DP4 Chemistry Significant Events
`
`March 30 ,200D
`
`Executive Summary: Pharrnacokinetic evaluation in cynos of our most potent and proprietary DP4
`inhibitor, BMS~405295, showed this compound to have good oral systemic bioavailability (79%) with a
`hah‘—life of 3.6 h. This compound had also previously demonstrated good PK in the rat (F = 77%, tw =
`2.8 h). Novartis’ clinicai candidate DPP T28 (BMS-428245) was recently synthesized and found to
`exhibit in vitro activity (Ki = 7 nM) comparabie to our current leads. Variation of the N-terminal amino
`acid of BMS405295 resulted several compounds with good in vitro potency, inciuding BMS—429G36.
`In vivo testing of these newer analogs, as well as the Novartls compound, are planned.
`
`V
`"2"
`
`/
`
`Me
`
`H2"
`
`M
`
`elllle
`
`N
`
`o
`
`cu
`
`BMS-405295
`DP4 Ki = 6 "M
`ED = 3.3
`Ik ,
`5°
`y
`mg 9 po
`
`"'9
`
`N
`
`S)
`CN
`o
`BMS-n_l-E9636
`DP4 KI — 4 HM
`
`CN
`
`l).
`
`/__\
`ENH
`E/\£:N\g
`DPP 728
`(ems-428245)
`DP4 Ki = 7 nM
`
`Competitive Update: Probiodrug has recently disclosed (patent application WO 99672798) dipeptide
`based DP4 inhibitors containing an activated alpha-pryridyl ketone. These compounds presumably
`irreversibly inactivate the enzyme via displacement of the pyrldyl functionality by the active site serine,
`forming a covalent intermediate. The specified compound caused an increase in time to DP4
`mediated degradation of substrate from 1 minute to 100 minutes at a concentration of 2.8 uM
`
`R2
`
`R1
`
`Me
`
`Me
`
`R3 x N
`H
`
`m
`N
`
`0
`
`‘g‘:\\)
`
`O
`
`cr
`
`Hzhl
`_
`
`N
`
`0
`
`Cr
`
`0
`
`+ N
`
`/_\
`
`Monthly Summary The goal of the program is to discover small molecule inhibitors of dipeptidyl
`peptidase IV (DP4) for use in the treatment and prevention of diabetes.
`Inhibition of DP4 should
`prevent the degradation of GLP-‘E and potentiate its action in vivo. Two new chemotypes were
`recently explored.
`In agreement with reported values, Novartis’ clinical candidate DPP 728 (EMS-
`428245) was found to be a 7 nM inhibitor of DP4 in vitro.
`In vivo testing of this compound is
`scheduted. A unique bicyclic pyrrolidide BMS428073, a hybrid between EMS-378738 and the
`Probiodrug compound Bil/lS—326430, was found to be poorly active against DP4. On the basis of this
`data, we believe that incorporation of an a|pha—cyano functionality in the ring structure would not
`sufficiently boost its activity to desired Eevels.
`
`Page 1 of 3
`
`Astraleneca Exhibit 2175
`Mylan V. Astraleneca
`IPR2015-013-l0
`
`AstraZeneca Exhibit 2175
`Mylan v. AstraZeneca
`IPR2015-01340
`
`Page 1 of 3
`
`

`

`Page 2
`
`CM
`A3
`l / N
`
`NH
`
`E
`
`f’‘\
`
`N
`
`DPP 728
`(BMS-428245)
`DP4 Ki = 7 nm
`
`Me
`Med‘ J
`
`I-{ZN
`
`’
`
`HI]?
`‘
`
`o
`BMS-378738
`
`DP4 Ki = 1400nM
`
`Me
`
`N’?
`
`Me‘, I
`
`HZN
`
`o
`BMS—326430
`
`DP4 Ki = 440 ntlll
`
`Me
`
`,
`
`H2",
`
`N»
`
`0
`
`BM$-428245
`DP4 KI — 5300 nM
`
`Pharmacokinetic evaluation in cynos of our most potent and proprietary DP4 inhibitor, BMS-405295,
`showed this compound to have good oral systemic bioavailability (79%) with a half—life of 3.6 h This
`compound had atso previously demonstrated good PK in the rat F
`= 2.8 ).
`h%
`
`
`
`
`9
`, ms--=th‘e,.inr;:,m<p
`allsfiw
`..
`4
`d;j5=ei3:rery=uof='rMS;-4@52=95= s ~
`fefiot
`
`
`
`
`
`metgsane .i-utfe ;aoy.aia_e: yrrelidieles.
`'
`oa‘;:h=ighly...,.,,
`'
`
`
`
`
`
`Some of the recent vana ions are
`own eow.
`he cyclo exylfrnethyl derivative BMS-429636 was
`only slightty more potent than BMS-405295. Introduction of more highiy branched and entropically
`disordered substituents led to a gradual
`reduction in inhibitory activity. Thus, although nearly
`isostructural with BMS-429636, the diethylmethyl and the triethyl derivatives BMS429806 and BlViS-
`42988O were 4 to 8 fold less active respectivel
`. Based on this observation, future derivatives will
`incorporate greater conformational restriction in t e alkyl group at the N-terminus.
`
`
`
`;
`
`5
`
`E
`
`
`
`H2"
`
`,N
`
`0
`
`on
`
`ems-405295
`DP4 Ki = 6 nM
`
`ED5D = 3.3 mglkg, po
`
`Variation at
`N-terminus
`l_____e.__fi> —
`
`R
`
`"2"
`
`N
`
`9
`
`°
`
`C"
`
`ems 1:
`
`405295
`
`429535
`
`429759
`
`429737
`
`" 429806
`
`R
`
`i,
`
`Q
`
`r
`
`Ft
`
`We
`
`DP4 Ki (nllll)
`
`6.2
`
`4.5
`
`6.2
`
`1 0.2
`
`19.9
`
`Page 2 of 3
`
`Page 2 of 3
`
`

`

`Page 3
`
`429380
`
`430143
`
`LP
`
`LP
`
`33.2
`
`37.3
`
`The first SAR analysis at the N-terminus of the related 3,4-methano cyano pyrrolidide series has been
`performed. Unlike the 4,5—methano series,
`limited variation of the alkyl group in the 3,4 series
`demonstrated essentially no differences with respect to inhibitory activity. This unexpected finding
`suggests a slight divergence of SAR between the two methano—substituted chemotypes and limits the
`predictability of activity within these series. Other variations are planned.
`.
`
`/I
`
`H N
`2
`
`N
`
`0
`
`CN
`
`EMS-3.39530
`D?‘ K‘ ' 15 "M
`
`Variation at
`N-terminus
`l
`
`R
`
`N
`
`H N
`2
`
`CN
`0
`BMS—4-30533 (R = cyciohexyl) Ki = 11.2
`BMS—43051 6 (R = t-butyl) Ki = 1 1.6 nM
`BMS-430526 (R = isopropyl) Ki = 12.1 nM
`
`In vivo studies: Metabolic Diseases Biology has successfully developed a GLP-1 (7-36) specific N-
`terminal directed antibody, enabling the quantification of intact GLP—1 at pM leveis in plasma.
`Application of this technique to DP4 inhibitor treated rats is reported below. Normal fasted rats were
`dosed with BMS-420597 (Ki = 2 nM, plasma DP4 ED50 = 3.8 mgfkg) at 30 mg/kg p.o., followed 30
`min later by glucose challenge and 20 min later with plasma sampling. This resulted in strong
`inhibition of plasma DP4 activity to an average of 12% of control, and a 270% increase (p = 0.107) in
`endogenous intact GLP-‘t(7-36) over basal values (Table below). These results, coupled with other
`recent studies, confirm the correlation between DP4 inhibition and GLP-1 potentiation in vivo and
`enable a good functional reaci—out on the activity of. our compounds in vivo.
`
`Effect of BMS—420597 on plasma DP4 actvity and endogenous GLP-1(7—C-J6} levels 20 min after
`glucose chaitenge. Drug or vehicle given p.o. 30 minutes before oral gtucose challenge.
`
`Vehicle
`
`BM5420597
`
`(39 ms/K9}
`
`* p = 0.107;
`
`n = 4 rats per group
`
`plasma DP4 act. (units)
`
`plasma GLP-1(7~36)
`
`32.1 x 2.5
`
`3.7 i 0.2
`
`5.5 11.0
`
`15.0 i 5.6*
`
`(12% of control)
`
`(2709; of controu
`
`Page 3 of 3
`
`Page 3 of 3
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`