h High-throughput Polymorph & Salt Screening Laboratory
`Memo
`
` 12 Nov 2002
`
`K. Hansen, J. Armstrong, E. Grabowski, Y. Xiao
`TO:
`K. Somerville, V. Vydra, J. Chilenski
`FROM:
`L. Crocker, D. Mathre, R. Sidler
`Cc:
`SUBJECT: L-000224715 Phosphate Salt Polymorph Screen
`Summary
`
`A polymorph screen of L-000224715 Phosphate salt was performed. Bulk compound
`was supplied as the crystalline phosphate salt. Very few crystalline solids were
`obtained from these experiments. Two notable forms identified were the hydrate
`evaporated
`from ethanol/water and
`the alcohol solvate precipitated
`from
`ethanol/ethylene glycol into butyl ether.
`Design
`
`F
`
`F
`
`F
`
`NH2
`
`O
`
`N
`
`N
`
`N
`
`N
`
`F
`F
`
`F
`
`Merck Exhibit 2137, Page 1
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`
`
`Procedure
`
`The substrate (L-000224715 Phosphate salt, K. Hansen) was dispensed as a powder
`(10mg +/-1 mg) using the Autodose Powdernium. After the substrate was dispensed to
`each of the wells, the 96-well plate was placed on the Cavro Liquid Handling Robot
`deck, with the wash solvent as methanol, for the solvent dispense (800 uL/well). The 96-
`well plate was mapped by quadrants.
`
`The solvent mapping of the 96-well plate is as follows:
`
`Quadrant 1: (upper left) 0-100% (v/v) Water by 20% increments
`Row A: Ethanol
`Row B: 1-Propanol
`Row C: Acetonitrile
`Row D: 1,4-Dioxane
`
`Quadrant 2 (upper right) 0-100% (v/v) Ethanol by 20% increments
`Row A: Ethylene Glycol
`Row B: Diisopropyleamine
`Row C: Trifluorotoluene
`Row D: 1,2-Dichloroethane
`
`Quadrant 3 (lower left) 0-100% (v/v) 1,2-Dichloroethane by 20% increments
`Row E: Nitromethane
`Row F: 1,4-Dioxane
`Row G: Toluene
`Row H: Perfluoroheptane
`
`Quadrant 4: (lower right) 0-100% (v/v) Cyclohexane by 20% increments
`Row E: 2-Butanone
`Row F: 1,2-Dimethoxyethane
`Row G: Trichloroethylene
`Row H: Isopropyl Acetate
`
`Once all the solvents were dispensed to each well, the 96-well plate was capped and
`placed on the crystallizer deck. The system was equilibrated at 65C for 2 hours (during
`which anti-solvents were added to the precipitation plate), filtered hot at 65C and
`daughtered to each of the three experiment plates (Evaporation 200uL, Precipitation
`100uL and Cooling 200uL). The cooling plate was thermal cycled with a cubic cool
`down temperature gradient of 65-10 C over 8 hours. The plate was equilibrated at 10
`C for 2 hours and then the supernatant was removed from the cooling and precipitation
`experiment plates. The following day, each experiment was wicked to dry the remaining
`solvent and the plates were removed for analysis.
`
`
`
`Merck Exhibit 2137, Page 2
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`
`
`Results
`The wells were inspected both visually and by polarized light microscopy for
`birefringence. Wells containing material were then scanned by XRPD.
`Evaporation (Library ID 100167)
`Birefringence
`
`X-ray Powder Diffraction
`
`
`
`Merck Exhibit 2137, Page 3
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`
`
`Precipitation (Library ID 100168)
`
`Anti-solvents for the precipitation plate were mapped as follows (200uL/well).
`
`Birefringence
`
`
`
`Merck Exhibit 2137, Page 4
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`
`
`X-ray Powder Diffraction
`
`Cooling (Library ID 100169)
`Birefringence
`
`
`
`Merck Exhibit 2137, Page 5
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`
`
`X-ray Powder Diffraction
`
`Sorting
`
`A single element library (100182, blue) was submitted as the bulk L-000224715
`Phosphate Salt and the XRPD was run to use for comparison. The screening XRPD
`data was then sorted using Spectra Studio and two possible new forms were identified.
`Well B02 from the evaporation plate (100167) was sorted with the bulk salt and
`therefore not repeated.
`
`Possible leads:
`
`Pink (Group 4)
`Element A02 from 100167 (Evaporation) scaled-up
`Element A02 from 100169 (Cooling)
`
`Light Blue (Group 2)
`Element A11 from 100168 (Precipitation) scaled-up
`
`
`
`Merck Exhibit 2137, Page 6
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`
`
`Scale-ups (Library ID 100200 Evaporation, 100201 Precipitation)
`
`Two experiments were scaled-up, A02 from Evaporation (100167) and A11 from
`Precipitation (100168). 50 mg of L-000224715 Phosphate Salt were manually charged
`to two vials and reaction volumes were scaled accordingly to 4 mL. The evaporation
`experiment (now well A01) was run in 20% water in Ethanol and the precipitation
`experiment (now well A02) was run in 20% ethylene glycol in ethanol. The mixtures we
`warmed to 65 C for 2 hours. The evaporation mixture was then transferred to a larger
`vial and allowed to evaporate. 500 uL of the precipitation experiment were charged
`subsurface to 1mL butyl ether and allowed to crystallize. Solids were retrieved from
`each of the experiments and analyzed by XRPD. Both scans sorted into Group 2 (from
`Precipitation plate).
`
`Results from the screen and scale-ups were compared with data from Physical
`Measurements (R. Ferlita). Group 2 matches the pattern for an alcohol solvate and
`Group 4 match for a hydrate form. Neither group was determined to be novel.
`
`
`
`Merck Exhibit 2137, Page 7
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`