Karl Hansen
`Weekly Report
`7 Jan 2002
`
`L-859,016
`
`Material produced in the prep lab was front run. Due to the termination of the project, only one 2 kg batch
`will be processed and responsibility for coupling will be transferred to M. Kubryk.
`
`L-224,715
`
`In order to support the delivery timeline, readily available starting materials needed to be identified. An
`analog of carboxylic acid 2 has been used to make the structurally similar L-822,869. This material is not
`readily available, however 500 g have been ordered by the medicinal scale-up group. Steps were taken to
`find this material and also purchase an additional 115g from the same lot from the supplier. This material
`will support the first 200g delivery of 715.
`
`For the second delivery, the most readily available starting point is arylbromide 3. 25kg of this compound
`were purchased at 130$/kg from interchem for a delivery in 3weeks. An additional 1kg was purchased
`from aldrich at ~2000$/kg to support research for the next three weeks.
`
`F
`
`F
`
`F
`
`Salt Form:
`
`NH2 O
`
`N
`
`1
`
`N
`
`N
`
`N
`
`CF3
`
`F
`
`F
`
`F
`
`CO2H
`
`2
`
`F
`
`F
`
`F
`
`Br
`3
`
`Salt studies have been performed by V. Vydra in Automation. The study has identified the phosphate,
`tartrate, sulfate and benzensulfonate as providing crystalline solids by powder diffraction. The bezylate
`and tartrate will probably be pursued first.
`
`Coupling:
`
`Initial experiments show the coupling of boc amino acid intermediate 4 to triazolopyrazine 5 to proceed in
`the absence of HOBT with isolation without chromatography.
`
`N
`
`N
`
`EDC
`DMF
`
`CF3
`
`F
`
`F
`
`F
`
`N
`HCl
`
`5
`
`Boc
`
`NH O
`
`1
`
`N
`
`N
`
`N
`
`N
`
`F
`
`F
`
`Boc
`
`NH
`
`+
`
`CO2H
`
`HN
`
`F
`
`4
`
`CF3
`70130-309 100mg of 4 was mixed with 1.0 equiv of 5 and 1.2 equiv. of EDC in 20ml/g DMF. After 18h
`75% conversion was observed to a later eluting peak. 1.0 equiv. of Cs2CO3 was charged, but no further
`conversion was observed. An additional 0.3 equiv of EDC was charged and 95% of 4 was observed after
`2h. The material was isolated by diluting with 10% NaCl (roughly 30ml/g). Solids present in the reaction
`mixture dissolved then new crystals formed. These were isolated at RT and washed with 10% NaCl and
`water. 84 mg (55%) of white solids were isolated. 1H-NMR was difficult to interpret due to Boc group,
`
`Merck Exhibit 2131, Page 1
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`but LCMS of the isolated solids gave m/z = 451 (M-tBu+1) for the major peak. Sample given to Dormer
`for confirmation. ML s contained product in comparable LCAP to unreacted starting material. Several
`impurities are present in both the MLs and product in the 1-5% range. One elutes very closely to the
`product with m/z = 530.
`
`70130-311 Coupling was performed as above with EDC and HOBT. Reaction stopped at 80% conversion
`and required a second charge of EDC. Solids were isolated as above (90% physical yield), but two major
`impurities were present one eluting early (3.4 min) and one at 14 min. LCMS is being performed to
`confirm identity. These impurities are the majority of the LCAP for this sample.
`
`Merck Exhibit 2131, Page 2
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Karl Hansen
`Weekly Report
`14 Jan 2002
`
`L-859,016
`
`2 kg of pyrrazolotriazine was coupled by M. Kubryk in prep lab. Material will be analyzed and then bagged
`next week.
`
`L-224,715
`
`Raw materials:
`
`615g of the trifluorophenyl acetic acid have been received.
`
`Salt Form:
`
`Besylate salt evaluated by Phys. Meas.
`Tartrate may have been prepares
`
`70130-321 Solutions of 715 and benzensulfonic acid in DCM were combined in 1:1 stoichiometry in 3
`vials giving ~35mg of salt in each. The DCM was removed in vacuo to give a white solidthat was
`amorphous under the microscope. The solids were then suspended in the crystallization solvents toluene,
`IPA and IPAC. The IPAC and toluene did not dissolve completely in 2ml while the IPA required 2
`volumes of Hexanes to afford solids. These were heated at 80C for 0.5h and then cooled to RT only IPA
`expt. dissolved completely. Of the three expts only the IPAC was judged to have crystals. These were
`isolated and then given to Physical Measurements along with a sample of besylate salt prepared by M.
`Palucki (72061-046). Both of these samples had identical HPLC traces, supporting a 1:1 salt formation.
`Physical measurements indicated that btoh samples had a MP of 179ºC, identical x-ray powder patterns and
`no appreciable loss oin the tg. Yield was 27mg (77%) for the IPAC conditions.
`
`70130-323 Dissolved 92mg of L224,715 in 4ml of IPA containing 1 equiv of H3PO4. Solids forme
`immediately and did not dissolve upon reheating. Removed IPA and was able to get solids to redissolve in
`MeOH. Held onto solution.
`
`70130-325 Attempted to make HBr salt of 715 as above. Only oily precitpitates formed in a variety of
`solvents.
`
`70130-327 Dissolved 50mg of 715 in 2ml of IPA. Added 1 equiv of (L)-tartaric acid dissolved in 0.2ml
`IPA. Solution was clear but after 10 minutes crystals began to form. The slurry was heated at 77ºC for
`30min but solids did not dissolve completely and appeared microcrystalline under scope before and after
`age. DSC gave an exothermic peak at 207ºC.
`
`70130-329 Repeated above expt with ethanol. Crystals formed quickly at RT in 2.2 ml of solvent. Diluted
`with 4ml of EtOH and achieved a clear soln at 75ºC. Temp was slowly lowered to RT and xtals formed at
`40ºC. Crystals looked much larger under scope. They were isolated at RT with an ethanol wash. 1H-NMR
`shows definite presence of 715 with shifting of peaks realtive to free base, although the tartrate peaks were
`buried. 58% recovery. Solids given to phys meas. For evaluation.
`
`70130-331 Attempted to form crystals as above in 2.2ml of MeOH-did not ppt even at 0C. Added 2ml of
`IPA and solids formed. Heated to clear soln and then cooled-gacve xtals similar to ethanol expt.
`
`70130-333 Performed crystallization in 1.2ml of MeOH (40mg scale, 25ml/g) Clear soln was aged over
`weekend and gave good size crystals. These were isolated and will be analyzed and compared to 329.
`
`Merck Exhibit 2131, Page 3
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Coupling:
`
`F
`
`F
`
`F
`
`Boc
`
`NH
`
`4
`
`+
`
`CO2H
`
`N
`
`N
`
`CF3
`
`N
`HCl
`
`HN
`
`5
`
`EDC
`DMF
`
`F
`
`F
`
`F
`
`Boc
`
`NH O
`
`1
`
`N
`
`N
`
`N
`
`N
`
`CF3
`
`70130-315 Scale-up to 500mg of the coupling (70130-313) gave 100% conv to the desired product
`however, two earlier eluting impurities are isolated with desired product. Samples of the ML s and product
`have been given to analytical for identification
`
`Merck Exhibit 2131, Page 4
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Karl Hansen
`Weekly Report
`21 Jan 2002
`
`L-859,016
`
`2 kg of L859,016 was prepared in the prep lab. This material was 99.9% pure with one impurity at 0.06%.
`Material will be sent to chem data on an a-sheet unmilled
`
`L224,715:
`
`Balsells: Racemic material has been brought through the beta-lactam intermediate. Noyori reduction of
`keto ester gave 89% ee material. This was brought throught to beta lactam and then upgraded to 99% ee in
`one crystallization.
`
`F
`
`F
`
`F
`
`CN
`
`1
`
`H2SO4, H2O,
`AcOH, reflux
`99%
`
`F
`
`F
`
`F
`
`1) (COCl)2
`2) Meldrum's acid
` 2,4,6-collidine
`
`77-84%
`
`F
`
`COOH
`
`2
`
`F
`
`F
`
`O
`
`O
`
`O
`
`OH
`
`O
`3
`
`OCH3
`
`OBn
`
`HN
`
`OCH3
`
`F
`
`F
`
`F
`
`F
`
`F
`
`OCH3
`
`(S)-BINAP(Ru)
`
`100%, 89% e.e.
`
`F
`
`O
`
`O
`
`4
`
`OH
`
`OH
`
`O
`
`6
`
`BnONH2, EDC
`80-95%
`
`F
`
`F
`
`F
`
`F
`
`F
`
`F
`
`OH
`
`OH
`
`O
`
`5
`
`O
`
`7
`
`O
`
`N
`BnO
`
`F
`
`8
`
`MeONa, MeOH
`99%
`
`F
`
`NH
`
`O
`
`9
`
`BnO
`
`F
`
`MeOH, reflux
`
`75-99%
`
`LiOH, THF
`
`100%
`
`DIAD, PPh3
`
`F
`
`F
`
`F
`
`-Lactam formation:
`85% yield, 89% e.e.
`
`Crystallization:
`50% yield, >99% e.e.
`
`Salt Form: Two salts (besylate and tartrate)have been scaled up and given to PR&D and Phy. Meas.
`Tartrate appears to form a hemi-hydrate while besylate forms a hemihydrate, only under very humid
`conditions. HCl salt appears to be crystalline as well
`
`Besylate salt
`
`70130-337, 339 Benzenesulfonic acid (0.2ml, 79mg/ml soln in IPAC, 1 equiv.) was added to solution of
`715 in IPAC (50ml/g). Solids did not form until the solution was heated at 55ºC. ML s were assayed to
`contain 1mg/g of 715. Cooled to RT, filtered, washed with IPA. Dried at 40ºC in vac oven w/ N2 sweep.
`85% isolated yield. Same form as previous samples. Solubility of Besylate salt is ~1.2 mg/g at RT by LC.
`
`Merck Exhibit 2131, Page 5
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`70130-347 Scaled up above procedure to 150mg of 715. Performed addition of benzenesulfonic acid at
`60ºC. Solids formed after 0.5 h . Cooled over 1 h to RT. Isolated as above. 87% yield. 100mg were
`given to PR&D and 50mg to physical measurements. Compound appears to form a hemihydrate at 85%
`humidity but then loses water upon drying.
`
`Tartrate Salt:
`
`70130-333 Added tartaric acid in MeOH to solution of 715 in MeOH. Crystals formed and slurry was
`thick. Diluted with more MeOH and then heated to 48ºC. Cooled to RT and filtered. DSC shows
`MP=206, Tg = 1.3% (hemihydrate?) , xray show crystalline pattern different from 329.
`
`70130-343,345 Two experiments performed by adding tartaric acid in MeOH (343) and water (345) to a
`solution of 715 in MeOH. In the aqueous system, solids formed within 5 minutes, while the methanol
`system required several hours before solids were observed. Isolated 345 to give 32% of the tartrate salt.
`Solution composition at isolation was 90%Meoh/water. [715] = 14 mg/g. Xray shows crystalline salt
`different from previous batches.
`
`70130-349 Added aq solution of L-tartaric acid to IPA solution of 715 (90% IPA/water, 55ml/g after
`addn.) Solids formed immediately at RT, which were microcrystalline. Heated at 60C fro 16h at which
`point slurry turned over and crystals were now needle. Isolated 23mg (84%) xray shows form similar to
`333. This result was scaled-up to further study system.
`
`70130-359 150mg sacle of 349. Yield was 87%. 1H-NMR shows a 1:1 stoichiometry (performed by P.
`Dormer). Hygroscopicity studies done by Phys. Measurements show that a hemihydrate forms reversibly
`and that the same form is recovered at the beginning and end of the experiment. 100mg given to PR&D.
`
`Succinate salt:
`
`70130-351 Attemted top generate salt in MeOH- too soluble no solids formed.
`
`70130-355 Attmepted to generate salt in iBuOH. Solids formed but these were sticky and not crystalline
`under microscope.
`
`70130-357 Formed salt in IPA. Solids formed but then re-dissolve and then reform sticky amorphous
`solids.
`
`HCl:
`
`70130-361 Added 1.0equiv. of HCl in Et2O(2.0M) to solution of 715 in IPA. Solids formed immediately
`which looked birefringent, but were not well-shaped. Warmed to 60C and solution was clear. Upon
`sooling solids formed quickly. Added small amt of MeoH and then repeated heat and cool. Small needles
`formed. Less than 20% yield.
`
`70130-369 Performed crystallization in 90%IPA/HCl. Isolated 13 mg of white solids which were
`crystalline. The DSC showed los of material at 60ºC followed by melting at 146ºC. Result will be scaled
`up to investigate further.
`
`Merck Exhibit 2131, Page 6
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Karl Hansen
`Weekly Report
`28 Jan 2002
`
`L224,715:
`
`Balsells: Racemic material has been brought through the beta-lactam intermediate. Noyori reduction of
`keto ester gave 89% ee material. This was brought throught to beta lactam and then upgraded to 99% ee in
`one crystallization.
`
`F
`
`F
`
`F
`
`CN
`
`1
`
`H2SO4, H2O,
`AcOH, reflux
`99%
`
`F
`
`F
`
`F
`
`1) (COCl)2
`2) Meldrum's acid
` 2,4,6-collidine
`
`77-84%
`
`F
`
`COOH
`
`2
`
`F
`
`F
`
`MeOH, reflux
`
`75-99%
`
`LiOH, THF
`
`100%
`
`DIAD, PPh3
`
`F
`
`F
`
`F
`
`-Lactam formation:
`85% yield, 89% e.e.
`
`Crystallization:
`50% yield, >99% e.e.
`
`Salt Form:
`
`Besylate:
`
`O
`
`O
`
`O
`
`OH
`
`O
`3
`
`OCH3
`
`OBn
`
`HN
`
`OCH3
`
`F
`
`F
`
`F
`
`F
`
`F
`
`OCH3
`
`(S)-BINAP(Ru)
`
`100%, 89% e.e.
`
`F
`
`O
`
`O
`
`4
`
`OH
`
`OH
`
`O
`
`6
`
`BnONH2, EDC
`80-95%
`
`F
`
`F
`
`F
`
`F
`
`F
`
`F
`
`OH
`
`OH
`
`O
`
`5
`
`O
`
`7
`
`O
`
`N
`BnO
`
`F
`
`8
`
`MeONa, MeOH
`99%
`
`F
`
`NH
`
`O
`
`9
`
`BnO
`
`F
`
`70130-373, 377, 379, 385,387- Several attempts were made to crystallize the besylate salt of 715 in IPAC
`containing water. 5 and 10% water resulted in phase separation and no solids formed. Using 5.5 equiv. of
`added water both in the crystallization and adding to a slurry out of dry IPAC at 60 ºC did not dissolve the
`solids. The mixtures were aged overnight, cooled to rt, isolated and given to Phys. Measurements to
`determine if the hydrate was formed.
`
`Tartarte:
`
`70130-389 Crystallized the L-tatrtate salt from dry n-PrOH and aq. n-ProH. Similar results were observed
`in the morphology of the crystals as is seen in dry and wet I-PrOH.
`
`Merck Exhibit 2131, Page 7
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Phosphate:
`
`70130-393 Generated phosphate salt by adding aq solution of phosphoric acid to IPA solution of 715.
`Solids were initially amorphous, but after aging at 60ºC seemed to turnover to crystalline solids. These
`were isolate and will be given to Phys. Meas.
`
`Sulphate:
`
`70130-395 Attemted to crystallize as above (adding aq soln of sulfuric acid to IPA soln of 715). Only
`amorphous solids were observed after aging at RT.
`
`Merck Exhibit 2131, Page 8
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Karl Hansen
`Weekly Report
`04 Feb 2002
`
`Balsells:
`
`Kubryk:
`
`Hansen summary: 1. 0.5 g of benzenesulfonate salt has been synthesized from phenyl acetic acid. End
`game yield is ~60%
`
`2. Phosphate salt has been isolated but is weakly Crystaline and not hygroscopic.
`
`Coupling
`
`F
`
`F
`
`F
`
`BnO
`
`NH O
`
`OH
`
`2
`
`HN
`
`+
`
`N
`
`N
`
`CF3
`
`N
`HCl
`
`3
`
`L224,715
`benzenesulfonate salt
`
`70316-001 500mg(1.47 mmol) of aminoacid 2 was dissolved in 5ml CAN. The flask was charged with
`0.5ml water and 500mg (1.5 equiv.) of heterocycle 3. The solution was cooled to 0ºC and then EDC was
`charged (1.5 equiv.) followed by 1 equiv. of N-methylmorpholine. The reaction was aged at 0ºC for 2 h at
`which point no aminoacid remained. 6ml/g of water was added along with 10ml/g EtOAc. The layers
`were cut and the organic washed with 5% NaHCO3. The Aq contained no product by HPLC. The volatiles
`were removed to give a thick oil.
`
`The oil was re-dissolved in 10 ml MeOH and then submitted for hydrogenation. Initially the solution was
`charged with 0.15g of 10% Pd/C and then shaken with 40PSI H2 at RT. After 12 h only ~30% conversion
`to 715 was observed (benzyl alcohol was observed in the crude reaction mix. The solution was filtered,
`concentrated and then re-dissolved in 10ml MeOH. This solution was subjected to 0.15 g 10% Pd/C,
`40PSI, 50º. After overnight, all of the starting material had been consumed. The mixture was filtered and
`then assayed to contain 422 mg of 715 (66% yield).
`
`The MeOH was removed and then oil was re-dissolved in IPAC(15ml). The solution was filtered to
`remove solids and the filter was washed with 5ml IPAC (20ml total volume). 1 equiv. of PhSO3H was
`dissolved in 2ml IPAC and this was added to the IPAC soln of the free base. This was heated at 60ºC for
`2h to give white crystalline solids, which were filtered and washed with IPAC. After drying 520mg (60%)
`of white solids were obtained. 1H-NMR and HPLC confirm that this is besylate salt. Samples were given
`to analytical. One late eluting peak is observed by HPLC and a small amount of IPAC was observed in the
`NMR.
`
`Salt Form:
`
`Phosphate:
`
`70130-393 Generated phosphate salt by adding aq solution of phosphoric acid to IPA solution of 715.
`Solids were initially amorphous, but after aging at 60ºC seemed to turnover to crystalline solids. These
`were isolate and will be given to Phys. Meas. Xray is weak, but crystalline. Material is not hygroscopic.
`Need to xtallize slower.
`
`70316-15 Attempted to crystallize phosphate salte from 8.3% water/IPA. Only powder obtained.
`
`Merck Exhibit 2131, Page 9
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`70316-17 Generated H3PO4 salt by adding 1.0 eq of acid to free base in ethanol. Powder solids formed
`immediately so 75 ul of water was added (Final composition: 12% water/EtOH, 31ml/g). Heating this
`slury to 75ºC gave clear soln which upon slow cooling gave crystals but also a lot of amorphous material.
`Slurry was placed in 70ºC oven and held for 1h at 70, then cooled to 10ºC over 18 h. Large plates were
`observed and these were isolated and given to phys. Meas. If this material has a better xray than
`previous and is still anhydrous, then phosphate will be pursued aggressively
`
`
`Merck Exhibit 2131, Page 10
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Karl Hansen
`Weekly Report
`11 Feb 2002
`
`Hansen summary:
`
`Coupling
`
`F
`
`F
`
`F
`
`BnO
`
`NH O
`
`OH
`
`2
`
`HN
`
`+
`
`N
`
`N
`
`CF3
`
`N
`HCl
`
`3
`
`L224,715
`benzenesulfonate salt
`
`70316-009 10.0g of beta-lactam was hydrolyzed to acid 2. This material was then redissolved in 10ml/g
`CH3CN. 1.5 equiv of triazole HCl salt was charged at 0ºC and then 1.5 equiv. of EDC and 1.0 equiv of N-
`methyl morpholine. After 4 h >99% of the acid had converted to product. The slurry was diluted with
`water and extracted 1.x10ml/g of EtOAc. The organic layer was washed with 5% NaHCO3 and 20% NaCl.
`The organic layer was concentrated into MeoH (10ml/g) and then submitted to Hyrdogenation conditions
`with 3.00 g of 10% Pd/C and 40PSI, 50ºC. After 18 h, all of the intermediate had converted to 715.
`
`The catalyst was filtered off through a celite pad and the MeoH solution was concentrated to an oil. The
`assay yield of the filtered methanol solution was assayed to contain 10.40 g of 715 (87% assay yield). The
`concentrated organic layer was dissolved in 50ml/g of IPAC and then 1.0 equiv of PhSO3H in 5ml/g IPAC
`was addd to this solution at 50ºC. After 20% of the addition 17mg of 70316-001 was adde as seed. The
`slurry was aged o/N and then cooled to RT, filtered and washed. 13.26 g of solids which were assayed to
`be 98.4 % PhSO3H salt were recovered. (78% overall yield 95% mass balance for crystallization.)
`
`Sample of solids were given to analytical:
`L-224,715 Besylate salt NB# 70316-009.
`
`RRT
`0.93
`0.95
`0.98
`1.00
`1.37
`1.43
`
`Area%
`0.15%
`0.20%
`0.17%
`99.06%
`0.18%
`0.17%
`
`Total imp = 0.87%
`
`LC Chiral
`
`Major = 99.98%
`Minor = 0.02%
`
`Coupling appears to be high yielding, PhSO3H salt gives a reasonable purity material. Yield is higher
`when no water is present during the coupling
`
`Salt Form:
`
`PhSO3H:
`
`Merck Exhibit 2131, Page 11
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`70316-001 Crystals formed in CD3CN NMR sample. These were given to Phys. Meas. And found to be a
`new crystal form.
`
`Phosphate:
`
`70316-011 Attempted to crystallize phosphate salt form THF water. Cryatals were small to amorphous-
`slurry given to phys. Meas.
`
`70316-013 Attempted to crystallize phosphate salt form ACN water. Cryatals were small to amorphous-
`slurry given to phys. Meas.
`
`70316-015 Crystallized Phosphate salt from IPA/water. Crystals were small and amorphous. Gave to
`Phys. Meas.
`
`70316-025 Crystallized phospahet salt from 27ml/g of 16% water/Ethanol. Clear solution at 75ºC was
`cooled at 5ºC/h. Once at RT, slurry was diluted to 52ml/g and filtered. Ml s were at 1.82mg/g 715 and
`199 mg ofphosphate salt were isolated. This material was highly crystalline by XRPD and had a shard dsc
`at 214ºC. The material is also not hygroscopic. 100mg was given to PR&D for stability measurements.
`
`70316-031 4.00g of PhSO3H salt was dissolved in EtoAc and washed with NaOH and sat bruine. The
`ethylacetate extratcs contained 99% of 715 and were concentrated to an oil. The oil was redissolved in
`28ml ethanol and then diluted with 2ml water. The solution was heated at 70ºC and a soln of H3PO4 in
`7ml ethanl was added slowly to the solution. A slurry formed and this was dissolved by adding 5.9ml of
`additional water. (15ml/g 18.4% water). This solution was cooled at 5ºC/hr to RT and then diluted with 57
`ml of ethanol. Isolated 3.03g (85%) of white solids.
`
`Tartrate
`
`70316-027 Scaled up formation of the tartrate salt. Isolated 2.10g (85% yield)
`
`Merck Exhibit 2131, Page 12
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Karl Hansen
`Weekly Report
`18 Feb 2002
`
`Hansen summary:
`
`Coupling
`
`F
`
`F
`
`F
`
`BnO
`
`NH O
`
`OH
`
`2
`
`HN
`
`+
`
`N
`
`N
`
`CF3
`
`N
`HCl
`
`3
`
`L224,715
`benzenesulfonate salt
`
`70316-035 10.54 g of amino acid was dissolved in 100ml CH3CN. The solution was charged with 1.5
`equiv. of 3 and then cooled to 0ºC. EDC and N-methyl morpholine were added sequentially to the
`heterogeneous mixture. The reaction was aged for 2h at which point 99% of 2 had converted to the amide.
`The slurry was diluted with 60ml water and 100ml EtOAc. The layers were cut and the organic was
`washed with saturated NaHCO3, then saturated NaCl. The organics were concentrated to an oil and then
`flushed twice with methanol and finally re-dissolved in 100ml MeOH. 3.00 g of 10% pd/C was charge and
`the mixture was subjected to 40PSI H2 and 50C for 18h. The benzyloxy group had been cleaved. The
`mixture was filtered though silkafloc, which was washed with methanol. The filtrate assayed to contain
`10.30g of 715 (91% assay yield).
`The methanol was removed in vacuo and the oil flushed twice with ethanol. The oil after flushing
`was re-dissolved in 35ml of ethanol and then filtered into a RBF. The filter was washed with 5ml ethanol
`(total 40ml, ~4ml/g). 2.91g (1.0 equiv.) of phosphoric acid was dissolved in 5ml water. This solution was
`added to the ethanol solution of 715, which was seeded with phosphate salt. Solids formed immediately at
`RT and these were thick and amorphous under scope. The slurry was heated to 72ºC but clear solution was
`not obtained. The addition of 10ml/g water resulted in clear solution and this was allowed to cool to RT at
`5ºC/h overnight. The slurry was then diluted with 145ml ethanol. The concentration of 715 was 2.22 mg/g
`after 0.5 h and 2.34 mg/g after 3h. The slurry was filtered and washed with 50ml EtOH.
`11.81g of white crystalline solids were recovered (75% overall yield). A 96% mass-balance was
`observed for crystallization. Solids are 80.6 wt% by HPLC (theoretical 80.6%) Analytical assayed solids
`to be 99.69% by HPLC with very little minor enantiomer and two other impurities at <0.2%. 1H-NMR
`shows no ethanol present and the methine a-to the amine is shifted downfield relative to free base.
`
`Merck Exhibit 2131, Page 13
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Karl Hansen
`Weekly Report
`25 Feb 2002
`
`Hansen summary:
`
`Coupling
`
`F
`
`F
`
`F
`
`BnO
`
`NH O
`
`OH
`
`2
`
`HN
`
`+
`
`N
`
`N
`
`CF3
`
`N
`HCl
`
`3
`
`L224,715
`H3PO4 salt
`
`70316-039 Followed procedure outlined in previous weekly using 90%ee 2. Assay yield was 71% before
`crystallization. Crystallization was carried out be slowly cooling (5ºC/h)a clear solution of phsophate salt
`in 30% aq Ethanol (5ml/g). Slurry was very thick and crystals were very thin needles under scope. A
`sample was filtered (Slow filtering) and given to Phys. Meas. The xray powder pattern of these solids
`showed mostly the anhydrous form of 715 phosphate, however several newe peaks were present. The
`slurry was heate to 65ºC and seeded with 35mg of 70316-035. The solids turned over after 1h to the
`expected plate like morphology. A sample of the hot slurry was taken and its xray was similar to the initial
`sample. The slurry was cooled and diluted to 20ml/g with ethanol and then filtered. The solids were found
`to be 92% ee and the ML s were 88%ee with comparable purity to earlier batches of phosphate. The final
`isolated solids were similar to earlier analyzed samples from this lot.
`
`70316-041 Racemic aminoacid was coupled using procedure for 035. Methanol filtrate after
`hydrogenation was assayed at 89% yield. The material was crystallized by concentrating into ethanol, then
`dissolving 4ml/g in ethanol and filtering off solids. The solution was heated to 60ºC and then seeded with
`1% of 70316-035. A solution of 1.05 equiv of phosphoric acid in 1.5ml/g water was then added at 60ºC
`over 1h. The slurry was then cooled to RT and then diluted with 14.5ml of ethanol. The slurry was filtered
`after 1h at RT and washed with 5ml/g ethanol. 5.25 g of solids were isolated (84% yield over 4 steps.
`[715] = 0.93 in ML s. X-ray shows that this is a unique form from 90% ee material and enantiomerically
`pure material.
`
`
`70316-045,47. Coupling was performed in ethyl acetate and IPAC. While reaction proceeded to nearly
`complete conversion of acid to amide after >8h, tacky solids were formed which would make the
`conditions difficult to scale-up.
`
`Merck Exhibit 2131, Page 14
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Karl Hansen
`Weekly Report
`11 March 2002
`
`Hansen summary:
`
`Coupling
`
`F
`
`F
`
`F
`
`BnO
`
`NH O
`
`OH
`
`2
`
`HN
`
`+
`
`N
`
`N
`
`CF3
`
`N
`HCl
`
`3
`
`L224,715
`H3PO4 salt
`
`Phosphate salt form: 99% ee samples prepared 2 weeks ago were found to have identical solid state NMR
`spectra, despite having significant differences in the x-ray powder diffraction.
`
`A standard of the benzyloxyamino adduct from the coupling was prepared and used for calibration
`
`70316-053 A stock soln of amino acid (94mg/ml, KF 300PPM, 10ml, 0.942g) was charged with EDC (1.5
`eq), then N-methyl morpholine (1.0Eq) at 0 ºC. After 3 h product concentration = 157 mg/g and <1 LCAP
`of aminoacid present. Quenched as previous. EtOAc organic layer assayed to contain 1.51g (106% of the
`desired product.
`
`70316-055 Ran as above, only 1.5 equiv of base used. Reaction was much slower achieving only 89%
`conversion after 3 h. 100% conv and [Prod] = 141mg/g observed after 18h at RT. Better rate crude assay
`yield with 1.0 equiv of base.
`
`70316-057 Ran identical to 053, except base added first at 0ºC with a 0.5 h age prior to EDC charge.
`Work up as previous. 1.53 g assay yield (107%). Base addition before EDC has no effect on yield.
`
`70316-059 coupled 4.54 g of amino acid using procedure as 053. Assayed organics for 6.98g product
`(102%). Concentrated and flushed with methanol
`
`70316-061 Oil from 059 was dissolved in 50ml methanol. 20ml of this solution was treated with 0.5 g
`DARCO G60 at RT for 1h and then filtered through silkfloc. The other 30ml was filtered through silka
`floc. The carbon treated soln was found to have a 14% drop in concentration, some of which might be due
`to dilution during wash. The carbon treated soln was split in two and the non carbon treated soln split in 3..
`each portion was charged with 10% pd/C and subjected to 40PSI H2 at 50C for 8 h. results in table below
`
`Run
`1
`2
`3
`4
`5
`
`Amt catalyst
`0.3 g
`0.2 g
`0.1 g
`0.2 g
`0.1 g
`
`Carbon treated
`N
`N
`N
`Y
`Y
`
`Conv.
`82
`69
`48
`77
`52
`
`The results show that carbon treatment can improve the efficiency of the catalyst by 10%(run 2 vs run 4).
`Run 1 roughly represents a 20 wt% catalyst loading relative to benzyloxy intermediate.
`
`70316-063 GMP triazole was used in a 14.1 g coupling identical procedure as 057. The triazole material
`was black in color. The coupling went as previous [Prod] = 155 mg/g at 3h, 99.4 LCAP conv. The
`
`Merck Exhibit 2131, Page 15
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`reaction was quenched and worked up as previous to give assay yield of 23.05 g in extracts (108%). The
`extracts were divided in 3 portions and used in 3 subsequent experiments.
`
`70316-065 Calibrated GC for MTBE, THF. Recalibrated HPLC for L224,715.
`
`70316-067 1/3 of ethyl acetate extracts from 70316-063 wconcentrated and flushed with methanol, then
`redissolved in 50 ml MeOH. The black soln was subjected to 40 PSI H2, 50ºC with 1.5g of 10% Pd/C
`(20wt% loading). After 18h, 97% conversion to L224,715 was observed. The solution was filtered
`through silkafloc and washed with methanol. Filtrate was clear-all of black color was gone. The filtrate
`was assayed to contain 5.28g of 715 (93% yield from amino acid). After 1 day at RT, the assay for 715
`was 1.3% lower. The material is being converted into phosphate salt to check impurity profile.
`
`70316-069 Performed as 67 on 1/3 of extracts from 63. Only 63% conv. as observed after same time as
`67. Catalyst was filtered off and soln was clear. Solution will be re-submitted for hydrogenation.
`
`Merck Exhibit 2131, Page 16
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`K. Hansen
`Weekly Report
`22 April 2002
`
`Coupling: Mode of addition of reagents was explored. A slow addn of the aminoacid substrate to the
`reaction mixture was investigated:
`
`70316-105 A solution of benzyloxyamino acid in 10ml/g acetonitrile was added over 2.5 h to a slurry of
`triazole, EDC and morpholine in CH3CN (5ml/g) at 0 ºC. After 20% of the addition only 50% of the acid
`added to the reaction was converted to product. Reaction did not finish until 1hr after all of substrate had
`been added. Normal work-up gave an assay yield of 103%-similar to normal procedure.
`
`70316-107 Performed identical to 105, however, base was mixed with aminoacid soln. Reaction was
`faster (after 20% of substrate added, 90% of it had converted to product). After quench and work-up assay
`was 103% yield. Identical to 105.
`
`Assay development for blocked (benzyloxy)
`
`V. Vydra screened for salts of benzyloxy intermediate and found the HBr salt is highly crystalline. This
`was scaled up into an isolation of pure material for standard. The MTBE extracts were concentrated and
`resuspended in IPAC at 75C. Ethanol was added slowly until complete soln was achieved. The soln was
`cooled to RT at 5C/hr and then filtered and washed. Material was crystalline and relatively pure by 1H-
`NMR. A second purification by crystallization from 20% ethanol/IPAC gave cleaner material which is
`99LCAP. Calibration of HPLC with this material is very close to that derived with free base oil,
`suggesting that the >100% yields observed may be due to a co-eluting of a peak. Assays will be
`incorporated into the process.
`
`Salt formation:
`
`An unseeded process, which is 10L/kg final volume, has been developed. Yield is 91% for the new
`conditions-identical to 30L/kg procedure. Oven-drying material at 40 ºC with N2 sweep seems to remove
`trace ethanol and water and gives consistent X-ray patterns. The effect of agitation on particle size and
`distribution is under study.
`
`70130-103 15.0 g of L-224715 freebase in ethanol (324mg/g, 422ml/g from hydrog) is charged to RBF
`and heated to 65 ºC. 1.1 equiv. of H3PO4 is dissolved in 1.25 ml/g water and this soln is added to the
`ethanol soln, keeping the temperature at 65 ºC. Upon complete addn of acid, the solution is clear and
`5ml/g of ethanol is added to the soln over 2.5 h by syringe pump. Upon completion of the addition the
`concentration of free base in solution is 34mg/g. The slurry is cooled to RT and then aged overnight. The
`slurry is filtered and washed with 5ml/g ethanol and dried under N2 (or in oven at 40C). 91.3% yield with
`6% loss to ML s and 97% mass balance overall.
`
`Merck Exhibit 2131, Page 17
`Mylan Pharmaceuticals Inc. v. Merck Sharp & Dohme Corp.
`IPR2020-00040
`
`

`

`Karl Hansen
`Weekly report
`11 Nov 2002
`
`70316-381 to 393 10 g of L224715 phosphate salt was crystallized from a series of solvent/water mixtures.
`A saturated solution was prepared at a temperature close to the maximum temperature of the system by
`adding water to the slurry of the solids in pure organic solvent. The saturated solution was then coole at
`5ºC/hr to RT and then filtered. The solids were assayed for mean particle size, Xray powder patternA
`summary of the conditions is in the table below.
`
`Notebook#
`381
`383
`385
`387
`389
`391
`393
`
`solvent % water
`t-BuOH
`24
`MeOH
`32
`Acetone
`16
`IPA
`24
`n-PrOH
`24
`EtOH
`28
`water
`100
`
`Temp mean PSD Mophology
`75
`43
`Rod
`65
`54
`Hex plate
`59
`13
`Hex plate
`75
`34
`Hex plate
`75
`54
`Hex plate
`75
`40
`Hex plate
`75
`37
`
`The only change in morphology observed was the material isolated from t-BuOH, which displayed rod-like
`morphology but was still slightly agglomerated. Solids state NMR showed the presence of solvent in the
`lattice of all the above solids when analyzed as a wet cake except for t-BuOH. The solids from water were
`poorly shaped and highly agglomerated.
`
`31878-001 Three ethanol/water solutions were saturated at 73ºC and then filtered hot. The ML s were
`then reheated to a clear solution then cooled to Rt at 3ºC/h. The slurries were then filtered.
`
`Morphology
`PSD
`[715] solution
`KF
`Expt
`Hex platew
`43
`