) as a colorless solid. IR(cm -1 )3022,1773,1717,1164,981; 1 H NMR (400MHz, CDCl3 / CD3OD) δ4.58(ddd,J=3.2,1.2,0.8Hz,1H),4.49(ddd,J=4.8,1.2,0.8Hz,1H),3.09(ddd,J=9.2,4.8,4.8Hz,1H),3. 00(dd,J=10.0,1.2Hz,1H),2.82-2.77(m,2H),1.15(ddd,J=6.8,4.0,3.6Hz,1H),1.12-1.03(m,2H),0.68(ddd,J=8.0,8.0,6.8Hz,1H); 13 C NMR (100 MHz, CDCl3 / CD3OD) δ 177.5, 173.5, 84.3, 46.4, 46.1, 41.9, 39.6, 35.7, 11.5, 8.0, 6.2; ES-API MS: m / z for C 11 H 12 BrO4 calculated value 287.0( 79 Br isotope), found 286.9[M+H]+ .

[0496] (1R,3aS,4S,5S,5aS,6aR,6bR,7R)-4-Bromo-N-(4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-2-oxooctahydro-2H-1,5-methanocyclopropyl[e]benzofuran-7-carboxamide (114).

[0497]

[0498] To a solution of bromolactone 113 (30 mg, 0.10 mmol) and HATU (40 mg, 0.10 mmol) in anhydrous DMF (0.2 mL) was added DIPEA (13.5 mg, 0.10 mmol). The reaction mixture was stirred at room temperature for 15 minutes until it became homogeneous. A solution of 2-(4-aminophenoxy)-1-(4-methoxyphenyl)ethan-1-one (26 mg, 0.10 mmol) in anhydrous DMF (0.2 mL) was added to the reaction mixture via a syringe, and the mixture was then stirred at room temperature until TLC showed complete consumption of aniline. The mixture was quenched with aqueous hydrochloric acid (1 N, 5 mL) and extracted with EtOAc (2×5 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→55% EtOAc in hexanes) to afford amide 114 (33 mg, 0.062 mmol, 63%) as a white amorphous solid. IR (cm -1 )3446,2934,1704,1694,1601,1513,1234,1172,834,729; 1 H NMR (400MHz, CDCl3) δ7.98(d,J=8.8Hz,2H),7.21(d,J=8.8Hz,2H),7.00(d,J=9.2Hz,2H),6 .96(d,J=9.2Hz,2H),5.20(s,2H),4.06(dd,J=6.4,3.2Hz,1H),3.88(s,3H),3.76(m,1H),3 .26(dd,J=9.6,4.0Hz,1H),3.13(dd,J=6.4,3.2Hz,1H),3.01(dd,J=10.0,3.2Hz,1H),2.90 (dd,J=6.8,3.2Hz,1H),2.68(d,J=2.8Hz,1H),1.39(m,1H),1.19-1.09(m,2H),0.74(m,1H); 13C NMR (100 MHz, CDCl3) δ 192.9, 178.5, 177.2, 164.3, 158.3, 130.8 (2C), 128.2 (2C), 127.7, 125.5, 115.6 (2C), 114.3 (2C), 76.3, 71.1, 55.8, 51.7, 47.4, 41.2, 38.0, 37.4, 13.7, 11.2, 8.6; ES-API MS: m / z for C 26 H 26 BrNO7 calculated value 527.1( 81 Br isotope), measured value 527.1[M] + .

[0499] (3aR,4R,4aR,5aS,6S,6aS,7S,8S)-7,8-Dihydroxy-2-(4-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)hexahydro-4,6-ethanocyclopropa[f]isoindole-1,3(2H,3aH)-dione (6).

[0500]

[0501] To a solution of lactone 114 (31 mg, 0.060 mmol) in anhydrous DMF (0.6 mL) was added NaH (2.6 mg, 0.065 mmol, 60% in disperse oil). The resulting mixture was sealed in a pressure sealed tube and heated to 100 ° C for 3 hours. The mixture was cooled to room temperature, quenched with H2O (5 mL), and extracted with EtOAc (2×10 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by preparative TLC (70% EtOAc in hexanes) to give diol 6 (18 mg, 0.038 mmol, 67%) as a white solid. IR (cm -1 )3343,2921,2851,1714,1694,1682,1506,1259,1221,1171,802; 1H NMR (400MHz, CDCl3) δ7.99(d,J=9.2Hz,2H),7.18(d,J=8.8Hz,2H),6.99(d,J=8.8 Hz,2H),6.96(d,J=8.8Hz,2H),5.19(s,2H),4.16(m,1H),3.88(s,3H),3.14(m,1H) ,3.00(d,J=4.8Hz,1H),2.84(ddd,J=8.8,3.2,1.6Hz,1H),1.89(dd,J=8.8,1.2Hz ,1H),1.80(d,J=1.6Hz,1H),1.58(br,2H),1.18(m,2H),0.58(m,1H),0.49(m,1H); 13 C NMR (100 MHz, CDCl3) δ 193.0, 176.2, 175.5, 164.3, 157.9, 130.8 (2C), 128.3 (2C), 127.7, 126.2, 115.5 (2C), 114.3 (2C), 71.9, 71.2, 55.8, 50.2, 40.6, 38.7, 36.1, 27.8, 14.9, 9.6, 8.0; ES-API MS: m / z for C 26 H 25 NO7 calculated value 463.2, measured value 463.2[M] + .

[0502] (4aS)-2-(4-(Hydroxy(4-methoxyphenyl)methyl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (14). To a solution of aldehyde 107 (67 mg, 0.23 mmol) in anhydrous THF (0.77 mL) at 0°C was added 4-methoxyphenylmagnesium bromide (0.7 mL, 0.34 mmol, 0.5 M solution in THF). The mixture was slowly warmed to room temperature and stirred overnight. An additional 1.5 equivalents of 4-methoxyphenylmagnesium bromide was added and the mixture was heated to 65°C. After cooling to room temperature, the reaction mixture was poured into a separatory funnel containing EtOAc and saturated aqueous NH4Cl. The aqueous phase was removed and the organic layer was neutralized with saturated aqueous NaHCO3 and dried over Na2SO4. The crude material was filtered, concentrated in vacuo, and purified by flash chromatography (gradient elution, 0-50% EtOAc / hexanes) to give compound 14 (45 mg, 0.11 mmol, 49%) to provide an amorphous yellow solid. 1H NMR (400MHz, CDCl3) δ7.43(d,J=8.4Hz,2H),7.26(d,J=8.4Hz,2H),7.13(d,J=8.4Hz,2H),6.85(d,J=8.8Hz,2H),5.84(dd,J=4.4,3.6 Hz,2H),5.80(br,1H),3.79(s,3H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),2.21(d,J=2.8Hz,1H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.7 (2C), 159.2, 144.3, 135.6, 130.8, 128.0 (2C), 127.8 (2C), 127.0 (2C), 126.4 (2C), 113.9 (2C), 75.4, 55.3, 45.3 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 25 H 22 NO3 calculated value 384.2, found value 384.1 [M-OH] + .

[0503] (4aS)-2-(4-(1-hydroxy-2-phenylethyl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcycloprop[f]isoindole-1,3(2H,3aH)-dione (21). To a solution of aldehyde 107 (50 mg, 0.17 mmol) in anhydrous THF (0.6 mL) at 0°C was added benzylmagnesium chloride (0.17 mL, 0.25 mmol, 2 M solution in THF). The mixture was slowly warmed to room temperature and stirred overnight. The reaction mixture was poured into a separatory funnel containing EtOAc and saturated aqueous NH4Cl. The aqueous phase was removed and the organic layer was neutralized with saturated aqueous NaHCO3 and dried over Na2SO4. The crude material was filtered, concentrated in vacuo, and purified by flash chromatography (gradient elution, 0-45% EtOAc / hexanes) to afford compound 21 (40 mg, 0.10 mmol, 62%) as an amorphous yellow solid. 1H NMR (400MHz, CDCl3) δ7.44 (d, J=8.8Hz, 2H), 7.32 (dd, J=7.6, 6.8Hz, 2H), 7.2 3(m,3H),7.16(d,J=8.4Hz,2H),5.87(dd,J=4.4,3.6Hz,2H),4.92(m,1H),3. 50(m,2H),3.14(dd,J=1.6,1.6Hz,2H),3.03(dd,J=13.6,4.4Hz,1H),2.93(d d,J=13.6,8.8Hz,1H),1.95(d,J=2.8Hz,1H),1.15(m,2H),0.36-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.7 (2C), 144.3, 137.8, 131.0, 129.5 (2C), 128.6 (2C), 127.8 (2C), 126.7, 126.50 (2C), 126.46 (2C), 74.8, 46.1, 45.3 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 25 H 24 NO3 Calculated value 386.2 Found value 386.1 [M+H] + .

[0504] (4aS)-2-(4-(1-hydroxy-3-phenylpropyl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropyl[f]isoindole-1,3(2H,3aH)-dione (22). To a solution of aldehyde 107 (50 mg, 0.17 mmol) in anhydrous THF (0.6 mL) at 0°C was added phenethylmagnesium chloride (0.34 mL, 0.34 mmol, 1 M solution in THF). The mixture was slowly warmed to room temperature and stirring was continued overnight. The reaction mixture was poured into a separatory funnel containing EtOAc and saturated aqueous NH4Cl. The aqueous phase was removed and the organic layer was neutralized with saturated aqueous NaHCO3 and dried over Na2SO4. The crude material was filtered, concentrated in vacuo, and purified by flash chromatography (gradient elution, 0-50% EtOAc / hexanes) to afford compound 22 (48 mg, 0.12 mmol, 71%) as an amorphous yellow solid. 1H NMR (400MHz, CDCl3) δ7.41(d,J=8.4Hz,2H),7.27(d,J=8.4Hz,2H),7.20-7.15(m,5H),5.86(dd,J=4.8,3.6Hz,2H),4.70(m,1H),3.4 9(m,2H),3.14(dd,J=1.6,1.6Hz,2H),2.73(m,2H),2.10(m,1H),2.00(m,1H),1.89(d,J=3.2Hz,1H),1.15(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.7 (2C), 145.0, 141.5, 131.0, 128.42 (2C), 128.40 (2C), 127.8 (2C), 126.6 (2C), 126.5 (2C), 125.9, 73.3, 45.3 (2C), 40.3, 33.8 (2C), 31.9, 9.9 (2C), 4.7; ES-API MS: m / z for C 26 H 24 NO2 calculated value 382.2, measured value 382.2 [M-OH] + .

[0505] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-ethynylphenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (115).

[0506]

[0507] Step A. Purification by flash chromatography on silica gel (gradient elution, 0→35% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded alkyne 115 (1.42 g, 4.9 mmol, 47%) as an orange solid. IR (cm -1 )3278,1703,1509,1391,1194; 1 H NMR (400MHz, CDCl3) δ7.52(d,J=8.4Hz,2H),7.15(d,J=8.8Hz,2H),5.83(dd,J=4.8,3.2Hz, 2H),3.47(m,2H),3.12(dd,J=1.6,1.6Hz,2H),3.08(s,1H),1.13(m,2H),0.33-0.24(m,2H); 13C NMR (100 MHz, CDCl3) δ 177.3 (2C), 132.7 (2C), 132.0, 127.8 (2C), 126.3 (2C), 122.3, 82.7, 78.2, 45.3 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 19 H 16 NO2 calculated value 290.1, found value 290.1 ​​[M+H] + .

[0508] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(3-(4-methoxyphenyl)-3-oxoprop-1-yn-1-yl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (23).

[0509]

[0510] A flame-dried round-bottom flask was charged with PdCl2(PPh3)2 (2.4 mg, 0.003 mmol), CuI (1.3 mg, 0.007 mmol) and anhydrous THF (0.6 mL). The mixture was degassed and purged with argon (×3). A degassed solution of Et3N (0.025 mL, 0.18 mmol) was added to the reaction mixture, followed by a syringe addition of a solution of 4-methoxybenzoyl chloride (31 mg, 0.18 mmol) and alkyne 115 (50 mg, 0.17 mmol) in anhydrous THF (0.5 mL). The resulting mixture was stirred at room temperature for 2 hours. It was noted that the mixture changed from yellow / orange to a green precipitate. The mixture was poured into a separation funnel containing a saturated NaHCO3 aqueous solution (5 mL) and EtOAc (10 mL). The organic phase was separated, washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→30% EtOAc in hexanes) and then recrystallized from CH2Cl2 / hexanes to afford the alkynone 23 (52 g, 0.12 mmol, 71%) as a yellow solid. IR (cm -1 )2201,1711,1633,1597,1509,1377,1292,1261,1163,736; 1H NMR (400MHz, CDCl3) δ8.17(d,J=8.8Hz,2H),7.72(d,J=8.4Hz,2H),7.29(d,J=8.4Hz,2H),6.99(d,J=9.2Hz,2H),5 .87(dd,J=4.8,3.6Hz,2H),3.90(s,3H),3.51(m,2H),3.16(dd,J=1.6,1.6Hz,2H),1.16(m,2H),0.37-0.27(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.1 (2C), 176.5, 164.6, 133.6, 133.5 (2C), 132.0 (2C), 130.2, 127.8 (2C), 126.5 (2C), 120.4, 113.9 (2C), 91.0, 87.4, 55.6, 45.4 (2C), 33.9 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 27 H 22 NO4 calculated value 424.2, found value 424.1 [M+H] + .

[0511] (1aS,2S,2aS,9aR,10R,10aR)-5-methoxy-1a,2,2a,9a,10,10a-hexahydro-2,10-vinylbenzo[4,5]imidazo[2,1-a]cyclopropa[f]isoindol-9(1H)-one (116).

[0512]

[0513] Step B (conventional heating). Purification by flash chromatography on silica gel (gradient elution, 0→70% EtOAc in hexanes) afforded an inseparable mixture of positional isomers (2:1 ratio favoring the desired isomer 116) (131 mg, 0.45 mmol, 85%) as a light yellow solid. IR (cm -1 )3458,3364,3007,2955,1768,1630,1515,1212,1188,735; 1H NMR (400MHz, CDCl3) 116: δ6.83 (d, J = 8.4Hz, 1H), 6.37 (dd, J = 8.4, 2.8Hz, 1H), 6.29 (d, J = 2.8Hz, 1H),5.94(dd,J=4.4,3.6Hz,2H),3.75(s,3H),3.50(m,2H),3.16(m,2H),1.15(m,2H),0.37-0.2 3 (m, 2H); Position isomers: δ 6.71 (d, J = 8.8 Hz, 1H), 6.37 (dd, J = 8.8, 2.4 Hz, 1H), 6.33 (d, J = 2.4 Hz, 1H), 5.87 (dd, J = 4.0, 3.6 Hz, 2H), 3.62 (m, 2H), 3.53 (s, 3H), 3.13 (m, 2H), 1.15 (m, 2H), 0.37-0.23 (m, 2H); 13 C NMR (100 MHz, CDCl3) 116: δ 177.7, 161.1, 143.8, 129.6, 128.6 (2C), 127.8, 110.8, 104.9, 102.78, 101.91, 55.3, 45.7, 33.9, 9.8, 4.8; Position isomers: δ 178.0, 161.1, 143.6, 129.2, 128.7, 127.8, 111.8, 110.0, 105.3, 102.80, 101.91, 55.30, 45.2, 33.8, 9.9, 4.6; ES-API MS: m / z for C 18 H 17 N2O2 calculated value 293.1, found value 293.1 [M+H] + .

[0514] (1aR,2R,2aR,9aS,10S,10aS)-5-Hydroxy-1a,2,2a,9a,10,10a-hexahydro-2,10-vinylbenzo[4,5]imidazo[2,1-a]cyclopropyl[f]isoindol-9(1H)-one (117).

[0515]

[0516] To a solution of the above mixture of positional isomeric methyl ethers 116 (873 mg, 3.00 mmol) in CH2Cl2 was added dropwise BBr3 (5 mL, 4.5 mmol, 1 M solution in CH2Cl2) at -78°C. The resulting mixture was stirred for 12 h while the temperature was slowly warmed to room temperature. The reaction was quenched by adding MeOH (5 mL) and H2O (2 mL) at 0°C, and the mixture was extracted with EtOAc (4×10 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→80% EtOAc in hexanes) to give a mixture of positional isomeric phenols 117 (680 mg, 2.44 mmol, 82%) as a white amorphous solid. IR (cm -1 )3360,1694,1514,1180; 1 HNMR(400MHz,CD3OD)δ6.70(d,J=8.4Hz,1H),6.26(m,1H),6.18(dd,J=8.8,2.8Hz,1H),5.92(d d,J=4.8,3.6Hz,2H),3.37(m,2H),3.20(dd,J=2.0,2.0Hz,2H),1.20(m,2H),0.35-0.22(m,2H); 13 C NMR (100MHz, CD3OD) δ179.6,158.7,145.6,128.9,128.3,127.5,109.7,105.3,104.7,102.3,45.6,45.3,33.7,9.3,9.2,3.8,3.5.

[0517] (1aR,2R,2aR,9aS,10S,10aS)-5-(2-(4-methoxyphenyl)-2-oxoethoxy)-1a,2,2a,9a,10,10a-hexahydro-2,10-vinylbenzo[4,5]imidazo[2,1-a]cyclopropyl[f]isoindol-9(1H)-one (87).

[0518]

[0519] To a solution of the above positional isomeric mixture of phenols 117 (50 mg, 0.18 mmol) in ethanol (1.8 mL) was added CsCO (176 mg, 0.54 mmol) and 2-bromo-4'-OMe-acetophenone (82 mg, 0.36 mmol). The resulting mixture was heated to 50 °C for 4 hours. The solvent was removed under reduced pressure and the residue was purified by flash chromatography on silica gel (gradient elution, 0→45% EtOAc in hexanes), then purified by HPLC (reverse phase, MeCN / H2O / 0.1% TFA) and recrystallized from CH2Cl2 / hexanes to give the desired product 87 as a single positional isomer (23 mg, 0.05 mmol, 30%) as a white solid. IR (cm -1 )3460,3365,3051,3009,2956,1704,1601,1514,1393,1308,1241,1173,735; 1 H NMR (400MHz, CDCl3) δ7.98(d,J=8.8Hz,2H),6.95(d,J=8.8Hz,2H),6.82(d,J=8.4Hz,1H),6.39(d,J=1.6Hz,1H),6.36(dd,J=6 .8,2.4Hz,1H),5.94-5.85(m,2H),5.11(s,2H),3.88(s,3H),3.50(m,2H),3.16-3.13(m,2H),1.15(m,2H),0.36-0.23(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.0, 177.6, 164.0, 159.5, 144.0, 130.7 (2C), 129.7, 128.6 (2C), 127.8, 127.6, 114.0 (2C), 111.6, 105.4, 102.9, 71.0, 55.5, 45.7, 45.2, 33.8, 9.9, 9.8, 4.8, 4.6; ES-API MS: m / z for C 26 H 23 N2O4 calculated value 427.2, found value 427.1 [M+H] + .

[0520] (5-Aminobenzofuran-2-yl)(4-methoxyphenyl)methanone (118).

[0521]

[0522] A mixture of N-(3-formyl-4-hydroxyphenyl)acetamide (200 mg, 1.12 mmol), K2CO3 (308 mg, 2.23 mmol) and 4'-MeO-2-bromoacetophenone (511 mg, 2.23 mmol) was dissolved in anhydrous DMF (3.72 mL). The resulting mixture was heated to 90°C and stirred for 4 hours. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→55% EtOAc in CH2Cl2) to give the intermediate acetamide (200 mg, 0.65 mmol, 58%) as a light yellow solid. Acetamide (200 mg, 0.65 mmol) was then dissolved in EtOH (6.5 mL), aqueous hydrochloric acid solution (3N, 3.2 mL) was added, and the mixture was refluxed overnight. The solvent was removed under reduced pressure, the residue was diluted with EtOAc, and neutralized with saturated aqueous NaHCO3. The organic layer was washed with brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (100% EtOAc) to give the free aniline 118 (168 mg, 0.63 mmol, 97%) as a bright yellow solid. IR (cm -1 )3445,3360,1634,1601,1573,1548,1510,1329,1261,1164,975,765; 1 H NMR (400MHz, CDCl3) δ8.08(d,J=8.8Hz,2H),7.40(d,J=8.8Hz,1H),7.35(s,1H),7.00(d,J= 9.2Hz,2H),6.91(d,J=2.0Hz,1H),6.87(dd,J=8.4,2.4Hz,1H),3.89(s,3H),3.71(br,2H); 13 C NMR (100 MHz, CDCl3) δ 183.0, 163.6, 153.3, 150.7, 143.2, 132.1 (2C), 130.1, 128.1, 118.3, 115.3, 113.9 (2C), 113.0, 106.5, 55.7; ES-API MS: m / z for C 16 H 14 NO3 calculated value 268.1, found value 268.1 [M+H] + .

[0523] (3aR,4R,4aR,5aS,6S,6aS)-2-(2-(4-methoxybenzoyl)benzofuran-5-yl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (88).

[0524]

[0525] A mixture of aniline 118 (175 mg, 0.92 mmol) and maleic anhydride 97 (175 mg, 0.92 mmol) was dissolved in acetic acid (3.0 mL), heated to 100 ° C and stirred for 3 hours. The mixture was cooled to room temperature, diluted with EtOAc, and quenched with saturated NaHCO3 aqueous solution. The organic layer was washed with brine, dried over anhydrous MgSO4, and filtered. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→60% EtOAc in hexanes) and then recrystallized in CH2Cl2 / hexanes to give the product 88 (227 mg, 0.52 mmol, 56%) as an orange-yellow solid. IR (cm -1 )2952,1714,1644,1601,1558,1392,1281,1259,1175,884,735; 1 H NMR (400MHz, CDCl3) δ8.09(d,J=8.8Hz,2H),7.68(d,J=8.8Hz,1H),7.54(d,J=2.0Hz,1H),7.50(s,1H),7.26(dd,J=8.8,2.0Hz,1H),7.0 2(d,J=8.8Hz,2H),5.90(dd,J=4.8,3.6Hz,2H),3.91(s,3H),3.52(m,2H),3.18(dd,J=1.6,1.6Hz,2H),1.17(m,2H),0.37-0.28(m,2H); 13 C NMR (100 MHz, CDCl3) δ 182.8, 178.0 (2C), 164.0, 155.2, 153.9, 132.2 (2C), 129.8, 128.1 (2C), 128.0, 127.7, 126.7, 121.7, 115.3, 114.1 (2C), 113.4, 55.8, 45.5 (2C), 34.1 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 27 H 22 NO5 calculated value 440.1, found value 440.1 [M+H] + .

[0526] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((E)-3-(2-bromo-4-methoxyphenyl)-3-oxoprop-1-en-1-yl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (119).

[0527]

[0528] Step E. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded enone 119 (333 mg, 0.66 mmol, 71%) as a pale yellow solid. IR (cm -1 )3008,2956,1709,1599,1378,1293,1233,1181,1032,732; 1 H NMR (400MHz, CDCl3) δ7.63(d,J=8.8Hz,2H),7.50(d,J=16.0Hz,1H),7.48(d ,J=8.8Hz,1H),7.25(d,J=8.8Hz,2H),7.18(d,J=2.4Hz,1H),7.17(d,J=16.0 Hz,1H),6.93(dd,J=8.4,2.4Hz,1H),5.86(dd,J=4.8,3.6Hz,2H),3.86(s,3H ),3.50(m,2H),3.15(dd,J=1.6,2.0Hz,2H),1.16(m,2H),0.36-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 193.1, 177.5 (2C), 161.9, 143.6, 135.0, 133.8, 133.4, 131.5, 129.2 (2C), 128.0 (2C), 127.2, 127.0 (2C), 121.4, 119.2, 113.5, 56.0, 45.6 (2C), 34.1 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 27 H 22 BrNO4Na calculated value 528.0( 81 Br isotope), found 528.0[M+Na] + .

[0529] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-(5-methoxy-1-oxo-1H-inden-3-yl)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (89).

[0530]

[0531] A mixture of enone 119 (20 mg, 0.040 mmol), PdCl2 (3.5 mg, 0.02 mmol), PPh3 (16 mg, 0.06 mmol) and K2CO3 (14 mg, 0.10 mmol) in DMF (0.5 mL) was heated to 110 °C and stirred for 3 h. The reaction was cooled to room temperature, filtered through a Celite pad, and washed with EtOAc. The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→45% EtOAc in hexanes) to give benzofuranone 89 (5.0 mg, 0.012 mmol, 31%) as a yellow solid. IR (cm -1 )3006,2925,2855,1711,1601,1384,1180,734; 1 H NMR (400MHz, CDCl3) δ7.68(d,J=8.4Hz,2H),7.49(d,J=8.0Hz,1H),7.35(d,J=8.4Hz,2H),6.88(d,J=2.0Hz,1H),6.69(dd,J=8. 0,1.6Hz,1H),6.01(s,1H),5.89(dd,J=4.4,4.0Hz,2H),3.85(s,3H),3.53(m,2H),3.19(m,2H),1.18(m,2H),0.38-0.28(m,2H); 13 C NMR (100 MHz, CDCl3) δ 195.6, 177.4, 164.0, 159.5, 146.3, 133.1, 128.0 (2C), 127.9 (2C), 126.9 (2C), 125.3, 124.8, 124.7, 110.9, 110.6, 55.8, 45.4 (2C), 33.9 (2C), 29.7, 9.9 (2C), 4.7; ES-API MS: m / z for C 27 H 22 NO4 calculated value 424.2, found value 424.1 [M+H] + .

[0532] (3aR,4R,4aR,5aS,6S,6aS)-2-(4-((6-methoxybenzo[d]isoxazol-3-yl)methoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (90).

[0533]

[0534] A mixture of phenol 102 (45 mg, 0.16 mmol) and K2CO3 (33 mg, 0.24 mmol) in anhydrous DMF (0.30 mL) was heated at 50°C for 30 min and cooled to room temperature before the addition of 3-(bromomethyl)-6-methoxy-benzisoxazole (120, 77 mg, 0.32 mmol). The resulting mixture was heated to 65°C and stirred overnight. The solvent was removed under reduced pressure and the residue was purified by flash chromatography on silica gel (gradient elution, 0→40% EtOAc in hexanes) and then recrystallized from CH2Cl2 / hexanes to afford benzisoxazole analog 90 (55 mg, 0.14 mmol, 77%) as a white solid. IR (cm -1 )3054,3006,2960,1705,1616,1511,1140,831; 1 H NMR (400MHz, CDCl3) δ7.65(dd,J=8.8,0.4Hz,1H),7.09(s,4H),7.00(d,J=2.0Hz,1H),6.92(dd,J=8.8,2.0Hz,1H),5.84(d d,J=4.8,3.6Hz,2H),5.42(s,2H),3.88(s,3H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 C NMR (100 MHz, CDCl3) δ 178.0 (2C), 165.7, 162.7, 158.0, 154.9, 128.1 (2C), 128.0 (2C), 125.6, 122.5, 115.4 (2C), 115.1, 114.2, 92.8, 62.3, 56.0, 45.5 (2C), 34.0 (2C), 10.1 (2C), 4.9; ES-API MS: m / z for C 26 H 23 N2O5 calculated value 443.2, found value 443.2 [M+H] + .

[0535] (3aR,4R,4aR,5aS,6S,6aS)-2-(3-Hydroxyphenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (121).

[0536]

[0537] Step B. Purification by flash chromatography on silica gel (gradient elution, 0→50% EtOAc in hexanes) followed by recrystallization from CH2Cl2 / hexanes afforded phenol 121 (90 mg, 0.32 mmol, 61%) as colorless crystals. IR (cm -1 )3310,1773,1686,1595,1389,1272,1179,733,714; 1 H NMR (400MHz, CDCl3) δ7.26 (dd, J=8.4, 7.6Hz, 1H), 6.79 (dd, J=8.4, 2.4Hz, 1H), 6.72 (dd, J=7.6, 0.8Hz, 1H), 6.62 (dd, J=2.0, 2. 0Hz,1H),5.85(dd,J=4.8,3.6Hz,2H),5.63(br,1H),3.49(m,2H),3.14(dd,J=2.0,1.6Hz,2H),1.15(m,2H),0.35-0.26(m,2H); 13 C NMR (100 MHz, CDCl3) δ 177.9 (2C), 156.2, 132.6, 130.0, 127.8 (2C), 118.6, 116.0, 113.8, 45.3 (2C), 33.8 (2C), 9.9 (2C), 4.6; ES-APIMS: m / z for C 17 H 16 NO3 calculated value 282.1, found value 282.1 [M+H] + .

[0538] (3aR,4R,4aR,5aS,6S,6aS)-2-(3-(2-(4-methoxyphenyl)-2-oxoethoxy)phenyl)-4,4a,5,5a,6,6a-hexahydro-4,6-vinylcyclopropa[f]isoindole-1,3(2H,3aH)-dione (91).

[0539]

[0540] To a solution of phenol 121 (37 mg, 0.13 mmol) in reagent grade acetone (1.3 mL) was added KCO (54 mg, 0.39 mmol), 18-crown-6 (17 mg, 0.07 mmol), tetrabutylammonium iodide (24 mg, 0.07 mmol) and 2-bromo-4'-OMe-acetophenone (60 mg, 0.26 mmol). The resulting mixture was refluxed at 65-70 °C for 48 h (note that the Rf of the product was the same as the starting material). The solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography (gradient elution, 0→50% EtOAc in hexanes) and then recrystallized from CHCl / hexanes to give the product 91 (40 mg, 0.09 mmol, 71%) as a light yellow solid. IR (cm -1 )3082,3051,3007,2841,1772,1709,1602,1493,1384,1262,1234,1181,971,839,733; 1 H NMR (400MHz, CDCl3) δ7.98(d,J=8.8Hz,2H),7.32(dd,J=8.0,8.0Hz,1H),6.96(d,J=8.8Hz,2H),6.93(m,1H),6.80(m,2H),5.8 4(dd,J=4.8,3.6Hz,2H),5.17(s,2H),3.88(s,3H),3.48(m,2H),3.12(dd,J=1.6,1.6Hz,2H),1.14(m,2H),0.34-0.25(m,2H); 13 CNMR (100 MHz, CDCl3) δ 192.6, 177.4 (2C), 164.1, 158.5, 132.9, 130.6 (2C), 129.9, 127.8 (2C), 127.5, 119.7, 114.8, 114.0 (2C), 113.6, 70.9, 55.5, 45.3 (2C), 33.8 (2C), 9.9 (2C), 4.7; ES-API MS: m / z for C 26 H 24 NO5 calculated value 430.2, found value 430.1 [M+H] + .

[0541] In vivo studies

[0542] Mice tolerated the long-term, once-daily oral administration of the analogs in Tables 1-9 well. The body weight of ICRSCID mice treated with analogs of a vehicle or 5, 20 or 60 mg / kg doses orally once a day for 14 consecutive days was evaluated. n = 5 per arm. Complete blood cell counts including total hemoglobin (Hb) and platelets (PLT) were evaluated on the 14th day. Serum chemistry analysis was also evaluated on the 14th day. Alanine (ALT) and aspartate (AST) transaminases were used as markers of liver damage. Creatinine was used as a marker of renal function. Triglycerides were used as systemic biomarkers for cholesterol synthesis inhibition. The analog levels of plasma were evaluated on the 14th day.

[0543] The results of these in vivo animal studies demonstrate that the analogs of Tables 1-9 are bioavailable and brain penetrant, do not cause weight loss, do not induce hematotoxicity, hepatotoxicity, or renal toxicity, and do not increase, but actually decrease, triglyceride levels in mice during chronic treatment.

[0544]

[0545]

[0546]

[0547]

[0548] Table 1. SAR of dicarboximide moiety

[0549]

[0550] Table 2. SAR of the middle benzene ring

[0551]

[0552] Compound R <![CDATA[Mut6 IC 50 (μM)]]> <![CDATA[S9 T 1 / 2 (minutes)]]> 7 3-F 0.928±0.066 >240 8 2-F 0.113±0.011 6.78 9 <![CDATA[3-CF3]]> 33.866±4.278 NT 10 2-Cl 0.100±0.015 NT 11 <![CDATA[2,6-Cl2]]> 0.210±0.020 NT

[0553] Table 3. SAR of linkers

[0554]

[0555]

[0556]

[0557] Table 4. SAR of the benzoyl moiety

[0558]

[0559]

[0560]

[0561]

[0562] Table 5. SAR of trans-benzoylcyclopropane analogs

[0563]

[0564] Compound R <![CDATA[Mut6 IC 50 (μM)]]> <![CDATA[S9 T 1 / 2 (minutes)]]> 65 3-Cl,4-OMe 0.030±0.002 157.5 66 3-Me,4-Cl 0.027±0.002 192.5 67 3-Cl,4-F 0.100±0.008 56.8 68 <![CDATA[3-CF3,4-Cl]]> 0.115±0.013 169 69 <![CDATA[3,4-Cl2]]> 0.034±0.003 47.1 70 <![CDATA[3,5-Cl2]]> 0.199±0.011 18.3

[0565] Table 6. SAR of cyclopropyl phenyl ketone analogs

[0566]

[0567]

[0568]

[0569] Table 7. Synthesis of Series II compounds was consistent with Synthetic Mechanisms 1-3 above; anti-GBM activity between 1-200 nm was demonstrated in the survival screen above using Mut 6—a mouse GBM stem cell-like cell (GSC) line from a genetically engineered mouse GBM model, in which spontaneous tumor formation was driven by GFAP-Cre-mediated silencing of tumor suppressors Trp53, Pten, and Nf1 in mouse astrocytes. Animal studies demonstrated that these analogs are bioavailable and brain penetrant, do not cause weight loss, do not cause hematotoxicity, hepatotoxicity, or renal toxicity, and do not increase, but actually reduce, triglyceride levels in mice during long-term treatment.

[0570]

[0571] Table 8. Synthesis of Series III compounds was consistent with Synthetic Mechanisms 1-3 above; Anti-GBM activity between 1-200 nm was demonstrated in the survival screen above using Mut 6—a mouse GBM stem cell-like cell (GSC) line from a genetically engineered mouse GBM model, in which spontaneous tumor formation was driven by GFAP-Cre-mediated silencing of tumor suppressors Trp53, Pten, and Nf1 in mouse astrocytes. Animal studies demonstrated that these analogs are bioavailable and brain penetrant, do not cause weight loss, do not cause hematotoxicity, hepatotoxicity, or renal toxicity, and do not increase, but actually reduce, triglyceride levels in mice during long-term treatment.

[0572]

[0573] Table 9. Synthesis of Series IV compounds was consistent with Synthetic Mechanisms 1-3 above; Anti-GBM activity between 1-200 nm was demonstrated in the survival screen above using Mut 6—a mouse GBM stem cell-like cell (GSC) line from a genetically engineered mouse GBM model, in which spontaneous tumor formation was driven by GFAP-Cre-mediated silencing of tumor suppressors Trp53, Pten, and Nf1 in mouse astrocytes. Animal studies demonstrated that these analogs are bioavailable and brain penetrant, do not cause weight loss, do not cause hematotoxicity, hepatotoxicity, or renal toxicity, and do not increase, but actually reduce, triglyceride levels in mice during long-term treatment.

[0574]

[0575] Table 10. Pharmacokinetic parameters

[0576]

[0577]

[0578] Calculated using MarvinSketch software b clogP, clogD, PSA and pKa. d BBB was calculated as AUC brain / AUC plasma.

Claims

1. A compound of formula I or a salt, hydrate or stereoisomer thereof: in: Q is selected from cyclopropane-1,2-diyl, CH2, (CH2)2, O; X, Y, Z and W are each independently selected from CR and N; R = H, halogen (eg, F, Cl, Br); CH3, optionally fluorinated (eg, CF3), optionally deuterated (eg, CD3); L is selected from: -OCHRC(O)- [R = H, Me, CF3, CHF2, CH2F], -NHCH2C(O)-, -NHCHMeC(O)-, -NMeCH2C(O)-, -NMeCHMeC(O)-, -SCH2C(O)-, -SCHMeC(O)-, -CH2OC(O)-, -CHMeOC(O)-, -CH2NHC(O)-, CHMeNHC(O)-, -OCH2NMeC(O)-, -CHMeNMeC(O)-, -CH2NHSO2-, -CHMeNHSO2-, -CH2NMeSO2-, -CMeNMeSO2-, -O(CH2)n- [n = 0 - 2], -OCHMe-, -OCHMeCH2-, -OCH2CHR- [R = Me, OH, OMe, OCF3, OCHF2, OCH2F], -OCHMeCHR- [R = OH, OMe, OCF3, OCHF2, OCH2F], -(CH2)nC(O)- [n = 0 - 2], -CHRCH2C(O)- [R = Me, OH, OMe, OCF3, OCHF2, OCH2F], -CH2CHRC(O)- [R = Me, OH, OMe, OCF3, OCHF2, OCH2F], -CHORCH2n- [n = 0 - 2; R = H, Me, CF3, CHF2, CH2F], -(CH2)nCHOR- [n = 1 - 2;R=H, Me, CF3, CHF2, CH2F], -CHORCH2CHOR'-[R, R' are independently selected from H, Me, CF3, CHF2, CH2F], -CHORCHMeCHOR'-[R, R' are independently selected from H, Me, CF3, CHF2, CH2F], -CHORCMeCH2-[R=H, Me, CF3, CHF2, CH2F], -CH(OR)CH=CH-[R=H, Me, CF3, CH HF2, CH2F], -(CH2)n-[n=0-3], -CH2NR-[R=H, Me]-, -CH=CHC(O)-, CH=CHCHR-[R=OH, OMe, OCF3, OCHF2, O CH2F], -CMe=CHC(O)-, CH=CMeC(O)-, CMe=CHCHOR-[R=H, Me, CF3, CHF2, CH2F], CH=CMeCHOR[R=H, Me, CF3 , CHF2, CH2F], -C□CC(O)-, -C□CCHR[R=H, Me, OH, OMe, OCF3, OCHF2, OCH2F], -(cyclopropane-1,2-diyl)C(O)-, -(cyclopropane-1,2-diyl)CHR-[R=H, Me, OH, OMe, OCF3, OCHF2, OCH2F], -(ethylene oxide-2,3-diyl)C(O)-, -(ethylene oxide-2,3-diyl)CHR-[R= H, Me, OH, OMe, OCF3, OCHF2, OCH2F], -C(O)(cyclopropane-1,2-diyl)-, -CHR(cyclopropane-1,2-diyl)-[R=H, Me, OH, OMe, OCF3, OCHF2, OCH2F], -C(O)(ethylene oxide-2,3-diyl)-, -CHR(ethylene oxide-2,3-diyl)-[R=H, Me, OH, OMe, OCF3, OCHF2, OCH2F]; and; Ar is selected from 2- or 3- or 4-monosubstituted phenyl or disubstituted phenyl or trisubstituted phenyl (preferred substituents are selected from halogen (e.g. F, Cl, Br), Me or OMe, cycloPr, CN, -NHCHO, each optionally fluorinated and optionally deuterated (e.g. OCF3, CF3, CD3); and optionally substituted phenyl and heteroaryl), monocyclic and fused bicyclic heteroaryl, fused heteroaryl [e.g. pyridine, pyrimidine, pyrazine , pyridazine, oxazole, benzoxazole, pyrazole, quinoline, quinoxaline, isoquinoline], biaryl and fused aryl (naphthyl); each optionally fluorinated and optionally deuterated (e.g. OCF3, CF3, CD3); optional substituents are selected from halogen (e.g. F, Cl, Br), CF3, Me or OMe, cPr, CN, aryl and heteroaryl, each optionally fluorinated and optionally deuterated (e.g. OCF3, CF3, CD3).

2. The compound of claim 1, wherein: Q is cyclopropane-1,2-diyl.

3. The compound of claim 1, wherein: X is N, and Y, Z and W are each CH; or, X, Y, Z and W are each CH.

4. The compound of claim 1, wherein: L is selected from: -OCHC(O)-, -NHCH2C(O)-, -NHCHMeC(O)-, -NMeCH2C(O)-, -NMeCHMeC(O)-, -CH2OC(O)-, -CHMeOC(O)-, -CH2NHC(O)-, CHMeNHC(O)-, -O CH2NMeC(O)-, -CHMeNMeC(O)-, -OCHMe-, -OCHMeCH2-, -OCH2CHMe-, -OCHMeCHOH-, -(CH2)C(O)-, -CHMeCH2C(O)-, -CH2CHMeC(O)-, -CHOHCH 2-、-CH2CHOH-、-CHOHCH2CHOH'-、-CHOHCHMeCHOH-、-CHOHCMeCH2-、-CH(OH)CH=CH2-、-CH2NH-、-(cyclopropane-1,2-diyl)C(O)-、-(cyclopropane-1,2-diyl)CH2-、-(ethylene oxide-2,3-diyl)C(O)-、-(ethylene oxide-2,3-diyl)CH2-、-C(O)(cyclopropane-1,2-diyl)-、-CH2(cyclopropane-1,2-diyl)-、-C(O)(ethylene oxide-2,3-diyl)-、-CH2(ethylene oxide-2,3-diyl)-.

5. The compound of claim 1, wherein: Ar is 2- or 3- or 4-monosubstituted phenyl or disubstituted phenyl or trisubstituted phenyl, the substituents being selected from halogen, Me or OMe, cycloPr, CN and -NHCHO, each optionally fluorinated and optionally deuterated.

6. The compound of claim 1, wherein: Q is cyclopropane-1,2-diyl, X is N, and Y, Z and W are each CH; or, X, Y, Z and W are each CH; and Ar is 2- or 3- or 4-monosubstituted phenyl or disubstituted phenyl or trisubstituted phenyl, the substituents being selected from halogen, Me or OMe, cycloPr, CN and -NHCHO, each optionally fluorinated and optionally deuterated.

7. The compound of claim 1, wherein: Q is cyclopropane-1,2-diyl, X is N, and Y, Z and W are each CH; or, X, Y, Z and W are each CH; L is selected from: -OCHC(O)-, -NHCH2C(O)-, -NHCHMeC(O)-, -NMeCH2C(O)-, -NMeCHMeC(O)-, -CH2OC(O)-, -CHMeOC(O)-, -CH2NHC(O)-, CHMeNHC(O)-, -O CH2NMeC(O)-, -CHMeNMeC(O)-, -OCHMe-, -OCHMeCH2-, -OCH2CHMe-, -OCHMeCHOH-, -(CH2)C(O)-, -CHMeCH2C(O)-, -CH2CHMeC(O)-, -CHOHCH2 -, -CH2CHOH-, -CHOHCH2CHOH'-, -CHOHCHMeCHOH-, -CHOHCMeCH2-, -CH(OH)CH=CH2-, -CH2NH-, -(cyclopropane-1,2-diyl)C(O)-, -(cyclopropane-1,2-diyl)CH2-, -(ethylene oxide-2,3-diyl)C(O)-, -(ethylene oxide-2,3-diyl)CH2-, -C(O)(cyclopropane-1,2-diyl)-, -CH2(cyclopropane-1,2-diyl)-, -C(O)(ethylene oxide-2,3-diyl)-, -CH2(ethylene oxide-2,3-diyl)-; and Ar is 2- or 3- or 4-monosubstituted phenyl or disubstituted phenyl or trisubstituted phenyl, the substituents being selected from halogen, Me or OMe, cycloPr, CN and -NHCHO, each optionally fluorinated and optionally deuterated.

8. The compound of claim 1 having the structure of Table 1 herein.

9. The compound of claim 1 having the structure of Table 2 herein.

10. The compound of claim 1 having the structure of Table 3 herein.

11. The compound of claim 1 having the structure of Table 4 herein.

12. The compound of claim 1 having the structure of Table 5 herein.

13. The compound of claim 1 having the structure of Table 6 herein.

14. The compound of claim 1 having the structure of Table 7 herein.

15. The compound of claim 1 having the structure of Table 8 herein.

16. The compound of claim 1 having the structure of Table 9 herein.

17. The compound of claim 1 which inhibits lanosterol synthase (LSS), is orally bioavailable and / or crosses the blood-brain barrier.

18. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

19. A method of inhibiting lanosterol synthase comprising administering the small molecule lanosterol synthase inhibitor of claim 1 to a person in need thereof.

20. A method for upregulating 24,25-epoxycholesterol (EPC), comprising administering the small molecule lanosterol synthase inhibitor of claim 1 to a person in need thereof.

21. A method for treating a neurological disease or disorder comprising administering the small molecule lanosterol synthase inhibitor of claim 1 to a person in need thereof.

22. The method of claim 21, wherein the neurological disease or disorder is glioblastoma (GBM) or a neurodegenerative disease such as amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, Alzheimer's disease, Huntington's disease.

23. The method of claim 21, further comprising a prior step of detecting or diagnosing a disease or condition and / or a subsequent step of detecting an improvement or delay in progression of the resulting disease or condition.

24. A method of screening for candidate therapeutics for treating a neurological disease or disorder, the method comprising identifying an inhibitor of lanosterol synthase (p75).

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