SUBSTITUTE for PTO/SB/05 (04-04) Utility Patent Application Transmittal
`Approved for use through 0713112006. OMB 0651-0032
`~~~----------------------------------------~----------u~·~s •. P.a.te.n-tan~~d·T•rn-~._mrr._k_O_ffi_c_e_·_u_.s~.-D.EP . . AR._TMENT._._._o.F•C•O•~~~~~CE
`21409Y
`,
`Attorney Docket No.
`CJ.:"
`UTILITY
`First Inventor or Application Identifier I Alex Minhua Chen, et al.
`pATENT APPLI CA TI QN
`~ ~
`TRANSMITTAL
`-
`i
`PHOSPHORIC ACID SALT OF A DIPEPTIDYL PEPTIDASO\'
`~ ~~
`t~
`(/) 0> -
`I
`v
`.~~:::::::::::::::::::::::::::::::::::::::EXp:::re:s~s:M:a:il:La::b:el:N:o:.:E:L:9:8:9:5:89:0:5:5U::S::::::::::::::::::~:)~;~~.~~~-~
`b;~ ~=-==--~
`Commissioner for Patents
`APPLICATION ELEMENTS
`ADDRESS TO: P.O. Box 1450
`~T""
`See M PEP clulpter 6()() concerning utility patent application contents.
`Alexandria, VA 22313-1450
`
`(Only for n.w nonprtJ\'isiona/ applications IJJUkr 37 CFR 1.53(b))
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`[ Total Pages I 25
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`15. CORRESPONDENCE ADDRESS
`~ Customer Number
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`NAME
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`ADDRESS
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`Philippe L. Durette
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`Merck & Co., Inc., P. 0. Box 2000- Patent Dept., RY60-30
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`Rahway
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`NJ
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`USA
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`TELEPHONE 732-594_4568
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`Philip~~
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`I Date
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`35,125
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`6/23n004
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`EXPRESS MAll.. CERTIFICATE
`:.;lu:::n""e'-'2""3'-'-"'2004"-"'-!....-------------------
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`ON THE AB~DA TE IN AN ENVELOPE ADDRESSED TO COMMISSIONER FOR PATENTS,
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`EXANt"UA.f\VA 2231~1450.
`~-J.. ~ 0V\Qfl (()J.
`DATE ~LlJu ~~ :J()QY
`MAILEDB
`f
`' \
`C~r generated form -rransmittal Form Non-PV· (Application Filing Folder) Merck & Co., Inc., 5/1812004
`~
`
`v
`
`DRL Ex. 1010, p. 001
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`

`

`IN THE UNITED STATES PATENT AND TRADEMARK OFFICE
`
`PATENT
`
`Commissioner for Patents
`P.O. Box 1450
`Alexandria, VA 22313-1450
`
`Sir:
`
`Deposit Acct. 13-2755
`MERCK & CO., INC.
`Our Case Docket No. =2.:..14.:...::0=9~Y __ _
`
`Transmitted herewith for filinQ under 37 C.F.R. §1.53(b) is the patent application of lnventor(s):
`Al~x Minhua Chen, Russell R. Ferlita, Karl Hansen, Ivan Lee, Stephen Howard Cypes, Vicky K. Vydra, Robert M Wenslow
`
`For: PHOSPHORIC ACID SALT OF A DIPEPTIDYL PEPTIDASE-IV INHIBITOR
`
`For
`
`Total Claims
`Independent Claims
`Multiple Dependent
`Claims*
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`Number
`Filed
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`35 -20 =
`2 - 3 =
`
`Number
`Extra
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`15 X
`0 X
`
`Rate
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`$18
`$86
`$290
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`Basic Fee
`$no
`$270
`$0
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`=
`=
`=
`
`* Add this fee if application contains any
`multiple dependent claims, regardless
`of number.
`
`TOTAL FILING FEE
`
`....
`,
`
`$1,040
`
`. The Commissioner is hereby
`Please charge my Deposit Account No. 13-2755 in the amount of $ 1 040
`authorized to charge any additional fees which may be required, or credit any overpayment to Account No. 13-2755.
`Duplicate copy of this sheet is enclosed.
`D Under the provisions of 37 C.F.R. §1.53, this application is being filed without the declaration of each inventor.
`
`EXPRESS MAIL CERTIFICATE
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`DATE OF DEPOSIT ~Ju::.:.n:.::.e..=2~3.o..:2::.:::0~04~-------
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`EXPRESS MAIL NO. =EL=9=8=95=8=9=0=55""'U=S.__ _____ _
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`I HEREBY CERTIFY THAT THIS CORRESPONDENCE IS BEING
`DEPOSITED WITH THE UNITED STATES POSTAL SERVICE AS
`EXPRESS MAIL "POST OFFICE TO ADDRESSEE" ON THE ABOVE DATE
`IN AN ENVELOPE ADDRESSED TO COMMISSIONER FOR PATENTS,
`P.O. BOX 1450, ALEXANDRIA, VA 22313-1450.
`
`MAILED aillo/u,kbep (~ i ~DATE ~II.AI..,j~....L.f'..a!..lii-L+
`
`pe L. Durette
`For Applicant(s)
`Attorney
`Reg. No. ""'"35,....'""'12=5"'----
`MERCK & CO., INC.
`Patent Dept., RY60-30
`P.O. Box 2000
`Rahway, N.J. 07065-0907
`(732) 594- .;;!;45~6::,:::8 __ _
`Date: June 23. 2004
`
`IN DUPLICATE
`
`Cclnlluter generated fonn "Transnlttal Lettlll" (Applicallon Fding Fdder), Merck & Co., InC., 1010112003
`
`DRL Ex. 1010, p. 002
`
`

`

`21409Y
`
`TITLE OF THE INVENTION
`PHOSPHORIC ACID SALT OF A DIPEPTIDYL PEPTIDASE-IV INHIBITOR
`
`5
`
`CROSS-REFERENCE TO RELATED APPLICATIONS
`The present invention is related to U.S. provisional application Serial No. 60/482,161,
`filed June 24, 2003, the contents of which are hereby incorporated by reference.
`
`FIELD OF THE INVENTION
`The present invention relates to a particular salt of a dipeptidyl peptidase-IV inhibitor.
`10 More particularly, the invention relates to a dihydrogenphosphate salt of 4-oxo-4-[3-(trifluoromethyl)-
`5,6-dihydro[ 1 ,2,4 ]triazolo[ 4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-amine, which is a
`potent inhibitor of dipeptidyl peptidase-IV. This novel salt and crystalline hydrates thereof are useful for
`the treatment and prevention of diseases and conditions for which an inhibitor of dipeptidyl peptidase-IV
`is indicated, in particular Type 2 diabetes, obesity, and high blood pressure. The invention further
`concerns pharmaceutical compositions comprising the dihydrogenphosphate salt and crystalline hydrates
`thereof useful to treat Type 2 diabetes, obesity, and high blood pressure as well as processes for
`preparing the dihydrogenphosphate salt and crystalline hydrates thereof and their pharmaceutical
`compositions.
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`BACKGROUND OF THE INVENTION
`Inhibition of dipeptidyl peptidase-IV (DP-IV), an enzyme that inactivates both glucose-
`dependent insulinotropic peptide (GIP) and glucagon-like peptide 1 (GLP-1), represents a novel approach
`to the treatment and prevention of Type 2 diabetes, also known as non-insulin dependent diabetes
`mellitus (NIDDM). The therapeutic potential of DP-IV inhibitors for the treatment of Type 2 diabetes
`has been reviewed: C. F. Deacon and J.J. Holst, "Dipeptidyl peptidase IV inhibition as an approach to the
`treatment and prevention of Type 2 diabetes: a historical perspective," Biochem. Biophys. Res.
`Commun., 294: 1-4 (2000); K. Augustyns, et al., "Dipeptidyl peptidase IV inhibitors as new therapeutic
`agents for the treatment of Type 2 diabetes," Expert. Opin. Ther. Patents, 13: 499-510 (2003); and D.J.
`Drucker, ''Therapeutic potential of dipeptidyl peptidase IV inhibitors for the treatment of Type 2
`diabetes," Expert Opin. Investig. Drugs, 12: 87-100 (2003).
`WO 03/004498 (published 16 January 2003), assigned to Merck & Co., describes a class
`of beta-amino tetrahydrotriazolo[4,3-a]pyrazines, which are potent inhibitors ofDP-IV and therefore
`useful for the treatment of Type 2 diabetes. Specifically disclosed in WO 03/004498 is 4-oxo-4-[3-
`( trifluoromethyl)-5,6-dihydro[ 1 ,2,4 ]triazolo[ 4,3-a ]pyrazin-7(8H)-yl]-1-(2,4,5-trifluoropheny l)butan-2-
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`- 1 -
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`DRL Ex. 1010, p. 003
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`

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`21409Y
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`amine. Pharmaceutically acceptable salts of this compound are generically encompassed within the
`scope of WO 03/004498.
`However, there is no specific disclosure in the above reference of the newly discovered
`monobasic dihydrogenphosphate salt of 4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[ 1 ,2,4 ]triazolo[ 4,3-
`a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-amine of structural formula I below.
`
`SUMMARY OF THE INVENTION
`The present invention is concerned with a novel dihydrogenphosphate salt of the
`dipeptidyl peptidase-IV (DP-IV) inhibitor 4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[ 1 ,2,4 ]triazolo[ 4,3-
`a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-amine and crystalline hydrates thereof, in particular
`a crystalline monohydrate. The dihydrogenphosphate salt and crystalline hydrates of the present
`invention have advantages in the preparation of pharmaceutical compositions of 4-oxo-4-[3-
`( trifluoromethyl)-5 ,6-dihydro[ 1 ,2,4 ]triazolo[ 4,3-a ]pyrazin-7 (SH)-yl]-1-(2,4,5-trifluorophenyl)butan-2-
`amine, such as ease of processing, handling, and dosing. In particular, they exhibit improved physical
`and chemical stability, such as stability to stress, high temperatures and humidity, as well as improved
`physicochemical properties, such as solubility and rate of solution, rendering them particularly suitable
`for the manufacture of various pharmaceutical dosage forms. The invention also concerns
`pharmaceutical compositions containing the novel salt and hydrates as well as methods for using them as
`DP-IV inhibitors, in particular for the prevention or treatment of Type 2 diabetes, obesity, and high blood
`pressure.
`
`BRIEF DESCRIPTION OF THE FIGURES
`FIG. 1 is a characteristic X-ray diffraction pattern of the crystalline monohydrate of the
`dihydrogenphosphate salt of structural formula n.
`FIG. 2 is a carbon-13 cross-polarization magic-angle spinning (CPMAS) nuclear
`magnetic resonance (NMR) spectrum of the crystalline monohydrate of the dihydrogenphosphate salt of
`structural formula n.
`FIG. 3 is a fluorine-19 magic-angle spinning (MAS) nuclear magnetic resonance (NMR)
`spectrum of the crystalline monohydrate of the dihydrogenphosphate salt of structural formula ll.
`FIG. 4 is a typical thermogravimetric analysis (TGA) curve of the crystalline
`monohydrate dihydrogenphosphate salt of structural formula ll.
`FIG. 5 is a typical differential scanning calorimetry (DSC) curve of the crystalline
`monohydrate of the dihydrogenphosphate salt of structural formula II.
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`DRL Ex. 1010, p. 004
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`21409Y
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`DET All..ED DESCRIPTION OF THE INVENTION
`This invention provides a new monobasic dihydrogenphosphate salt of 4-oxo-4-[3-
`( trifluoromethyl)-5 ,6-dihydro[ 1 ,2,4 ]triazolo[ 4,3-a ]pyrazin-7 (8H)-yl] -1-(2,4,5-trifluorophenyl)butan-2-
`amine of the following structural formula I:
`
`5
`
`F
`
`or a crystalline hydrate thereof. In particular, the instant invention provides a crystalline monohydrate of
`the dihydrogenphosphate salt of formula I.
`The dihydrogenphosphate salt of the present invention has a center of asymmetry at the
`stereogenic carbon atom indicated with an* and can thus occur as a racemate, racemic mixture, and
`single enantiomers, with all isomeric forms being included in the present invention. The separate
`enantiomers, substantially free of the other, are included within the scope of the invention, as well as
`mixtures of the two enantiomers.
`One embodiment of the present invention provides the dihydrogenphosphate salt of (2R)-
`4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-
`trifluorophenyl)butan-2-amine of structural formula II:
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`10
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`F
`
`F
`
`or a crystalline hydrate thereof.
`A second embodiment of the present invention provides the dihydrogenphosphate salt of
`(2S)-4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[ 1 ,2,4 ]triazolo[ 4,3-a ]pyrazin-7(8H)-yl]-1-(2,4,5-
`trifluorophenyl)butan-2-amine of structural formula ill:
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`20
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`DRL Ex. 1010, p. 005
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`21409Y
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`F
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`or a crystalline hydrate thereof.
`More specifically, the dihydrogenphosphate salt of the present invention is comprised of
`one molar equivalent of mono-protonated 4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-
`a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-amine cation and one molar equivalent of
`dihydrogenphosphate (biphosphate) anion.
`In a further embodiment of the present invention, the dihydrogenphosphate salt of
`structural formulae I-ll is a crystalline hydrate. In one class of this embodiment, the crystalline hydrate
`is a crystalline monohydrate.
`A further embodiment of the present invention provides the dihydrogenphosphate salt
`drug substance of structural formulae l-ID that comprises the crystalline monohydrate present in a
`detectable amount. By "drug substance" is meant the active pharmaceutical ingredient ("API"). The
`amount of crystalline monohydrate in the drug substance can be quantified by the use of physical
`methods such as X-ray powder diffraction, solid-state fluorine-19 magic-angle spinning (MAS) nuclear
`magnetic resonance spectroscopy, solid-state carbon-13 cross-polarization magic-angle spinning
`(CPMAS) nuclear magnetic resonance spectroscopy, solid state Fourier-transform infrared spectroscopy,
`and Raman spectroscopy. In a class of this embodiment, about 5% to about 100% by weight of the
`crystalline monohydrate is present in the drug substance. In a second class of this embodiment, about
`10% to about 100% by weight of the crystalline monohydrate is present in the drug substance. In a third
`class of this embodiment, about 25% to about 100% by weight of the crystalline monohydrate is present
`in the drug substance. In a fourth class of this embodiment, about 50% to about 100% by weight of the
`crystalline monohydrate is present in the drug substance. In a fifth class of this embodiment, about 75%
`to about 100% by weight of the crystalline monohydrate is present in the drug substance. In a sixth class
`of this embodiment, substantially all of the dihydrogenphosphate salt drug substance is the crystalline
`monohydrate of the present invention, i.e., the dihydrogenphosphate salt drug substance is substantially
`phase pure monohydrate.
`The crystalline dihydrogenphosphate salt of the present invention exhibits pharmaceutic
`advantages over the free base and the previously disclosed hydrochloride salt (WO 03/004498) in the
`preparation of a pharmaceutical drug product containing the pharmacologically active ingredient. In
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`DRL Ex. 1010, p. 006
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`21409Y
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`particular, the enhanced chemical and physical stability of the crystalline dihydrogenphosphate salt
`monohydrate constitute advantageous properties in the preparation of solid pharmaceutical dosage forms
`containing the pharmacologically active ingredient.
`The dihydrogenphosphate salt of the present invention, which exhibits potent DP-N
`inhibitory properties, is particularly useful for the prevention or treatment of Type 2 diabetes, obesity,
`and high blood pressure.
`Another aspect of the present invention provides a method for the prevention or
`treatment of clinical conditions for which an inhibitor of DP-N is indicated, which method comprises
`administering to a patient in need of such prevention or treatment a prophylactically or therapeutically
`effective amount of the dihydrogenphosphate salt of structural formula I or a hydrate thereof, in
`particular the crystalline monohydrate thereof. Such clinical conditions include diabetes, in particular
`Type 2 diabetes, hyperglycemia, insulin resistance, and obesity.
`The present invention also provides the use of the dihydrogenphosphate salt of structural
`formula I or a hydrate thereof, in particular the crystalline monohydrate, for the manufacture of a
`medicament for the prevention or treatment of clinical conditions for which an inhibitor of DP-N is
`indicated.
`
`The present invention also provides pharmaceutical compositions comprising the
`dihydrogenphosphate salt of structural formula I or a hydrate thereof, in particular the crystalline
`monohydrate, in association with one or more pharmaceutically acceptable carriers or excipients. In one
`embodiment the pharmaceutical composition comprise a therapeutically effective amount of the active
`pharmaceutical ingredient in admixture with pharmaceutically acceptable excipients wherein the active
`pharmaceutical ingredient comprises a detectable amount of the crystalline monohydrate of the present
`invention. In a second embodiment the pharmaceutical composition comprise a therapeutically effective
`amount of the active pharmaceutical ingredient in admixture with pharmaceutically acceptable excipients
`wherein the active pharmaceutical ingredient comprises about 5% to about 100% by weight of the
`crystalline monohydrate of the present invention. In a class of this second embodiment, the active
`pharmaceutical ingredient in such compositions comprises about 10% to about 100% by weight of the
`crystalline monohydrate. In a second class of this embodiment, the active pharmaceutical ingredient in
`such compositions comprises about 25% to about 100% by weight of the crystalline monohydrate. In a
`third class of this embodiment, the active pharmaceutical ingredient in such compositions comprises
`about 50% to about 100% by weight of the crystalline monohydrate. In a fourth class of this
`embodiment, the active pharmaceutical ingredient in such compositions comprises about 75% to about
`100% by weight of the crystalline monohydrate. In a fifth class of this embodiment, substantially all of
`the active pharmaceutical ingredient is the crystalline dihydrogenphosphate salt monohydrate of the
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`DRL Ex. 1010, p. 007
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`21409Y
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`present invention, i.e., the active pharmaceutical ingredient is substantially phase pure
`dihydrogenphosphate salt monohydrate.
`The compositions in accordance with the invention are suitably in unit dosage forms
`such as tablets, pills, capsules, powders, granules, sterile solutions or suspensions, metered aerosol or
`liquid sprays, drops, ampoules, auto-injector devices or suppositories. The compositions are intended for
`oral, parenteral, intranasal, sublingual, or rectal administration, or for administration by inhalation or
`insufflation. Formulation of the compositions according to the invention can conveniently be effected by
`methods known from the art, for example, as described in Remington's Pharmaceutical Sciences, 17th ed.,
`1995.
`
`The dosage regimen is selected in accordance with a variety of factors including type,
`species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated;
`the route of administration; and the renal and hepatic function of the patient. An ordinarily skilled
`physician, veterinarian, or clinician can readily determine and prescribe the effective amount of the drug
`required to prevent, counter or arrest the progress of the condition.
`Oral dosages of the present invention, when used for the indicated effects, will range
`between about 0.01 mg per kg of body weight per day (mgfkg/day) to about 100 mgfkg/day, preferably
`0.01 to 10 mglkg/day, and most preferably 0.1 to 5.0 mglkg/day. For oral administration, the
`compositions are preferably provided in the form of tablets containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0,
`10.0, 15.0, 25.0, 50.0, 100, 200, and 500 milligrams of the active ingredient for the symptomatic
`adjustment of the dosage to the patient to be treated. A medicament typically contains from about 0.01
`mg to about 500 mg of the active ingredient, preferably, from about 1 mg to about 200 mg of active
`ingredient. Intravenously, the most preferred doses will range from about 0.1 to about 10 mglkg/minute
`during a constant rate infusion. Advantageously, the crystalline forms of the present invention may be
`administered in a single daily dose, or the total daily dosage may be administered in divided doses of
`two, three or four times daily. Furthermore, the crystalline forms of the present invention can be
`administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes,
`using those forms of transdermal skin patches well known to those of ordinary skill in the art. To be
`administered in the form of a transdermal delivery system, the dosage administration will, of course, be
`continuous rather than intermittent throughout the dosage regimen.
`In the methods of the present invention, the dihydrogenphosphate salt and crystalline
`hydrates herein described in detail can form the active pharmaceutical ingredient, and are typically
`administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively
`referred to herein as 'carrier' materials) suitably selected with respect to the intended form of
`administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional
`pharmaceutical practices.
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`DRL Ex. 1010, p. 008
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`21409Y
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`For instance, for oral administration in the form of a tablet or capsule, the active
`pharmaceutical ingredient can be combined with an oral, non-toxic, pharmaceutically acceptable, inert
`carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium
`phosphate, calcium sulfate, mannitol, sorbitol and the like; for oral administration in liquid form, the
`active phamiaceutical ingredient can be combined with any oral, non-toxic, pharmaceutically acceptable
`inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable
`binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture.
`Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, com sweeteners,
`natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose,
`polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate,
`sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.
`Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the
`like.
`
`\
`
`The dihydrogenphosphate salt of structural formula I and the crystalline monohydrate
`have been found to possess a high solubility in water, rendering it especially amenable to the preparation
`of formulations, in particular intranasal and intravenous formulations, which require relatively
`concentrated aqueous solutions of active ingredient. The solubility of the crystalline
`dihydrogenphosphate salt monohydrate of formula I in water has been found to be about 72 mg/mL.
`According to a further aspect, the present invention provides a process for the
`preparation of the dihydtogenphosphate salt of formula I, which process comprises reacting 4-oxo-4-[3-
`(trifluoromethyl)-5,6-dihydro[ 1 ,2,4 ]triazolo[ 4,3-a ]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-
`amine of structural formula IV below:
`
`F
`
`F
`
`with approximately one equivalent of phosphoric acid in a suitable Ct-C5 alkanol, such as methanol,
`ethanol, isopropyl alcohol (IPA), and isoamyl alcohol (IAA) or aqueous Ct-C5 alkanol. The reaction is
`carried out at a temperature range of about 25 oc to about 80 oc. The phosphoric acid solution can be
`added to a solution of the amine, or the addition can be performed in the reverse direction. The
`crystalline dihydrogenphosphate salt monohydrate is obtained by crystallization from an aqueous Ct-C5
`alkanol solution of the dihydrogenphosphate salt as described below.
`
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`DRL Ex. 1010, p. 009
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`

`

`21409Y
`
`GENERAL METHODS FOR CRYSTALLIZING THE MONOHYDRATE OF THE
`DIHYDROGENPHOSPHATE SALT OF STRUCTURAL FORMULA I:
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`(a) In ethanol/water system at 25 °C:
`(1) crystallization from a mixture of compound I in ethanol and water, such that the water concentration
`is above 31 weight percent,
`(2) recovering the resultant solid phase, and
`(3) removing the solvent therefrom.
`
`(b) In isoamyl alcohol (IAA)/water system at 25 °C:
`(1) crystallization from a mixture of compound I in IAA and water, such that the water concentration is
`above 2.9 weight percent;
`(2) recovering the resultant solid phase; and
`(3) removing the solvent therefrom.
`
`(c) In IAN water system at 40 °C:
`(1) crystallization from a mixture of compound I in IAA and water, such that the water concentration is
`above 3.6 weight percent;
`(2) recovering the resultant solid phase; and
`(3) removing the solvent therefrom
`
`(d) In IANwater system at 60 °C:
`(1) crystallization from a mixture of compound I in IAA and water, such that the water concentration is
`above 4.5 weight percent;
`(2) recovering the resultant solid phase; and
`(3) removing the solvent therefrom.
`
`(e) In Isopropyl alcohol (IPA)/water system at 25 °C:
`(1) crystallization from a mixture of compound I in IPA and water, such that the water concentration is
`above 7.0 weight percent;
`(2) recovering the resultant solid phase; and
`(3) removing the solvent therefrom
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`(f) In IPA/water system at 40 °C:
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`-8-
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`DRL Ex. 1010, p. 010
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`21409Y
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`(1) crystallization from a mixture of compound I in IPA and water, such that the water concentration is
`above 8.1 weight percent;
`(2) recovering the resultant solid phase; and
`(3) removing the solvent therefrom.
`
`(g) In IPA/water system at 75°C:
`( 1) crystallization from a mixture of compound I in IP A and water, such that the water concentration is
`above about 20 weight percent;
`(2) recovering the resultant solid phase; and
`(3) removing the solvent therefrom.
`
`The starting compound of structural formula IV can be prepared by the procedures
`detailed in Schemes 1-3 and Example 1 below.
`In a still further aspect, the present invention provides a method for the treatment and/or
`prevention of clinical conditions for which a DP-IV inhibitor is indicated, which method comprises
`administering to a patient in need of such prevention or treatment a prophylactically or therapeutically
`effective amount of the salt of Formula I as defined above or a crystalline hydrate thereof.
`The following non-limiting Examples are intended to illustrate the present invention and
`should not be construed as being limitations on the scope or spirit of the instant invention.
`Compounds described herein may exist as tautomers such as keto-enol tautomers. The
`individual tautomers as well as mixtures thereof are encompassed with compounds of structural formula
`I.
`
`The term"% enantiomeric excess" (abbreviated "ee") shall mean the % major
`enantiomer less the % minor enantiomer. Thus, a 70% enantiomeric excess corresponds to formation of
`85% of one enantiomer and 15% of the other. The term "enantiomeric excess" is synonymous with the
`term "optical purity."
`
`EXAMPLE
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`-9-
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`DRL Ex. 1010, p. 011
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`21409Y
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`C2R)-4-oxo-4-f3-( trifluoromethyll-5.6-dihydro[ 1 ,2,4ltriazolof 4.3-a lpyrazin-7 (8ffi-yll-1-(2.4.5-
`trifluorophenyl)butan-2-amine dihydrogenphosphate monohydrate
`
`Preparation of 3-(trifluoromethyl)-5,6, 7 ,8-tetrahydrof 1 ,2.4ltriazolof 4.3-a]pyrazine hydrochloride ( 1-4)
`
`Scheme 1
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`1. CF3COOEt, CH3CN
`
`2. CICOCH2CI, NaOH
`
`POCI3
`CH3CN
`
`N-N
`F3C_.{(0 )l_CH2CI
`1-2
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`MeOH
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`MeOH, HCI, 55 °C
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`Step A:
`
`Preparation of his hydrazide ( 1-1)
`Hydrazine (20.1 g, 35 wt% in water, 0.22 mol) was mixed with 310 mL of acetonitrile.
`31.5 g of ethyl trifluoroacetate (0.22 mol) was added over 60 min. The internal temperature was
`increased to 25 oc from 14 oc. The resulting solution was aged at 22-25 oc for 60 min. The solution
`was cooled to 7 oc. 17.9 g of 50 wt% aqueous NaOH (0.22 mol) and 25.3 g of chloroacetyl chloride
`(0.22 mol) were added simultaneously over 130 min at a temperature below 16 °C. When the reaction
`was complete, the mixture was vacuum distilled to remove water and ethanol at 27 - 30 oc and under 26
`- 27 in Hg vacuum. During the distillation, 720 mL of acetonitrile was added slowly to maintain
`constant volume (approximately 500 mL). The slurry was filtered to remove sodium chloride. The cake
`was rinsed with about 100 mL of acetonitrile. Removal of the solvent afforded his-hydrazide H ( 43.2 g,
`96.5% yield, 94.4 area% pure by HPLC assay).
`1H-NMR (400 MHz, DMSO-d6): 8 4.2 (s, 2H), 10.7 (s, 1H), and 11.6 (s, 1H) ppm.
`13C-NMR (100 MHz, DMSO-d6): 8 41.0, 116.1 (q, J =362Hz), 155.8 (q, J =50 Hz), and 165.4 ppm.
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`-10-
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`DRL Ex. 1010, p. 012
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`Step B:
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`Preparation of 5-(trifluoromethyl)-2-(chloromethyl)-1.3.4-oxadiazole
`(1-2)
`Bishydrazide 1::1 from Step A (43.2 g, 0.21 mol) in ACN (82 mL) was cooled to 5 °C.
`Phosphorus oxychloride (32.2 g, 0.21 mol) was added, maintaining the temperature below 10 °C. The
`mixture was heated to 80 °C and aged at this temperature for 24 h until HPLC showed less than 2 area%
`of 1::1. In a separate vessel, 260 mL of IP Ac and 250 mL of water were mixed and cooled to 0 °C. The
`reaction slurry was charged to the quench keeping the internal temperature below 10 °C. After the
`addition, the mixture was agitated vigorously for 30 min, the temperature was increased to room
`temperature and the aqueous layer was cut. The organic layer was then washed with 215 mL of water,
`215 mL of 5 wt% aqueous sodium bicarbonate and finally 215 mL of 20 wt% aqueous brine solution.
`HPLC assay yield after work up was 86-92%. Volatiles were removed by distillation at 75-80 mm Hg,
`55 oc to afford an oil which could be used directly in Step C without further purification. Otherwise the
`product can be purified by distillation to afford 1-2 in 70-80% yield.
`1H-NMR (400 MHz, CDCh): B 4.8 (s, 2H) ppm.
`13C-NMR (100 MHz, CDCh): B 32.1, 115.8 (q, J =337Hz), 156.2 (q, J =50 Hz), and 164.4 ppm.
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`Step C:
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`Preparation of N-[(2Z>-piperazin-2-ylideneltrifluoroacetohydrazide
`!..U)
`To a solution of ethylenediamine (33.1 g, 0.55 mol) in methanol (150 mL) cooled at -20
`°C was added distilled oxadiazole 1-2 from Step B (29.8 g, 0.16 mol) while keeping the internal
`temperature at -20 °C. After the addition was complete, the resulting slurry was aged at -20 °C for 1 h.
`Ethanol (225 mL) was then charged and the slurry slowly warmed to -5 °C. After 60 min at -5 °C, the
`slurry was filtered and washed with ethanol (60 mL) at -5 °C. Amidine 1-3 was obtained as a white solid
`in 72% yield (24.4 g, 99.5 area wt% pure by HPLC).
`1H-NMR (400 MHz, DMSO-d6): B 2.9 (t, 2H), 3.2 (t, 2H), 3.6 (s, 2H), and 8.3 (b, 1H) ppm. 13C-NMR
`(100 MHz, DMSO-d6): B 40.8, 42.0, 43.3, 119.3 (q, J =350Hz), 154.2, and 156.2 (q, J =38Hz) ppm.
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`Step D:
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`Preparation of 3-(trifluoromethyl)-5.6. 7 .8-tetrahydro[ 1.2.4ltriazolo[ 4.3-alpyrazine
`hydrochloride (1-4)
`A suspension of arnidine 1-3 (27 .3 g, 0.13 mol) in 110 mL of methanol was warmed to
`55 °C. 37% Hydrochloric acid (11.2 mL, 0.14 mol) was added over 15 min at this temperature. During
`the addition, all solids dissolved resulting in a clear solution. The reaction was aged for 30 min. The
`solution was cooled down to 20 °C and aged at this temperature until a seed bed formed ( 10 min to 1 h).
`300 mL of MTBE was charged at 20 °C over 1 h. The resulting slurry was cooled to 2 °C, aged for 30
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`- 11 -
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`DRL Ex. 1010, p. 013
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`min and filtered. Solids were washed with 50 mL of ethanol:MTBE (1:3) and dried under vacuum at 45
`°C. Yield oftriazole 1-4 was 26.7 g (99.5 area wt% pure by HPLC).
`1H-NMR (400 MHz, DMSO-d6): o 3.6 (t, 2H), 4.4 (t, 2H), 4.6 (s, 2H), and 10.6 (b, 2H) ppm; Bc-NMR
`(100 MHz, DMSO-d6): o: 39.4, 39.6, 41.0, 118.6 (q, J =325Hz), 142.9 (q, J =50 Hz), and 148.8 ppm.
`Scheme 2
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`F 2-1
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`0~~
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`OH
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`tBuCOCI, 1Pr2NEt,
`DMAP, DMAc
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`F
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`F
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`0
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`0
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`0 o)(
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`2-2
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`F
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`F
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`HCI
`HN~N.
`-~N-{N
`1-4
`CF3
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`F