Discovery of a Novel Oral Glucocorticoid Receptor Modulator

Feb 9, 2018 - Synthetic glucocorticoids (GC) are essential for the treatment of a broad range of inflammatory diseases. However, their use is limited ...
3 downloads 21 Views 3MB Size
Subscriber access provided by MT ROYAL COLLEGE

Drug Annotation

Discovery of a Novel Oral Glucocorticoid Receptor Modulator (AZD9567) with Improved Side Effect Profile. Lena Ripa, Karl Edman, Matthew Dearman, Goran Edenro, Ramon Hendrickx, Victoria Ullah, HuiFang Chang, Matti Lepistö, Dave Chapman, Stefan Geschwindner, Lisa Wissler, Petter Svanberg, Karolina Lawitz, Jesper Malmberg, Antonios Nikitidis, Roine I Olsson, James Bird, Antonio Llinas, Tove Hegelund-Myrbäck, Markus Berger, Philip Thorne, Richard Harrison, Christian Köhler, and Tomas Drmota J. Med. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jmedchem.7b01690 • Publication Date (Web): 09 Feb 2018 Downloaded from http://pubs.acs.org on February 11, 2018

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

Journal of Medicinal Chemistry is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 57 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Journal of Medicinal Chemistry

Discovery of a Novel Oral Glucocorticoid Receptor Modulator (AZD9567) with Improved Side Effect Profile. Lena Ripa,*a Karl Edman,b Matthew Dearman,a Goran Edenro,a Ramon Hendrickx,a Victoria Ullah,a Hui-Fang Chang,a Matti Lepistö,a Dave Chapman,a Stefan Geschwindner,b Lisa Wissler,b Petter Svanberg,a Karolina Lawitz,c Jesper Malmberg,a Antonios Nikitidis,a Roine I. Olsson,a James Bird,a Antoni Llinas,a Tove Hegelund-Myrbäck,a Markus Berger,d Philip Thorne,e Richard Harrison,e Christian Köhler,b Tomas Drmota,a a

Respiratory, Inflammation and Autoimmunity, IMED Biotech Unit, AstraZeneca, Gothenburg,

Pepparedsleden 1, Mölndal 43183, Sweden. bDiscovery Sciences, IMED Biotech Unit, AstraZeneca, Gothenburg, Pepparedsleden 1, Mölndal 431 83, Sweden. cAstraZeneca R&D Lund, Scheelevägen 1, Lund 22187, Sweden. dMedicinal Chemistry Berlin, Drug Discovery, Pharmaceuticals, Bayer AG, Berlin 13353, eAstraZeneca R&D Charnwood, Bakewell Road, Loughborough, Leicestershire, LE11 5RH, UK Abstract Synthetic glucocorticoids (GC) are essential for the treatment of a broad range of inflammatory diseases. However, their use is limited by target related adverse effects on e.g. glucose homeostasis and bone metabolism. Starting from a non-steroidal GR ligand (4) that is a full agonist in reporter gene assays, we exploited key functional triggers within the receptor, generating a range of structurally diverse partial agonists. Of these, only a narrow subset exhibited full anti-inflammatory efficacy and a significantly reduced impact on adverse effect markers in human cell assays compared to prednisolone. This led to the discovery of AZD9567 (15) with excellent in vivo efficacy when dosed orally in a rat model of joint inflammation.

ACS Paragon Plus Environment

Journal of Medicinal Chemistry 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Compound 15 is currently being evaluated in clinical trials comparing the efficacy and side effect markers with those of prednisolone. Introduction The glucocorticoid receptor (GR) is a ligand activated transcription factor and a member of the steroid hormone receptor family comprising the estrogen receptors (ERα and ERβ), the androgen receptor (AR), the progesterone receptor (PR) and the mineralocorticoid receptor (MR). GR is activated by the glucocorticoid (GC) hormone cortisol. It is of fundamental importance for regulating development, metabolism, inflammation and stress response. GCs were introduced to treat patients with rheumatoid arthritis (RA) nearly 70 years ago1. Since then, the use of synthetic GCs has transformed the treatment of inflammatory and autoimmune diseases2, but the chronic use of GCs at optimal efficacy doses is still limited by severe adverse effects including hyperglycemia, muscle wasting, hypertension and osteoporosis. Several programs to discover safer GR ligands have been reported and new classes of oral non-steroidal ligands have been identified.3 However, only limited progress has been reported for the most advanced compounds 13e, f and 23g (Figure 1) thus the need for safer GR modulators remains.4 In the resting state, GR predominantly resides as a chaperone complex in the cytoplasm. Ligand activation leads to nuclear translocation where GR binds to the DNA response elements and recruits co-regulators to modulate the transcriptional activity in a context dependent manner. Several genomic mechanisms have been described.5 The typical activation process is transactivation (TA) where GR binds as a dimer to the consensus glucocorticoid DNA response element (GRE) to upregulate gene transcription (e.g. tyrosine aminotransferase;TAT and glucocorticoid-induced leucine zipper; GILZ). However, GR can also bind to negative GRE

ACS Paragon Plus Environment

Page 2 of 57

Page 3 of 57 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Journal of Medicinal Chemistry

(nGRE) resulting in repression of transcription6 (e.g. thymic stromal lymphopoietin; TSLP). In addition, GR may also interfere with other transcription factors such as activator protein 1 (AP-1) and nuclear factor κΒ (NF-κB), through a mechanism called transrepression (TR) resulting in repression of gene products such as TNFα and IL-67 Early observations indicated that while many genes controlling adverse effects are regulated by TA, several key pro-inflammatory genes could be suppressed through TR. However, several exceptions have been identified (e.g. GILZ, IL-10) and drug discovery campaigns based upon full agonists with separation of TR over TA alone have been unsuccessful.5b, 8 Looking across the steroid receptor family, partial agonism was key in the development of selective ER modulators9 (SERMs) to achieve tissue and/or pathway specific pharmacology. In accordance to this, we adopted a strategy hypothesizing that a potent GR modulator with partial efficacy in the TA assay and full efficacy in the TR assay would be well positioned to provide differentiation. Compounds with such a mechanistic profile would then be progressed to functional human cell assays measuring markers of anti-inflammatory and adverse effects focusing on hyperglycemia and bone resorption.

A

2 R=PO3 H 3 R=H

1

B

4

5

ACS Paragon Plus Environment

Journal of Medicinal Chemistry 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Figure 1 (A) Oral GR modulators that have entered clinical trials: BI 653,0483e, f (1) and fosdagrocorat3g (2) (prodrug of 3) (B) tool compounds used in design: AZD290610 (4) and mifepristone (5)

Compound design and hypothesis for introducing partial agonist/antagonist activity. GR ligands bind in a fully occluded pocket in the GR ligand binding domain (LBD)11 and allosterically modulate the receptor conformation. The resulting structural state controls the coregulator recruitment and DNA interaction pattern and ultimately the functional response. A large number of GR-ligand complex structures have been published to date.12 They provide a solid foundation for understanding the molecular drivers of receptor activation. We have previously described the discovery of phenyl indazole based non-steroidal GR modulators, where compound 4 (Figure 1), displayed excellent in vivo efficacy in a rat model of joint inflammation after oral dosing.10 With respect to its side effect profile 4 was found to have limited advantage compared to prednisolone and was not progressed further. However, this chemical series provided a good starting point for designing compounds with a potential for a differentiated functional profile. The X-ray structure of GR LBD in complex with 4 clearly indicates that it maintains key pharmacophore elements for receptor activation (Figure 2). The amide functionality interacts with Asn564 and Gln642 and the p-fluoro phenyl residue extends through the gatekeeper region by rearranging Gln570 and Arg611, creating non-polar contacts to helices 3 and 5 (Figure 2A). Compound 4 exhibits full efficacy in the reporter gene assay measuring transrepression13 (TR) (Table 1), full efficacy in the transactivation assay14 (TAag), and was completely inactive in concentrations up to 1 µM in the reporter gene assay measuring inhibition of dexamethasone induced transactivation15 (TAantag). Following the project hypothesis, namely to identify

ACS Paragon Plus Environment

Page 4 of 57

Page 5 of 57 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Journal of Medicinal Chemistry

compounds with partial efficacy for TA (defined as 50% efficacy in TAantag), but full efficacy in the TR assay (>80% efficacy), we needed to modulate the mechanistic profile of 4.

Figure 2. (A) The X-ray structure of GR LBD (green) in complex with compound 4 (magenta). Key residues in the ligand binding pocket are highlighted in orange stick representation. Putative hydrogen bonds are marked with dashed lines. (B) Superimposition of GR LBD in complex with 5 (white, PDB accession code 3H52) onto GR LBD in complex with 4 (magenta).

The structural overlay of GR LBD in complex with 4 onto the structure of GR LBD in complex with the antagonist 516 (Mifepristone, Figure 1) is shown in Figure 2B. While both compounds occupy the central volume of the ligand binding pocket, they extend into different sub-pockets. Specifically, the dimethyl-amino phenyl residue of 5 pushes towards Leu753 in helix 12. Helix 12 is part of the activation-function 2 (AF-2), which is a key element for interaction with coregulator proteins and ultimately the genomic response. The presence of the dimethyl-amino phenyl residue results in a considerable rearrangement observed in this region16 that largely

ACS Paragon Plus Environment

Journal of Medicinal Chemistry 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

accounts for the antagonistic properties of 5.12, 17 The structural comparison showed that the methyl group of 4 is positioned along the same vector as the dimethyl-amino phenyl residue in 5. Expansion of the compound along this axis could potentially modulate the functional response in a similar manner. Indeed, replacement of the methyl group in compound 4 by benzyl (6), i-propyl (7) and t-butoxymethyl (8) residues led to an increased efficacy in the TAantag assay, while, at the same time, the efficacy in the TAag assay was decreased (Table 1). Unfortunately, the introduction of more lipophilic groups towards helix 12 reduced the aqueous solubility to below 0.01 µM. At the same time, replacing the central methoxy pyridine group of compound 7 with a phenyl group 9 and 10 had little or no effect on the agonist/antagonist profile, and therefore phenyl or fluoro substituted phenyls were used for further SAR exploration. We have previously reported that indazole ether based ligands bearing heterocyclic amides capable of making bidentate interactions to Asn564 are potent and efficacious compounds in the TR assay.10 These amides also render the compounds less lipophilic. Introduction of a medium sized helix 12 push group, namely the 1-hydroxy ethyl residue shown in 11, yielded a compound with favorable potency in both TAag and TAantag assays, 3.5 nM and 270 nM, respectively. Moreover, compound 11 had a potency of 1.6 nM and an efficacy of 88% in the TR assay, being stable in rat hepatocytes although the solubility was still low (0.9 µM).

ACS Paragon Plus Environment

Page 6 of 57

Page 7 of 57 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Journal of Medicinal Chemistry

Table 1.

6-8

9-10

GRa IC50 nM

compound

11

Cell TAa Agonistb EC50 nM

(eff%)

Cell TRa,d

Antagonistc IC50 nM (eff %)

IC50 nM

cLogP / Sol (µM) (eff %)

7.0

110 (98)

NV

40 (99)

1.4 / 310

1

19

54%@1 µM

18%@1 µM

84 (75)

2.7 / 160

3