Brief Article pubs.acs.org/jmc
A Highly Selective Hydantoin Inhibitor of Aggrecanase‑1 and Aggrecanase‑2 with a Low Projected Human Dose Timothy B. Durham,* Jothirajah Marimuthu, James L. Toth, Chin Liu, Lisa Adams, Daniel R. Mudra, Craig Swearingen, Chaohua Lin, Mark G. Chambers, Kannan Thirunavukkarasu, and Michael R. Wiley Eli Lilly and Company, Lilly Corporate Center, Indianapolis, Indiana 46285, United States S Supporting Information *
ABSTRACT: Aggrecanase-1 and -2 (ADAMTS-4 and ADAMTS-5) are zinc metalloproteases involved in the degradation of aggrecan in cartilage. Inhibitors could provide a means of altering the progression of osteoarthritis. We report the identification of 7 which had good oral pharmacokinetics in rats and showed efficacy in a rat chemical model of osteoarthritis. The projected human dose required to achieve sustained plasma levels ≥10 times the hADAMTS-5 IC50 is 5 mg q.d.
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able to demonstrate that a ≥50% reduction in synovial biomarkers of aggrecan cleavage was achieved at doses capable of sustaining plasma levels of the inhibitors ≥10-fold their IC50 in rat plasma.5b This suggested that selective inhibitors able to achieve Cmin ≥ 10-fold the plasma IC50 with a minimal Cmax at reasonable doses would be attractive candidates for clinical OA investigation.5b In this report, we share the further optimization of this hydantoin scaffold culminating in the identification of a molecule with an improved projected human dose and lower risk of reactive metabolite formation compared to our previous leading compound (1, Figure 1)
INTRODUCTION ADAMTS-4 and ADAMTS-5 (aggrecanase-1 and aggrecanase2) are zinc metalloproteases known to have a specific and primary role in aggrecan degradation.1 Aggrecan is the component of cartilage that provides compressibility. This compressibility is derived from the highly glycosylated nature of the protein that results in high levels of hydration. Aggrecan loss would be expected to increase the mechanical trauma the joint undergoes during movement, leading to structural damage and inflammation. Thus, inhibition of ADAMTS-4 and ADAMTS-5 action has been suggested as a potential opportunity for therapeutic intervention in osteoarthritis (OA).2 This hypothesis is supported by data generated in genetically modified mice and human chondrocytes.3 A key challenge to clinical testing of aggrecanase inhibitors is the need to achieve high levels of selectivity against the related matrix metalloprotease (MMP) enzymes.4 It has been clinically established that broad spectrum inhibition of MMPs leads to musculoskeletal syndrome in patients. The active sites of the MMPs have high degrees of structural homology to the aggrecanase enzymes. In previous reports, we disclosed a class of hydantoins (i.e., 1, Figure 1) found to be dual inhibitors of ADAMTS-4 and ADAMTS-5.5 These compounds displayed high levels of selectivity against a number of MMPs. We also showed that compounds that were active in an acute monoiodoacetate (MIA) pharmacodynamic model of OA were also active in the much longer and resource intensive meniscal-tear (MT) surgical model.5a These inhibitors were efficacious in the MT model at the MIA ED50 dose. Using osmotic pumps, we were © 2017 American Chemical Society
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CHEMISTRY The hydantoin core was prepared according to our previously described approach.5b The chiral amides were synthesized by acylation of auxiliary 9 with commercially available acid chlorides 10−13 (Scheme 1). The resulting imide was then alkylated at −78 °C using 4-trifluoromethylbenzyl bromide. Removal of the chiral auxiliary gave acids 22−25. Amide formation with hydantoin 265b completed the synthesis of inhibitors 2 and 5−7. Diastereomer 3 (Figure 1) was prepared by the same route from ent-9. Dimethylamide 4 was prepared starting from commercial phosphonate 27 (Scheme 2). Olefination6 followed by hydrogenation and subsequent alkylation with methyl iodide7 provided the geminally substituted ester. This was hydrolyzed Received: May 2, 2017 Published: June 14, 2017 5933
DOI: 10.1021/acs.jmedchem.7b00650 J. Med. Chem. 2017, 60, 5933−5939
Journal of Medicinal Chemistry
Brief Article
Scheme 1a
a Conditions: (a) DMAP, Et3N, CH2Cl2, 0 °C to rt, 50−85%; (b) LDA, 4-trifluoromethylbenzyl bromide, THF, −78 °C, 40−50%; (c) H2O2, LiOH, THF/water, 0 °C, quant; (d) HATU, i-Pr2NEt, DCM/ DMF, rt, 50−80%.
Figure 1. Inhibitors of aggrecanase.
Scheme 2a
to give carboxylic acid 29.8 Carbodiimide mediated amide bond formation provided hydantoin 4.
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RESULTS AND DISCUSSION In our previous report, we identified 1 as a lead compound for development of a clinically useful aggrecanase inhibitor based on its potency and projected human PK profile (Figure 1).5b We conducted a bile duct cannulated rat study to understand metabolism of 1. This experiment revealed glutathione conjugation was the sole metabolic clearance pathway (see Supporting Information). The glutathione conjugates were arising via reactive oxidation products derived from the benzofuran moiety. Significant glutathione depletion could result in contraindication to acetaminophen. As we further developed this series, we wanted to minimize glutathione conjugation and lower the projected human dose to 100000 >100000 3
9 70 5 9 7 71 26 3400 270 100000 47000 1
7600 >50000 3400 3900 1600 15000 8000 >100000 50000 150
Values are an average of n = 3 or more data points. 5935
DOI: 10.1021/acs.jmedchem.7b00650 J. Med. Chem. 2017, 60, 5933−5939
Journal of Medicinal Chemistry
Brief Article
Figure 5. Projected human PK for 7. Solid red line = simulated human PK at steady state following 5 mg po q.d. Black lines = projection ± 30% CV. Dashed line = hADAMTS-5 IC90.
Figure 3. Model of 6 overlaid with ADAMTS-4, MMP-12, and MMP14: 6 (orange); ADAMTS-4 (gray with solid surface), Met395 (stick); MMP2 (1QIB)9 = magenta with wire surface, TYR223 (line); MMP12 (1JK3)10 = cyan with wire surface, TYR240 (line). Overlays were generated using the sequence overlay tool in MOE.
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EXPERIMENTAL SECTION
General. Protocols for the in vitro assays are included in the Supporting Information. All animals were maintained in accordance with the Institutional Animal Care and Use Committee of Eli Lilly and the National Institutes of Health Guide for the Care and Use of Laboratory Animals. MIA Assay. This assay has been described in detail in the literature.11 Briefly, 7- to 8-week-old male Lewis rats are anesthetized and injected intra-articularly with either 3 mg of freshly prepared MIA (sodium salt) in 50 μL of 0.9% saline into the right knee or 0.9% saline alone in the left knee on day 0. This induced aggrecanase activity and release of aggrecanase specific fragments into the synovial fluid. Aggrecanase inhibitor or vehicle [1% hydroxyethyl cellulose; 0.25% Tween 80; 0.05% antifoam] is dosed po, b.i.d. starting from day 3. A single dose of compound is given on day 7, the animals are sacrificed 4 h later, and the knee joints are lavaged with 200 μL of saline. The synovial lavage is assayed for aggrecanase-cleaved fragments of aggrecan using the NITEGE/ARGNV sandwich ELISA (see Supporting Information). The amounts of aggrecan fragments present in the synovial lavage are determined based on a standard curve generated with aggrecanase-digested rat aggrecan. Statistical analysis is performed using Dunnett’s test. Chemistry: General Methods. 1H and 13C NMR spectra were measured in deuterated solvents at 400 and 100 MHz, respectively, on a Varian or Bruker spectrometer. Data are given as δ. All solvents were dry. Reagents were obtained from commercial sources and used as received. Purity of final compounds used in biological assays was determined by electrospray LCMS to be ≥95% (see Supporting Information for method and data). Compounds 2, 3, 5, and 6 were prepared according to the same general procedure as 7. (R)-N-(((R)-4-Cyclopropyl-2,5-dioxoimidazolidin-4-yl)methyl)-2-methyl-3-(4-(trifluoromethyl)phenyl)propanamide (2). [α]D23 −14.10° (c 1.0, MeOH). 1H NMR (DMSO): 10.58−10.56 (m, 1H), 7.91−7.86 (m, 1H), 7.59 (d, J = 8.1 Hz, 2H), 7.44 (s, 1H), 7.35 (d, J = 7.9 Hz, 2H), 3.43−3.35 (m, 2H), 2.95−2.91 (m, 1H), 2.65−2.61 (m, 1H), 2.48−2.46 (m, 1H), 1.03−0.99 (m, 1H), 0.90 (d, J = 6.8 Hz, 3H), 0.42−0.39 (m, 4H). 13C (DMSO) 176.76, 175.81 157.41, 145.62, 129.99, 127.31, 127.00, 125.54, 125.50 (m), 125.47, 65.93, 43.67, 78, 14.61, 0.78, −0.70 (S)-N-(((R)-4-Cyclopropyl-2,5-dioxoimidazolidin-4-yl)methyl)-2-methyl-3-(4-(trifluoromethyl)phenyl)propanamide (3). Prepared according to the same sequence as 2 beginning with (R)4-benzyl-2-oxazolidinone. 1H NMR (DMSO): 10.56−10.54 (m, 1H), 7.93−7.89 (m, 2H), 7.60−7.57 (m, 2H), 7.38−7.35 (m, 3H), 3.57− 3.52 (m, 1H), 3.27−3.26 (m, 1H), 2.97−2.93 (m, 1H), 2.66−2.65 (m, 1H), 2.56−2.54 (m, 1H), 1.00−0.98 (m, 1H), 0.93−0.91 (d, J = 6.8 Hz, 3H), 0.52−0.46 (m, 3H), 0.13−0.08 (m, 1H). 13C (DMSO): 176.91, 175.87, 157.38, 145.64, 130.06, 127.31, 127.00, 126.28, 125.40 (m), 123.57, 65.95, 43.55, 41.18, 39.26, 18.17, 14.39, 0.78, −0.72. (R)-N-((4-Cyclopropyl-2,5-dioxoimidazolidin-4-yl)methyl)2,2-dimethyl-3-(4-(trifluoromethyl)phenyl)propanamide (4).
Compound 7 was evaluated in our rat MIA pharmacodynamics model (Figure 4).11 This model simulates OA in an
Figure 4. Rat MIA assay: (∗) p ≤ 0.05.
acute time frame through chemical injury to the joint, resulting in release of large amounts of proteases, including aggrecanases. We have developed ELISA assays to detect both fragments (NITEGE or ARGNV) of aggrecan cleavage by ADAMTS-5.12 With oral b.i.d. dosing, 7 lowered aggrecan degradation products in synovial fluid at all doses tested, performing as robustly as the broad spectrum metalloprotease inhibitor 8.13 Further, consistent with the PK/PD relationship defined using 1, ED50 effects were delivered by doses capable of sustaining ≥10 times the plasma ADAMTS-5 IC50. We estimated the human PK of 7 (see Supporting Information for details). On the basis of our previous work,5b we modeled the dose at which the sustained human plasma concentration would be ∼10 times the hADAMTS-5 IC50 (a concentration equivalent to the ED50 in the MIA model). As shown in Figure 5, a 5 mg q.d. dose is projected to achieve this plasma level in humans, a meaningful improvement from the projected dose of 1 (45 mg, q.d.).
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CONCLUSIONS The hydantoin scaffold5 has proven to be a good platform for development of potent and selective ADAMTS-4 and ADAMTS-5 inhibitors. Compound 7 has been identified as a molecule with an attractive preclinical efficacy and projected human PK profile. 5936
DOI: 10.1021/acs.jmedchem.7b00650 J. Med. Chem. 2017, 60, 5933−5939
Journal of Medicinal Chemistry
Brief Article
The mixture was cooled to 0 °C. Et3N (3 equiv, 147.00 mmol, 20.49 mL) was added followed by dropwise addition of 2-cyclopropylacetyl chloride (49 mmol, 5.47 mL). The reaction was allowed to warm to ambient temperature overnight. The reaction was diluted with CH2Cl2, washed with water, saturated NaCl solution, dried over MgSO4, filtered, and concentrated. The residue was purified by flash chromatography (gradient 0−30% THF/hexanes) to give the compound as a yellow oil (6.48 g, 51%) which was used directly in the next reaction. To a dry flask under nitrogen was added i-Pr2NH (4.6 mL, 33 mmol). To the amine was slowly added 2.5 M n-BuLi in hexanes (12.5 mL, 31 mmol). The LDA reagent was diluted with dry THF (20 mL). In a separate flask under N2, imide 17 (6.5 g, 25 mmol) was dissolved in THF (160 mL) and cooled to −78 °C. The LDA solution was slowly added via syringe such that the internal temperature did not rise above −70 °C. After the addition, the mixture was stirred for 1 h. To the resulting anion was added dropwise 4-(trifluoromethyl)bromomethylbenzene (6.6 g, 27.5 mmol) in THF (20 mL) such that the internal temperature did not rise above −70 °C. The cooling bath was allowed to expire overnight. The reaction was quenched with saturated NH4Cl solution and extracted with EtOAc. The extracts were combined, washed with saturated NaCl solution, dried over MgSO4, filtered, and concentrated. The residue was purified by chromatography (gradient 0−70% EtOAc/hexanes) to give 21 as a yellow oil (4.8 g, 46%) which was used directly in the next reaction. Compound 21 (4.82 g, 11.6 mmol) was dissolved in 1:2 THF/water (300 mL) and cooled to 0 °C. To the resulting solution was added 30% aqueous H2O2 (15 mL, 150 mmol) followed by 1 M LiOH solution (20 mL, 20 mmol). The resulting mixture was stirred at 0 °C for 2 h. The reaction was quenched via dropwise addition of Na2S2O3 (5.82 g, 46 mmol) in water (40 mL). The reaction was slowly warmed to ambient temperature. The THF was removed on a rotary evaporator. The aqueous solution was washed with dichloromethane (×2). The solution was acidified with 2 M HCl and extracted with EtOAc. The extracts were combined, washed with saturated NaCl, dried (MgSO4), filtered, and concentrated to give the title compound as an oil (3 g, quantitative). 1H NMR (DMSO): 12.14−12.07 (bs, 1H), 7.59 (d, J = 8.0 Hz, 2H), 7.39 (d, J = 7.9 Hz, 2H), 2.99−2.88 (m, 2H), 1.89−1.83 (m, 1H), 0.92−0.89 (m, 1H), 0.42−0.40 (m, 2H), 0.27−0.25 (m, 1H), 0.13−0.08 (m, 1H). (R)-N-((4-Cyclopropyl-2,5-dioxoimidazolidin-4-yl)methyl)-3(4-(trifluoromethyl)phenyl)propanamide (31). Prepared according to the same general procedure as 7 from the commercially available carboxylic acid and amine 26.5b 1H NMR (DMSO): 10.59 (s, 1H), 7.93 (t, J = 6.1 Hz, 1H), 7.62 (d, J = 8.1 Hz, 2H), 7.50 (s, 1H), 7.41 (d, J = 8.0 Hz, 2H), 3.49−3.37 (m, 2H), 2.88 (at, J = 7.6 Hz, 2H), 2.45− 2.41 (m, 2H), 1.08−1.01 (m, 1H), 0.46−0.37 (m, 3H), 0.10 (td, J = 9.6, 5.3 Hz, 1H). 13C (DMSO): 176.81, 172.19, 157.37, 146.81, 129.43, 127.59, 127.28, 126.96, 126.65, 126.27, 125.60 (m), 123.57, 65.76, 43.76, 36.58, 31.19, 14.63, 0.78, −0.73.
Hydantoin 265b (205 mg, 1 mmol) was dissolved in 15 mL of dry CH3CN under nitrogen. To the resulting solution was added i-Pr2NH (387 mg, 3 mmol), 2,2-dimethyl-3-[4-(trifluoromethyl)phenyl]propanoic acid14 (246 mg, 1 mmol), EDCI (229 mg, 1.2 mmol), and HOAT (163 mg, 1.2 mmol). The resulting mixture was stirred for 12 h at ambient temperature. Solvents were removed on a rotary evaporator and the residue was purified by reverse phase flash chromatography to give the title compound (282 mg, 70%). 1H NMR (DMSO): 10.63 (s, 1H), 7.62 (d, J = 8.1 Hz, 2H), 7.47 (t, J = 6.1 Hz, 1H), 7.37 (s, 1H), 7.33 (d, J = 8.0 Hz, 2H), 3.60 (dd, J = 6.9, 13.6 Hz, 1H), 3.34 (m, 1H), 2.92−2.89 (m, 1H), 2.86−2.83 (m, 1H), 1.05 (s, 3H), 1.04 (s, 3H), 1.02 (m, 1H), 0.42−0.33 (m, 3H), 0.08 (td, J = 9.4, 5.3 Hz, 1H). 13C (DMSO): 177.04, 157.53, 143.81, 131.25, 127.45, 127.14, 126.29, 125.10 (m), 123.59, 65.90, 45.17, 44.39, 43.23, 25.38, 25.24, 14.32, 0.96, −0.87. (R)-N-(((R)-4-Cyclopropyl-2,5-dioxoimidazolidin-4-yl)methyl)-2-(4-(trifluoromethyl)benzyl)butanamide (5). [α]D23 −22.10° (c 1.0, MeOH). 1H NMR (DMSO): 10.56−10.54 (m, 1H), 7.89−7.87 (m, 2H), 7.59 (d, J = 8.2 Hz, 3H), 7.43−7.41 (m, 1H), 7.35−7.33 (d, J = 8.0 Hz, 2H), 3.56−3.54 (m, 1H), 3.29−3.28 (m, 1H), 2.90−2.87 (m, 2H), 2.65−2.64 (m, 1H), 1.53−1.48 (m, 1H), 1.33−1.30 (m, 1H), 1.03−1.01 (m, 1H), 0.72 (t, J = 7.4 Hz, 3H), 0.42−0.40 (m, 3H), 0.14−0.10 (m, 1H). 13C (DMSO): 176.70, 174.89, 157.39, 145.75, 129.96, 127.26, 126.94, 125.47, 123.59, 65.87, 48.61, 43.52, 38.16, 25.39, 14.76, 11.99, 0.74, −0.68. (S)-N-(((R)-4-Cyclopropyl-2,5-dioxoimidazolidin-4-yl)methyl)-3-methyl-2-(4-(trifluoromethyl)benzyl)butanamide (6). [α]D23 −38.90° (c 1.0, MeOH). 1H NMR (DMSO): 10.50 (s, 1H), 7.68 (t, J = 6.0 Hz, 1H), 7.55 (d, J = 8.1 Hz, 2H), 7.37 (d, J = 0.8 Hz, 1H), 7.31 (d, J = 7.9 Hz, 2H), 3.59−3.54 (m, 1H), 3.10−3.05 (m, 1H), 2.90−2.87 (m, 1H), 2.71−2.67 (m, 1H), 2.37−2.32 (m, 1H), 1.73−1.71 (m, 1H), 0.97−0.92 (m, 1H), 0.88 (d, J = 6.8 Hz, 2H), 0.83 (d, J = 6.7 Hz, 2H), 0.37−0.32 (m, 3H), 0.09−0.05 (m, 1H). 13C (DMSO): 176.64, 174.18, 157.32, 146.31, 129.82, 127.09, 126.77, 126.31, 125.39, 123.61, 65.69, 53.57, 43.43, 35.07, 31.02, 20.55, 20.50, 14.89, 0.69, −0.71. (S)-2-Cyclopropyl-N-(((R)-4-cyclopropyl-2,5-dioxoimidazolidin-4-yl)methyl)-3-(4-(trifluoromethyl)phenyl)propanamide (7). Salt 265b (2.33 g, 11.3 mmol), acid 25 (2.66 g, 10.3 mmol), and HATU (4 g, 10.3 mmol) were dissolved in dry 5:1 CH2Cl2/DMF under nitrogen. To the resulting mixture was added iPr2NEt amine (5.6 mL, 51.5 mmol). After 48 h, solvents were removed and the residue was purified by chromatography (C18, gradient 10−70% CH3CN/water + 0.1% TFA) to give 7 as a white solid (3.1 g, 73%). [α]D23 3.30° (c 1.0, MeOH). 1H NMR (DMSO): 10.56−10.54 (m, 1H), 7.71−7.66 (m, 1H), 7.57−7.55 (d, J = 8.4 Hz, 2H), 7.41−7.37 (m, 1H), 7.34−7.32 (d, J = 8.0 Hz, 2H), 3.51−3.46 (dd, J = 6.0, 13.6 Hz, 1H), 3.28−3.24 (dd, J = 1.2, 13.6 Hz, 1H), 3.05− 3.03 (dd, J = 8.0, 13.6 Hz, 1H), 2.79−2.76 (dd, J = 5.6, 13.6 Hz, 1H), 1.87−1.84 (m, 1H), 1.02−0.98 (m, 1H), 0.85−0.83 (m, 1H), 0.39− 0.37 (m, 7H), 0.11−0.09 (m, 1H), −0.08--0.10 (m, 1H). 13C (DMSO) δ: 176.63, 174.41, 157.38, 145.70, 129.96, 127.18, 126.87, 126.31, 125.34 (m), 65.77, 51.76, 43.54, 38.18, 14.93, 14.71, 4.99, 3.64, 0.75, −0.70. Carboxylic acids 22−24 were made via the same general procedure as compound 25. (R)-2-Methyl-3-(4-(trifluoromethyl)phenyl)propanoic Acid (22). 1H NMR (DMSO): 12.19 (s, 1H), 7.60 (d, J = 8.1 Hz, 2H), 7.39 (d, J = 8.1 Hz, 2H), 2.97−2.91 (m, 1H), 2.72−2.63 (m, 2H), 1.03 (d, J = 6.8 Hz, 3H). (R)-2-(4-(Trifluoromethyl)benzyl)butanoic Acid (23). 1H NMR (DMSO): 12.18−12.12 (bs, 1H), 7.60 (d, J = 8.2 Hz, 2H), 7.38 (s, 2H), 2.88−2.74 (m, 2H), 2.53−2.48 (m, 1H), 1.52−1.46 (m, 2H), 0.86−0.83 (m, 3H). (S)-3-Methyl-2-(4-(trifluoromethyl)benzyl)butanoic Acid (24). 1H NMR (DMSO): 12.06 (s, 1H), 7.59 (d, J = 8.1 Hz, 2H), 7.38 (d, J = 8.0 Hz, 2H), 2.87−2.74 (m, 2H), 2.41−2.36 (m, 1H), 1.87−1.78 (m, 1H), 0.96 (d, J = 6.8 Hz, 3H), 0.92 (d, J = 6.7 Hz, 3H). (S)-2-Cyclopropyl-3-(4-(trifluoromethyl)phenyl)propanoic Acid (25). To a flask under nitrogen were added 9 (49 mmol, 8.68 g), CH2Cl2 (160 mL), and DMAP (0.1 equiv, 4.90 mmol, 598.63 mg).
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ASSOCIATED CONTENT
S Supporting Information *
The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.jmedchem.7b00650. Bile duct cannulated rat study, assay protocols, human PK modeling (PDF) Molecular formula strings and some data (CSV) PDB information for compounds in Figure 2 (PDB) PDB information for compounds in Figure 3 (PDB)
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AUTHOR INFORMATION
Corresponding Author
*E-mail:
[email protected]. Phone: 317-433-8447. ORCID
Timothy B. Durham: 0000-0003-4124-1188 5937
DOI: 10.1021/acs.jmedchem.7b00650 J. Med. Chem. 2017, 60, 5933−5939
Journal of Medicinal Chemistry
Brief Article
Author Contributions
Neeb, M.; Thompson, M. M.; Lohr, T.; Gross, J. W.; Centrella, P. A.; O’Donovan, G. K.; Bedard, K. L.; van Vloten, K.; Mataruse, S.; Skinner, S. R.; Belyanskaya, S. L.; Carpenter, T. Y.; Shearer, T. W.; Clark, M. A.; Cuozzo, J. W.; Arico-Muendel, C. C.; Morgan, B. A. Discovery of highly potent and selective small molecule ADAMTS-5 inhibitors that inhibit human cartilage degradation via encoded library technology (ELT). J. Med. Chem. 2012, 55, 7061−7079. (d) Ding, Y.; O’Keefe, H.; DeLorey, J. L.; Israel, D. I.; Messer, J. A.; Chiu, C. H.; Skinner, S. R.; Matico, R. E.; Murray-Thompson, M. F.; Li, F.; Clark, M. A.; Cuozzo, J. W.; Arico-Muendel, C.; Morgan, B. A. Discovery of potent and selective inhibitors for ADAMTS-4 through DNA-encoded library technology (ELT). ACS Med. Chem. Lett. 2015, 6, 888−893. (5) (a) Durham, T. B.; Klimkowski, V. J.; Rito, C. J.; Marimuthu, J.; Toth, J. L.; Liu, C.; Durbin, J. D.; Stout, S. L.; Adams, L.; Swearingen, C.; Lin, C.; Chambers, M. G.; Thirunavukkarasu, K.; Wiley, M. R. Identification of potent and selective hydantoin inhibitors of aggrecanase-1 and aggrecanase-2 that are efficacious in both chemical and surgical models of osteoarthritis. J. Med. Chem. 2014, 57, 10476− 10485. (b) Wiley, M. R.; Durham, T. B.; Adams, L. A.; Chambers, M. G.; Lin, C.; Liu, C.; Marimuthu, J.; Mitchell, P. G.; Mudra, D. R.; Swearingen, C. A.; Toth, J. L.; Weller, J. M.; Thirunavukkarasu, K. Use of osmotic pumps to establish the pharmacokinetic-pharmacodynamic relationship and define desirable human performance characteristics for aggrecanase inhibitors. J. Med. Chem. 2016, 59, 5810−5822. (6) (a) Wang, Q.; Liu, X.; Liu, X.; Li, B.; Nie, H.; Zhang, S.; Chen, W. Highly enantioselective hydrogenation of 2-substituted-2-alkenols catalysed by a chenphos-rh complex. Chem. Commun. (Cambridge, U. K.) 2014, 50, 978−980. (b) Schomaker, J. M.; Bhattacharjee, S.; Yan, J.; Borhan, B. Diastereomerically and enantiomerically pure 2,3disubstituted pyrrolidines from 2,3-aziridin-1-ols using a sulfoxonium ylide: A one-carbon homologative relay ring expansion. J. Am. Chem. Soc. 2007, 129, 1996−2003. (c) Malkov, A. V.; Czemerys, L.; Malyshev, D. A. Vanadium-catalyzed asymmetric epoxidation of allylic alcohols in water. J. Org. Chem. 2009, 74, 3350−3355. (7) Wang, Z.; Kuninobu, Y.; Kanai, M. Copper-mediated direct C(SP3)-H and C(SP2)-H acetoxylation. Org. Lett. 2014, 16, 4790− 4793. (8) Durham, T. B.; Marimuthu, J.; Wiley, M. R. Preparation of N(imidazolidinylmethyl)propanamides as aggrecanase inhibitors. WO2014066151 A1, 2014. (9) Dhanaraj, V.; Williams, M. G.; Ye, Q.-Z.; Molina, F.; Johnson, L. L.; Ortwine, D. F.; Pavlovsky, A.; Rubin, J. R.; Skeean, R. W.; White, A. D.; Humblet, C.; Hupe, D. J.; Blundell, T. L. X-ray structure of gelatinase A catalytic domain complexed with a hydroxamate inhibitor. Croat. Chem. Acta 1999, 72, 575−591. (10) Lang, R.; Kocourek, A.; Braun, M.; Tschesche, H.; Huber, R.; Bode, W.; Maskos, K. Substrate specificity determinants of human macrophage elastase (MMP-12) based on the 1.1 Å crystal structure1. J. Mol. Biol. 2001, 312, 731−742. (11) Swearingen, C. A.; Chambers, M. G.; Lin, C.; Marimuthu, J.; Rito, C. J.; Carter, Q. L.; Dotzlaf, J.; Liu, C.; Chandrasekhar, S.; Duffin, K. L.; Mitchell, P. G.; Durham, T. B.; Wiley, M. R.; Thirunavukkarasu, K. A short-term pharmacodynamic model for monitoring aggrecanase activity: Injection of monosodium iodoacetate (MIA) in rats and assessment of aggrecan neoepitope release in synovial fluid using novel elisas. Osteoarthr. Cartil. 2010, 18, 1159−1166. (12) Swearingen, C. A.; Carpenter, J. W.; Siegel, R.; Brittain, I. J.; Dotzlaf, J.; Durham, T. B.; Toth, J. L.; Laska, D. A.; Marimuthu, J.; Liu, C.; Brown, D. P.; Carter, Q. L.; Wiley, M. R.; Duffin, K. L.; Mitchell, P. G.; Thirunavukkarasu, K. Development of a novel clinical biomarker assay to detect and quantify aggrecanase-generated aggrecan fragments in human synovial fluid, serum and urine. Osteoarthr. Cartil. 2010, 18, 1150−1158. (13) Noe, M. C.; Natarajan, V.; Snow, S. L.; Wolf-Gouveia, L. A.; Mitchell, P. G.; Lopresti-Morrow, L.; Reeves, L. M.; Yocum, S. A.; Otterness, I.; Bliven, M. A.; Carty, T. J.; Barberia, J. T.; Sweeney, F. J.; Liras, J. L.; Vaughn, M. Discovery of 3-oh-3-methylpipecolic hydroxamates: Potent orally active inhibitors of aggrecanase and MMP-13. Bioorg. Med. Chem. Lett. 2005, 15, 3385−3388.
T.B.D. wrote the manuscript. All authors reviewed and approved the manuscript. Design of inhibitors: T.B.D., J.M., M.R.W. Synthesis: J.M., J.L.T., C.L. Design of in vivo experiments: K.T., M.G.C., M.R.W., T.B.D., L.A. Biochemical and biomarker assays: C.S., K.T. MIA experiments: C.L. Human PK projection: D.R.M. Modeling: T.B.D. Notes
The authors declare the following competing financial interest(s): All the authors are employees and shareholders of Eli Lilly and Company.
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ABBREVIATIONS USED ADAMTS, a disintegrin and metalloproteinase with thrombospondin motifs; MMP, matrix metalloprotease; OA, osteoarthritis; MIA, monoiodoacetate; MT, meniscal tear
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DOI: 10.1021/acs.jmedchem.7b00650 J. Med. Chem. 2017, 60, 5933−5939
