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Effect of BAY 1217389 in monotherapy and combination therapy with paclitaxel on total KNL1 in A2780Cis tumors in nude mice

Multinuclearity in U2OS cells induced by Mps1 inhibition

H&E stained A2780cis tumors after treatment with vehicle paclitaxel alone BAY 1161909 alone or in combination with BAY 1161909 and paclitaxel

Additional data showing pharmacological characterization of Mps1 Inhibitors BAY 1161909 and BAY 1217389; Suppl. Table 1: Kinase selectivity profile of BAY 1161909; Suppl. Table 2: Kinase selectivity profile of BAY 1217389; Suppl. Table 3: Kinase selectivity profiles of BAY 1161909 compared to SP600125 and Reversine; Suppl. Table 4: Inhibition of tumor cell proliferation by BAY 1161909 or BAY 1217389; Suppl. Table 5: Summary of in vivo activity of BAY 1161909 in human xenograft tumor models in mice; Suppl. Table 6: Summary of in vivo activity of BAY 1217389 in human xenograft tumor models in mice.

Effect of BAY 1161909 in monotherapy on the frequencies of peripheral blood reticulocytes and micronuclei-harboring reticulocytes in nude mice

Supplementary Data from Comparative Profiling of the Novel Epothilone, Sagopilone, in Xenografts Derived from Primary Non–Small Cell Lung Cancer

Purpose: Characterization of new anticancer drugs in a few xenograft models derived from established human cancer cell lines frequently results in the discrepancy between preclinical and clinical results. To take the heterogeneity of tumors into consideration more thoroughly, we describe here a preclinical approach that may allow a more rational clinical development of new anticancer drugs.Experimental Design: We tested Sagopilone, an optimized fully synthetic epothilone, in 22 well-characterized patient-derived non–small cell lung cancer models and correlated results with mutational and genome-wide gene expression analysis.Results: Response analysis according to clinical trial criteria revealed that Sagopilone induced overall responses in 64% of the xenograft models (14 of 22), with 3 models showing stable disease and 11 models showing partial response. A comparison with response rates for established drugs showed the strong efficacy of Sagopilone in non–small cell lung cancer. In gene expression analyses, Sagopilone induced tubulin isoforms in all tumor samples, but genes related to mitotic arrest only in responder models. Moreover, tumors with high expression of genes involved in cell adhesion/angiogenesis as well as of wild-type TP53 were more likely to be resistant to Sagopilone therapy. As suggested by these findings, Sagopilone was combined with Bevacizumab and Sorafenib, drugs targeting vascular endothelial growth factor signaling, in Sagopilone-resistant models and, indeed, antitumor activity could be restored.Conclusion: Analyses provided here show how preclinical studies can provide hypotheses for the identification of patients who more likely will benefit from new drugs as well as a rationale for combination therapies to be tested in clinical trials. Clin Cancer Res; 16(5); 1452–65

Supplementary Figure 1 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Figure 4 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Table 4 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Sagopilone (ZK-EPO) is the first fully synthetic epothilone undergoing clinical trials for the treatment of human tumors. Here, we investigate the cellular pathways by which sagopilone blocks tumor cell proliferation and compare the intracellular pharmacokinetics and the in vivo pharmacodynamics of sagopilone with other microtubule-stabilizing (or tubulin-polymerizing) agents. Cellular uptake and fractionation/localization studies revealed that sagopilone enters cells more efficiently, associates more tightly with the cytoskeleton, and polymerizes tubulin more potently than paclitaxel. Moreover, in contrast to paclitaxel and other epothilones [such as the natural product epothilone B (patupilone) or its partially synthetic analogue ixabepilone], sagopilone is not a substrate of the P-glycoprotein efflux pumps. Microtubule stabilization by sagopilone caused mitotic arrest, followed by transient multinucleation and activation of the mitochondrial apoptotic pathway. Profiling of the proapoptotic signal transduction pathway induced by sagopilone with a panel of small interfering RNAs revealed that sagopilone acts similarly to paclitaxel. In HCT 116 colon carcinoma cells, sagopilone-induced apoptosis was partly antagonized by the knockdown of proapoptotic members of the Bcl-2 family, including Bax, Bak, and Puma, whereas knockdown of Bcl-2, Bcl-XL, or Chk1 sensitized cells to sagopilone-induced cell death. Related to its improved subcellular pharmacokinetics, however, sagopilone is more cytotoxic than other epothilones in a large panel of human cancer cell lines in vitro and in vivo. In particular, sagopilone is highly effective in reducing the growth of paclitaxel-resistant cancer cells. These results underline the processes behind the therapeutic efficacy of sagopilone, which is now evaluated in a broad phase II program. [Cancer Res 2008;68(13):5301–8]

Supplementary Methods, Legends for Videos and Figures from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Figure 3 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Table 3 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Figure 2 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Video 4 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Figure 6 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Table 2 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Video 3 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Video 2 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Video 5 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Figure 5 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Supplementary Table 1 from Improved Cellular Pharmacokinetics and Pharmacodynamics Underlie the Wide Anticancer Activity of Sagopilone

Inhibition of monopolar spindle 1 (MPS1) kinase represents a novel approach to cancer treatment: instead of arresting the cell cycle in tumor cells, cells are driven into mitosis irrespective of DNA damage and unattached/misattached chromosomes, resulting in aneuploidy and cell death. Starting points for our optimization efforts with the goal to identify MPS1 inhibitors were two HTS hits from the distinct chemical series "triazolopyridines" and "imidazopyrazines". The major initial issue of the triazolopyridine series was the moderate potency of the HTS hits. The imidazopyrazine series displayed more than 10-fold higher potencies; however, in the early project phase, this series suffered from poor metabolic stability. Here, we outline the evolution of the two hit series to clinical candidates BAY 1161909 and BAY 1217389 and reveal how both clinical candidates bind to the ATP site of MPS1 kinase, while addressing different pockets utilizing different binding interactions, along with their synthesis and preclinical characterization in selected in vivo efficacy models.

Progesterone plays an important role in the female reproductive system. However, there is also evidence that gynecologic disorders/diseases such as uterine fibroids and endometriosis are progesterone‐dependent. Steroidal and non‐steroidal selective progesterone receptor modulators (SPRMs) have shown potential for the treatment of such diseases. Steroidal SPRMs, including mifepristone and ulipristal acetate, have proven effective in clinical trials. However, several steroidal SPRMs containing a dimethylamino substituent have been associated with elevated liver enzymes in patients. An earlier drug discovery program identified lonaprisan as a highly selective SPRM that did not show drug‐related change in liver enzyme activity. Building on data obtained from that work, here we describe the research program that culminated in the discovery of a novel steroidal SPRM, vilaprisan, which combines an extremely high potency with very favorable drug metabolism and pharmacokinetic properties. Vilaprisan has entered clinical development and is currently undergoing phase 3 clinical trials.

The presented investigation focuses on the determination of unsteady aerodynamic loads for different kinds of wake vortex encounters. The wake-generating aircraft is a delta–canard configuration, whereas for the penetration, just a delta wing is considered. Based on extensive experimental and numerical investigations of the wake vortex system of a preceding aircraft, the wake-induced angle of attack on an encountering aircraft is determined. Therefore, the induced flowfield of the wake-generating aircraft is transformed into the coordinate system of the penetrating aircraft. Because the unsteadiness results on the one hand from the velocity fluctuations inside the wake, and on the other hand from the relative movement of the two aircraft, the wake problem is split into a discrete and a continuous wake problem. Both problems lead to an unsteady distribution of the angle of attack, which is variable in the chord and spanwise directions. The resulting lift and pitching moment distribution is calculated by an...

We describe our successful efforts for the translation of our research strategy that was focused on a broad structural modification of the natural product epothilone B, to the production of the distinct drug candidate sagopilone in a kg scale. By a direct comparison of the research and development procedures we address some important aspects to be considered for an up-scaling process. 1 Introduction and Concept 2 Preparation of Building Block A 2.1 Research Sequence A1––A3–A4–A5–A6–A7–A8–A9– A10–(A12–)A13 2.2 Development Sequence A11–A12–A13 3 Preparation of Building Block B 3.1 Research Sequence B1–B2–B3–B4–(B6–B7)–B9–B10 3.2 Development Sequence B1–B2–B3–B4–B5–B8–B10 4 Preparation of Building Block C 4.1 Research Sequence C1–C2–C3–C4–C6–C7–C9–C10–C11–C12 4.2 Development Sequence C1–C2–C5–C8–C9–C10–C11–C12 5 Preparation of Building Block BC 5.1 Research 5.2 Development 6 Preparation of the Final Sequence ABC 6.1 Research 6.2 Development 7 Summary

The presented investigation includes a combined experimental–numerical approach to quantify the wake vortex system of a high-agility aircraft from the near field up to the far field. Detailed near-field data are obtained by lowspeed wind-tunnel tests on a delta-canard configuration of 1:15 scale. The measurements are performed at several angles of attack applying advanced hot-wire anemometry. For a wake distance of up to 16 wing spans, mean and turbulentvelocity fieldsaremeasured.Theupstreamdataareusedtoinitializeimplicitlarge-eddysimulationsaimed tocomputethevelocity fieldsofthewakevortexsystemoverawakedistanceofupto50spans.Here,avalidationcase is shown comparing measured and calculated wake data over a distance from 4 to 16 spans, with the implicit largeeddy simulations initialized by the measured quantities at a position of two wing spans. Compared with the experimental data,the numericalresults showthe expectedlateral andvertical movementof thewakevortexsystem due to the interaction of the single vortices. The distributions of axial vorticity, crossflow velocities, and turbulence intensities match well with the experimental data. Inaddition, the dissipation process can be observed, resulting in a reduction of circulation. In the context of this study, the measured and computed velocity fields will be used to determine unsteady aerodynamic loads acting on a fighter aircraft encountering the wake. This is of great importance as wake induction may result in critical structural dynamic loads.

Sagopilone, a fully synthetic epothilone and very potent anti-tumor agent, has proved to be efficient in inhibiting bone destruction and tumor burden in a mouse model of breast cancer bone metastasis. In addition to its antiproliferative effects, this study shows direct effects of sagopilone on bone resorption and osteoclast activity.Sagopilone, a novel fully synthetic third-generation epothilone, has proved to be efficient in inhibiting bone destruction and tumor burden in a mouse model of breast cancer bone metastasis. The aim of this study was to investigate whether the effect was primarily due to sagopilone's antiproliferative effect and consequent inhibition of tumor cell growth, or if sagopilone exerts direct effects on bone resorption and osteoclast activity.Sagopilone was studied and compared to paclitaxel in vitro in human osteoclast differentiation and activity cultures. For studying the potential of sagopilone for inhibiting bone resorption in vivo, a mouse model of ovariectomy (ovx)-induced osteoporosis was utilized.Sagopilone inhibited osteoclast differentiation and activity more efficiently than paclitaxel and showed less cytotoxicity. Whereas sagopilone showed inhibitory effects on human osteoclast differentiation and activity already at 5 and 15 nM, respectively, paclitaxel started to show effects only at 20 and 100 nM concentrations, respectively. Sagopilone treatment increased BMD In the mouse ovx model even though a non-optimized dose was used which is effective in tumor-bearing mice.This is the first study to evaluate sagopilone's effects on bone resorption in non-cancerous situation. The evidence that sagopilone is beneficial for bone will strengthen the status of sagopilone as an anti-cancer compound compared to other microtubule stabilizing agents.

Purpose: Bone metastases have a considerable impact on quality of life in patients with breast and other cancers. Tumors produce osteoclast-activating factors, whereas bone resorption promotes the growth of tumor cells, thus leading to a “vicious cycle” of bone metastasis. Sagopilone, a novel, fully synthetic epothilone, inhibits the growth of breast cancer cells in vitro and in vivo, and here we report its activity in the MDA-MB-231(SA) breast cancer bone metastasis mouse model. Experimental Design: The potency of sagopilone was determined in treatment models simulating the adjuvant (preventive) and metastatic (therapeutic) settings in the clinic. Results: We showed that sagopilone inhibited tumor burden and bone destruction, in addition to reducing tumor-induced cachexia and paraplegia. The reduction in osteolytic lesions, tumor growth in bone, and weight loss was statistically significant in the preventive model compared with the vehicle group. In the therapeutic model, sagopilone treatment significantly lowered the number of activated osteoclasts and significantly reduced the osteolytic lesion area, bone volume loss, and bone resorption compared with vehicle treatment while simultaneously inhibiting tumor burden. An in vitro assay confirmed that sagopilone inhibited osteoclast activation without cytotoxic effects, whereas paclitaxel resulted in lower inhibition and high levels of cytotoxicity. Conclusions: Sagopilone seems to inhibit the vicious cycle at both the tumor growth and bone resorption stages, suggesting the possibility for substantial benefit in the treatment of patients with breast cancer at risk from bone metastases or with bone lesions already present. Phase II clinical trials with sagopilone in patients with breast cancer are ongoing.

Tubulin is among the most established and clinically validated targets in oncology. The taxanes, paclitaxel and docetaxel, stabilize microtubules and have shown significant clinical activity, but factors such as the development of resistance can limit their clinical use. The epothilones are a novel class of natural microtubule-stabilizing products with potential activity in an expanded spectrum of tumour indications.In an extensive lead optimization programme, we selected sagopilone from 350 compounds produced by total synthesis because of its combination of potent activity and good tolerability in tumour models. It is the first fully synthetic epothilone in clinical development.Here we review the directed optimization of the natural product epothilone B to produce sagopilone, along with its mechanism of action, preclinical data and emerging clinical results.We show how this optimization process translated into superior preclinical activity, coupled with a favourable tolerability profile. Activity has been determined in a number of animal models, including those from tumours resistant to other systemic treatments. The approach used to develop sagopilone may become more common as structure-driven research is increasingly employed to exploit the enormous potential of natural products, in parallel with other targeted approaches, heralding a new era of anticancer therapy.

Sagopilone (ZK-EPO) is the first fully synthetic epothilone undergoing clinical trials for the treatment of human tumors. Here, we investigate the cellular pathways by which sagopilone blocks tumor cell proliferation and compare the intracellular pharmacokinetics and the in vivo pharmacodynamics of sagopilone with other microtubule-stabilizing (or tubulin-polymerizing) agents. Cellular uptake and fractionation/localization studies revealed that sagopilone enters cells more efficiently, associates more tightly with the cytoskeleton, and polymerizes tubulin more potently than paclitaxel. Moreover, in contrast to paclitaxel and other epothilones [such as the natural product epothilone B (patupilone) or its partially synthetic analogue ixabepilone], sagopilone is not a substrate of the P-glycoprotein efflux pumps. Microtubule stabilization by sagopilone caused mitotic arrest, followed by transient multinucleation and activation of the mitochondrial apoptotic pathway. Profiling of the proapoptotic signal transduction pathway induced by sagopilone with a panel of small interfering RNAs revealed that sagopilone acts similarly to paclitaxel. In HCT 116 colon carcinoma cells, sagopilone-induced apoptosis was partly antagonized by the knockdown of proapoptotic members of the Bcl-2 family, including Bax, Bak, and Puma, whereas knockdown of Bcl-2, Bcl-XL, or Chk1 sensitized cells to sagopilone-induced cell death. Related to its improved subcellular pharmacokinetics, however, sagopilone is more cytotoxic than other epothilones in a large panel of human cancer cell lines in vitro and in vivo. In particular, sagopilone is highly effective in reducing the growth of paclitaxel-resistant cancer cells. These results underline the processes behind the therapeutic efficacy of sagopilone, which is now evaluated in a broad phase II program. [Cancer Res 2008;68(13):5301–8]

Perfluoroalkanesulfonyl fluorides (C n F 2n+1 SO 2 F) decompose slowly even at -20 °C in the presence of tertiary heteroaromatic or aliphatic amines to form the corresponding perfluorinated alkanes (C n F 2n+2 ) and amine-sulfur dioxide adducts.

Monopolar spindle 1 (Mps1) has been shown to function as the key kinase that activates the spindle assembly checkpoint (SAC) to secure proper distribution of chromosomes to daughter cells. Here, we report the structure and functional characterization of two novel selective Mps1 inhibitors, BAY 1161909 and BAY 1217389, derived from structurally distinct chemical classes. BAY 1161909 and BAY 1217389 inhibited Mps1 kinase activity with IC50 values below 10 nmol/L while showing an excellent selectivity profile. In cellular mechanistic assays, both Mps1 inhibitors abrogated nocodazole-induced SAC activity and induced premature exit from mitosis (“mitotic breakthrough”), resulting in multinuclearity and tumor cell death. Both compounds efficiently inhibited tumor cell proliferation in vitro (IC50 nmol/L range). In vivo, BAY 1161909 and BAY 1217389 achieved moderate efficacy in monotherapy in tumor xenograft studies. However, in line with its unique mode of action, when combined with paclitaxel, low doses of Mps1 inhibitor reduced paclitaxel-induced mitotic arrest by the weakening of SAC activity. As a result, combination therapy strongly improved efficacy over paclitaxel or Mps1 inhibitor monotreatment at the respective MTDs in a broad range of xenograft models, including those showing acquired or intrinsic paclitaxel resistance. Both Mps1 inhibitors showed good tolerability without adding toxicity to paclitaxel monotherapy. These preclinical findings validate the innovative concept of SAC abrogation for cancer therapy and justify clinical proof-of-concept studies evaluating the Mps1 inhibitors BAY 1161909 and BAY 1217389 in combination with antimitotic cancer drugs to enhance their efficacy and potentially overcome resistance. Mol Cancer Ther; 15(4); 583–92. ©2016 AACR.

A novel, highly potent progesterone receptor antagonist, ZK 230211, has been identified. Introduction of a 17α-pentafluoroethyl side chain into the D-ring of the steroidal skeleton resulted in a compound that combines a pure anti-progestagenic endocrinological profile with high potency. Additions of perfluoroalkyl groups to steroidal 17-ketones following modified Prakash and Gassman protocols are described.

An efficient chiral pool synthesis of the C1-C6 fragment of epothilones starting from readily available (-)-pantolactone is described.

For the antitumour agent sagopilone, an epothilone analogue, a large-scale synthesis was developed to synthesise the active pharmaceutical ingredient for clinical trials, exploring enzymatic and microbial methods to produce chiral building blocks on a multi-kilogram scale. The three building blocks were identified as key intermediates in the synthesis and needed to be produced with high optical purity in yields higher than those previously published. The improved syntheses of two of these building blocks are detailed herein. For building block A, the chemical research synthesis was abandoned, and a novel chemical route was developed leading to building block A via an enzymatic hydrolysis process. For building blocks C, replacement of a chemical catalytic procedure by a microbial process meant that the development of a new starting material could be avoided, thereby accelerating the development process. For the clinical development process, a human metabolite of sagopilone was required as a reference. To accelerate the synthesis of the metabolite, no chemical synthesis was investigated; rather, we relied solely on oxidoreductases. The human metabolite of sagopilone was synthesised on a multi-gram scale in a single-step process using genetically engineered E. coli expressing human cytochrome P450 enzyme 2C19. The integration of enzymatic and microbial processes provided tools that enable the synthesis of highly functionalised intermediates and metabolites.

The synthesis of carbacyclin analogues I based on the introduction of an α-chain to a suitable substituted bicyclo[3.3.0]octan-3-one derivative like 5 by a Wittig reaction leads generally to nearly 1:1-mixtures of E/Z-isomers. This lack of selectivity will change dramatically, if an acidic sulfonamide in the bicyclic framework is present. The observed Z-selectivity can be explained by an interaction of charges of the deprotonated sulfonamide and the electrophilic phosphorous atom in the Wittig reagent.

The syntheses of 11β-arylandrost-4-en-3-one 24 and the corresponding 9β,19-cyclo derivative 8 are described. Steric interaction between C-19 and the aryl residue effects conformational changes of the steroid ring system that result in reduced affinity for the progesterone receptor. The conformation of 11β-arylandrostenes is discussed in comparison with known antiprogestational steroids.

The chemistry and in vitro pharmacology of novel prostanoid TXA 2 (TP)-receptor antagonists is described. (5Z)-(9R)-12-(4-Chlorobenzenesulfonamido)-9-fluoro-13,14,15,16,17,18,19,20-octanor-5-prostenoic acid was identified as a potent TP-receptor antagonist.

The preparation and biological characterisation of prostanoid TXA2(TP)-antagonists which are modified at position 9 to add a PGI2-like quality is described. Although we obtained highly potent TP-receptor antagonists, the synthesized compounds do not inhibit the ADP induced platelet aggregation and thus do not have any PGI2(IP)-mimetic properties.

We describe our successful efforts for the translation of our research strategy that was focused on a broad structural modification of the natural product epothilone B, to the production of the distinct drug candidate sagopilone in a kg scale. By a direct comparison of the research and development procedures we address some important aspects to be considered for an up-scaling process. 1 Introduction and Concept 2 Preparation of Building Block A 2.1 Research Sequence A1––A3–A4–A5–A6–A7–A8–A9– A10–(A12–)A13 2.2 Development Sequence A11–A12–A13 3 Preparation of Building Block B 3.1 Research Sequence B1–B2–B3–B4–(B6–B7)–B9–B10 3.2 Development Sequence B1–B2–B3–B4–B5–B8–B10 4 Preparation of Building Block C 4.1 Research Sequence C1–C2–C3–C4–C6–C7–C9–C10–C11–C12 4.2 Development Sequence C1–C2–C5–C8–C9–C10–C11–C12 5 Preparation of Building Block BC 5.1 Research 5.2 Development 6 Preparation of the Final Sequence ABC 6.1 Research 6.2 Development 7 Summary

A novel HBr elimination method for the introduction of a 13.14-triple bond into the prostanoid framework starting from readily available vinyl bromides is described. The transformation can be performed even on sensitive compounds in the presence of various functional groups in synthetically useful yields.

Syntheses of novel TXA2PGH2 receptor antagonists possessing the bicyclo[3.3.0]octane carbacyclin skeleton as described and structure activity relationships are discussed. It is shown that highly potent compounds can be obtained, when the lower prostanoid side chain is replaced by substituted aryl semicarbazones.