The discovery of BMS-457, a potent and selective CCR1 antagonist

Article abstract of DOI:10.1016/j.bmcl.2013.04.079

A series of compounds which exhibited good human CCR1 binding and functional potency was modified resulting in the discovery of a novel series of high affinity, functionally potent antagonists of the CCR1 receptor. Issues of PXR activity, ion-channel pote

Full text of DOI:10.1016/j.bmcl.2013.04.079

Accepted Manuscript
The discovery of BMS-457, a potent and selective CCR1 antagonist
Daniel S. Gardner, Joseph B. Santella, John V. Duncia, Percy H. Carter, T.G.
Murali Dhar, Hong Wu, Weiwei Guo, Cullen Cavallaro, Katy Van Kirk, Melissa
Yarde, Stephanie W. Briceno, R. Robert Grafstrom, Richard Liu, Sima R. Patel,
Andrew J. Tebben, Dan Camac, Javed Khan, Andrew Watson, Guchen Yang,
Anne Rose, William R. Foster, Mary Ellen Cvijic, Paul Davies, John Hynes
PII:
S0960-894X(13)00569-6
DOI:
http://dx.doi.org/10.1016/j.bmcl.2013.04.079
BMCL 20444
Reference:
To appear in:
Bioorganic & Medicinal Chemistry Letters
Received Date:
Revised Date:
Accepted Date:
22 February 2013
24 April 2013
29 April 2013
Please cite this article as: Gardner, D.S., Santella, J.B., Duncia, J.V., Carter, P.H., Murali Dhar, T.G., Wu, H., Guo,
W., Cavallaro, C., Van Kirk, K., Yarde, M., Briceno, S.W., Robert Grafstrom, R., Liu, R., Patel, S.R., Tebben, A.J.,
Camac, D., Khan, J., Watson, A., Yang, G., Rose, A., Foster, W.R., Cvijic, M.E., Davies, P., Hynes, J., The discovery
of BMS-457, a potent and selective CCR1 antagonist, Bioorganic & Medicinal Chemistry Letters (2013), doi: http://
dx.doi.org/10.1016/j.bmcl.2013.04.079
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The discovery of BMS-457, a potent and selective CCR1 antagonist
Daniel S. Gardner*, Joseph B. Santella, III, John V. Duncia, Percy H. Carter, T. G. Murali Dhar, Hong Wu, Weiwei Guo,
Cullen Cavallaro, Katy Van Kirk, Melissa Yarde, Stephanie W. Briceno, R. Robert Grafstrom, Richard Liu, Sima R. Patel,
Andrew J. Tebben, Dan Camac, Javed Khan, Andrew Watson, Guchen Yang, Anne Rose, William R. Foster, Mary Ellen
Cvijic, Paul Davies, and John Hynes, Jr.
Bristol-Myers Squibb Company, R&D, PO Box 4000, Princeton, NJ 08543-4000, USA
*Corresponding author. Tel.: +1 609 252 4233; Fax: +1 609 252 3993; email: daniel.gardner@bms.com
Abstract: A series of compounds which exhibited good human CCR1 binding and functional potency was modified
resulting in the discovery of a novel series of high affinity, functionally potent antagonists of the CCR1 receptor. Issues of
PXR activity, ion-channel potency, and poor metabolic stability were addressed by the addition of a hydroxyl group to an
otherwise lipophilic area in the molecule resulting in the discovery of preclinical candidate BMS-457 for the treatment of
rheumatoid arthritis.
The CCR1 receptor (CC Chemokine Receptor-1) is a 7 trans-membrane G-protein coupled receptor (7
TM GPCR) that is expressed on the surface of monocytes/macrophages, osteoclasts, T-cells, and neutrophils.1
This receptor interacts with eleven different chemokines, including MIP-1 (CCL3), RANTES (CCL5), and
Leukotactin-1 (CCL15), resulting in leukocyte activation and migration (chemotaxis), processes that are critical
for the progression of inflammatory diseases.2
Preclinical studies in rodents demonstrated that CCR1 receptor blockade, as with other chemokine
receptors,3,4,5 represents a viable strategy for the development of therapies for inflammatory disorders.6 Several
small molecule antagonists of CCR1 have appeared in the literature,7,8 and a number have advanced to human
clinical trials9 for inflammatory disorders such as multiple sclerosis,10,11 rheumatoid arthritis (RA),9,12-16 and
chronic obstructive pulmonary disease.17 Unfortunately, these molecules failed to achieve their targeted clinical
endpoints in Phase II studies. It has been suggested that the Phase II clinical experiments failed due to
inadequate blockade of the CCR1 receptor over the course of the trial.2b,5,18 In spite of these clinical failures, it

remains clear that CCR1 plays an important role in the migration of cells involved in the pathogenesis of human
RA.13,18 In a recent clinical study, CCR1 antagonist CCX354-C20 (structure unknown) showed clinical efficacy
in a three month Phase II RA study.19-22 This new data supports the further investigation of CCR1 antagonists to
treat RA.
We have also been working towards the discovery of a small-molecule CCR1 receptor antagonist for the
treatment of RA. In a previous publication,23 we described the discovery of a novel series of CCR1 antagonists
such as 1. In this series the preferred scaffold consisted of a 4-(4-chlorophenyl)piperidine “headpiece,” D-valine
“linker,” and terminal amide “tailpiece,” as shown in Figure 1. After extensive SAR development in which the
terminal amide tailpiece was varied it became evident that while single-digit nanomolar potency in our primary
binding assay could be achieved, functional potency (chemotaxis) meeting our criteria for advancement (IC50 < 25
nM) was difficult to attain (Table 1, compounds 1, 4a, 5a, 6a, 7a).
In our prior efforts to develop antagonists of chemokine receptors CCR324 and CCR2,25 a common feature
in preferred scaffolds was a basic nitrogen. Furthermore, many of the CCR1 antagonists previously disclosed in
the literature also incorporated a basic nitrogen, often within a piperidine or piperazine moiety.10,15,17b We
theorized that the functional potency of these antagonists could be due to a salt bridge interaction of this basic
nitrogen with a glutamic acid residue (E287) on CCR1,23 and that a similar effect could be achieved in our series
by converting the piperidine amide to a basic amine via the removal of the central amide carbonyl. However, due
to the reported neurotoxicity of N-alkyl-4-phenylpiperidines such as MPPP and its metabolites MPTP and MPP,26
we chose not to synthesize these targets. We reasoned that gem-dimethyl substitution on our piperidine would
prevent the formation of N-alkyl-pyridinium compounds and eliminate this potential toxicity, a strategy
employed by Sayre and coworkers in their studies of MPTP metabolites.27 Alkylation at the piperidine’s 3-
position was chosen for our initial targets due to ease of synthesis from N-BOC-4-piperidinone. The 4-hydroxy
substituent resulting from 4-chlorophenyllithium addition to the alkylated piperidone initially proved difficult to
remove and was thus maintained in our target (Scheme 1). Unfortunately, target compound 2 was significantly

less potent than 1. We were pleasantly surprised to find that reintroducing the central amide carbonyl (3) into
compound 2 resulted in enhanced binding and chemotaxis potency (Figure 2).
Following up on these results, a series of compounds utilizing the (S)-4-(4-chlorophenyl)-3,3-
dimethylpiperidin-4-ol headpiece, D-valine linker, and a variety of amide tailpieces was prepared. In most cases
the change resulted in improved binding and a substantial improvement in chemotaxis potency (Table 1, 1 vs. 3,
then a vs. b compound pairs). Strikingly, several of these new compounds showed single-digit nanomolar
inhibition of both CCR1 binding and MIP-1 induced chemotaxis (Table 1, 6b and 7b).
The results of further SAR development around the amide tailpiece are shown in Table 2. Amides with
alkyl substituents larger than a methyl group tended to be more potent than aryl amides in both binding and
chemotaxis, though single-digit nanomolar binding and chemotaxis activity could be achieved in both series. It
was particularly interesting to note the wide variety of terminal amides which were tolerated in the CCR1
receptor. For example, aryl substituents (8–10, 6b, 13, 15, and 16), alkyl substituents (17–19, 21–30), acidic
substituents, (13) and basic substituents (15, 16, and 30) all yielded compounds with single-digit nanomolar
binding affinity. Furthermore, there did not seem to be a significant steric constraint in this region, as large
substituents also yielded compounds with single-digit nanomolar binding affinity (29, 30). This led us to believe
that the terminal amide substituent was an area of the molecule which could be modified to address safety and
pharmacokinetics concerns.
Results from an effort to further optimize the piperidine headpiece are shown in Table 3. Entries 31–37
illustrate modifications at the piperidine’s 4-position. The (R)-enantiomer was 545-fold less potent (31 vs. 10).
Methylation of the oxygen resulted in the complete loss of activity (32). Replacing the oxygen with nitrogen
resulted in a 59-fold loss in potency, and further functionalization of this nitrogen resulted in the complete loss of

activity (33–36 vs. 10). Removal of the oxygen resulted in a compound with equivalent binding affinity of 3.7
nM, but 5-fold lower chemotaxis inhibition (37 vs. 15). We had hoped that the chiral piperidine-4-ol could be
replaced by an analogous but achiral piperazine. Unfortunately, this modification resulted in a 7-fold loss in
chemotaxis inhibition (38 vs. 27). Modification of the piperidine’s 3-position are illustrated in entries 39 and 40.
Expansion of the gem-dimethyl substitution pattern into a 5,6-spirocyclic system resulted in a 63-fold loss in
binding potency (39 vs. 10). Finally, contraction of the gem-dimethyl substituents into a 3,6-spirocyclic system
resulted in reasonable binding affinity, but a 3-fold loss in chemotaxis potency (40 vs. 15). The narrow SAR
shown in these examples suggested that the 4-arylpiperidine moiety occupies a tight binding pocket in the CCR1
receptor. Since none of these modifications yielded results superior to the parent compounds, we moved forward
with the gem-dimethylpiperidinol headpiece.
As a result of this SAR effort and subsequent liability profiling, several common issues were noted within
this series: 1) activation of the pregnane X receptor (PXR), potentially leading to upregulation of cytochrome
P450 enzyme CyP3A428; 2) inhibition of the calcium and sodium ion channels, which can lead to undesirable
cardiac events; 3) poor liver microsomal stability.
Comparing compound 3 (R = Ph) to compounds 6b and 13 it was noted that these liabilities were greatly
reduced or eliminated via the addition of a polar substituent to the terminal aryl ring (Table 4). Neither 6b nor 13
could be advanced due to poor pharmacokinetic properties (data not shown). Subsequent efforts to discover a
compound in the aryl series which met our criteria for advancement were unsuccessful due to loss in potency,
poor pharmacokinetics, or failure to address any of the concerns noted above.
We theorized that as seen in the aryl series, the addition of a polar substituent to an alkyl
tailpiece could result in similar improvements to the poor counter screen selectivity and metabolism
profile of an alkyl-amide while maintaining binding and chemotaxis potency. Compound 27 (R = c-
pentyl) was of great interest to us due to its excellent potency and long half-life on the receptor (t1/2 >
200 minutes at 25 oC, data not shown), so the cyclopentane scaffold was chosen for modification. The
results of these modifications are summarized in Table 4. When the cyclopentane’s 3-position was

substituted with a primary amine, three diastereomers (41–43) were significantly less potent than 27.
When the cyclopentane’s 2-position was substituted with a hydroxyl-group, the cis-isomer pair was
equipotent to the unsubstituted cyclopentane and ion-channel selectivity was improved, but PXR and
metabolic stability were unaffected (44 vs. 27). The trans-isomer pair was 14-fold less potent (45 vs.
27). Two of the isomers derived from 2-aminocyclopentanoic acid maintained reasonable CCR1
binding potency (46–47), and cis-isomer 46 maintained good chemotaxis potency (the absolute
stereochemistry of 46 was not determined). This modification improved metabolic stability in both
cases. PXR and ion-channel selectivity for 46 were improved compared to 27, but did not meet our
criteria for advancement (PXR EC50 > 50 M, sodium and calcium ion-channel IC50 > 80 M). The
four diastereomers of 3-hydroxycyclopentanoic acid were coupled to our scaffold, and we were
delighted to find that in all four cases the addition of this simple hydroxyl group yielded substantial
improvements in PXR selectivity, sodium and calcium channel activity, and metabolic stability while
maintaining potency in both binding and chemotaxis (48–51). Though improved vs. 27, the two cis-
diastereomers (50 and 51) still did not meet our criteria for advancement due to PXR activity. However,
in the case of both trans-diastereomers (48 and 49), PXR activity was essentially eliminated. Since R,R-
isomer 48, BMS-457, was more potent than S,S-isomer 49 in both binding (8-fold) and chemotaxis (6-
fold), it was chosen for advanced profiling evaluation.
BMS-457 displays single-digit nanomolar inhibition of chemotaxis induced by a number of CCR1
ligands, including MIP-1 (2.1 nM), Leukotactin-1 (4.4 nM),29 RANTES (1.0 nM), MPIF-1 (2.7 nM),29 and
HCC-1 (4.0 nM). The molecule was not highly protein bound in human serum, with a free fraction of 16%. In
whole blood it blocked the ligand-stimulated upregulation of the 2-integrin CD11b, with IC50 values of 46 nM
(MIP-1) and 54 nM (LKN-1).29 This measurement defines an early critical event in cellular arrest at the
endothelial surface. BMS-457 displayed greater than 1000-fold selectivity against other CC family receptors
including CCR2, CCR3, CCR4, CCR5, CXCR2, CXCR3, and CXCR5. It was also selective against a broad

panel of other G-protein coupled receptors and biogenic amine transporters. No inhibition of cytochrome P450
isozymes CyP 1A2, 2C19, 2C9, 2D6, and 3A4 was observed. Preliminary counter screen profiling did not reveal
any significant safety (hERG, Na+, Ca2+ channel) issues. Finally, the molecule exhibited good metabolic stability,
and good pharmacokinetics across four species: Mouse, rat, cynomolgus monkey, and dog (Figure 3). Oral
bioavailability was measured at 100%, 37%, 54%, and 72%, respectively.
We were intrigued by the difference in PXR activity of trans-3-hydroxycyclopentane 48 (EC20 = 41 M)
vs. cis-3-hydroxycyclopentane 51 (EC20 = 5.3 M). Insight into this was gained when X-ray co-crystal structures
of compounds 52 and 53 bound to the PXR receptor were obtained (Figure 4). These crystal structures indicate
that the piperidine-valine scaffold provides an excellent template for PXR substrates due to hydrogen bonding
interactions of the piperidine’s hydroxyl-substituent and the carbonyl group of the piperidine’s amide, as well as
hydrophobic interactions of the valine isopropyl group and the piperidine gem-dimethyl substituents.
Furthermore, the methylene-cyclopropyl tailpiece of compound 52 fills another hydrophobic pocket in the
receptor, resulting in the compounds observed potency in PXR. Somewhat surprisingly, compound 53, which
incorporates a polar hydroxyl-substituent in the tailpiece, was also a significant PXR inducer. However, in the
crystal structure it is seen that instead of projecting into the receptor’s hydrophobic region, the hydroxyl group
forms an intramolecular hydrogen bond with the valine N-H, resulting in a conformation which allows the
neighboring methyl substituents to occupy this hydrophobic pocket.
Docking compounds 48 and 51 into this model provides rationale for the observed difference in
their PXR selectivity (Figure 5). The cyclopentane tailpiece of these compounds provides a more rigid
scaffold than the conformationally mobile tailpiece of 53. The cis-orientation of compound 51 places
the hydroxyl-group and the amide on the same side of the cyclopentane ring, allowing for a
intramolecularly H-bonded conformation similar to that seen in compound 53. In contrast, the trans-
orientation of 48 prevents the hydroxyl group from forming this intramolecular H-bond, and instead

forces it into a repulsive steric interaction with the PXR receptor’s hydrophobic pocket, resulting in the
elimination of PXR activity.
The synthesis of key intermediate 57 is outlined in Scheme 1. Bis-alkylation of N-BOC-4-piperidone
with iodomethane, followed by crystallization from hexanes yielded 54 in 35% yield. In this reaction,
iodomethane is added to a stirring mixture of the piperidinone and sodium hydride in THF at 5 oC, and the
mixture is stirred at this temperature for 1 hour before the ice-bath is removed and the reaction mixture is allowed
to come to room temperature. It should be noted that this reaction produces a significant exotherm and vigorous
gas evolution at anywhere from 30 minutes to 2 hours after the ice bath is removed. This can be controlled by
returning the reaction vessel to an ice/water bath once the exotherm begins. Due to the somewhat unpredictable
nature of these events, the reaction should be monitored carefully. 4-Chlorophenyllithium addition to 54 in THF
at -78 oC followed by chromatography over silica gel yielded 55 in 74% yield. Removal of the BOC-protecting
group with HCl in dioxane, followed by co-crystallization of the free-based amine with L-(+)-tartaric acid from
9:1 methylethylketone (MEK)/water yielded 56 in 32% yield and 98.6% ee after free-basing. A single
recrystallization of the tartarate from 9:1 MEK/water increases the ee to >99.9%. EDC/HOBT mediated coupling
of 56 and BOC-D-Val-OH, followed by aqueous acid/base workup and chromatography over silica gel yielded
tert-butyl (R)-1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-
ylcarbamate in 95% yield, and removal of the BOC-protecting group with HCl in dioxane yielded 57 in
quantitative yield and of sufficient purity to take on to the final coupling step.
Scheme 1. Synthesis of penultimate intermediate 57 and final products.
Two synthetic routes to enantiopure BMS-457 are shown in Schemes 2 and 3. The key step in our
original route (Scheme 2) was based on the work of H. C. Brown, and co-workers.30 The asymmetric
hydroboration of 58 with chiral dialkylborane ((S)-ipc)2-BH at -45 oC for 48 hours, followed by oxidation with

aqueous hydrogen peroxide gave a 85:15 mixture of cis- and trans-diastereomers which was enriched in the
desired R,R-isomer (55% ee). Subsequent resolution by preparative SFC chromatography yielded enantiopure
59a in 50% yield. Removal of the benzyl ester via catalytic hydrogenation yielded 60 in quantitative yield.
EDC/HOBT mediated coupling of 60 and 57 followed by silica gel chromatography yielded BMS-457 in 56%
yield and >99% ee. While this route produced satisfactory results, the asymmetric hydroboration step was not
practical for scale-up due to several factors: The tedious preparation and handling of enantiopure ((S)-ipc)2-BH;
the long reaction time at low temperature; and poor atom-economy of the process due to the molecular weight
ratio of ((S)-ipc)2-BH vs. the desired product. Thus, a second route was developed.
Scheme 2. First-generation synthesis of enantiopure BMS-457.
The second-generation route also involved the hydroboration of 58, this time without the use of
enantioselective reagents (Scheme 3). Thus, hydroboration of 58 with catecholborane in the presence of
Wilkinson’s catalyst in THF followed by treatment with hydrogen peroxide yielded 59b in 79% yield and 12:1
trans/cis ratio.31 DCC mediated coupling of 59b with BOC-L-Pro-OH, followed by supercritical fluid
chromatography resolution yielded (S)-2-((1R,3R)-3-(benzyloxycarbonyl)cyclopentyl) 1-tert-butyl pyrrolidine-
1,2-dicarboxylate in 27% yield (2 steps) and >99.9% ee. Removal of the benzyl ester via catalytic hydrogenation
yielded 61 in 82% yield. EDC/HOBT mediated coupling of 57 and 61 yielded (S)-1-tert-butyl 2-((1R,3R)-3-((R)-
1-((S)-4-(4-chlorophenyl)-4-hydroxy-3,3-dimethylpiperidin-1-yl)-3-methyl-1-oxobutan-2-
ylcarbamoyl)cyclopentyl) pyrrolidine-1,2-dicarboxylate in 88% yield. The proline ester was hydrolyzed with 2.1
eq. of aqueous lithium hydroxide in THF. Workup (ethyl acetate/1 N NaOH) removed all traces of BOC-proline,
and yielded BMS-457 in 82% yield and 99.2% ee.
Scheme 3. Second-generation synthesis of enantiopure BMS-457.

By modifying a series of compounds which exhibited good binding and functional potency we have
discovered preclinical candidate molecule BMS-457. Improved potency was achieved via the elaboration of a 4-
(4-chlorophenyl)piperidine moiety to (S)-4-(4-chlorophenyl)-3,3-dimethylpiperidin-4-ol. Metabolism and
selectivity issues were addressed by adding a polar substituent to the molecule’s cycloalkyl tailpiece. The
molecule’s observed selectivity versus PXR was rationalized by docking the structure into a model generated
from X-ray co-crystal structures of analogous compounds bound to PXR.
BMS-457 is a potent and selective CCR1 antagonist which inhibits leukocyte chemotaxis induced by
multiple ligands at single-digit nanomolar levels. Due to its excellent potency, selectivity, and pharmacokinetic
properties, it was chosen for advanced preclinical studies. Despite a clean in vitro cardiac safety profile (hERG
IC50 > 80 M, hERG patch clamp: 3% inhibition @ 10 M), significant QT prolongation was observed in both
rabbits and dogs during advanced safety studies. Dose escalation studies showed this response to be robust with
increasing drug exposures, and further development of BMT-457 was halted. While the failure of BMS-457 was
disappointing, lessons learned from the development of this molecule were vital to the later discovery of more
successful compounds. These compounds will be the subject of a future report.
Acknowledgements
The authors wish to thank Dauh-Rurng Wu for chiral separations in the synthesis of BMS-457, and
Rodney Vickery for formulations support.
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Watson, P. S.; Clark, C. M.; Ko, S. S.; Welch, P.; Covington, M.; Stowell, N.; Wadman, E.; Davies, P.; Solomon,
K.; Newton, R. C.; Trainor, G. L.; Decicco, C. P.; Wacker, D. A.; Bioorg. Med. Chem. Lett. 2004, 14, 1645. (c)
Santella, J. B.; Gardner, D. S.; Yao, W.; Shi, C.; Reddy, P.; Tebben, A.; Delucca, G. V.; Wacker, D. A.;
Watson, P. S.; Welch, P. K.; Wadman, E. A.; Davies, P.; Solomon, K. A.; Graden, D. M.; Yeleswaram, S.;
Mandlekar, S.; Kariv, I.; Decicco, C. P.; Ko, S. S.; Carter, P. H.; Duncia, J. V.Bioorg. Med. Chem. Lett. 2008, 18,
576; (d) Gardner, D. S.; Santella III, J. B.; Tebben, A. J.; Batt, D. G.; Ko, S. S.; Welch, P. K.; Wadman, E. A.;
Davies, P.; Carter, P. H.; Duncia, J. V.; Bioorg. Med. Chem. Lett. 2008, 18, 586.
25. (a) Cherney, R. J.; Nelson, D. J.; Lo, Y. C.; Yang, G.; Scherle, P. A.; Jezak, H.; Solomon, K. A.; Carter, P. H.;
Decicco, C. P.; Bioorg. Med. Chem. Lett. 2008, 18, 5063; (b) Cherney, R. J.; Brogan, J. B.; Mo, R.; Lo, Y. C.;
Yang, G.; Miller, P. B.; Scherle, P. A.; Molino, B. F.; Carter, P. H.; Decicco, C. P.; Bioorg. Med. Chem. Lett. 2009,

19, 597; (c) Cherney, R. J.; Mo, R.; Meyer, D. T.; Voss, M. E.; Lo, Y. C.; Yang, G.; Miller, P. B.; Scherle, P. A.;
Tebben, A. J.; Carter, P. H.; Decicco, C. P.; Bioorg. Med. Chem. Lett. 2009, 19, 3418; (d) Cherney, R. J.; Mo, R.;
Meyer, D. T.; Voss, M. E.; Yang, M. G.; Santella, J. B., III; Duncia, J. V.; Lo, Y. C.; Yang, G.; Miller, P. B.;
Scherle, P. A.; Zhao, Q.; Mandlekar, S.; Cvijic, M. E.; Barrish, J. C.; Decicco, C. P.; Carter, P. H.; Bioorg. Med.
Chem. Lett. 2010, 20, 2425; (e) Yang, M. G.; Xiao, Z.; Shi, Q.; Cherney, R. J.; Tebben, A. J.; De Lucca, G. V.;
Santella, J. B.; Mo, R.; Cvijic, M. E.; Zhao, Q.; Barrish, J. C.; Carter, P. H.; Bioorg. Med. Chem. Lett. 2012, 22,
1384; (f) Carter, P. H.; Brown, G. D.; King, S. R.; Voss, M. E.; Tebben, A. J.; Cherney, R. J.; Mandlekar, S.; Lo, Y.
C.; Yang, G.; Miller, P. B.; Scherle, P. A.; Zhao, Q.; Decicco, C. P.; Bioorg. Med. Chem. Lett. 2012, 22, 3311.
26. MPTP is the acid-mediated dehydration product of MPPP. In-vivo, MPTP is metabolized to alkyl piperidinium
species MPP, which destroys dopaminergic neurons in the substantia nigra of the brain, resulting in severe
Parkinsonism. For further reading, see (a) Langston, J. W.; Palfreman, J.; The Case of the Frozen Addicts. Pantheon
Books. 1995, ISBN 0-679-42465-2; (b) Fahn, S.; "Book Review -- The Case of the Frozen Addicts: How the
Solution of an Extraordinary Medical Mystery Spawned a Revolution in the Understanding and Treatment of
Parkinson's Disease". The New England Journal of Medicine 1996, 335, 2002.
27. Sayre, L. M.; Arora, P. K.; Feke, S. C.; Urbach, F. L.; J. Am. Chem. Soc. 1986, 108, 2464.
28. For further reading on the topic of PXR activation, see (a) Lehmann, J.M.; McKee, D.D.; Watson, M.A.; Willson,
T.M.; Moore, J.T.; Kliewer, S.A.;. J. Clin. Invest. 1998, 102, 1016; (b) Bertilsson, G.; Heidrich, J.; Svensson, K.;
Asman, M.; Jendeberg, L.; Sydow-Bäckman, M.; Ohlsson, R.; Postlind H.; Blomquist, P.; Berkenstam, A.; Proc.
Natl. Acad. Sci. U.S.A. 1998, 95, 12208; (c) Kliewer, S. A.; Goodwin, B.; Willson, T. M.; Endocrine Reviews 2002,
23, 687; and the references contained therein.
29. Truncated forms of leukotactin-1 and MPIF-1 were used for the determination of chemotaxis inhibition in these
assays. See Schall, T. J.; Berahovich, R. D.; Miao, Y. W.; Premack, B.; and Howard, M. C.; J. Immunol., 2005, 174,
7341.
30. (a) Brown, H. C.; Ayyangar, N. R.; Zweifel, G.; J. Am. Chem Soc. 1964, 86, 397; (b) Brown, H. C.; Jadhav, P. K.;
Mandal, M. K.; Tetrahedron 1981, 37, 3547; (c) Brown, H. C.; Desai, C. M.; Jadhav, P. K.; J. Org. Chem. 1982, 47,
5065; (d) Brown, H. C.; Joshi, N. N.; J. Org. Chem. 1988, 53, 4059; (e) Brown, H.C.; Dhokte, U.P.; Tet. Lett. 1996,
37, 9021.
31. Because the desired R, R-diastereomer was not enriched with respect to the undesired S,S-diastereoisomer in this
mixture, resolution via preparative SFC was not possible.

Table 1
Table 1. Comparison of binding and chemotaxis potencies of compounds in the lead series (A) vs. compounds in the new
series (B).
A
B
CCR1
IC50 (nM)a
Chemotaxis
IC50 (nM)a,b
78±10 (11)
10±5 (2)
Series
A
R
1
Ph
Ph
30±15(209)
3
B
18
4a
4b
5a
5b
6a
6b
7a
7b
A
4-F-5-OH-Ph
10
48±10 (2)
13±1 (2)
B
4-F-5-OH-Ph
2.9
6.8
3.4
40
A
3-(2-(NH(CO)NH2)Ph)Ph
3-(2-(NH(CO)NH2)Ph)Ph
3-(SO2NH2)Ph
153
B
48±9 (3)
A
123±3 (2)
9.9±2.8 (2)
107±7 (2)
3.5±0.6 (2)
B
3-(SO2NH2)Ph
3±1 (5)
9.4
1.2
A
CH2-4-pyridyl
B
CH2-4-pyridyl
(a)Values without standard deviations represent a single determination.
Values in parentheses represent the number of determinations. (b) inhibition
of MIP-1induced chemotaxis.

Table 2
Table 2. Binding and chemotaxis potencies of a selection of CCR1 antagonists.
CCR1
IC50 (nM)a
18
Chemotaxis
IC50 (nM)a,b,c
10±5 (2)
4.7±0.0 (2)
10±0.4 (2)
5.5±1.6 (2)
n/a
Entry
R
3
Ph
2-Cl-Ph
8
6.6
3-Cl-Ph
9
9.9±5.6 (3)
3.3
4-Cl-Ph
10
11
12
13
14
15
16
17
18
19
20
21
22
4-(SO2NH2)Ph
2-(CO2H)Ph
3-(CO2H)Ph
2-pyridyl
3-pyridyl
4-pyridyl
CH2Ph
233
2940
7.1±0.7 (2)
15
n/a
8.9±1.6 (2)
n/a
4.3
6.1±2.8 (2)
n/a
7.6
4.1
1.8±0.1 (2)
n/a
CH2CH2Ph
CH2CH2CH2Ph
CH3
5.8
1.7
n/a
13
15±3 (2)
3.6±0.8 (2)
4.4±1.0 (2)
CH2CH3
4.5
CH(CH3)2
4.6±2.2(2)
23
24
25
26
2.9
1.9±0.5 (2)
1.2±1.1 (2)
n/a
1.7±0.1 (3)
3
1.8
n/a
27
28
29
1.4±0.5 (177)
2.5±2.2 (72)
2.2
2.6
n/a
n/a
30
2.3±0.5 (3)
8.3±1.6 (2)
(a) Values without standard deviations represent a single
determination. Values in parantheses represent the number of
determinations. (b) Inhibition of MIP-1 induced
chemotaxis. (c) n/a = no data available.

Table 3_R3
Table 3. SAR of piperidine modifications.
CCR1 IC50
(nM)a
Chemotaxis
IC50 (nM)a,b,c
R1
R2
R3
R4
31 (R)-OH
Me
Me
Me
Me
Me
Me
Me
Me
Me
Me
Me
Me
Me
Me
4-Cl-Ph
4-Cl-Ph
4-Cl-Ph
4-Cl-Ph
4-Cl-Ph
4-Cl-Ph
3-Pye
1800
25000
n/a
n/a
32
33
O-Me
NH2
196
n/a
34 NH-Ac
35 NH-Me
>8333
>8333
>8333
3.7±0.5 (3)
n/a
n/a
36
37
NMe2
H
n/a
29±8 (4)
38
c-pentyl
4.1±0.9 (2)
18±10 (3)
39
40
OHd
OH
spiro-c-pentyl
spiro-c-propyl
4-Cl-Ph
3-Pye
208
7.2
n/a
21±13 (5)
(a) Values without standard deviations represent a single determination.
Values in parentheses represent the number of determinations. (b) Inhibition
of MIP-1 induced chemotaxis. (c) n/a = no data available. (d) Racemic
piperidine headpiece. (e) Py = pyridyl.

Table 4
Table 4. Binding, chemotaxis, PXR, and ion channel selectivity results from cyclopentane modifications.
PXR
EC20/EC60
(M)c
Ion Channels
Binding
IC50
Chemotaxis
IC50
Metabolic Stability
% Remainingc
RLM MsLM
(% inhibition @ 10 M)c
R
(nM)a
(nM)a,b,c
HLM
MkLM
Na
Ca
3
Ph
18
10±5.0 (2)
9.9±2.8 (2)
8.9±1.6 (2)
1.1, 2.9
>50, >50
>50, >50
n/a
96
n/a
100
97
n/a
99
n/a
100
100
48
64
31
0
6b
13
3-(SO2NH2)Ph
3-(CO2H)Ph
3.0±1.1 (5)
7.1±0.7 (2)
39
3
100
100
27
41
42
43
44d
45d
46
47
48
49
50
51
1.7±0.6 (31)
341
4.6±1.5 (40)
n/a
0.4, 1.4
n/a
47
n/a
n/a
n/a
33
n/a
94
97
98
95
69
43
50
n/a
n/a
n/a
20
n/a
93
84
87
74
67
75
26
n/a
n/a
n/a
49
n/a
92
96
88
90
65
60
4
52
n/a
50
n/a
n/a
n/a
n/a
1
1603
n/a
n/a
n/a
n/a
194
n/a
n/a
n/a
n/a
1.4e
>80 Mf
n/a
61 Mf
n/a
2.3±0.6 (3)
20
3.1±0.4 (2)
n/a
n/a
n/a
93
n/a
66
59
21
22
8.5e
39 Mf
n/a
21 Mf
n/a
2.2±1.0 (3)
17
5.6±0.1 (2)
n/a
n/a
0.8±0 (2)
6.3
2.1±0.1 (2)
12±3.0 (2)
1.1±0.1 (2)
2.9±1.6 (2)
41, >50
>50, >50
1.8, 7.4
5.3, >50
14
11
4
-3
1.6
19
9
0.8±0.2 (2)
5
-4
(a) Values without standard deviations represent a single determination. Values in parantheses represent the number of determinations. (b)
Inhibition of MIP-1 induced chemotaxis. (c) n/a = no data available. (d) Mixture of relative stereoisomers. (e) Data reported as EC50.due to a
mid-program change in the PXR assay format. (f) Data reported as IC50 due to a mid-program change in the ion-channel assay format .

Figure 1. Lead compound 1, bracketed to indicate our short-hand references to three regions of the molecule.
Figure 2. CCR1 binding and MIP-1 chemotaxis data for the lead piperidine amide (1), the modified piperidine
incorporating a basic nitrogen (2), and the modified piperidine amide (3).
Figure 3. Eight-hour oral exposure profile of BMS-457 in mouse, rat, cynomolgus monkey, and dog.
Figure 4. X-ray co-crystal structures of compounds 52 (green) and 53 (yellow) bound to the PXR receptor. The
binding site is shown as a surface view with the surface of the front most residues turned off for clarity.
Figure 5. Compounds 48 (orange) and 51 (blue) superposed into the X-ray co-crystal structures of compounds
52 and 53 bound to the PXR receptor. The binding site is shown as a surface view with the surface of the front most
residues turned off for clarity.

Figure 1
“Headpiece” “Linker” “Tailpiece”
1
CCR1 IC50 – 30 nM
MIP-1 Chemotaxis IC50 = 78 nM

Figure 2

PLEASE print this figure in color both on the web and in print.

Figure 4
52 (green)
53 (yellow)
PXR EC60 = 0.5 M
CCR1 IC50 = 1.0 nM (n=1)
PXR EC50 = 1.8 M
CCR1 IC50 = 5.2 nM (n=1)
Figure 4. X-ray co-crystal structures of compounds 52 (green) and 53 (yellow) bound to the PXR receptor. The binding
site is shown as a surface view with the surface of the front most residues turned off for clarity.
Please print this figure in color both in print and on the web.

PLEASE print this figure in color both on the web and in print.

Scheme 1. Synthesis of penultimate intermediate 57 and final products.
SCHEME
o
o
Reagents and Conditions: (a) NaH, CH3I, THF, 0 to 25 C, 35%; (b) 4-Cl-PhBr, n-BuLi, THF- 78 C, 74%; (c) HCl,
dioxane, 25 oC, quant., crude; (d) 1. L-(+)- tartaric acid, crystallize twice from 9:1 2-butanone/water, 2. Na2CO3 (aq), EtOAc,
o
o
32%, >99.9% ee; (e) BOC-(D)-Val-OH, EDC, HOBT, Hunig’s base, DCM, 25 C, (95%); (f) HCl, dioxane, 25 C, quant.,
crude; (g) R-COOH, EDC, HOBT, Hunig’s base, DCM, 25 oC.
Scheme 2. First-generation synthesis of enantiopure BMS-457.
SCHEME
Reagents and Conditions: (a) Bn-OH, EDC, DMAP, DCM, 77%; (b) 1. ((S)-ipc)2-BH, 2. Acetaldehyde, 3. NaOAc/H2O2,
THF, -45 oC, 48 h; (c) resolution via chiral SFC chromatography, 50% yield, >99% ee; (d) H2, Pd(OH)2, EtOAc, quantitative
yield; (e) 57 (Scheme 1), EDC, HOBT, Hunig’s base, DCM, 56%.
Scheme 3. Second-generation synthesis of enantiopure BMS-457.
SCHEME
Reagents and Conditions: (a) Catecholborane, Wilkinson’s catalyst, then H2O2, THF, 79%, 12:1 trans/cis; (b) BOC-L-Pro-
OH, DCC, DMAP, DCM, 81%; (c) Chiral SFC on Chiralpak AD-H (3 X 25 cm) CO2/IPA 90/10 75mL/min, 35 oC, 100 bars,
UV 215 nm, loading: 1.4cc (100mg/cc)/8min, 34%, >99.9% ee; (d) H2, Pd(OH)2, EtOAc, 83%; (e) 57 (Scheme 1), EDC,
HOBT, Hunig’s base, DCM, 88%; (f) 2.1 eq. LiOH, THF/H2O, 82%, 99.2% ee.

Scheme 1

Scheme 2

Scheme 3

*Graphical Abstract (for review)