DP4 Chemistry Significant Events
`
`November 4 ,1999
`
`Competitive Update: Nothing to report
`
`Current FTE’s: 3
`
`Monthly Summary: The goal of the program is to discover small! molecule inhibitors of dipeptidyi
`peptidase IV (DP4) for use in the treatment and prevention of diabetes. Inhibition of DP4 should
`prevent the degradation of GLP—1 and potentiate its action in vivo. in a past report, BMS-3563?9 was
`identified as a potent and proprietary inhibitor of DP4. in vltro and MAP Characterization has recently
`shown this compound to possess 53% oral biavailability in the rat (t‘1f2 = 4.4 hr) and tow propensity for
`in vitro metabolism and P450 inhibition. Scale-up for in vivo testing is in progress. Recently,
`two
`unique lead compounds have been identified: I) the 3,4—methano cyanopyrollidide BMS-383680 (Ki =
`15 nlvl), the most potent proprietary DP4 inhibitor identified to date and ii) BMS-382436 (Ki = 36 nM)
`which contains a cyano-substituted dihydropyrazole. in recent in vivo studies, the non-proprietary DP4
`inhibitor BMS—32820‘l {lie-Thia, Ki = 110 nM) exhibited glucose lowering i) acutely in the normal rat
`upon oral glucose loading and ii) in the fasting high fat fed mouse when dosed chronically. The in vivo
`responses are consistant with GLP-1(T-36) potentiation via DP4 inhibition.
`
`Me
`
`HEN
`
`Me
`
`0
`
`N
`
`A
`V
`
`CM
`
`'
`
`Me‘
`
`l-I2N
`
`Me
`
`N
`
`0
`
`CN
`
`HZN
`
`0
`
`14...
`9
`
`N
`
`CN
`
`BMS-356379
`DP4 Ki = 28 nllli
`
`BNES-383680
`DP4 Ki =15 nM
`
`BMS-382436
`DP4 Ki = 36 nM
`
`Full Text: The goal of the program is to discover smali molecule inhibitors of dipeptidyl peptidase lV
`(DP4) for use in the treatment and prevention of diabetes.
`Inhibition of DP4 should prevent the
`degradation of GL.F’«‘l and potentiate its action in vivo.
`l3MS—356379 was previously identified as a
`novel and proprietary inhibitor with an inhibitory constant of 28 nM vs pig kindey DP4. Recent MAP
`data indicates this Compound exhibits 53% orai systemic bioavailability in the rat with a t1/2 of 4.4 hr.
`in vitro, BMS—356379 was not metabolized in human liver microsomes after 10 min exposure. This
`data coupled with its low propensity for P450 inhibition (>10 uM for all isozymes) makes BMS—356379
`an exceilent
`iead for further development. Scale—up of this compound is nearing completion for
`evaluation in our in vivo assays. Work is progressing in determining the SAR about the amino
`terminus in order to optimize in vitro potency.
`
`Bl\/iS—382436 has also recently been identified as a potent and proprietary DP4 inhibitor. This
`compound (Ki = 36 nM) possesses a unique cyano-substituted dihydropyrazole. Since this structural
`chernotype has not been reported in the literature. efforts will be made to determine the solution and
`metabolic stability of this compound prior to future work.
`
`Page 1 of 3
`
`Astraleneca Exhibit 2186
`Mylan V. Astraleneca
`IPR2015-013-l0
`
`AstraZeneca Exhibit 2186
`Mylan v. AstraZeneca
`IPR2015-01340
`
`Page 1 of 3
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`

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`Page 2
`
`Me
`
`H2N
`
`Me
`
`N
`
`0
`
`CN
`
`E”.i‘f“.?,52?:£,9nM
`rat oral bioavail. 53%
`t-“'2 =
`hr
`
`/i
`
`H2N
`
`'N_.
`N
`
`0
`
`CN
`
`BMS-382436
`DP4 Ki = 36 nM
`
`
`
`to expand the SAR a ound BMS—3563"/"9 and to identify additional proprietary
`In an attempt
`chemotypes, several novel cyclopropanat
`cyanoproyl derivatives have been explored. Incorporation
`
`of the methane group at the 2,3-position,
`ffording BMS-378736, resulted in a dramatic loss in
`potency (Ki = 76,400 nM). The des—cyano
`, -methane derivative BIVES-378738 exhibited modest
`
`activity vs DP4 (Ki = 1,400 nM) but activity was
`hanced 100—fold by incorporation of the (S-)—cyano
`functionality, affording BMS-383680 (Ki = 15 nM). T '
`is the most potent proprietary DP4 inhibitor yet
`
`discovered in the program. Subsequent SAR has show
`4 is exquisitely sensitive to the correct
`stereochemical configuration of
`the substituents on the five—
`bered ring. The related trans-
`methano isomer of BMS-383680, BMS-380845 (Ki = 1,300 nM), ex '
`'
`arked reduction in
`potency against DP4. its stereoisomer BMS—384189 was essentially inactive (0% inhibi I
`Stereoselective generation of the core cyano-cyclopropanated pyrrolide in BMS—383E380 has
`difficult, requiring further efforts before SAR studies in this series can be pursued.
`
`M
`
`Me
`
`_
`
`H2N
`
`Heterocycle
`
`o
`
`,/I
`
`.
`
`gr
`
`roved
`
`
`fr
`
`
`
`
`
`
`
`
`
`
` BMS# 378735 378733 333630 330345 384139
`
`
`
`Heterocycle
`
`i/
`
`ifN
`
`C”
`
`DP4 Ki (HM)
`
`76,400
`
`1,400
`
`N
`
`CN
`
`15
`
`57/5/1?
`
`cu
`
`LL’/N\;)
`
`"EN
`
`1,300
`
`0°/o @ 10 }.LM
`
`In vivo results: P32/98 (|le—Thia, Bil/IS-328201) is a weak and reportedly short-acting DP4 inhibitor
`being developed by Probiodrug for the treatment of diabetes. Due to its ease of large scale synthesis
`and previous reports of efficacy in animal models,
`this compound was used to refine our in vivo
`models in—house. A dose response versus plasma DP4 inhibition is depicted below. BMS-328201
`demonstrates >70“/o inhibition @ 100 mpk and an ED50 of ~30 mpk in this assay. The simplicity of
`this assay should allow plasma DP4 inhibition to serve as a potential acute in vivo assay for the
`discrimination of future analogs. allowing a comparison based on ED50 values and potentially duration
`of action.
`
`Page 2 of 3
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`Page 2 of 3
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`

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`Page 3
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`d__
`/O\
`g 100 0/ g
`0
`30
`33
`+
`EL
`D
`“,
`E
`5;:
`Q
`
`40 —
`20
`0
`
`‘I
`1
`
`“"“““‘““”'
`
`\\\
`
`I
`U\
`
`Oral dose-response in rats
`for inhibition of plasma
`DP-4 activity at 30 min
`M
`Me
`r-—-S
`NV}
`
`e
`
`\O HZN
`
`0
`P32l98 (lle-Thia)
`‘ ' BMS-328201
`DP4Ki=11DnM
`100
`
`'
`
`10
`
`dose, mglkg
`
`Based on these results, Bil/ES-328201 (100 mpk) or vehicle were orally administered to normal rats (n
`= 4) and subject to a standard oral glucose challenge 30 min later. DP4 activity, plasma glucose, and
`insulin were measured at 10 minute intervals. During the course of the study. Bl\/!S—32820‘l treated
`animals reduced plasma DP4 activity to ~30% of control while plasma glucose concentrations were
`significantly attenuated, Plasma insulin exhibited a more rapid rise and fall in mean levels. All of these
`results are consistent with the expected biological responses of GLP-1(7-36) potentiation.
`
`Acute effects of BMS-328201 (ile-thiazolidicle) in normal rats given a glucose challenge
`
`DP4 activity Ulmi
`
`500
`
`glucose mgldl
`
`insulin nglml
`
`180
`
`150
`
`120
`
`90
`
`60
`
`30
`
`O
`
`
`
`400
`
`300
`
`200
`
`100
`
`BM3328201
`
`BMS—32820‘l
`
`BMS—32820’l
`
`G
`
`10
`
`20
`
`30
`
`O
`
`10
`
`20
`
`30
`
`0
`
`10
`
`20
`
`30
`
`minutes
`
`minutes
`
`minutes
`
`The effect of Bil/ES—3282D’l on plasma glucose, food intake, and body weight upon chronic (‘l4 days)
`dosing of ElMS—32820'l
`(0.1% in diet, ~ 80 mg/kg/d) in high fat fed (3 month feeding) mice was
`recently studied. At day 15, fasting plasma glucose levels were significantly and dramatically reduced
`versus control (49%). Minimal effect was observed on food intake but a trend towards lower body
`weight in the treated mice was observed. This study is still
`in progress and the effects of the DP4
`inhibitor will be ascertained at later time points.
`
`Compound
`
`Vehicle control
`
`BMS-328201
`
`(0.1% in diet)
`n = '10 per group, *p < 0.05
`
`Plasma glucose
`(mgImL)
`
`Total food intake over
`15 days (grns)
`
`175 i 13
`
`89 t 2*
`
`387
`
`377
`
`Avg. body weight
`gain
`Per mouse (gms)
`
`1.3
`
`0
`
`Page 3 of 3
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`Page 3 of 3