Improved Manufacturing Route and Polymorphic Control of a Potent

Article pubs.acs.org/OPRD

Cite This: Org. Process Res. Dev. XXXX, XXX, XXX−XXX

Improved Manufacturing Route and Polymorphic Control of a Potent and Selective Anaplastic Lymphoma Kinase (ALK) Inhibitor ASP3026 Yuji Takahama,* Kazuyoshi Obitsu, Kazuhiro Takeguchi, Shun Hirasawa, Koji Kobayashi, Takahiro Akiba, Norihiro Ueda, Ryoki Orii, Atsushi Ohigashi, Takumi Takahashi, Minoru Okada, and Shigeru Ieda Downloaded via WEBSTER UNIV on February 21, 2019 at 02:08:33 (UTC). See https://pubs.acs.org/sharingguidelines for options on how to legitimately share published articles.

Process Chemistry Laboratories, Astellas Pharma Inc., 160-2 Akahama, Takahagi-shi, Ibaraki 318-0001, Japan S Supporting Information *

ABSTRACT: Our effort toward the process improvement of anaplastic lymphoma kinase (ALK) inhibitor ASP3026 (1) is described. A cost-effective and practical synthesis of 1 was accomplished as a result of the change of starting material from 2,4dichloro-1,3,5-triazine (6) to cyanuric chloride (9) and late-stage introduction of a highly reactive N-methyl piperazine moiety by reductive amination of intermediate ketone 13. The modified process avoided the challenges with the original synthesis and furnished the several hundred kilograms of high-quality API with high economic efficiency, operability, and reproducibility. Furthermore, a sequence of investigation of polymorphic control in the second-generation synthetic route to obtain the thermodynamically desired, most stable polymorph Form A04 is also discussed. KEYWORDS: ALK inhibitor, SNAr reaction, 1,3,5-triazine, polymorph

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INTRODUCTION Anaplastic lymphoma kinase (ALK) is a validated therapeutic target for treating echinoderm microtubule-associated proteinlike 4 (EML4)-ALK positive non-small cell lung cancer (NSCLC).1−5 ASP3026 (1) (Figure 1) is a potent and

the presence of DBU and by following hydrogenation of the nitro group utilizing Pd/C catalyst and salt formation with trifluoroacetic acid, which gave the best result in the subsequent SNAr reaction of 5 and intermediate 8, furnishing aniline 5·TFA. Aside from the synthetic stream of intermediate 5·TFA, 1,3,5-triazine derivative 8 was obtained by SNAr reaction between 6 and aniline 7. Various conditions of consecutive substitution reactions of 8 and 5 were examined (Table 1). First, a series of protic acids were screened (entries 1−4). While MsOH and TFA gave 1 in a high HPLC ratio (entries 1,2), the use of formic acid and acetic acid led to poor conversion of 5 (entries 3,4). Eventually, TFA was selected as a suitable acid considering a possible generation of genotoxic alkyl mesylates in the MsOH condition. Also, the conditions of Lewis acids gave 1 in moderate to low yield (entries 5−9), and screening of solvent with TFA revealed that various solvents are applicable (entries 10−13). With respect to the stoichiometry of TFA, it was found that 3 equiv. is the best condition (entries 14−18). Also, we examined the effects of temperature (entry 19) and H2O (entry 20), though both did not provide significant change. Although we learned that 2-PrOH is the best solvent, we finally chose 2-butanone as the reaction solvent (entry 13). A reaction with 8 and 5·TFA had to be conducted continuously from a reaction of 6 and 7 without any isolation of 8, since 8 was unstable under air. 8 was obtained as 2-butanone solution after workup of the previous reaction, and a subsequent solvent switch to 2-PrOH generated gummy solid, which seems to

Figure 1. ASP3026, a potent ALK inhibitor.

selective ALK inhibitor that Astellas designed and synthesized through detailed structure−activity relationship (SAR) studies.6,7 We describe herein our effort toward the development of an improved synthetic route for a commercial stable supply of 1 and method to stably obtain desired polymorph Form A04.

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RESULTS AND DISCUSSION First-Generation Manufacturing Route. During the preclinical and early clinical phases of the development, 1 was supplied in kilogram quantities through the firstgeneration synthetic route, which was established by introducing minor changes into the discovery route using 2,4-dichloro-1,3,5-triazine (6) and 5-fluoro-2-nitroanisole (2) as starting material (Scheme 1). Anisole 2 was converted to intermediate 4 by a substitution reaction with piperidine 3 in © XXXX American Chemical Society

Special Issue: Japanese Society for Process Chemistry Received: December 6, 2018

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DOI: 10.1021/acs.oprd.8b00427 Org. Process Res. Dev. XXXX, XXX, XXX−XXX

Organic Process Research & Development

Article

Scheme 1. First-Generation Synthetic Route

synthetic scheme by obtaining 5 as a TFA salt after a hydrogenation of 4 and using such salt directly in a substitution reaction with 8. Since crude 1 was obtained as a crystal of a metastable polymorph (Form A02), solvent-mediated polymorphic transformation with aqueous acetone was employed to convert 1 to thermodynamically desired most stable polymorph, Form A04.8,9 By this synthetic protocol, multiple kilograms of 1 were delivered for GLP/GMP studies. Necessity of Process Improvement. While the firstgeneration synthetic route was able to be applied to the API supply in discovery and the early clinical stage, the process was not fully satisfactory as the manufacturing method for future commercial production because of the following points: (1) starting 1,3-dichlorotriazine (6) was considerably expensive and had low commercial availability, (2) aniline 5·TFA was unstable under air and had difficulties in quality control, storage, and transportation, and (3) the efficiency of an SNAr reaction between 8 and 5 was low despite of the thorough investigation shown above because of significant side-reaction. In order to establish a cost-effective, easy-to-operate, commercially sustainable manufacturing process, we decided to develop an another synthetic route of 1. Unsuccessful Route Using Cyanuric Chloride 9. At first, we tried a synthetic method utilizing cyanuric chloride (9) as the starting material instead of 6 (Scheme 2). Although an additional hydrogenation to remove the extra chloro group would be required in a later stage of synthetic stream, the higher cost efficiency and commercial availability of 9 were considered to be vital to develop an economically efficient manufacturing process. 9 was smoothly reacted with 7 to give intermediate 10. Next, we investigated various solvents in the SNAr reaction of 10 and 5·TFA (Table 2, entries 1−9).10 When THF was used, the reaction proceeded with the highest conversion, and the generation of byproduct was comparatively inhibited (entry 3). As a result of examination of the lower reaction temperature (entries 10, 11), it was found that 40 °C is enough to ensure higher conversion (entry 11). However, despite the sequence of investigation above, the efficiency of the SNAr reaction was not improved sufficiently. One of the reasons for this was thought to be the extreme instability of resulting 11, and this nature not only led to poor yield but also made it difficult to isolate 11 in high purity. Furthermore, it was found that 11 easily decomposed under a condition of

Table 1. Screening of Reaction Conditions: SNAr between 8 and 5a

HPLC area% entry

acid

equiv

solvent

1

5

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19b 20c

MsOH TFA formic acid acetic acid BF3·Et2O AlCl3 FeCl3 ZnCl2 LiCl TFA TFA TFA TFA TFA TFA TFA TFA TFA TFA TFA

3 3 3 3 3 3 3 3 3 3 3 3 3 0 1 2 3 4 3 3

EtOH EtOH EtOH EtOH EtOH EtOH EtOH EtOH EtOH MeCN DMF EtOAc 2-butanone 2-PrOH 2-PrOH 2-PrOH 2-PrOH 2-PrOH 2-PrOH 2-PrOH

73.4 77.6 58.2 39.3 56.5 10.1 1.0 17.7 13.5 79.9 33.9 73.7 72 9.9 39.8 73.9 81.1d 78.6 77.1 81.3

4.6 5.7 14.4 18.8 8.3 36.6 2.2 0.9 6.0 1.1 37.2 3.1 3.8 3.0 7.8 2.7 3.4 3.8 6.0 3.0

a

8 (0.30 mmol), 5 (0.30 mmol), and solvent (4.5 mL) were used. Reaction was conducted at 40 °C. cH2O (1.0 equiv) was added. d Isolated yield was 49%. b

indicate that 2-PrOH cannot be applied to manufacturing. From this context, we selected 2-butanone considering manufacturability, although the yield with it was slightly inferior than that with 2-PrOH. TFA was embedded into the B



Article

Scheme 2. Unsuccessful Route from Cyanuric Chloride (9)

and 15. We expected that this new protocol could resolve all of the issues in first synthetic route mentioned above and embarked on the investigation based on this hypothesis. Second-Generation Synthetic Route. A synthetic route designed on the basis of the retrosynthetic study is shown below (Scheme 4). A substitution reaction of 2 and piperizine 16·HCl using DBU proceeded well and furnished intermediate 17 in good yield, and following hydrogenation with Pd/C, aniline 15 was yielded. In keeping with our strategy, we reacted 15 with aniline 10. As we expected, a reaction proceeded in excellent yield to give intermediate 14 owing to the absence of a reactive N-methyl piperazine moiety. A residual chloro group on 14 was easily removed by hydrogenation with Pd/C under a basic condition and further converted to ketone 13 by subsequent deprotection with aqueous HCl in good yield with high reproducibility. A reductive amination of 13 to introduce N-methylpiperazine under a condition of NaBH(OAc)3 and AcOH gave crude 1 in good yield. However, a crystal was obtained as a mixed polymorph of the desirable Form A04 and metastable Form A02 and A06; furthermore, the ratio of polymorph in obtained crystal was unstable. Hence, recrystallization with 2-butanone was incorporated in a process to consolidate polymorphic mixture into Form A04. This secondgeneration synthetic route was applied to plural manufacturing to provide over 200 kg of API of 1. The overall yield was significantly improved in comparison with the first-generation synthetic route, and the manufacturing cost has been reduced to approximately half (Table 3). Furthermore, although the number of overall steps and reactions were increased due to additional dechlorination and late-stage introduction of piperazine, the production lead time was not affected by these changes as a result of the optimization of workup and isolation protocol. Polymorph Control of ASP3026 (1). Polymorphic control is also a crucial issue to a stable API supply of 1,8,9 since (1) 1 has multiple anhydrous polymorphs (Forms A01− A06) as shown in the following XRD and DSC charts (Figures 2 and 3);12 (2) a thermodynamic stability between the most stable Form A04 (desired polymorph) and metastable Form A03 is considerably close (Figure 4);13 (3) Moreover, it was impossibly difficult to convert Form A03 to Form A04 by solvent-mediated polymorphic transformation within a realistic operation time, once a concomitant polymorph of Form A04/ A03 is obtained.9a

Table 2. Screening of Reaction Conditions: SNAr between 10 and 5·TFAa

HPLC area% entry

solvent

temp (°C)

time (h)

11

5

1 2 3 4 5 6 7 8 9 10 11

2-butanone 2-PrOH THF MeCN toluene MeOH DMF EtOAc CPME THF THF

60 60 60 60 60 60 60 60 60 20 40

2 2 3 14 14 14 14 14 14 18 18

34.9 30.7 56.8 28 10.6 18 12.6 26.2 7.4 59.5 69.9

18.4 21.3 11.6 31.5 17.5 29.2 30.9 25.2 17.2 8.7 11.1

a

10 (0.30 mmol), 5 (0.30 mmol), and solvent (3 mL) were used.

following hydrogenation, and the subsequent yield of these two steps fell 17% eventually. From these results, we relinquished this synthetic route. Retrosynthetic Analysis. We assumed that the low yield of the SNAr reaction between 10 and 5·TFA and the instability of intermediate 11 comes from the highly nucleophilic tertiary amine group on the piperazine moiety, based on the analysis of a byproduct generated in a substitution reaction.11 From this reasoning, we turned our attention to late-stage introduction of an N-methylpiperazine moiety. As a result of a retrosynthetic analysis (Scheme 3), we expected that the N-methylpiperazine moiety could be introduced by reductive amination of ketone 13, which forms by a hydro-dechlorination and acidic deprotection of 14. Furthermore, 14 was thought to be obtained through a subsequent substitution reaction of 7, 9, C



Article

Scheme 3. Retrosynthetic Analysis

Scheme 4. Second-Generation Synthetic Route

Table 3. Comparison of the Synthetic Routes starting material overall yielda manufacturing cost overall steps production lead time

first generation

second generation

low availability expensive 31% six (four reactions) -

high availability inexpensive 45% approximately half eight (seven reactions) comparable

a

From 6 or 9.

Polymorph Control in the Second-Generation Synthetic Route. In the first-generation synthetic route, as we previously reported, desired Form A04 was reproducibly obtained by solvent-mediated polymorphic transformation of crude 1 (Form A02) in aqueous acetone.9b,c In the second

Figure 2. XRD chart of polymorphs of 1 (Forms A01−A04 and A06).

D



Article

Scheme 5. Recrystallization of Crude 1

unexpected change in quality of crude 1 during manufacturing might lead to this uncontrollability; in other words, some impurity could prominently inhibit a formation of Form A04 in a recrystallization process.14 In regard to manufacturing, an activated carbon treatment was introduced in a recrystallization process to improve purity (Scheme 6), and this enabled consolidation of polymorph into Form A04. While we met our obligations to the API supply, we ended up facing the following challenges for future commercial production: (1) the yield of recrystallization step was decreased from 90 to 75% because of adsorption loss on activated carbon; (2) purified 1 was colored yellow through the activated carbon process; and (3) another new impurity was generated possibly because of oxidation by the activated carbon−molecular oxygen system.15 Through careful investigation, we found that the contents of a specific impurity (18, Scheme 7) in crude 1 show correlation with the polymorph ratio of purified 1 (Figure 6). This is an impurity that was generated by “over-reaction” of an extra C− Cl bond on the triazine ring of 10 with 15, i.e., this impurity is derived from the change of starting material from 6 to 9. The analytical result indicating the possibility that 18 would exert a negative impact to polymorphic control prompted us to implement further investigation. To prove this hypothesis, we conducted two types of verification experiments: (1) spike test with impurity 18 to recrystallization process of crude 1 and (2) complete elimination of 18 from a synthetic stream by modification of the reaction condition of SNAr between 10 and 15 and subsequent confirmation of the polymorph in the final crystal. Spike Test with 18. For preparation of impurity 18, we first reacted an intermediate 14 with 1 equiv of aniline 15 under basic conditions to give 19, and resulting 19 was converted to precursor 20 by an acidic deprotection with aqueous HCl. Finally, 18 was obtained by reductive amination of 20 with N-methylpiperazine. To confirm a polymorphic inhibitory effect of 18, we added 1.0 wt % (0.75 area% in HPLC) of synthesized 18 into a solution of 1 after a hot filtration Scheme 5 and then checked the polymorphic ratio of resulted crystal. As we expected, the crystal to which 18 was added in the process clearly contained 45% (ratio of amount of heat in DSC) of Form A03, while pure Form A04 was obtained in the control experiment in which no 18 was added (Figure 7). This result strongly supports that 18 was a root cause of contamination of Form A03. Elimination of 18 by Reaction Optimization. To verify our hypothesis and to ensure a solid polymorphic control to Form A04, next we attempted to completely eliminate 18 from a synthetic stream (Table 5). As shown in Scheme 7, impurity 19, a source of 18, pertains to the SNAr reaction between 10

Figure 3. DSC chart of polymorphs of 1 (Forms A01−A04 and A06)

Figure 4. Solubility of polymorphs of 1 in 2-butanone (Forms A01− A04)

route, on the contrary, a crude 1 was obtained as a mixed polymorph comprising Forms A02, A04, and A06 without reproducibility possibly because of a difference in quality. Given an instability in solvent-mediated polymorphic transformation by using a mixed polymorph, we decided to turn our attention to recrystallization. 2-Butanone was selected from these following reasons: (1) solvent-mediated polymorphic transformation from Form A02 to A04 was best accelerated in 2-butanone9b and (2) 1 was soluble enough in 2-butanone to conduct recrystallization within a practical solvent volume (Table 4). On the basis of the fact that Form A04 is preferably Table 4. Solubility of 1 (Form A04) to Various Solvents Form A04 solubility

2butanone

acetone

ethyl acetate

MIBK

EtOH

2PrOH

25 °C (g/L) reflux (g/L)

5.8 53.8

4 13

3.8 -

3.2 -

3.1 -

0.8 -

nucleated at a higher temperature,9 we established a procedure shown as Scheme 5 and obtained several hundred grams of purified 1 as desirable Form A04 in a premanufacturing study. However, the in-laboratory recrystallization of the manufacturing lot of crude 1 gave a concomitant polymorph of Forms A04 and A03 with a ratio of 85:15 (ratio of amount of heat in DSC), although the same recrystallization protocol as that used in the premanufacturing study was applied (Figure 5, SEM; VE-8800, Keyence Corp., Japan). We assumed that an E



Article

Figure 5. SEM image (500×) of Form A04 (left) and mixture of Forms A04 and A03 (right).

solution of this unstable polymorph. As we expected, a decrease of 15 to 0.95 equiv. and change of the reaction temperature to 0 °C effectively inhibited 19 (Table 5, entries 2,3). When −10 °C was adopted, 19 was best controlled to the level of ND (Table 5, entry 4). As per the second-generation synthetic route, resulting 14, in which 19 was completely eliminated, was led to crude 1 which does not contain impurity 18. Finally, it was confirmed that such crude 1 can be reproducibly converted to pure Form A04 by same recrystallization condition as manufacturing without an activated carbon process. From the result of these two types of experiments, we concluded that impurity 18 was the root cause of contamination of Form A03 in manufacturing. This finding and modification of a reaction condition made it possible to conduct robust polymorphic control of 1 even in the secondgeneration synthetic route and eliminate problematic activated carbon process.

Scheme 6. Recrystallization of Crude 1 with Activated Carbon

and 15. In an original protocol, which was adopted in the manufacturing, the substitution reaction was performed with a slight excess of 1.05 equiv of 15 (against 10) at 25 °C and furnished intermediate 14 containing 0.21 area% of 19 (Table 5, entry 1). Importantly, in the actual manufacturing, 19 was further increased to 0.40% during the overnight storage of the workup solution. We therefore expected that the use of a smaller amount of 15 and lowering the reaction temperature would suppress generation of 19 and eventually lead to a

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CONCLUSIONS We have developed an improved practical manufacturing process for ALK inhibitor ASP3026 (1). A change of starting material from 2,4-dichloro-1,3,5-triazine (6) to cyanuric chloride (9) significantly improved the economic efficiency

Scheme 7. Generation Path of 18

F



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Article

EXPERIMENTAL SECTION General. Starting materials (2, 3, 6, 7, 9, 12, and 16·HCl), reagents, and solvents were purchased from suppliers and used without further purification. Shimadzu LC-2010 AHT and Hitachi Lachrom Elite L-2000 were used for HPLC analysis. HPLC Analysis. For 1, 4, 5, 11, 18, 19, 20: L-column2 ODS (Chemical Evaluation and Research Institute, Japan) 150 × 4.6 mm at a flow rate of 1.0 mL/min; peaks were monitored at 225 nm. For 8, 14: Inertsil ODS-4 (GL Sciences, Japan) 150 × 4.6 mm at a flow rate of 1.0 mL/min; peaks were monitored at 225 nm. For 10, 13: YMC-Pack ODS-A (YMC, Japan) 150 × 4.6 mm at a flow rate of 1.0 mL/min; peaks were monitored at 225 nm. For 15, 17: YMC-Pack Pro C18 RS (YMC, Japan) 150 × 4.6 mm at a flow rate of 1.0 mL/min; peaks were monitored at 225 nm. Improved condition to detect 18 in 1: CAPCELL PAK C18 AQ (Shiseido, Japan) 150 × 4.6 mm at a flow rate of 1.2 mL/min; peaks were monitored at 238 nm. Purities are reported by relative HPLC area. NMR Measurement. 1H and 13C NMR spectra were acquired with a JEOL ECA600 NMR spectrometer at 600 and 150 MHz, respectively. Chemical shifts (δ) of 1H NMR spectra are reported in parts per million (ppm) referenced to TMS at 0.00 or acetone-d at 2.09. Chemical shifts of 13C NMR spectra are reported in parts per million (ppm) referenced to solvent peak (chloroform-d at 77.0; DMSO-d at 39.5; acetone-d at 30.6). Coupling constants (J) are reported in hertz (Hz). Mass Measurement. Mass spectra data were acquired on a Waters Xevo TQ MS, triple quadrupole mass spectrometer, in ESI positive mode. Column chromatography was performed with silica gel (63-210 mesh). PXRD Measurement. ASP3026 polymorphs were evaluated with Miniflex (Rigaku Corp., Japan) under the following conditions: X-ray tube, copper; tube current, 15 mA; tube voltage, 30 kV; sampling width, 0.010°; scanning speed, 2°/ min; wavelength, 11.49 mm; and range of measurement diffraction angles (2θ), 3−35°. DSC Measurement. DSC analysis was conducted with a TA Instruments Q-2000 calorimeter (TA Instruments, U.S.A.) under the following conditions: temperature range of measurement, ambient temperature to 493 K or higher; rate of temperature increase, 30 °C/min; nitrogen flow rate, 50 mL/ min; and sample pan material, aluminum. Solubility Measurement. An excess quantity of each polymorph was added to the solvent. The temperature of the slurry was controlled with a thermostat bath, and the slurry was agitated over 2 h and then filtrated with a 0.45 μm membrane filter. The concentration of a supernatant was measured by the HPLC condition above. First-Generation Synthetic Route. 1-[1-(3-Methoxy-4nitrophenyl)piperidin-4-yl]-4-methylpiperazine (4). A 1000 L reactor was charged with 2 (23.0 kg, 134 mol), 3 (29.6 kg, 161 mol), and THF (230 L). 1,8-Diazabicyclo[5.4.0]undec-7-ene (24.6 kg, 162 mol) was added, and the mixture was stirred at 50 °C for 8 h and then cooled to room temperature. Ethyl acetate (460 L) and NaCl aqueous solution (46 kg in 230 L of water) were added to the reaction mixture, and the organic layer was washed twice with NaCl aqueous solution (46 kg in 230 L of water) and concentrated to 46 L under vacuum. The resulting residue was added to EtOH (115 L) and concentrated to 46 L under vacuum, and this protocol was repeated again. n-Heptane (138 L) was added to the residue, and the mixture was stirred at 40 °C for 1 h. To the resulting

Figure 6. HPLC charts of crude 1: relationship between 18 and polymorph ratio of purified 1.

Figure 7. DSC charts of resulting crystal (control experiment vs spike test of 18).

of the process owing to its high commercial availability. Furthermore, late-stage introduction of N-methyl piperazine by reductive amination was effective in avoiding undesirable sidereactions in the SNAr reaction. In regard to polymorphic control in the second-generation synthetic route, we found that a small amount of impurity 18 behaves in a way that inhibits formation of desired polymorph Form A04 in a recrystallization process. An improved process, in which 18 is completely eliminated, enabled stable control to Form A04 with high practicality and reproducibility. G



Article

Table 5. Screening of Reaction Condition of SNAr of 10 and 15a

a

10 (1.00 mmol), 15, DIPEA (1.10 mmol), and THF (3.5 mL) were used. bIn actual manufacturing, 19 was further increased to 0.40% during overnight storage after the reaction.

(150 MHz, DMSO-d) δ 159.2, 159.0, 158.8, 158.5, 119.6, 117.6, 115.7, 113.7, 108.0, 108.0, 100.9, 60.9, 55.9, 50.9, 48.2, 45.5, 42.1, 25.9. LC−MS (ESI): calcd for C17H30N4O ([M + H]+) 305.2; found 305.1. 4-Chloro-N-[2-(propane-2-sulfonyl)phenyl]-1,3,5-triazin2-amine (8) and N2-{2-Methoxy-4-[4-(4-methylpiperazin-1yl)piperidin-1-yl]phenyl}-N4-[2-(propane-2-sulfonyl)phenyl]1,3,5-triazine-2,4-diamine (Crude 1, Form A02). A 1000 L reactor was charged with 6 (18.0 kg, 134 mol), 7 (26.3 kg, 132 mol), diisopropylethylamine (17.1 kg, 132 mol), and toluene (180 L). After 3 h of stirring at 70 °C, the mixture was cooled to 45 °C. To the resulting mixture 2-butanone (180 L) was added and was concentrated under vacuum. 2-Butanone (90 L) was added to the residue and concentrated under vacuum. To the residue was added 2-butanone (360 L) and K2CO3 aqueous solution (18 kg in 180 L of water), and the residue was separated. The reactor was washed twice with 2-butanone (18 L). 8 was able to be isolated by concentration of this combined organic phase and further purified by column chromatography to obtain analytical sample. HPLC purity: 97.0A%. 1H NMR (600 MHz, chloroform-d) δ 9.88 (bs, 1 H), 8.61 (s, 1 H), 8.50 (d, J = 8.3, 1 H), 7.93 (dd, J = 8.3, 2.1 Hz, 1 H), 7.74−7.69 (m, 1 H), 7.36−7.31 (m, 1 H), 3.27−3.18 (m, 1 H), 1.32 (d, J = 6.9, 6 H). 13C NMR (150 MHz, chloroform-d) δ 170.9, 167.5, 163.9, 136.8, 135.0, 131.4, 124.9, 124.4, 123.0, 56.1, 15.2. LC−MS (ESI): calcd for C12H14ClN4O2S ([M + H]+) 313.1; found 313.0. A 1000 L reactor was charged with the combined 2butanone solution and 5·TFA (57.9 kg, 89.5 mol), and the mixture was stirred at 65 °C for 3 h and then cooled to 20 °C. NaOH aqueous solution (17.2 kg in 364 L of water) premixed with NaCl (36.4 kg) was added to the reaction mixture, and the organic layer was washed twice with NaCl aqueous solution (72.8 kg in 364 L of water). The mixture was transferred to a 1000 L reactor through a 0.6 μm filter, and the reactor was washed with 2-butanone (42 L). The solution was concentrated to 168 L under vacuum, seed crystals (28 g,

slurry was added n-heptane (276 L) at 40 °C, and this mixture was stirred at 40 °C for 1 h and then cooled to 0 °C over 2 h. The crystals were collected by centrifugation, washed with the precooled mixture of EtOH/n-heptane (6.9 L/62.1 L), and dried at 50 °C under vacuum to give 4 (40.5 kg, 121 mol, 90% yield) as a dark purple solid. HPLC purity: 99.2 A%. 1H NMR (600 MHz, chloroform-d) δ 8.00 (d, J = 8.9 Hz, 1 H), 6.42 (dd, J = 8.9, 2.1 Hz, 1 H), 6.31 (d, J = 2.1 Hz, 1 H), 3.97−3.92 (m, 2 H), 3.95 (s, 3 H), 3.01−2.93 (m, 2 H), 2.75−2.55 (m, 4 H), 2.55−2.34 (m, 5 H), 2.29 (s, 3 H), 2.00−1.94 (m, 2 H), 1.66−1.57 (m, 2 H). 13C NMR (150 MHz, chloroform-d) δ 156.4, 155.2, 128.9, 128.8, 105.4, 96.8, 61.1, 56.1, 55.3, 49.0, 46.8, 45.9, 27.7. LC−MS (ESI): calcd for C17H27N4O3 ([M + H]+) 335.2; found 335.1. 2-Methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]aniline Tris(trifluoroacetate) (5·TFA). A 1000 L autoclave was charged with 4 (40.0 kg, 120 mol) and THF (400 L), and the mixture was stirred at 25 °C until the solid was dissolved. 10% Pd/C (AD-type, 50% wet, 6.0 kg dry-based, Kawaken Fine Chemicals, Japan) was added, and the mixture was stirred under H2 atmosphere (0.10 MPa) at 25 °C for 5 h. The reaction mixture was warmed to 35 °C and transferred to the another 1000 L reactor through a 1 μm filter, and the autoclave was washed with THF (80 L). To the mixture was added TFA (42.3 kg, 371 mol) at 35 °C. Seed crystals (4 g) were added at 35 °C and stirred at the same temperature for 1 h and then stirred at 20 °C for 3 h. n-Heptane (320 L) was added to the reactor, and the mixture was stirred at 20 °C for 3 h. The crystals were collected by centrifugation, washed with the mixture of THF/n-heptane (72 L/48 L), and dried at 50 °C under vacuum to give 5·TFA(69.6 kg, 108 mol, 90% yield) as a dark purple solid. HPLC purity: 99.9A%. 1H NMR (600 MHz, MeOH-d) δ 7.11 (d, J = 8.9 Hz, 1 H), 6.81 (d, J = 2.8 Hz, 1 H), 6.70 (dd, J = 8.9, 2.8 Hz, 1 H), 3.94 (s, 3 H), 3.87 (d, J = 13.1 Hz, 2 H), 3.52−3.35 (m, 4 H), 3.35−3.18 (m, 4 H), 3.12−3.05 (m, 1 H), 3.01 (t, J = 11.7 Hz, 2 H), 2.89 (s, 3 H), 2.14 (d, J = 12.4 Hz, 2 H), 1.89−1.79 (m, 2 H). 13C NMR H



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mixture of Acetone and Isopropyl acetate (0.1 mL/0.1 mL) and Isopropyl acetate (0.2 mL), and dried at 50 °C under vacuum to give crude 1 (29.6 mg, 0.051 mmol, 17% yield) as pale yellow solid. HPLC purity: 97.1A%. Second-Generation Synthetic Route. 4,4-Dimethoxy-1(3-methoxy-4-nitrophenyl)piperidine (17). A 1000 L reactor was charged with 16·HCl (70.1 kg, 386 mol), DMF (150 L), and 1,8-diazabicyclo[5.4.0]undec-7-ene (117.4 kg, 771 mol). To the mixture was added a solution of 2 (60.0 kg, 351 mol) in DMF (90 L), and this was stirred at 25 °C for 5 h. Seed crystal (6 g) was added at 25 °C, and the resulting slurry was stirred at 25 °C for 3 h. Water (240 L) was added to the mixture, with continued stirring at 25 °C for 3 h. The crystals were collected by centrifugation, washed twice with a mixture of DMF/water (60 L/60 L), and dried at 50 °C under vacuum to give 17 (98.7 kg, 333 mol, 95% yield) as dark purple solid. HPLC purity: 99.9A%. 1H NMR (600 MHz, chloroform-d) δ 8.01 (d, J = 8.9 Hz, 1 H), 6.43 (dd, J = 8.9, 2.8 Hz, 1 H), 6.32 (d, J = 2.8 Hz, 1 H), 3.95 (s, 3 H), 3.48−3.43 (m, 4 H), 3.24 (s, 6 H), 1.90−1.85 (m, 4 H). 13C NMR (150 MHz, chloroform-d) δ 156.4, 155.2, 129.1, 128.8, 105.6, 98.1, 97.1, 56.1, 47.6, 44.6, 31.9. LC−MS (ESI): calcd for C14H21N2O5 ([M + H]+) 297.2; found 297.1. 4-(4,4-Dimethoxypiperidin-1-yl)-2-methoxyaniline (15). A 1000 L autoclave was charged with 17 (90.0 kg, 304 mol), THF (450 L) and 10% Pd/C (M-type, 50% wet, 4.5 kg drybased, Kawaken Fine Chemicals, Japan). The mixture was stirred under H2 atmosphere (0.30 MPa) at 25 °C for 3 h. The reaction mixture was transferred to another 1500 L reactor through a 1 μm filter, and the autoclave was washed with THF (180 L). The filtrate was concentrated to 180 L under vacuum, and seed crystal (9 g) was added. The slurry was stirred at 40 °C, and then, n-heptane (270 L) was added. After 1 h of stirring at 40 °C, the mixture was cooled to 0 °C over 4 h and stirred at 0 °C for 2 h. To the mixture was added n-heptane (810 L) at 0 °C, and the mixture was stirred at 0 °C for 2 h. The crystals were collected by centrifugation, washed with a mixed solution of THF/n-heptane (18 L/108 L), and dried at 50 °C under vacuum to give 15 (72.8 kg, 273 mol, 90% yield) as dark purple solid. HPLC purity: 99.9A%. 1H NMR (600 MHz, chloroform-d) δ 6.64 (d, J = 8.3 Hz, 1 H), 6.54 (d, J = 2.8 Hz, 1 H), 6.44 (dd, J = 8.3, 2.8 Hz, 1 H), 3.84 (s, 3 H), 3.53 (bs, 2 H), 3.23 (s, 6 H), 3.08−3.04 (m, 4 H), 1.94−1.88 (m, 4 H). 13C NMR (150 MHz, chloroform-d) δ 147.9, 145.0, 130.0, 115.4, 110.0, 102.9, 98.3, 55.4, 48.7, 47.4, 32.5. LC−MS (ESI): calcd for C14H23N2O3 ([M + H]+) 267.2; found 267.1. 4,6-Dichloro-N-[2-(propane-2-sulfonyl)phenyl]-1,3,5-triazin-2-amine (10). A 1000 L reactor was charged with 9 (40.0 kg, 217 mol), NaHCO3 (21.9 kg, 261 mol) and Acetone (320 L). The mixture was added 7 (47.5 kg, 238 mol), and stirred at 25 °C for 21 h. Water (320 L) was added, and the mixture was stirred at 25 °C for 21 h. The crystals were collected by centrifugation, washed with a mixture of Acetone/water (80 L/ 80 L), and dried at 50 °C under vacuum to give 10 (71.5 kg, 206 mol, 95% yield) as white solid. HPLC purity: 99.9A%. 1H NMR (600 MHz, ACETONE-d) δ 8.41 (d, J = 7.6 Hz, 1 H), 7.97 (dd, J = 7.6, 1.4 Hz, 1 H), 7.89−7.85 (m, 1 H), 7.55−7.50 (m, 1 H), 3.52−3.45 (m, 1 H), 1.26 (d, J = 6.2 Hz, 6 H). − 1 H (NH) 13C NMR (150 MHz, chloroform-d) δ 165.6, 137.1, 136.1, 132.4, 127.7, 126.6, 125.2, 100.9, 56.6, 15.5. LC−MS (ESI): calcd for C12H13Cl2N4O2S ([M + H]+) 347.0; found 346.9.

Form A02) were added, and the mixture was stirred for 1 h. The mixture was concentrated to 56 L under vacuum. Acetone (98 L) was added, and the mixture was concentrated to 56 L under vacuum, and this procedure was repeated two more times. To the slurry acetone (36 L) was added, and the mixture was stirred at 45 °C for 1 h. Isopropyl acetate (91 L) was added to the slurry, which was cooled to 0 °C over 4 h and stirred at 0 °C for 5 h. The crystal was obtained by filtration, washed with a mixture of acetone and isopropyl acetate (18 L/ 18 L) and isopropyl acetate (56 L), and dried at 50 °C under vacuum to give crude 1 (25.0 kg, 43.0 mol, 48% yield) as a pale yellow solid in Form A02. HPLC purity: 97.9A%. 1H NMR (600 MHz, chloroform-d) δ 9.28 (s, 1H), 8.47−8.61 (bs, 1H), 8.30−8.48 (m, 1H), 8.10 (bs, 1H), 7.88 (d, J = 8.2 Hz, 1H), 7.49−7.72 (m, 2H), 7.23 (t, J = 8.2 Hz, 1H), 6.45−6.60 (m, 2H), 3.88 (s, 3H), 3.63−3.75 (m, 2H), 3.18−3.32 (m, 1H), 2.31−2.78 (m, 11H), 2.30 (s, 3H), 1.92−2.01 (m, 2H), 1.65− 1.79 (m, 2H), 1.31 (d, J = 6.9 Hz, 6H). 13C NMR (150 MHz, chloroform-d) δ (ppm) 166.3, 163.6, 149.6, 148.6, 138.4, 134.5, 131.1, 124.4, 123.6, 123.0, 122.1, 121.4, 119.6, 108.0, 100.4, 61.6, 55.6, 55.5, 55.4, 50.0, 49.0, 46.0, 28.1, 15.3. LC− MS (ESI): calcd for C29H41N8O3S ([M + H]+) 581.3; found 581.2. N2-{2-Methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1yl]phenyl}-N4-[2-(propane-2-sulfonyl)phenyl]-1,3,5-triazine2,4-diamine (1, Form A04). A 1000 L reactor was charged with crude 1 (26.0 kg, 44.7 mol), and hot acetone/water (78 L/78 L) and seed crystals (26 g of 1, Form A04) were added. The mixture was stirred at 62 °C for 6 h, and hot acetone/ water (52 L/52 L) was added. After 18 h of stirring at 62 °C, the mixture was cooled to 25 °C over 8 h and was stirred at 25 °C for 8 h. The crystals were filtered, washed with a mixture of acetone/water (78 L/78 L), and dried at 50 °C under vacuum to give 1 (22.1 kg, 38.0 mol, 85% yield) as a pale yellow solid in Form A04. HPLC purity: 99.9A%. Unsuccessful Route from Cyanuric Chloride (9). 6Chloro-N 2 -{2-methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]phenyl}-N4-[2-(propane-2-sulfonyl)phenyl]1,3,5-triazine-2,4-diamine (11) and N2-{2-Methoxy-4-[4-(4methylpiperazin-1-yl)piperidin-1-yl]phenyl}-N4-[2-(propane2-sulfonyl)phenyl]-1,3,5-triazine-2,4-diamine (1). Procedure for Table 2, entry 11: A test tube was charged with 10 (104 mg, 0.30 mmol) and THF (3 mL). To the mixture was added 5·TFA (194 mg, 0.30 mmol) at room temperature. After stirring at 40 °C for 18 h, the precipitated solid was collected by filtration to give 11, which was used for the next step without characterization by NMR for stability issues. LC−MS (ESI): calcd for C29H40ClN8O3S ([M + H]+) 615.3; found 615.2. An autoclave was charged with above solid, ammonium formate (95 mg, 1.5 mmol), MeOH (3 mL) and 10% Pd/C (NX-type, 21 mg, N.E. CHEMCAT, Japan), and the mixture was stirred under H2 atmosphere (0.10 MPa) at 50 °C for 3 h. The reaction mixture was transferred to the another flask through a 1 μm filter, and the autoclave was washed with MeOH (5 mL). The filtrate was concentrated under vacuum, and then diluted with 2-butanone (10 mL), NaOH aqueous solution (47 mg in 10 mL of water) premixed with NaCl (1 g) was added, then organic layer was separated. The organic layer was washed twice with NaCl aqueous solution (2 g in 10 mL of water). The 2-butanone solution was concentrated to dryness, and Acetone (0.2 mL) and Isopropyl acetate (0.2 mL) were added. The crystal was collected by filtration, washed with the I



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6-Chloro-N2-[4-(4,4-dimethoxypiperidin-1-yl)-2-methoxyphenyl]-N4-[2-(propane-2-sulfonyl)phenyl]-1,3,5-triazine2,4-diamine (14). A 1000 L reactor was charged with 10 (60.0 kg, 173 mol) and THF (600 L). To the mixture was added 15 (48.3 kg, 181 mol), followed by diisopropylethylamine (24.6 kg, 190 mol) at 25 °C. After stirring at 25 °C for 3 h, isopropyl acetate (60 L) and K2CO3 aqueous solution (3.0 kg in 60 L of water) were added to the mixture. After the phase separation, the organic layer was concentrated under vacuum. Seed crystal (6 g) was added at 25 °C, and the mixture was stirred at 25 °C for 1 h. To the slurry was added 2-propanol (120 L) and nheptane (300 L), and the mixture was stirred at 25 °C for 2 h. The mixture was cooled to 0 °C over 2 h and was stirred at 0 °C for 5 h. The crystal was collected by centrifugation, washed with a mixture of THF/2-propanol/n-heptane (72 L/48 L/120 L), and dried at 50 °C under vacuum to give 14 (93.7 kg, 162 mol, 94% yield) as white solid. HPLC purity: 99.9A%. 1H NMR (600 MHz, chloroform-d) δ 9.48−9.31 (m, 1 H), 8.46 (d, J = 8.3 Hz, 1 H), 8.16−8.00 (m, 1 H), 7.90 (d, J = 7.6 Hz, 1 H), 7.72 (s, 1 H), 7.68−7.54 (m, 1 H), 7.30−7.23 (m, 1 H), 6.61−6.48 (m, 1 H), 6.54 (d, J = 2.8 Hz, 1 H), 3.87 (s, 3 H), 3.31−3.24 (m, 1 H), 3.24 (s, 6 H), 3.23−3.18 (m, 4 H), 1.94− 1.89 (m, 4 H), 1.31 (d, J = 6.9 Hz, 6 H). 13C NMR (150 MHz, chloroform-d) δ 169.3, 164.3, 163.5, 149.8, 148.7, 137.7, 134.5, 131.1, 124.9, 123.6, 121.6, 119.0, 108.1, 100.2, 98.3, 55.8, 55.6, 47.5, 47.1, 32.1, 15.3. LC−MS (ESI): calcd for C26H34ClN6O5S ([M + H]+) 577.2; found 577.0. 1-[3-Methoxy-4-({4-[2-(propane-2-sulfonyl)anilino]-1,3,5triazin-2-yl}amino)phenyl]piperidin-4-one (13). A 1500 L autoclave was charged with 14 (90.0 kg, 156 mol), THF (810 L), and 2-propanol (90 L), and the mixture was stirred at 25 °C until the solid was dissolved. 10% Pd/C (AD-type, 50% wet, 9.0 kg dry-based, Kawaken Fine Chemicals, Japan) and diisopropylethylamine (24.2 kg, 187 mol) were added, and the mixture was stirred under H2 atmosphere (0.25 MPa) at 40 °C for 7 h. The reaction mixture was transferred to another 1000 L reactor through a 1 μm filter, and the autoclave was washed with THF (180 L). HCl aqueous solution (32.5 kg of 36 wt % hydrochloric acid in 180 L of water) was added, and the mixture was stirred at 25 °C for 10 h. To the reaction mixture was added K2CO3 aqueous solution (53.9 kg in 180 L of water), and this was separated. To the organic layer was added activated carbon (9.0 kg), and the mixture was stirred at 25 °C for 12 h. The reaction mixture was transferred to the another 1000 L reactor through 1 μm filter, and the used reactor was washed with THF (90 L). The mixture was concentrated to 315 L under vacuum, and acetone (270 L) was added. Seed crystals (9 g) were added at 25 °C and stirred at 25 °C for 1 h. To the residue was added water (720 L), and the mixture was stirred at 25 °C for 1 h. The crystal was collected by centrifugation, washed with a mixture of acetone/water (54 L/ 144 L), and dried at 50 °C under vacuum to give 13 (71.2 kg, 143 mol, 92% yield) as white solid. HPLC purity: 99.9A%. 1H NMR (600 MHz, chloroform-d) δ 9.31 (s, 1 H), 8.53 (bs, 1 H), 8.50−8.31 (m, 1 H), 8.18 (bs, 1 H), 7.89 (dd, J = 8.3, 1.4 Hz, 1 H), 7.63 (bs, 1 H), 7.23 (t, J = 7.6 Hz, 1 H), 6.67−6.56 (m, 2 H), 3.91 (s, 3 H), 3.58 (t, J = 5.6 Hz, 4 H), 3.31−3.22 (m, 1 H), 2.60 (t, J = 5.6 Hz, 4 H), 1.31 (d, J = 6.2 Hz, 6 H). 13 C NMR (150 MHz, chloroform-d) δ 207.8, 166.4, 163.6, 163.5, 150.0, 146.5, 138.5, 134.5, 131.1, 124.4, 123.4, 122.9, 121.7, 120.2, 107.9, 100.1, 55.7, 55.6, 49.6, 40.8, 15.3. LC−MS (ESI): calcd for C24H29N6O4S ([M + H]+) 497.2; found 497.0.

N2-{2-Methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1yl]phenyl}-N4-[2-(propane-2-sulfonyl)phenyl]-1,3,5-triazine2,4-diamine (Crude 1, Polymorphic Mixture). A 1000 L reactor was charged with 13 (50.0 kg, 101 mol), 12 (20.2 kg, 202 mol), and toluene (500 L). To the mixture was added acetic acid (22.5 L). The mixture was stirred at 20 °C for 1 h, and NaBH(OAc)3(42.7 kg, 201 mol) was added. After 16 h of stirring at 20 °C, MeOH (50 L) and water (150 L) were added and separated. The organic layer was extracted with water (50 L), and the combined aqueous layer was washed twice with isopropyl acetate (500 L). The aqueous layer was charged into a 2000 L reactor using water (25 L). To the aqueous solution, MeOH (550 L), NaOH aqueous solution (24.2 kg in 120 L of water), and water (25 L) were added. Seed crystals (5 g, Form A04) were added at 25 °C, and the mixture was stirred at 25 °C for 1 h. To the mixture, water (550 L) was added and stirred at 25 °C for 2 h. The crystal was collected by centrifugation, washed with a mixture of MeOH/water (100 L/100 L), and dried at 50 °C under vacuum to give crude 1 as pale yellow solid comprising polymorphs of Form A02/A04/ A06 (42.7 kg, 73.5 mol, 73% yield). HPLC purity: 99.6A%. N2-{2-Methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1yl]phenyl}-N4-[2-(propane-2-sulfonyl)phenyl]-1,3,5-triazine2,4-diamine (1, Form A04). Procedure using activated carbon: A 2000 L reactor was charged with crude 1 (50.0 kg, 64.6 mol), activated carbon (5.0 kg), and 2-butanone (1200 L). After 1 h of stirring at 70 °C, the mixture was transferred to a 2000 L reactor through a filter paper and 1 μm cartridge filter, and the reactor was washed with 2-butanone (100 L). To the filtrate was added activated carbon (5.0 kg), which was stirred at 70 °C for 1 h. The mixture was transferred to a 2000 L reactor through a filter paper and 1 μm cartridge filter, and the reactor was washed with 2-butanone (100 L). Again, to the filtrate was added activated carbon (5.0 kg), which was stirred at 70 °C for 1 h. The mixture was transferred to a 2000 L reactor through a filter paper and 1 μm cartridge filter, and the reactor was washed with 2-butanone (100 L). The filtrate was concentrated to 400 L. Seed crystal (5 g, Form A04) was added at 70 °C and stirred for 3 h at same temperature. The slurry was cooled to 0 °C over 4 h, the crystal was collected by centrifugation, washed with 2-butanone (200 L), and dried at 50 °C under vacuum to give 1 (37.5 kg, 64.6 mol, 75% yield) as a pale yellow solid in Form A04. HPLC purity: 99.7A%. Synthesis of Impurity 18. N2,N4-Bis[4-(4,4-dimethoxypiperidin-1-yl)-2-methoxyphenyl]-N 6 -[2-(propane-2sulfonyl)phenyl]-1,3,5-triazine-2,4,6-triamine (19) and 1,1′({6-[2-(Propane-2-sulfonyl)anilino]-1,3,5-triazine-2,4-diyl}bis[azanediyl(3-methoxy-4,1-phenylene)])di(piperidin-4one) (20). A 100 mL three-neck round-bottom flask was charged with 14 (5.0 g, 8.66 mmol), 15 (2.3 g, 8.66 mmol), and THF (35 mL). To the mixture was added 1 M NaOH aqueous solution (17.3 mL), and the mixture was stirred at 63 °C for 32 h. The mixture was transferred to a separating funnel with THF (5.0 mL) and separated. The organic layer was washed three times with K2CO3 aqueous solution (2.0 g in 38 mL water) and charged into a 100 mL three-neck roundbottom flask. 19 was able to be isolated as a white solid by concentrating this organic layer in 99.9A% of HPLC purity. 1H NMR (600 MHz, chloroform-d) δ 9.03 (s, 1 H), 8.60 (bs, 1 H), 8.20 (bs, 2 H), 7.85 (dd, J = 7.6, 1.4 Hz, 1 H), 7.63−7.56 (m, 1 H), 7.27 (bs, 2 H), 7.16 (t, J = 7.6 Hz, 1 H), 6.61−6.52 (m, 4 H), 3.88 (s, 6 H), 3.33−3.27 (m, 1 H), 3.24 (s, 12 H), 3.22−3.18 (m, 8 H), 1.95−1.91 (m, 8 H), 1.32 (d, J = 6.9 Hz, J



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6 H). 13C NMR (150 MHz, chloroform -d) δ 164.3, 164.2, 149.5, 147.7, 139.5, 134.3, 130.9, 123.8, 122.0, 121.6, 121.5, 121.1, 108.6, 100.8, 98.4, 55.6, 55.4, 47.6, 47.5, 32.3, 15.4. LC−MS (ESI): calcd for C40H55N8O8S ([M + H]+) 807.4; found 807.1. 6 M HCl aqueous solution (7.2 mL) was added to the mixture and stirred at 25 °C for 21 h. To the mixture was added 5 M NaOH aqueous solution (7.2 mL), and the aqueous layer was extracted with THF (35 mL). The combined organic layer was concentrated under vacuum, and the residue was purified by silica gel column chromatography. The resulting crystal was further purified by washing with mixed solvent of acetone/water (30 mL/20 mL) and dried at 50 °C under vacuum to give 20(4.1 g, 5.74 mmol, 66% yield) as a white solid. HPLC purity: 99.9A%. 1H NMR (600 MHz, chloroform-d) δ 9.09 (s, 1 H), 8.60 (bs, 1 H), 8.27 (bs, 2 H), 7.88 (dd, J = 7.6, 1.4 Hz, 1 H), 7.65−7.57 (m, 1 H), 7.32 (bs, 2 H), 7.19 (t, J = 7.6 Hz, 1 H), 6.64−6.55 (m, 4 H), 3.91 (s, 6 H), 3.57 (t, J = 5.5 Hz, 8 H), 3.34−3.26 (m, 1 H), 2.60 (t, J = 5.5 Hz, 8 H), 1.33 (d, J = 6.9 Hz, 6 H). 13C NMR (150 MHz, DMSO-d) δ 207.7, 164.6, 163.5, 152.3, 146.8, 139.3, 134.7, 130.6, 125.2, 122.2, 122.1, 121.7, 119.1, 106.5, 99.8, 55.5, 54.9, 48.0, 40.0, 14.8. LC−MS (ESI): calcd for C36H43N8O6S ([M + H]+) 715.3; found 715.1. N 2 ,N 4 -Bis{2-methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]phenyl}-N6-[2-(propane-2-sulfonyl)phenyl]1,3,5-triazine-2,4,6-triamine (18). A 300 mL autoclave was charged with 20 (2.0 g, 2.8 mmol), 12 (1.12 g, 11.2 mmol), and THF (150 mL), and the mixture was stirred at 25 °C until the solid was dissolved. 10% Pd/C (EB-type, 50% wet, 0.2 g dry-based, Kawaken Fine Chemicals, Japan) was added, and the mixture was stirred under H2 atmosphere (0.20 MPa) at 50 °C for 18 h. The mixture was transferred to another flask through a Celite pad. The filtrate was concentrated under vacuum, and the residue was purified by column chromatography to give 18 (1.8 g, 2.04 mmol, 73% yield) as white solid. HPLC purity: 97.1A%. 1H NMR (600 MHz, chloroform-d) δ 9.03 (s, 1 H), 8.60 (bs, 1 H), 8.20 (bs, 2 H), 7.86 (dd, J = 8.3, 1.4 Hz, 1 H), 7.64−7.56 (m, 1 H), 7.27 (bs, 2 H), 7.17 (t, J = 7.6 Hz, 1 H), 6.58−6.49 (m, 4 H), 3.88 (s, 6 H), 3.68 (d, J = 12.4 Hz, 4 H), 3.34−3.26 (m, 1 H), 2.71 (t, J = 12.4 Hz, 4 H), 2.68−2.42 (m, 12 H), 2.42−2.35 (m, 2 H), 2.30 (s, 6 H), 1.96 (d, J = 11.7 Hz, 4 H), 1.77−1.66 (m, 8 H), 1.31 (d, J = 6.9 Hz, 6 H). 13C NMR (150 MHz, DMSO-d) δ 164.6, 163.5, 151.9, 148.9, 139.3, 134.6, 130.5, 124.9, 122.1, 122.0, 121.6, 119.0, 106.9, 100.1, 60.8, 55.4, 55.1, 54.9, 48.6, 48.5, 45.6, 27.7, 14.8. LC−MS (ESI): calcd for C46H67N12O4S ([M + H]+) 883.5; found 883.2.

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ACKNOWLEDGMENTS We thank Hodogaya JRF Contract Laboratories for their support on documentation of the manufacturing procedure as well as NMR and MS measurement. We thank Dr. Yasuhiro Kondo for the support on NMR measurement.

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REFERENCES

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S Supporting Information *

The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.oprd.8b00427. 1 H and 13C NMR spectra (PDF)

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AUTHOR INFORMATION

Corresponding Author

*E-mail: [email protected]. ORCID

Yuji Takahama: 0000-0002-3863-1997 Notes

The authors declare no competing financial interest. K



Article

M.; Hamano, M.; Igarashi, K.; Ooshima, H. The Effects of Impurities on Crystallization of Polymorphs of a Drug Substance AE1−923. J. Chem. Eng. Jpn. 2004, 37, 1224−1231. (c) Gong, Y.; Collman, B. M.; Mehrens, S. M.; Lu, E.; Miller, J. M.; Blackburn, A.; Grant, D. J. W. Stable-Form Screening: Overcoming Trace Impurities That Inhibit Solution-Mediated Phase Transformation to the Stable Polymorph of Sulfamerazine. J. Pharm. Sci. 2008, 97, 2130−2144. (d) Machiya, K.; Ieda, S.; Hirano, M.; Ooshima, H. Effects of Impurities on Crystal Polymorphism of an Imidazopyridine Derivative Developed as a Drug Substance for Osteoporosis. J. Chem. Eng. Jpn. 2009, 42, 147−152. (15) When crude 1 was treated with an activated carbon in a recrystallization step, less than 0.2% of new impurity was observed in purified 1. LC−MS analysis indicated that this impurity is derived from an oxidation of the methylene group on 1 (ESI calcd for C29H41N8O3S (1) [M+H]+ 581.3, found 595.2). For examples of oxidation by an activated carbon−molecular oxygen system, please see: (a) Nakamichi, N.; Kawabata, H.; Hayashi, M. Oxidative Aromatization of 9,10-Dihydroanthracenes Using Molecular Oxygen Promoted by Activated Carbon. J. Org. Chem. 2003, 68, 8272−8273. (b) Hayashi, M. Oxidation Using Activated Carbon and Molecular Oxygen System. Chem. Rec. 2008, 8, 252−267.

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Improved Manufacturing Route and Polymorphic Control of a Potent

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