Remibrutinib active pharmaceutical ingredient and formulations that are substantially free of nitrosamine impurities.

Abstract

The present invention relates to a novel process for preparing the N-(3-(6-amino-5-(2-(N-methylacrylamide)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide active pharmaceutical ingredient that is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. The present invention further relates to a pharmaceutical composition comprising N-(3-(6-amino-5-(2-(N-methylacrylamide)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, wherein the composition is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. Pharmaceutical products containing the above active pharmaceutical ingredient and composition, as well as methods for preparing the above active pharmaceutical ingredient, pharmaceutical composition, and product, are also disclosed.

Description

[Technical Field]

[0001] The present invention relates to a pharmaceutical active pharmaceutical ingredient (API) of N-(3-(6-amino-5-(2-(N-methylacrylamide)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide that is substantially free of nitrosamine impurities, and to a novel method for preparing the same. The present invention further relates to a pharmaceutical composition comprising N-(3-(6-amino-5-(2-(N-methylacrylamide)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, wherein the composition is substantially free of nitrosamine impurities. The present invention also relates to a method for preparing the API, the pharmaceutical composition thereof, and the product. [Background technology]

[0002] Nitrosamines are organic compounds containing an -NO functional group bonded to an amine group. Some members of this family of compounds have been identified as potentially carcinogenic to humans. Nitrosamines can form in pharmaceuticals and medicinal products during manufacture, storage, or further use.

[0003] A major concern is that prolonged exposure to high levels of nitrosamines may increase the risk of cancer. Several types of nitrosamines are classified as known or likely human carcinogens by regulatory bodies such as the International Agency for Research on Cancer (IARC). To ensure patient safety, regulatory authorities such as the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) impose strict limits on the amount of nitrosamines permissible in pharmaceutical products. Pharmaceutical manufacturers must implement rigorous quality control to minimize the formation of these undesirable compounds and ensure compliance with established safety standards. Continuous monitoring of drug quality is therefore crucial to prevent any potential risks to patient health.

[0004] N-(3-(6-amino-5-(2-(N-methylacrylamide)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide (IUPAC name: N-[3-(6-amino-5-{2-[methyl(prop-2-enoyl)amino]ethoxy}pyrimidine-4-yl)-5-fluoro-2-methylphenyl]-4-cyclopropyl-2-fluorobenzamide), also known as remibrutinib, is a highly potent and selective oral Bruton's tyrosine kinase (BTK) inhibitor. [ka]

[0005] Remibrutinib (also known as "LOU064") was first disclosed in Example 6 of International Publication No. 2015 / 079417, filed on November 28, 2014. International Publication No. 2015 / 079417 is incorporated in its entirety by reference. "LOU064" and "remibrutinib" are used without distinction. In Example 6(2) of International Publication No. 2015 / 079417, remibrutinib is prepared by cross-coupling "INT5" with "INT8" to obtain "INT9". [ka]

[0006] INT9 is then deprotected with TFA (Example 6(3)), reacted with acrylic acid, and purified to obtain remibrutinib (Example 6(4)). The preparation of INT5 is described in Example 1(5) of International Publication Brochure 2015 / 079417. INT5 is prepared by amide coupling of INT3 and INT4. [ka]

[0007] However, INT3 (5-fluoro-2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline) is a potentially mutagenic compound and has therefore been found to be a harmful intermediate in pharmaceutical synthesis. The genotoxicity of INT3 is reported for the first time in this application.

[0008] Therefore, a novel synthetic route for the preparation of remibrutinib that avoids the use of 5-fluoro-2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline is disclosed in PCT / IB2023 / 059664 (agent reference number (PAT059209-WO-PCT)). PCT / IB2023 / 059664 is incorporated herein by reference in its entirety.

[0009] It has now been discovered that the process disclosed in PCT / IB2023 / 059664 provides remibrutinib with a nitrosamine impurity at a level of approximately 1.6 ppm. The nitrosamine impurity is N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide (IUPAC name: N-[3-(6-amino-5-{2-[methyl(nitroso)amino]ethoxy}pyrimidine-4-yl)-5-fluoro-2-methylphenyl]-4-cyclopropyl-2-fluorobenzamide) and has the following structure: [ka] This also exists in another stable form. [ka]

[0010] It was observed that these nitrosamine impurities were formed during the last two steps of the process. [ka]

[0011] Now further, N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide has been found to be positive in the improved Ames Test (EAT), and therefore, it has been established that the health authorities consider it likely to be mutagenic under metabolically susceptible conditions. Even though the in vivo relevance of such positive EAT has not yet been established, it would be preferable to provide a drug substance that does not contain or substantially contains no nitrosoamine impurities that give positive EAT results, for example, does not contain N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. The presence of nitrosoamine impurities in the regorafenib drug substance and its mutagenicity in the EAT are first reported in this application.

[0012] For example, therefore, an object of the present invention is to provide a regorafenib drug substance that substantially contains no nitrosoamine impurities while avoiding the use of, for example, the 5-fluoro-2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline intermediate. Further, an object of the present invention is to provide a pharmaceutical composition comprising regorafenib or a pharmaceutically acceptable salt thereof and one or more excipients, wherein the composition substantially contains no nitrosoamine impurities, for example, substantially contains no N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. Summary of the Invention

[0013] The present invention relates to reducing the amount of nitrosoamine impurities, for example, the amount of nitrosoamine impurities that are positive in EAT. For example, reducing the amount of N-(3-(6-Amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide in the LOU064 drug substance to a certain level. For example, the content of nitrosoamine in the LOU064 drug substance is less than about 1000 ppb (for example, less than about 550 ppb, for example, less than about 530 ppb; less than about 400 ppb, for example, less than about 360 ppb; less than about 150 ppb, for example, less than about 130 ppb; less than about 100 ppb, for example, less than about 90 ppb); less than about 50 ppb; or less than about 25 ppb. All of these nitrosoamine impurity levels are determined based on the total amount of the LOU064 drug substance.

[0014] In another aspect, the present invention relates to a vemurafenib drug substance or formulation that is substantially free of impurities that give a positive EAT, for example, nitrosoamine impurities; for example, substantially free of the nitrosoamine impurity N-(3-(6-Amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

[0015] In another aspect, the present invention relates to a vemurafenib drug substance and formulation, and the vemurafenib drug substance and formulation comply with the safety or pharmaceutical product regulations of the EMA and FDA (for example, the new EMA guidance on nitrosoamine impurities). In one embodiment, the vemurafenib drug substance is substantially free of genotoxic impurities, for example, 5-Fluoro-2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline, and substantially free of nitrosoamine impurities, for example, the nitrosoamine impurity N-(3-(6-Amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

[0016] In one embodiment, the present invention relates to a remibrutinib active pharmaceutical ingredient that is substantially free of nitrosamine impurities, such as N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. In one embodiment, the present invention relates to a remibrutinib active pharmaceutical ingredient in which the level of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide is less than about 1000 ppb (e.g. less than about 550 ppb, e.g. less than about 530 ppb; less than about 400 ppb, e.g. less than about 360 ppb; less than about 150 ppb, e.g. less than about 130 ppb; less than about 100 ppb, e.g. less than about 90 ppb); less than about 50 ppb; or less than about 25 ppb.

[0017] In another embodiment, the level of nitrosamines in the remibrutinib API, for example, N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is between about 25 ppb and about 550 ppb, for example between about 25 ppb and about 530 ppb; or between about 25 ppb and about 400 ppb, for example between about 25 ppb and about 360 ppb; or between about 25 ppb and about 300 ppb; or between about 25 ppb and about 200 ppb, for example between about 25 ppb and about 90 ppb; or between about 25 ppb and about 100 ppb, for example between about 25 ppb and about 90 ppb.

[0018] In yet another embodiment, the level of nitrosamines in the remibrutinib active pharmaceutical ingredient, for example, N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is between about 100 ppb and about 650 ppb; or between about 100 ppb and about 550 ppb, for example between about 100 ppb and about 530 ppb; or between about 100 ppb and about 400 ppb, for example between about 100 ppb and about 360 ppb; or between about 100 ppb and about 350 ppb, for example between about 100 ppb and about 320 ppb; between about 100 ppb and about 250 ppb; or between about 100 ppb and about 150 ppb, for example between about 100 ppb and about 130 ppb.

[0019] In another embodiment, the present invention provides an improved process that enables the preparation of a remibrutinib API that is substantially free of nitrosamine impurities, such as N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, a. [ka] To provide a suspension containing a base, water, and a solvent; b. The present invention relates to a process comprising reacting a suspension with acrylic anhydride to provide a LOU064 active pharmaceutical ingredient that is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

[0020] In one embodiment, the process described above is carried out while limiting the amount of nitrite in any solvent used during the preparation of the active pharmaceutical ingredient, for example, by distilling the solvent before use and / or passing the solvent through an ion exchange resin capable of adsorbing nitrite.

[0021] In another embodiment, the process is carried out in the presence of about 1.1 to about 1.2 molar equivalents of base. In another embodiment, the base has a low nitrite content, for example, less than about 250 ppb or less than about 120 ppb. In one embodiment of this embodiment, the nitrite content in the base can be determined using a Griess test, for example, as described in Example 16. In one embodiment, the process is carried out using at least about 12 molar equivalents per mole of F8, for example at least about 25 molar equivalents per mole of F8, for example at least about 25 molar equivalents, for example at least about 35 molar equivalents, at least about 125 molar equivalents or at least about 150 molar equivalents.

[0022] In one embodiment, the process is carried out in ethyl acetate as the solvent. In further embodiments, details regarding the reaction conditions between the steps described above are provided herein.

[0023] In one embodiment, the process further includes a step for the preparation of F8. In another embodiment, the present invention relates to a pharmaceutical composition comprising remibrutinib or a pharmaceutically acceptable salt thereof, wherein the composition is substantially free of nitrosamine impurities, such as N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. In one embodiment of this design, the amount of nitrosamine impurities in the active pharmaceutical ingredient (e.g., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) is less than about 1000 ppb (e.g., less than about 550 ppb, e.g., less than about 530 ppb; less than about 400 ppb, e.g., less than about 360 ppb; less than about 150 ppb, e.g., less than about 130 ppb; less than about 100 ppb, e.g., less than about 90 ppb); less than about 50 ppb; or less than about 25 ppb, all based on the total amount of LOU064 in free or salt form.

[0024] In another embodiment of the above embodiment, the amount of nitrosamine in the composition, for example N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is between about 25 ppb and about 550 ppb, for example between about 25 ppb and about 530 ppb; or between about 25 ppb and about 400 ppb, for example between about 25 ppb and about 360 ppb; or between about 25 ppb and about 300 ppb; or between about 25 ppb and about 200 ppb, for example between about 25 ppb and about 90 ppb; or between about 25 ppb and about 100 ppb, for example between about 25 ppb and about 90 ppb, all based on the total amount of LOU064 in free or salt form.

[0025] In yet another embodiment of the above embodiment, the amount of nitrosamine in the composition, for example, N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is between about 100 ppb and about 650 ppb; or between about 100 ppb and about 550 ppb, for example, between about 100 ppb and about 530 ppb. ;or between approximately 100 ppb and approximately 400 ppb, for example between approximately 100 ppb and approximately 360 ppb; or between approximately 100 ppb and approximately 350 ppb, for example between approximately 100 ppb and approximately 320 ppb; or between approximately 100 ppb and approximately 250 ppb; or between approximately 100 ppb and approximately 150 ppb, for example between approximately 100 ppb and approximately 130 ppb, all based on the total amount of LOU064 in free or salt form.

[0026] In other embodiments, the present invention provides a method for producing a pharmaceutical composition comprising (ii) remibrutinib or a pharmaceutically acceptable salt thereof, and (ii) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition is substantially free of nitrosamines, in particular N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. In one embodiment of this present invention, the method is carried out while limiting the amount of nitrite in the solvent and excipient, for example, the nitrite level is less than 1.5 ppm, for example less than 1 ppm, less than 0.5 ppm or less than 0.2 ppm.

[0027] The present invention also provides a pharmaceutical product comprising the composition and documentation, for example, in the form of packaging or accompanying leaflets. For example, the pharmaceutical product may include documentation providing instructions to a patient on how to administer the composition and / or documentation certifying that the composition is substantially free of nitrosamines or at least N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

[0028] These pharmaceutical compositions are suitable for the treatment of BTK-related diseases as disclosed herein. [Modes for carrying out the invention]

[0029] Remibrutinib active pharmaceutical ingredient that is substantially free of nitrosamine impurities, particularly N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. In one embodiment, the present invention relates to a remibrutinib active pharmaceutical ingredient that is substantially free of nitrosamine impurities, such as N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. In one embodiment, the present invention relates to a remibrutinib active pharmaceutical ingredient in which the level of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide is less than about 1000 ppb (e.g. less than 550 ppb, e.g. less than about 530 ppb; less than about 400 ppb, e.g. less than about 360 ppb; less than about 150 ppb, e.g. less than about 130 ppb; less than about 100 ppb, e.g. less than about 90 ppb); less than about 50 ppb or less than about 25 ppb.

[0030] In some embodiments, the nitrosamine content in the remibrutinib active pharmaceutical ingredient is between about 25 ppb and about 550 ppb, for example between about 25 ppb and about 530 ppb; or between about 25 ppb and about 400 ppb, for example between about 25 ppb and about 360 ppb; or between about 25 ppb and about 300 ppb; or between about 25 ppb and about 200 ppb, for example between about 25 ppb and about 90 ppb; or between about 25 ppb and about 100 ppb, for example between about 25 ppb and about 90 ppb.

[0031] In another embodiment, the nitrosamine content in the remibrutinib active pharmaceutical ingredient is between about 100 ppb and about 650 ppb; or between about 100 ppb and about 550 ppb, for example between about 100 ppb and about 530 ppb; or between about 100 ppb and about 400 ppb, for example between about 100 ppb and about 360 ppb; or between about 100 ppb and about 350 ppb, for example between about 100 ppb and about 320 ppb; between about 100 ppb and about 250 ppb; or between about 100 ppb and about 150 ppb, for example between about 100 ppb and about 130 ppb.

[0032] In some further embodiments of the previous embodiments, remibrutinib is substantially pure (e.g., substantially chemically pure) as defined herein. In another embodiment, remibrutinib is in crystalline form as disclosed in International Publication No. 2020 / 234779, for example, in anhydrous crystalline form A as disclosed in Example 1 of International Publication No. 2020 / 234779. In one embodiment of this embodiment, remibrutinib is in the form of crystalline form A and is substantially phase-pure. In another embodiment, remibrutinib is substantially chemically pure and substantially phase-pure. As provided herein, remibrutinib is also substantially free of nitrosamine N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

[0033] The present invention is also useful in the preparation of LOU064 active pharmaceutical ingredient that is substantially free of nitrosamine impurities, such as nitrosamine N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. [ka]

[0034] In one embodiment, remibrutinib or any other compound described herein may be provided as a salt. As used herein, the term “salt” means an acid addition salt or base addition salt of a compound disclosed herein. “Salt” includes, in particular, “pharmaceutically acceptable salt.” The term “pharmaceutically acceptable salt” means a salt that retains the biological effects and properties of a compound disclosed herein and is not typically biologically or otherwise harmful. In many cases, the compounds disclosed herein can form salts of acids and / or bases in the presence of an amino group and / or a carboxyl group or similar group. Pharmacologically acceptable acid addition salts can be formed from inorganic and organic acids, such as acetate, aspartate, benzoate, besilate, bromide / hydrobromide, bicarbonate / carbonate, bisulfate / sulfate, camphor sulfonate, chloride / hydrochloride, chlortheophyllonate, citrate, ethandisulfonate, fumarate, gluceptate, gluconate, glucuronate, hippurate, hydroiodide / iodide These include salts, isethionates, lactates, lactobionates, lauryl sulfates, malates, maleates, malons, mandelates, mesylates, methyl sulfates, naphthoates, napsylates, nicotinates, nitrates, octadecanoates, oleates, oxalates, palmitates, pamoates, phosphates / hydrogen phosphates / dihydrogen phosphates, polygalactuloses, propions, stearates, succinates, sulfosalicylates, tartrates, tosylates, and trifluoroacetates. Inorganic acids that can derive salts include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and others of the same kind. Organic acids that can induce salt formation include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, and other similar substances.pharmaceutically acceptable base addition salts can be formed from inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, ammonium salts and metals of columns I to XII of the periodic table. In some embodiments, salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper, with ammonium, potassium, sodium, calcium, and magnesium salts being particularly suitable. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and others of the same kind. Certain organic amines include isopropylamine, benzathine, corinate, diethanolamine, diethylamine, lysine, meglumine, piperazine, and trimethamine. pharmaceutically acceptable salts of the compounds disclosed herein can be synthesized from basic or acidic components by conventional chemical methods. Generally, such salts can be prepared by reacting the free acidic form of these compounds with a theoretical amount of a suitable base (such as hydroxide, carbonic acid, sodium bicarbonate, Ca, Mg or K, or other similar substances) or by reacting the free base form of these compounds with a theoretical amount of a suitable acid. Such reactions are typically carried out in water, in organic solvents, or in mixtures thereof. Generally, the use of non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile is preferred when feasible. A list of additional suitable salts can be found, for example, in "Remington's Pharmaceutical Sciences," 20th ed., Mack Publishing Company, Easton, Pa., (1985); and "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).

[0035] In some embodiments, remibrutinib is in a crystalline form as described in International Publication No. 2020 / 234779, which is thus incorporated in whole by reference. In one embodiment, remibrutinib is in an anhydrous crystalline form A as described in Example 1 of International Publication No. 2020 / 234779. In another embodiment of this embodiment, remibrutinib is substantially pure phase as defined herein.

[0036] Many organic solvents are suitable for the chemical reactions described herein. For example, the reactions described herein may be carried out in aprotic organic solvents. Suitable examples include: acetonitrile; dimethyl sulfoxide (DMSO); dimethylformamide (DMF); halogenated alkanes such as dichloromethane (DCM); aromatic compounds such as benzene, toluene, xylene, mesitylene, and naphthalene; alkanes such as hexane, heptane, and octane; ketones such as acetone; ether compounds such as diethyl ether, tetrahydrofuran (THF), and THF derivatives such as methyl THF; ester compounds such as ethyl acetate and isopropyl acetate; amines such as pyridine; polyethylene glycol (PEG); in particular PEG having an average molecular weight of about 100 g / mol to about 2000 g / mol, such as PEG200, PEG600, PEG1000, and PEG2000; mono- or dialkyl PEG, especially its derivatives such as mono- or dimethyl PEG, mono- or diethyl PEG, and mono- or dipropyl PEG; and polypropylene glycol (PPG). Protic solvents may also be used in the reactions described herein. Protic solvents include: water; C 1~10Alcohols such as aliphatic branched or linear alcohols, particularly C1-C6 alcohols; and carboxylic acids such as methaneic acid, acetic acid, and propanoic acid. In one embodiment, the solvent includes toluene, ethanol, ethyl acetate, isopropyl acetate, methyl THF, heptane, and isopropanol. In one embodiment, the reactions described herein are carried out in a manner that avoids undesirable solvents such as DCM, DME, DMF, dioxane, and 1,2-dichloroethane, or other carcinogenic or teratogenic solvents. In certain embodiments, the amount of solvent in the reaction mixture is in the range of 0.1%-99% (v / v), 0.1%-80% (v / v), 0.1%-75% (v / v), 0.1%-50% (v / v), 1%-40% (v / v), 2%-30% (v / v), 4%-25% (v / v), or 5%-20% (v / v).

[0037] Some of the chemical reactions described herein can be carried out under acidic conditions, for example, at a pH of less than 7, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 or less. Acids suitable for the chemical reactions described are known to those skilled in the art. Commonly used acids include inorganic acids, such as sulfuric acid, phosphoric acid and nitric acid, and boric acid; halo acids such as hydrofluoric acid, hydrochloric acid, hydrobromic acid and hydroiodic acid; organic acids, such as carboxylic acids and acetic acid, benzoic acid and its derivatives; and halogenated acetic acids such as trifluoroacetic acid and dichloroacetic acid. In one embodiment, the acid is HF, HCl, or H2SO4. In another embodiment, fluorinated acids such as TFA are avoided in order to avoid the generation of fluorinated waste.

[0038] Some of the chemical reactions described herein can be carried out under basic conditions, for example, at a pH greater than 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14. Basic compounds suitable for the chemical reactions described herein are known to those skilled in the art. Commonly used bases include inorganic bases, such as alkali metal and alkaline earth metal hydroxides, such as lithium hydroxide, sodium hydroxide, potassium hydroxide, magnesium hydroxide, and calcium hydroxide. Stronger bases can be made from the addition of alkaline earth metals to hydrocarbons, amines, alcohols, and dihydrogens. Examples include butyllithium, lithium diisopropylamide (LDA), lithium diethylamide (LDEA), sodium amide, sodium ethanolate, sodium hydride (NaH), and lithium bis(trimethylsilyl)amide. Weaker bases include ammonia and amines, such as trialkylamines, such as triethylamine and diisopropylethylamine, and anions of weak acids, such as acetates (e.g., sodium acetate), potassium acetate, and carbonates (e.g., sodium carbonate, potassium carbonate). In some embodiments, the base is inorganic and has a low nitrite content (e.g., less than about 250 ppb, less than about 120 ppb, less than about 100 ppb, e.g., about 60 ppb). The nitrite content can be determined using a Griess test, as described in Example 16, for example.

[0039] The reactions described herein may be continued for as long as necessary to complete the reaction or at least to achieve an acceptable yield of the product. For example, the duration of the reaction may be less than 1 minute, less than 5 minutes, less than 10 minutes, less than 30 minutes, less than 1 hour, less than 2 hours, less than 3 hours, less than 5 hours, less than 10 hours, less than 20 hours, less than 30 hours, less than 40 hours, less than 50 hours, or less than 60 hours. The reaction time may depend, in particular, on the scale of the reaction. Those skilled in the art can monitor the progress of the reaction in many different ways, including by monitoring physical changes such as a change in color, or by monitoring the reaction using analytical methods such as NMR, FT-IR, XRPD, or chromatography, such as thin-layer chromatography (TLC) or liquid chromatography (LC-MS) connected to mass spectrometry.

[0040] Upon completion of the reactions described herein, the reaction mixture is optionally purified. Purification techniques known to those skilled in the art include: chromatography (e.g., HPLC, which may be reversed-phase or normal-phase); liquid-liquid separation using, for example, several immiscible solvents; and / or liquid-solid separation using, for example, filtration, decantation, (re)crystallization, trituration, evaporation, or freeze-drying.

[0041] The reactions described herein may be carried out on any suitable scale. In one embodiment, the reaction mixture is on an industrial scale. The reaction mixture may have a volume of, for example, at least 1 liter, particularly at least 10 liters, at least 100 liters, or at least 1000 liters. In another embodiment, the reaction mixture is on a microscale. The reaction mixture may have a volume of, for example, 10 ml or less, particularly 1 ml or less, 100 μl or less, 10 μl or less, or 1 μl or less.

[0042] The reactions described herein may be part of a series of reactions, including synthesis. Where multiple reactions are described, they may be carried out sequentially or in a one-pot manner. Sequential reactions typically involve the completion of a first reaction, followed by work-up and purification of that reaction before the second reaction is carried out, and further reactions are continued until the desired product is produced. In contrast, in a one-pot reaction, the first reaction may be completed, and then the second reaction may be carried out using one or more products of the first reaction without isolation. One-pot reactions are advantageous because they avoid unnecessary purification steps and save time and materials. In the synthesis of the remibrutinib drug substance described herein, some or all of the reactions may be carried out in a one-pot manner, or instead, some or all of the reactions may be carried out sequentially.

[0043] As used herein, the expression “includes” also includes, in addition to its literal meaning, the expressions “essentially consist of” and “consist of.” Thus, the expression “includes” refers to embodiments in which the subject “includes” the specifically enumerated elements may and / or actually include further elements, as well as embodiments in which the subject “includes” the specifically enumerated elements does not include further elements.

[0044] Numerical ranges described herein include the number defining the range. Headings provided herein are not limitations on the various aspects or embodiments of the invention as can be read by reference to the entire specification. In one embodiment, a subject described herein as including a particular step in the case of a method, or including a particular component in the case of a composition, refers to a subject consisting of the respective step or component. It is preferable to select and combine specific aspects and embodiments described herein, and the specific subjects resulting from each combination of specific embodiments also exist as part of this disclosure.

[0045] For the purposes of interpreting this specification, the following definitions shall apply, and whenever appropriate, a term used in the singular also includes the plural, and vice versa.

[0046] The term "substantially free" of nitrosamine impurities means that the nitrosamine content in the formulation is less than about 1,000 ppb (parts per billion) (e.g., less than about 550 ppb, less than about 530 ppb, less than about 400 ppb, less than about 360 ppb, less than about 150 ppb, less than about 130 ppb, less than about 100 ppb, less than about 90 ppb), less than about 50 ppb, or less than about 25 ppb. In one embodiment

[0047] The terms "ppm" and "ppb" represent parts per million and parts per billion, respectively. These terms indicate the weight ratio used to describe the concentration. For example, parts per billion (ppb) in an active pharmaceutical ingredient (API) is the number of unit masses of contaminants (e.g., nitrosamine impurities (e.g., nitrosamine N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) per 100 million units of the total mass of the API. The mass of the API is the sum of the mass of LOU064 (free base) and the mass of all impurities (including nitrosamine impurities).

[0048] This ppb ratio can be expressed as follows:

number

[0049] Similarly, the ppm ratio can be expressed as follows:

number

[0050] For example, regarding the "ppm" and "ppb" content of nitrosamines in a formulation, the nitrosamine content is expressed based on the amount of LOU064 in the formulation.

[0051] The amount of nitrosamine impurities (e.g., nitrosamine N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) is measured using mass spectrometry (MS) connected to a separation method (e.g., liquid chromatography or high-performance liquid chromatography (HPLC)). In one embodiment, HPLC is performed using a C18 column (e.g., porous or non-porous silica or ceramic microparticles, 1.5-10 μm in diameter, or octadecylsilane chemically bonded to a monolithic rod, e.g., a column listed in USP "L1", e.g., YMC-Triart C18, 100 × 3.0 mm, particle size 1.9 μm, 12 nm). In one embodiment, the detection level sensitivity can be improved by using selected ion monitoring (SIM) in mass spectrometry, and the scan mode is adjusted to detect only the selected mass (i.e., MH+483) of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. In one embodiment, the quantification of the nitrosamine content (i.e., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) in the active pharmaceutical ingredient is determined according to the method described in Example 1b or Example 15. Those skilled in the art will readily understand the use of these methods and other (or alternative) methods for determining the amount of impurities in the low ppm range.

[0052] As used herein, the term "approximately" means that any value may be "slightly above" or "slightly below" the endpoint, and is intended to provide a degree of freedom in the endpoint of a numerical range, insofar as it describes the variability that may be observed in measurements taken between various instruments, samples, and sample preparations. This term usually means within 5%, e.g., 1%, of any value or range. For example, 500 ppb means 500 ppb + / - 25 ppb (between 475 ppb and 525 ppb) or 500 + / - 5 ppb (i.e., between 495 ppb and 505 ppb).

[0053] The term "API" refers to a substance containing an active pharmaceutical ingredient intended for use in the manufacture of a pharmaceutical product (i.e., a pharmaceutical composition, such as a tablet or capsule). The active pharmaceutical ingredient (i.e., LOU064 (remibrutinib)) is present in the API at varying degrees of purity. The terms "API," "LOU064 API," and "remibrutinib API" are used interchangeably. Therefore, "LOU064 API" is LOU064 at varying degrees of purity. In one embodiment, remibrutinib is substantially pure (e.g., substantially chemically pure) as defined below. In another embodiment, remibrutinib is in crystalline form, as disclosed in International Publication No. 2020 / 234779, for example, in anhydrous crystalline form A, as disclosed in Example 1 of International Publication No. 2020 / 234779. In one aspect of this embodiment, remibrutinib is in crystalline form A and the phase is substantially pure. In another embodiment, remibrutinib is substantially chemically pure and substantially phase-pure. As provided by this disclosure, remibrutinib is also substantially free of nitrosamine N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

[0054] The terms “crystal form,” “crystal variety,” “polymorph,” or “polymorph” are used interchangeably herein. As used herein, “polymorph” refers to a crystal form having the same chemical composition but different stereochemistry of the molecules, atoms, and / or ions forming the crystal. Each polymorph differs with respect to thermodynamic stability, physical parameters, X-ray structure, and preparation method.

[0055] As used herein, “substantially pure,” when used in reference to LOU064, means a compound having remibrutinib of a purity higher than 90% by weight, higher than 90, 91, 92, 93, 94, 95, 96, 97, 98, and 99% by weight, and even equivalent to about 100% by weight, based on the weight of the active pharmaceutical ingredient. The remaining material includes other forms of the compound and / or reaction impurities and / or processing impurities resulting from its preparation. For example, the crystalline form of remibrutinib may be considered substantially pure in that it has a purity higher than 90% by weight, as measured by means currently known and generally accepted in the art, and the remaining less than 10% by weight of the material includes other forms of remibrutinib and / or reaction impurities and / or processing impurities.

[0056] As used herein, “substantially phase-pure” means, when used with respect to the crystalline form of LOU064, remibrutinib having a phase purity higher than about 90 wt% based on the weight of LOU064 on an anhydrous basis, higher than about 90, 91, 92, 93, 94, 95, 96, 97, 98 and about 99 wt%, and even equivalent to about 100 wt%. The terms “phase-pure” or “phase purity” herein refer to the homogeneity of the phase with respect to a particular solid state form of remibrutinib and do not necessarily mean high chemical purity unless that intention is explicitly stated. Phase purity may be determined according to methods known in the art, for example, using XRPD for quantitative phase analysis, using one or more approaches known in the art, for example, by external standard methods, by direct comparison of line (peak) characteristics attributable to various phases in a particular spectrum, or by internal standard methods. However, the quantification of phase purity by XRPD can be complicated in the presence of amorphous materials. Therefore, other methods that may be useful for determining phase purity include, for example, solid-state NMR spectroscopy, Raman spectroscopy, and / or infrared spectroscopy. Those skilled in the art will readily understand these methods and how to use these other (or alternative) methods for determining phase purity.

[0057] As used herein, “substantially chemically pure,” when used in reference to remibrutinib, means remibrutinib having a chemical purity higher than about 90% by weight, higher than about 90, 91, 92, 93, 94, 95, 96, 97, 98 and about 99% by weight, and even equivalent to about 100% by weight, based on the weight of the active pharmaceutical ingredient. The remaining material generally includes other compounds such as reaction impurities, starting materials, reagents, by-products and / or other processing impurities resulting from the preparation and / or isolation and / or purification of remibrutinib. For example, remibrutinib may be considered substantially chemically pure if it is determined to have a chemical purity higher than about 90% by weight, as measured by standard and generally accepted methods known in the art, with the remaining less than about 10% by weight constituting other material such as other stereoisomers of the compound of formula (I), reaction impurities, starting materials, reagents, by-products and / or processing impurities. Chemical purity may be determined by methods known in the art, such as high-performance liquid chromatography (HPLC), LC-MS (liquid chromatography-mass spectrometry), nuclear magnetic resonance (NMR) spectroscopy, or infrared spectroscopy. Those skilled in the art will readily understand the use of these methods and these other (or alternative) methods for determining chemical purity.

[0058] Manufacturing of remibrutinib active pharmaceutical ingredient that is substantially free of nitrosamine impurities. In one embodiment, the present invention relates to a novel synthetic route for the remibrutinib API that is substantially free of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. For example, the synthetic route also avoids the formation of the genotoxic intermediate INT-3. Furthermore, the process minimizes the purification step, improves the overall yield, and provides a more efficient process. The process can also be carried out in green solvent. In one embodiment, the present invention relates to a synthetic method for preparing the LOU064 API that is substantially free of nitrosamine impurities, for example, the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. [ka] It is reacted with acrylic anhydride in a solvent in the presence of a base. [ka] This includes generating, The present invention provides a synthesis method in which water is added before the addition of acrylic anhydride.

[0059] In one embodiment, the present invention is a process for preparing a LOU064 drug substance that is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, c. [ka] To provide a suspension containing a base, water, and a solvent; d. A process comprising reacting a suspension with acrylic anhydride to provide a LOU064 active pharmaceutical ingredient that is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

[0060] The above process of converting F8 to the LOU064 drug substance can be achieved in the presence of an organic base (e.g., trialkylamines such as triethylamine and diisopropylethylamine, N-methylmorpholine, and N-methylpyrrolidine; aromatic heterocyclic compounds such as pyridine and N-methylimidazole; or hydroxides of quaternary ammonium cations such as tetrabutylammonium hydroxide).

[0061] The above process for converting F8 to the LOU064 drug substance can be achieved in the presence of alkali metal and alkaline earth metal hydroxides such as Na2CO3, K2CO3, lithium hydroxide, sodium hydroxide, potassium hydroxide, magnesium hydroxide, and calcium hydroxide; or inorganic bases such as alkaline earth metal alkoxides or alkaline earth metal hydrides. In one embodiment of this design, the above reaction for converting F8 to the remibrutinib drug substance is achieved in the presence of Na2CO3.

[0062] In one embodiment, the base used in the process of converting F8 to the remibrutinib API is a base with a low nitrite content. In some embodiments, the nitrite content is less than about 250 ppb, for example less than about 120 ppb, for example about 60 ppb. In some embodiments, the level of nitrite in the base is determined using a Griess test, as described in Example 16. For example, bases with a low nitrite content are Na2CO3.10H2O or K2CO3.7H2O. In another embodiment, a base with a low nitrite content is Na2CO3 with a low nitrite content, for example less than about 250 ppb or less than about 120 ppb, for example about 60 ppb. In yet another embodiment, a base with a low nitrite content is K2CO3 with a low nitrite content, for example less than about 250 ppb or less than about 120 ppb, for example about 60 ppb.

[0063] In one embodiment, the above process of converting F8 to remibrutinib drug substance is achieved with approximately 1.1 to 1.2 molar equivalents of base per mole of intermediate F8.

[0064] The above process for converting F8 to remibrutinib API can be achieved in the presence of any suitable solvent. In one embodiment, the solvent can be selected from MeTHF, THF, alcohol (isopropanol), DCM, toluene, ethyl acetate, isopropyl acetate, acetonitrile, acetone, and tert-butyl methyl ether (TBME). In one aspect of this embodiment, the solvent has a low nitrite content, for example, less than 1.5 ppm, less than 1 ppm, less than 0.5 ppm, or less than 0.25 ppm.

[0065] In one embodiment, the above process of converting F8 to remibrutinib active pharmaceutical ingredient is carried out in ethyl acetate.

[0066] In one embodiment, the above process of converting F8 to remibrutinib API can be achieved using an amount of water sufficient to at least partially dissolve the base. In one embodiment, the above reaction of converting F8 to remibrutinib API can be achieved, for example, using at least about 12 molar equivalents of water, or using at least about 25 molar equivalents of water before adding acrylic anhydride, or using at least about 35 molar equivalents, or using at least about 125 molar equivalents of water before adding acrylic anhydride (step b), for example, between at least about 120 and about 150 molar equivalents of water. In one embodiment, the above process of converting F8 to LOU064 API is achieved with at least 150 molar equivalents of water before adding acrylic anhydride (step b). In one embodiment, the water used in the process (step a) is purified water (e.g., distilled water), for example, purified water with a nitrite level of less than about 50 ppb, for example, less than 20 ppb.

[0067] In some embodiments, the above process for converting F8 to remibrutinib API can be carried out at room temperature or heated at a temperature below the boiling point of the chosen solvent. For example, when ethyl acetate is used as the solvent, the reaction mixture can be heated to a temperature of about 50°C to about 65°C.

[0068] In some embodiments, F8 and the base are suspended in a solvent, and water is added to form a suspension. The suspension can be kept at room temperature or heated before the addition of acrylic anhydride. For example, F8 and Na2CO3 are suspended in ethyl acetate, water is added, and the suspension is heated to a temperature of about 50 to about 65°C before the addition of acrylic anhydride. In one embodiment, the acrylic anhydride is added in a solution in a suitable solvent, e.g., the same solvent used to dissolve or suspend F8 and the base. In one embodiment, the acrylic anhydride is gradually added to a solution or suspension of F8, the base and water in the solvent.

[0069] In one embodiment, acrylic anhydride is dissolved in ethyl acetate, and the solution of acrylic anhydride in ethyl acetate is gradually added to a suspension of F8, Na2CO3 in ethyl acetate, and water.

[0070] In one embodiment, the active pharmaceutical ingredient obtained by the above process is substantially free of nitrosamines, such as N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. In another embodiment of this embodiment, the content of nitrosamines, such as N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide in the active pharmaceutical ingredient obtained by the above process is less than about 550 ppb, for example less than about 530 ppb; less than about 400 ppb, for example less than about 360 ppb; less than about 300 ppb; less than about 200 ppb; less than about 150 ppb, for example less than about 130 ppb. In another embodiment of this design, the content of nitrosamines, such as N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide in the active pharmaceutical ingredient obtained by the above process is between about 100 ppb and about 650 ppb; or between about 100 ppb and about 550 ppb, for example between about 100 ppb and about 530 ppb; or between about 100 ppb and about 400 ppb, for example between about 100 ppb and about 360 ppb; or between about 100 ppb and about 350 ppb, for example between about 100 ppb and about 320 ppb; or between about 100 ppb and about 250 ppb; or between about 100 ppb and about 150 ppb, for example between about 100 ppb and about 130 ppb.

[0071] In one embodiment, the present invention further includes the step of forming an intermediate F8.

[0072] Preparation of F8 by deprotecting F7: [ka] P is an amine protecting group.

[0073] In one embodiment, F7 is deprotected using acidic conditions (for example, in the presence of HCl). F8 can then be isolated after the neutralization step.

[0074] It was found that the nitrosamine content of intermediate F8 increased upon exposure to air and / or during storage (Example 5, concerning the stability investigation of F8).

[0075] In one embodiment of the present invention, the nitrosamine content in F8 is reduced by utilizing purified water (e.g., distilled water) and a pure NaOH base (e.g., NaOH with a low nitrite content, e.g., less than about 250 ppb, less than about 120 ppb (e.g., by using a fresh bottle of NaOH, determined e.g., using a Griess test - Example 16) during the neutralization step (Example 5). In one embodiment, the nitrosamine content in F8 is reduced to, for example, less than about 100 ppb, less than about 80 ppb, less than about 50 ppb, less than about 40 ppb, for example, about 30 ppb.

[0076] In another embodiment of the present invention, intermediate F8 is isolated as described above but used directly for conversion step F8->API without drying (i.e., F8 is added to the API without any drying step as described above (step a)-Example 6). Drying and further exposure of F8 to air may increase the nitrosamine content. In one embodiment of this embodiment, the nitrosamine content in the API is reduced to, for example, less than about 100 ppb, less than about 50 ppb, less than about 30 ppb, for example, about 25 ppb.

[0077] The present invention further includes the step of forming an intermediate F7. A method for producing F7 is described in PCT / IB2023 / 059664, which is incorporated herein by reference in its entirety.

[0078] Preparation of F7 by reacting compound X6b and compound F6: [ka] X and Y are independently Cl, Br, or I (e.g., Br), and P is an amine protecting group.

[0079] In some embodiments, X is Cl or Br. In some embodiments, Y is Cl or Br. In some embodiments, X and Y are Cl or Br, respectively. In some embodiments, X is Br. In some embodiments, Y is Cl. In some embodiments, X is Br and Y is Cl. These embodiments apply to all examples of X and Y described herein, including the X and Y groups present in the synthetic precursors of X6b and F6, respectively.

[0080] Protecting group P can be any suitable amine protecting group that is stable during any of the chemical changes described herein (except for the deprotection step). The amine protecting group may be removed by conditions such as acid, base, hydrogenation, light, or heat. Examples of suitable amine protecting groups include carbamate protecting groups such as 9-fluorenylmethyl carbamate (Fmoc), t-butyl carbamate (Boc), or benzyl carbamate (Cbz); acetamide protecting groups such as acetamide, trifluoroacetamide, or benzylamide; and sulfonamide protecting groups such as p-toluenesulfonamide.

[0081] X6b and F6 can be converted to F7 according to coupling conditions suitable for forming a carbon-carbon bond. For example, the coupling of X6b and F6 can be achieved using an organometallic cross-coupling reaction, thereby linking the two fragments together with the help of a metal catalyst. Cross-coupling conditions that may be used in the coupling of X6b and F6 include: Kumada coupling; Negishi coupling; Still coupling; Suzuki-Miyaura coupling; and Hiyama coupling. In a typical cross-coupling reaction, a compound of type RM (R = first organic fragment, M = metal or typical compound) reacts with an organohalide of type R'-X (R' = second organic fragment, X = halide) to form a new carbon-carbon bond in the product R-R'.

[0082] Therefore, in some embodiments, the preparation of F7 involves the conversion of F6 to its precursor F6' by substituting Y's "M" with a metal-containing component or a main-group element-containing component, for example, M containing Zn (Negishi), B (Suzuki-Miyaura), Mg (Kumada), Sn (Still), or Si (Hiyama): [ka] P is an amine protecting group, such as Boc.

[0083] F6' reacts with X6b under cross-coupling conditions to obtain F7. In some embodiments, the conversion of F6 to F6' and the cross-coupling of F6' with X6b occur in a one-pot reaction. In some embodiments, the conversion of F6 to F6' and the cross-coupling of F6' with X6b occur in a sequential reaction.

[0084] Instead, the preparation of F7 involves the conversion of X6b to its precursor compound X6b' by substituting "M" in X with a metal-containing component or a main-group element-containing component, for example, M containing Zn (Negishi), B (Suzuki-Miyaura), Mg (Kumada), Sn (Still), or Si (Hiyama): [ka]

[0085] The precursor compound X6b' reacts with F6 under cross-coupling conditions to obtain F7. In some embodiments, the conversion of X6b to X6b' and the cross-coupling of X6b' with F6 occur in a one-pot reaction. In some embodiments, the conversion of X6b to X6b' and the cross-coupling of X6b' with F6 occur in a sequential reaction.

[0086] Preparation of X6a - Boration reaction The present invention further provides a synthesis method comprising the borylation of X6b to obtain X6a: [ka] X is F, Cl, Br, or I, n is 0 or 1, and R is F, Cl, Br, or I, OH, OC1-C6 alkyl, N(C1-C6 alkyl)2, aryl, or two or three R groups other than F, Cl, Br, I, or OH can together form a cyclic boronic acid ester, such as pinacolboronic acid or N-methyliminodiacetic acid (MIDA) boronate.

[0087] Boration of X6b can be achieved using one or more catalysts, one or more ligands, one or more boronating agents, one or more bases, and / or one or more additives. In some embodiments, the boronation comprises one or more catalysts, one or more ligands, one or more boronating agents, and one or more bases. In some embodiments, the boronation further comprises one or more additives.

[0088] A boronating agent is a boron-containing compound that can convert organic halogen compounds into boronic acids or boronic acid esters under typical metal-catalyzed cross-coupling conditions. In some embodiments, the boronating agent is selected from the group consisting of diborone compounds, boronic acids, boranes, boron trihalides, and borates. In some embodiments, the boronating agent is selected from the group consisting of bis(pinacolate)diborone, B2(NMe2)4, B2F4, B2Cl4, B2Br4, B214, bisboronic acid, pinacolborane, HB(NMe2)2, B(OH)3, BF3, BCl3, BBr3, BI3, C1-C6 borate mono, di, or trialkyl, borate mono, di, or trimethyl, borate mono, di, or triethyl, and borate mono, di, or tritripropyl, for example, bis(pinacolate)diborone or bisboronic acid. Compared to pinacolborane or bis(pinacolate)diboron, bisboronic acids may be attractive because they allow for less catalyst use, milder reaction conditions, and avoid the formation of pinacol-related impurities. Bisboronic acids also allow for the use of green solvents such as alcohol solvents and milder reaction conditions (e.g., lower temperatures).

[0089] The metal catalyst used in the boronation reaction may contain palladium, nickel, copper, or a combination thereof, for example, palladium.

[0090] In some embodiments, the metal catalyst is provided as a complex of the catalyst precursor, for example, a Buchwald G1, G2, G3, or G4 catalyst precursor complexed with a phosphine ligand. The Buchwald catalyst precursor is used to generate active Pd(0) in insights via rapid deprotonation and reductive elimination. The catalyst precursor is useful because it allows for low catalyst usage, is stable to air, moisture, and heat, and has good solubility. These catalyst precursors have been optimized from the first to fourth generations (G1 to G4) to further improve functionality and solubility. The catalyst precursor consists of a Parada cycle (shown below) having a phenyl or 1,1-biphenyl skeleton, where L represents a bonded phosphine ligand, e.g., XPhos, SPhos, etc. (see below), and the bonded amine substituent and leaving group (Cl, OMs) differ by generation.

[0091] Examples of Buchwald catalyst precursors that form complexes with palladium and exemplary XPhos ligands are shown below.

[0092] [Table 1]

[0093] Any other phosphine ligand described herein may be used as L instead of XPhos in the table above.

[0094] Other catalyst precursors for borylation may include Pd(TFA)2, PdBr2, or Pd(MeCN)2Cl2. These catalyst precursors can be used in the presence of ligands such as Ph2P(t-Bu);Cy3P-HBF4;RuPHOS;S-PHOS, Cy-BIPHEP;SPHOS-SO3Na.

[0095] In some embodiments, the borylation of X6b involves a further ligand in addition to ligand L, which forms part of the complex of the catalyst precursor. In other embodiments, the further ligand is not required. In some embodiments, the borylation of X6b uses a catalyst and ligand without a catalyst precursor (Pd(0) catalyst; e.g., Pd(PPh3)4).

[0096] A wide range of ligands can be used in boration reactions, and these ligands can influence the reactivity of the reagents. For example, ligands can increase the electron density at the metal center of a metal complex, which can improve the oxidative addition step. In addition, bulky ligands are useful in the reductive elimination step. In some embodiments, the ligands used in the boration of X6b are selected from the group consisting of organophosphines, N-heterocyclic carbenes, diazabutadienes, dibenzylideneacetone, and combinations thereof.

[0097] In one embodiment, the ligand is an organophosphine selected from the group consisting of organophosphine ligands such as XPhos, APhos, CPhos, RuPhos, SPhos, cataCXium, DavePhos, JohnPhos, MePhos, XantPhos, Cy3P-HBF4, Cy-BIPHEP, SPhos-SO3Na, PPh3, tBuPPh2 and combinations thereof, such as XPhos, APhos, CPhos, RuPhos, SPhos, cataCXium, and tBuPPh2. The phosphine ligands are shown in the table below.

[0098] [Table 2]

[0099] [Table 3]

[0100] The borylation of X6b may include a base. In some embodiments, the base is an organic or inorganic salt such as NaOH, Ca(OH)2, Na2CO3, K2CO3, K3PO4, Cs2CO3, KOAc, KOPh or NaOAc, diisopropylethylamine (DIPEA), triethylamine or other tertiary amines or combinations thereof. In one embodiment, the base is DIPEA, KOAc or KOH, for example, KOAc.

[0101] The boration of X6b may include additives, such as alcohols like ethylene glycol. In some embodiments, the boration of X6b does not include additives.

[0102] Boration of X6b can be carried out in any suitable solvent. Examples of suitable organic solvents include polar solvents, nonpolar solvents, protic solvents, aprotic solvents, polar protic solvents, and polar aprotic solvents. In one embodiment, boration can be carried out in alcohol solvents including t-amyl alcohol, hexanol, pentanol, butanol (terto-butanol, isobutanol, and n-butanol), propanol (isopropanol and n-propanol), ethanol, and / or methanol. In one embodiment, boration can be carried out in methanol, toluene, and / or MeTHF, for example, in MeTHF. Other solvents, such as halogenated alkane solvents such as dichloromethane, can also be used. Ether-based solvents such as dioxane, MeTHF, THF, and dialkyl ethers such as diethyl ether can also be used. Boration can also be carried out in an aqueous environment, including a micelle environment. In some embodiments, mixtures of solvents are used.

[0103] The borylation of X6b can be achieved using one or more catalysts, one or more ligands, one or more boronating agents, one or more bases, and / or optionally one or more additives. Those skilled in the art can determine appropriate amounts of these reagents. Nevertheless, in some embodiments of the borylation reaction: i) The catalyst or catalyst precursor is present in an amount of 0.01 mol% to 3 mol%, 0.05 mol% to 2 mol%, 0.1 mol% to 2 mol%, 0.1 to 1 mol%, preferably 0.25 mol%, and more preferably 0.5 mol%, relative to the number of moles of X6b; ii) The amount of ligand is 0.02 mol% to 6 mol%, 0.1 mol% to 2 mol%, 0.2 mol% to 1 mol%, 0.5 mol%, or 1 mol% compared to the number of moles of X6b; iii) The number of moles of ligand is 2 or 3 times the number of moles of catalyst or catalyst precursor; preferably 2 times; iv) The amount of the boronating agent is 1 to 3 molar equivalents compared to X6b, preferably 1 to 2 molar equivalents, more preferably 1.05 or 1.5 molar equivalents compared to X6b; v) The amount of base is 2 to 5 molar equivalents, preferably 2 to 3 molar equivalents, most preferably 2.5 or 3 molar equivalents, relative to the number of moles of X6b; and / or vi) Additives are optional and, if present, in an amount of 2 to 5 molar equivalents compared to X6b; preferably, no additives are present.

[0104] A boration reaction may be characterized by any one of the above i) to vi). A boration reaction may be characterized by any two of the above i) to vi). A boration reaction may be characterized by any three of the above i) to vi). A boration reaction may be characterized by any four of the above i) to vi). A boration reaction may be characterized by any five of the above i) to vi). A boration reaction may be characterized by all of the above i) to vi).

[0105] The boration reaction may be characterized by the above i) and ii). The boration reaction may be characterized by the above i) and iii). The boration reaction may be characterized by the above i) and iv). The boration reaction may be characterized by the above i) and v). The boration reaction may be characterized by the above i) and vi). The boration reaction may be characterized by the above ii) and iii). The boration reaction may be characterized by the above ii) and iv). The boration reaction may be characterized by the above ii) and v). The boration reaction may be characterized by the above ii) and vi). The boration reaction may be characterized by the above iii) and iv). The boration reaction may be characterized by the above iii) and v). The boration reaction may be characterized by the above iii) and vi). The boration reaction may be characterized by the above iv) and v). The boration reaction may be characterized by the above iv) and vi). The boration reaction may be characterized by the above v) and vi).

[0106] In one example, a borylation reaction having excellent yield and minimal byproducts may be as follows: [ka]

[0107] In one embodiment, a boration reaction having excellent yield and minimal byproduct formation is characterized by at least one of the following: i) The catalyst is Pd(MeCN)2Cl2 in an amount of 0.1 mol% to 2 mol% or 0.1 mol% to 1.5 mol%, preferably 0.25 mol%, or more preferably 0.5 mol%, relative to the number of moles of X6b; ii) The ligand is tBuPPh2 in an amount of 0.2 mol% to 4% relative to the number of moles of X6b, preferably 0.5 mol%, or more preferably 1 mol%. iii) The catalyst is Pd(MeCN)2Cl2, the ligand is tBuPPh2, and the number of moles of tBuPPh2 is 2 or 3 times the number of moles of Pd(MeCN)2Cl2, preferably 2 times the number of moles of Pd(MeCN)2Cl2; iv) The boronating agent is bis(pinacolate)diboron in an amount of 1 to 2 molar equivalents compared to X6b, preferably about 1.05 molar equivalents compared to X6b; v) The base is KOAc in an amount of 2 to 5 molar equivalents compared to X6b, preferably 2.5 equivalents compared to X6b; and vi) No additives are present; and / or vii) The reaction temperature is 30°C to 120°C, for example 40°C to 50°C, preferably 60°C or 70°C.

[0108] A boration reaction may be characterized by any one of the above i) to vii). A boration reaction may be characterized by any two of the above i) to vi). A boration reaction may be characterized by any three of the above i) to vii). A boration reaction may be characterized by any four of the above i) to vii). A boration reaction may be characterized by any five of the above i) to vii). A boration reaction may be characterized by any six of the above i) to vii). A boration reaction may be characterized by all of the above i) to vii).

[0109] The boration reaction may be characterized by the above i) and ii). The boration reaction may be characterized by the above i) and iii). The boration reaction may be characterized by the above i) and iv). The boration reaction may be characterized by the above i) and v). The boration reaction may be characterized by the above i) and vi). The boration reaction may be characterized by the above i) and vii). The boration reaction may be characterized by the above i) and vii). The boration reaction may be characterized by the above ii) and iii). The boration reaction may be characterized by the above ii) and iv). The boration reaction may be characterized by the above ii) and v). The boration reaction may be characterized by the above ii) and vii). The boration reaction may be characterized by the above iii) and iv). The boration reaction may be characterized by iii) and v) above. The boration reaction may be characterized by iii) and vi) above. The boration reaction may be characterized by iii) and vii) above. The boration reaction may be characterized by iv) and v) above. The boration reaction may be characterized by iv) and vi) above. The boration reaction may be characterized by iv) and vii) above. The boration reaction may be characterized by v) and vi) above. The boration reaction may be characterized by v) and vii) above. The boration reaction may be characterized by vi) and vi) above.

[0110] In one embodiment, a boration reaction having good yield and minimal byproduct formation is characterized by at least one of the following: i) The catalyst is a catalyst precursor which is Pd-XPhos-2G in an amount of 0.05 mol% to 0.5 mol%, preferably 0.25 mol%, relative to the number of moles of X6b; ii) The ligand is XPhos in an amount of 0.1 mol% to 1 mol% relative to the number of moles of X6b; preferably 0.5 mol% relative to the number of moles of X6b; iii) The catalyst is Pd-XPhos-2G, the ligand is XPhos, and the number of moles of XPhos is twice the number of moles of Pd-XPhos-2G; iv) The boronating agent is bisboronic acid in an amount of 1 to 3 molar equivalents compared to X6b, preferably 1.5 molar equivalents compared to X6b; v) The base is potassium acetate in an amount of 2 to 5 molar equivalents, preferably 3 molar equivalents, compared to X6b; vi) The additive is ethylene glycol in an amount of 2 to 5 molar equivalents compared to X6b, preferably 3 molar equivalents compared to X6b; and vii) The reaction temperature is 30°C to 70°C, preferably 40°C to 50°C, and more preferably 50°C.

[0111] A boration reaction may be characterized by any one of the above i) to vii). A boration reaction may be characterized by any two of the above i) to vi). A boration reaction may be characterized by any three of the above i) to vii). A boration reaction may be characterized by any four of the above i) to vii). A boration reaction may be characterized by any five of the above i) to vii). A boration reaction may be characterized by any six of the above i) to vii). A boration reaction may be characterized by all of the above i) to vii).

[0112] The boration reaction may be characterized by the above i) and ii). The boration reaction may be characterized by the above i) and iii). The boration reaction may be characterized by the above i) and iv). The boration reaction may be characterized by the above i) and v). The boration reaction may be characterized by the above i) and vi). The boration reaction may be characterized by the above i) and vii). The boration reaction may be characterized by the above i) and vii). The boration reaction may be characterized by the above ii) and iii). The boration reaction may be characterized by the above ii) and iv). The boration reaction may be characterized by the above ii) and v). The boration reaction may be characterized by the above ii) and vii). The boration reaction may be characterized by the above iii) and iv). The boration reaction may be characterized by iii) and v) above. The boration reaction may be characterized by iii) and vi) above. The boration reaction may be characterized by iii) and vii) above. The boration reaction may be characterized by iv) and v) above. The boration reaction may be characterized by iv) and vi) above. The boration reaction may be characterized by iv) and vii) above. The boration reaction may be characterized by v) and vi) above. The boration reaction may be characterized by v) and vii) above. The boration reaction may be characterized by vi) and vii) above.

[0113] In another embodiment, the boration reaction may be as follows: [ka]

[0114] In one embodiment, a boration reaction having good yield and minimal byproduct formation is characterized by at least one of the following: i) The catalyst is Pd-cataCXium-3G in an amount of 0.001 mol% to 0.5 mol%, preferably 0.05 mol%, relative to the number of moles of X6b; ii) The ligand is cataCXium in an amount of 0.02 mol% to 1% relative to the number of moles of X6b, preferably 0.1 mol% relative to the number of moles of X6b; iii) The catalyst is Pd-cataCXium-3G, the ligand is cataCXium, and the number of moles of cataCXium is twice the number of moles of Pd-cataCXium-3-3G; iv) The boronating agent is bisboronic acid in an amount of 1 to 3 molar equivalents, preferably 1.5 molar equivalents, compared to X6b; v) The base is N,N-diisopropylethylamine in an amount of 2 to 5 molar equivalents compared to X6b, preferably an equivalent amount compared to X6b; and vi) No additives are present; and / or vii) The reaction temperature is 30°C to 70°C, preferably 40°C to 50°C, and more preferably 50°C.

[0115] A boration reaction may be characterized by any one of the above i) to vii). A boration reaction may be characterized by any two of the above i) to vi). A boration reaction may be characterized by any three of the above i) to vii). A boration reaction may be characterized by any four of the above i) to vii). A boration reaction may be characterized by any five of the above i) to vii). A boration reaction may be characterized by any six of the above i) to vii). A boration reaction may be characterized by all of the above i) to vii).

[0116] The boration reaction may be characterized by the above i) and ii). The boration reaction may be characterized by the above i) and iii). The boration reaction may be characterized by the above i) and iv). The boration reaction may be characterized by the above i) and v). The boration reaction may be characterized by the above i) and vi). The boration reaction may be characterized by the above i) and vii). The boration reaction may be characterized by the above i) and vii). The boration reaction may be characterized by the above ii) and iii). The boration reaction may be characterized by the above ii) and iv). The boration reaction may be characterized by the above ii) and v). The boration reaction may be characterized by the above ii) and vii). The boration reaction may be characterized by the above iii) and iv). The boration reaction may be characterized by iii) and v) above. The boration reaction may be characterized by iii) and vi) above. The boration reaction may be characterized by iii) and vii) above. The boration reaction may be characterized by iv) and v) above. The boration reaction may be characterized by iv) and vi) above. The boration reaction may be characterized by iv) and vii) above. The boration reaction may be characterized by v) and vi) above. The boration reaction may be characterized by v) and vii) above. The boration reaction may be characterized by vi) and vii) above.

[0117] For example, the reaction may be as follows: [ka]

[0118] Coupling of X6a and F6 In some embodiments of the present invention, the borylation of X6b to obtain X6a is used in a method for synthesizing compound F7. In such embodiments, X6b is converted to X6a, and then X6a is reacted with F6 under cross-coupling conditions to produce F7. In one embodiment, the conversion of X6b to X6a and the cross-coupling of X6a with F6 are carried out in a one-pot reaction. In some embodiments, the conversion of X6b to X6a and the cross-coupling of X6a with F6 are carried out in a sequential reaction.

[0119] According to the present invention, a borated compound X6a can react with an aryl halide in a cross-coupling reaction. In one embodiment, the coupling reaction is carried out using one or more catalysts, one or more ligands, one or more bases and / or one or more additives. In one embodiment, the coupling reaction is carried out using one or more catalysts, one or more ligands and one or more bases. In some embodiments, the coupling further includes one or more additives.

[0120] The metal catalyst used in the cross-coupling reaction may contain palladium, nickel, copper, or a combination thereof, for example, palladium.

[0121] A wide range of ligands can be used in the cross-coupling of X6a and F6, and the ligands can influence the reactivity of the coupling reagent. For example, ligands can increase the electron density at the metal center of the metal complex, which can improve the oxidative addition step. In addition, bulky ligands are useful in the reductive elimination step. In some embodiments, the ligand used in the coupling of X6a and F6 is selected from the group consisting of organophosphines, N-heterocyclic carbenes, diazabutadienes, dibenzylideneacetone, and combinations thereof. In one embodiment, the ligand is an organophosphine selected from the group consisting of organophosphine ligands, e.g., XPhos, APhos, CPhos, RuPhos, SPhos, cataCXium, DavePhos, JohnPhos, MePhos, XantPhos, PPh3, tBuPPh2, and combinations thereof. In another embodiment, the ligand is XPhos, APhos, CPhos, RuPhos, SPhos, cataCXium, for example XPhos, cataCXium and tBuPPh2. In yet another embodiment, the ligand is tBuPPh2.

[0122] In the coupling of X6a and F6, the metal catalyst and ligand may be provided as a Buchwald G1, G2, G3, or G4 catalyst precursor, preferably G2, which is a complex of a catalyst precursor, for example, an organic phosphine ligand selected from the group consisting of XPhos, APhos, CPhos, RuPhos, SPhos, cataCXium, DavePhos, JohnPhos, MePhos, XantPhos, Cy3P-HBF4, Cy-BIPHEP, SPHOS-SO3Na, PPh3, tBuPPh2, and combinations thereof. In one embodiment, the organic phosphine is XPhos, APhos, CPhos, RuPhos, SPhos, cataCXium, for example, XPhos, cataCXium, and tBuPPh2. In another embodiment, the organic phosphine is tBuPPh2.

[0123] In some embodiments, a catalyst precursor containing a phosphine ligand is used, and no additional phosphine ligand is used. Alternatively, a catalyst precursor containing a phosphine ligand is used, and additional phosphine ligands are also used. Examples of catalyst precursors for cross-coupling reactions may include Pd(TFA)2, PdBr2, or Pd(MeCN)2Cl2. These catalyst precursors can be used in the presence of ligands such as Ph2P(t-Bu);Cy3P-HBF4;RuPHOS;S-PHOS, Cy-BIPHEP;SPHOS-SO3Na.

[0124] The coupling of X6a and F6 may include a base. In some embodiments, the base is an organic or inorganic salt such as KOH, NaOH, Ca(OH)2, Na2CO3, K2CO3, K3PO4, Cs2CO3, KOAc, KOPh or NaOAc, diisopropylethylamine (DIPEA), triethylamine or other tertiary amines or combinations thereof. In one embodiment, the base is triethylamine or KOH, for example, KOH.

[0125] The coupling of X6a and F6 may optionally include additives, such as alcohols, for example, ethylene glycol, if a PdXPhos-2G / XPhos complex is used.

[0126] The coupling of X6a and F6 can be carried out in any suitable solvent. Examples of suitable organic solvents include polar solvents, nonpolar solvents, protic solvents, aprotic solvents, polar protic solvents, and polar aprotic solvents. In one embodiment, the cross-coupling reaction can be carried out in alcohol solvents including t-amyl alcohol, hexanol, pentanol, butanol (terto-butanol, isobutanol, and n-butanol), propanol (isopropanol and n-propanol), ethanol, and / or methanol. Other solvents, such as halogenated alkane solvents such as dichloromethane, can also be used. Ether solvents such as dioxane, MeTHF, THF, and dialkyl ethers such as diethyl ether can also be used. The coupling can also be carried out in an aqueous environment, including a micelle environment. In some embodiments, a mixture of solvents, such as MeTHF and water, is used. When methanol is used, the reaction mixture may be precipitated to simplify purification.

[0127] The coupling of X6a and F6 can be achieved using one or more catalysts, one or more ligands, one or more boronating agents, one or more bases, and / or one or more additives. Those skilled in the art can use well-known general knowledge to determine appropriate amounts of these reagents.

[0128] In one embodiment, a coupling reaction having excellent yield and minimal byproduct formation is characterized by at least one of the following: i) The catalyst or catalyst precursor is present in an amount of 0.1 mol% to 5 mol%, 0.25 mol% to 3 mol%, 0.5 mol% to 1.5 mol%, preferably 0.5 mol%, or more preferably 1 mol%, relative to the number of moles of F6 or X6a; ii) The number of moles of ligand, if present, is 2 or 3 times the number of moles of catalyst or catalyst precursor, preferably 2 times; iii) The molar ratio of F6:X6a is 2:1 to 1:2, i.e., 1.5:1 to 1:1.5, 1.2:1 to 1:1.2, or 1:1; iv) Additives are optional and, if present, in an amount of 2 to 5 molar equivalents compared to F6 or X6a; and / or v) The amount of base is 2 to 5 molar equivalents, preferably 2 to 3 molar equivalents, and most preferably 3 molar equivalents, compared to the number of moles of F6 or X6a.

[0129] A coupling reaction may be characterized by any one of the above i) to v). A coupling reaction may be characterized by any two of the above i) to v). A coupling reaction may be characterized by any three of the above i) to v). A coupling reaction may be characterized by any four of the above i) to v). A coupling reaction may be characterized by all of the above i) to v).

[0130] The coupling reaction may be characterized by i) and ii) above. The coupling reaction may be characterized by i) and iii) above. The coupling reaction may be characterized by i) and iv) above. The coupling reaction may be characterized by i) and v) above. The coupling reaction may be characterized by ii) and iii) above. The coupling reaction may be characterized by ii) and iv) above. The coupling reaction may be characterized by ii) and v) above. The coupling reaction may be characterized by iii) and iv) above. The coupling reaction may be characterized by iii) and v) above. The coupling reaction may be characterized by iv) and v) above.

[0131] In one embodiment, a coupling reaction having good yield and minimal byproduct formation is characterized by at least one of the following: i) The catalyst and ligand are provided as catalyst precursor-ligand complexes, with Pd and X-Phos-2G in amounts of 0.5 mol% to 2 mol% compared to the number of moles of F6 or X6a; ii) The base is triethylamine in an amount of 2 to 5 molar equivalents, preferably 3 molar equivalents, compared to F6 or X6a; iii) The additive is ethylene glycol in an amount of 2 to 5 molar equivalents, preferably 3 molar equivalents, compared to F6 or X6a; iv) The reaction is carried out in an alcohol solvent, preferably methanol; and v) The reaction temperature is 30°C to 70°C, preferably 40°C to 50°C, and more preferably 50°C.

[0132] A coupling reaction may be characterized by any one of the above i) to v). A coupling reaction may be characterized by any two of the above i) to v). A coupling reaction may be characterized by any three of the above i) to v). A coupling reaction may be characterized by any four of the above i) to v). A coupling reaction may be characterized by all of the above i) to v).

[0133] The coupling reaction may be characterized by i) and ii) above. The coupling reaction may be characterized by i) and iii) above. The coupling reaction may be characterized by i) and iv) above. The coupling reaction may be characterized by i) and v) above. The coupling reaction may be characterized by ii) and iii) above. The coupling reaction may be characterized by ii) and iv) above. The coupling reaction may be characterized by ii) and v) above. The coupling reaction may be characterized by iii) and iv) above. The coupling reaction may be characterized by iii) and v) above. The coupling reaction may be characterized by iv) and v) above.

[0134] In one embodiment, a coupling reaction having excellent yield and minimal byproduct formation is characterized by at least one of the following: i) The catalyst is Pd(MeCN)2Cl2 in an amount of 0.25 mol% to 2 mol%, preferably 0.5 mol%, or more preferably 1 mol%, relative to the number of moles of X6b; (the conversion of X6b to X6a is approximately 98%). ii) The ligand is tBuPPh2 in an amount of 0.5 mol% to 4% compared to the number of moles of X6b, preferably 1 mol% or 2 mol% compared to the number of moles of X6b; in particular, the catalyst is Pd(MeCN)2Cl2, the ligand is tBuPPh2, and the number of moles of tBuPPh2 is twice the number of moles of Pd(MeCN)2Cl2; iii) The base is 2 to 5 molar equivalents, preferably 3 molar equivalents, of KOH compared to X6b; iv) The reaction is carried out in a mixture of MeTHF and water; and v) The reaction temperature is 30°C to 70°C, preferably 60°C.

[0135] A coupling reaction may be characterized by any one of the above i) to v). A coupling reaction may be characterized by any two of the above i) to v). A coupling reaction may be characterized by any three of the above i) to v). A coupling reaction may be characterized by any four of the above i) to v). A coupling reaction may be characterized by all of the above i) to v).

[0136] The coupling reaction may be characterized by i) and ii) above. The coupling reaction may be characterized by i) and iii) above. The coupling reaction may be characterized by i) and iv) above. The coupling reaction may be characterized by i) and v) above. The coupling reaction may be characterized by ii) and iii) above. The coupling reaction may be characterized by ii) and iv) above. The coupling reaction may be characterized by ii) and v) above. The coupling reaction may be characterized by iii) and iv) above. The coupling reaction may be characterized by iii) and v) above. The coupling reaction may be characterized by iv) and v) above.

[0137] In one embodiment, the borylation of X6b to X6a and the cross-coupling of X6a and F6 are carried out in a one-pot reaction.

[0138] Preparation of X6b X6b is a key intermediate in the novel synthesis described herein. Therefore, the present invention further provides the synthetic intermediate, X6b: [ka] X is F, Cl, Br, or I. In one embodiment, X is Br.

[0139] X6b can be synthesized by any suitable means. The present invention relates to a method for preparing the synthetic intermediate X6b: [ka] The present invention provides a method in which X is F, Cl, Br, or I. In one embodiment, X is Br.

[0140] In some embodiments, the method includes reacting compound X6d with compound N6a: [ka] X is Cl, Br, or I. In one embodiment, X is Br.

[0141] Carboxylic acid coupling reactions, including amidation reactions, are well known to those skilled in the art and typically involve reacting an amine with a carboxylic acid under coupling conditions or converting a carboxylic acid group to an activated group that can react more readily with an amine.

[0142] Therefore, in one embodiment, the synthesis of X6b involves using the conversion of the carboxylic acid group of X6d to an activated carboxylic acid group. For example, the method may include the conversion of compound X6d to compound X6c: [ka] R 10 is an activated carboxylic acid group, such as an acyl anhydride, acyl halogen, or acyl phosphoric acid, and X is Cl, Br, or I. For example, the conversion of X6d to the corresponding acyl chloride can be achieved using thionyl chloride. The solvent may be an aromatic solvent such as toluene. The base may be pyridine. X6c can then be reacted with N6a to form compound X6b. These reactions can be carried out as a one-pot synthesis or sequentially. The formation of N6a from N6b may also be carried out in this one-pot synthesis, where X6c and N6a are prepared separately but then coupled.

[0143] Alternatively, X6b is prepared directly from X6d and N6a by using a carboxylic acid activating reagent. Well-known carboxylic acid activating reagents include HBT, HATU, HBTU, TBTU, HOBt, PyAOP, HCTU, PyClocK, TFFH, carbodiimides (e.g., DCC), carbonyldiimidazole (CDI), and phosphonium salts (e.g., BOP, PyBOP).

[0144] The coupling of X6d or X6c and N6a can be carried out in the presence of a base, such as a tertiary alkylamine base like triethylamine or DIPEA, or an arylamine base like pyridine. The coupling of X6d or X6c and N6a can be carried out in isopropyl acetate, toluene, or a mixture thereof.

[0145] X6d can be prepared from X6e: [ka]

[0146] In one embodiment, X6d is prepared by contacting X6e with a base that converts cyano groups to carboxylic acid groups, such as sodium hydroxide.

[0147] X6e can be prepared from X6f: [ka] X is Cl, Br, or I.

[0148] X6e is prepared by contacting X6f with X6g under cross-coupling conditions: [ka] X is F, Cl, Br, or I, m is 2 or 3, and R is F, Cl, Br, or I, OH, OC1-C6 alkyl, N(C1-C6 alkyl)2, aryl, or two or three R groups other than F, Cl, Br, I, or OH can together form a cyclic boronic acid ester, such as pinacolboronic acid or N-methyliminodiacetic acid (MIDA) boronate. The coupling of organoboron and aryl halide compounds is described above in relation to the coupling of X6b and F7, and similar conditions can be used for the formation of X6e.

[0149] X6f can be prepared from X6h: [ka] X is Cl, Br, or I.

[0150] X6f can be prepared, for example, by diazotizing X6h with nitrite or sodium nitrite under acidic conditions, followed by cyanosis of a diazonium compound using, for example, CuCN and / or NaCN.

[0151] The X6h can be prepared from the X6i: [ka]

[0152] X6h can be prepared by contacting X6i with a halogenating agent, such as a chlorinating agent such as AlCl3 or N-chlorosuccinimide, N-bromosuccinate, 1,3-dibromo-5,5-dimethylhydantoin (DBDMH), N-bromosuccinimide, TBAB, phosphorus tribromide, bromine chloride, aluminum tribromide, Br2 and FeBr3, HBr, tribromoisocyanuric acid, ozone and ammonium bromide, TBBDA, and combinations thereof, or an iodinating reagent such as N-iodosuccinimide. X6h can also be prepared via the Sandmeyer reaction.

[0153] Preparation of N6a N6a is used in the preparation of X6b. Therefore, the present invention further provides the preparation of N6a. N6a can be prepared from N6b: [ka] Y is either Cl, Br, or I.

[0154] N6a can be prepared by contacting N6b with a reducing agent, for example, a reducing agent selected from the group consisting of: H2 and Pt(V) / C; Raney nickel catalyst and H2; Urushihara nickel catalyst and H2; Adams catalyst (PtO2) and H2; TiCl3 and H2; HCl and iron; NH4Cl and iron; HCl and SnCl2; samarium and NH4Cl; FeCl3, hydrazine hydrate; sodium hydrosulfite; hydrogen sulfide and base; hydroiodic acid; 1,3-dimethyl-2-imidazolidinone and sodium triethylsilantholate; and combinations thereof. In some embodiments, this reaction is carried out under micelle conditions.

[0155] N6b can be prepared from N6c: [ka]

[0156] N6b can be prepared by contacting X6h with a halogenating agent, such as a chlorinating agent such as AlCl3 or N-chlorosuccinimide, a brominating agent selected from the group consisting of N-bromosuccinate, N-bromosuccinimide, 1,3-dibromo-5,5-dimethylhydantoin (DBDMH), TBAB, phosphorus tribromide, bromine chloride, aluminum tribromide, Br2 and FeBr3, HBr, tribromoisocyanuric acid, ozone and ammonium bromide, TBBDA, and combinations thereof, or an iodinating reagent such as N-iodosuccinimide. X6h can also be prepared via the Sandmeyer reaction.

[0157] N6c can be prepared from N6d: [ka]

[0158] N6c can be prepared by contacting N6d with a nitrating agent, for example, a nitrating agent selected from the group consisting of: nitric acid and sulfuric acid; nitric acid and acetic anhydride; tetrachloromethane, nitric acid and phosphorus pentoxide; isopentyl nitrate, trifluoromethanesulfonic acid and 1-ethyl-3-methylimidazolium triflate; H-beta zeolite catalyst and N2O5; acetyl nitrate; and combinations thereof.

[0159] N6d can be prepared from N6e: [ka]

[0160] N6d can be prepared by contacting N6e with a diazotizing agent such as nitrite or sodium nitrite under acidic conditions, followed by a fluorinating agent such as HF.

[0161] Preparation of F6 F6 is used in the preparation of F7 and can itself be prepared by any suitable method. Therefore, the present invention further provides a method for preparing F6. F6 is prepared from F2 and F3: [ka] Y is independently Cl, Br, or I.

[0162] In some embodiments, the preparation of F6 involves reacting compound F2 with compound F3 to obtain compound F4: [ka]

[0163] The reactions of F2 and F3 can be carried out under Mitsunobu conditions in the presence of a phosphine compound (optionally on a resin support), such as PPh3, and an azodicarbocylate, such as DIAD or DEAD. In one embodiment, the reaction is carried out in an aromatic solvent such as toluene. In one embodiment, the solvent is dried to have a water content of less than 0.5 wt%, for example, 0.1 wt%.

[0164] The preparation of F6 may include the conversion of F4 to F6: [ka]

[0165] The conversion of F4 to F6 may be carried out using any suitable amination reagent, such as ammonium hydroxide or water and ammonia. In one embodiment, the solvent is an alcohol solvent such as iPrOH.

[0166] The reaction of F2 with F3 to obtain F4, and the conversion of F4 to compound F6, may be carried out in a sequential reaction or in a one-pot reaction.

[0167] Alternatively, F2 can be converted to F2' via amination. The amination reagent comprises water and ammonia or ammonium hydroxide, and this reaction may be carried out in a polar solvent such as an alcohol solvent such as iPrOH. F2 can then optionally be reacted with F3 under Mitsunobu conditions in the presence of a phosphine compound such as PPh3 and an azodicarbosylate such as DIAD or DEAD to obtain F6: [ka]

[0168] These reactions can be carried out sequentially or in a single pot.

[0169] Products prepared according to the processes described herein and their use In one embodiment, the present invention provides a synthetic route for a LOU064 API that is substantially pure with respect to nitrosamines, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. Thus, the protection granted by the patent arising from this application may extend to the direct product of the process herein, which is a remibrutinib API that is substantially pure with respect to the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

[0170] In another embodiment, the present invention comprises LOU064 active pharmaceutical ingredient substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. For example, the amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide is less than about 1000 ppb, less than about 550 ppb, e.g., less than about 530 ppb; less than about 400 ppb, e.g., less than about 360 ppb; less than about 300 ppb; less than about 200 ppb; less than about 150 ppb, e.g., less than about 130 ppb; less than about 100 ppb, e.g., less than about 90 ppb; less than about 50 ppb or less than about 25 ppb.

[0171] Furthermore, the present invention includes a LOU064 API prepared or prepareable by the process described herein, which does not contain INT3 at any stage. Accordingly, in one embodiment, the remibrutinib API prepared or prepareable by the process described herein also substantially does not contain INT3 (5-fluoro-2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline). For example, the amount of INT3 may be less than about 100 ppm (parts per million), less than about 10 ppm, less than about 1 ppm, less than about 100 ppb (parts per billion), less than about 10 ppb, or less than about 1 ppb. In one embodiment, the remibrutinib prepared or prepareable by the process described herein does not contain INT3 (5-fluoro-2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline). Alternatively, or in addition, the remibrutinib API prepared or prepareable by the processes described herein is substantially free of (3-amino-5-fluoro-2-methylphenyl)boronic acid. For example, the amount of (3-amino-5-fluoro-2-methylphenyl)boronic acid may be less than about 100 ppm (parts per million), less than about 10 ppm, less than about 1 ppm, less than about 100 ppb (parts per billion), less than about 10 ppb, or less than about 1 ppb. In one embodiment, the remibrutinib prepared or prepareable by the processes described herein is free of 3-amino-5-fluoro-2-methylphenyl)boronic acid.

[0172] Pharmaceutical composition of the present invention In one embodiment, the present invention also includes a pharmaceutical composition comprising a remibrutinib active pharmaceutical ingredient prepared or prepareable by a process described herein, and therefore substantially free of INT3.

[0173] In another embodiment, the present invention also includes a pharmaceutical composition comprising a remibrutinib API prepared or prepareable by a process described herein, which may therefore be substantially free of nitrosamines, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. In one embodiment, the composition also contains at least one pharmaceutically acceptable excipient, and often contains at least two or more pharmaceutically acceptable excipients. Several suitable excipients are disclosed herein. Other excipients known in the art may be used without departing from the intent and scope of this application.

[0174] As used herein, the term “pharmaceutically acceptable excipients” includes all solvents, carriers, diluents, dispersions, coatings, surfactants, antioxidants, preservatives (e.g., antimicrobials, antifungals, antioxidants), isotonic agents, absorption retarders, salts, drug stabilizers, binders, additives, bulking agents, disintegrants, lubricants, sweeteners, fragrances, pigments, and other similar substances and combinations thereof (see, for example, Remington's Pharmaceutical Sciences, 18th Ed., Mack Printing Company, 1990, pp. 1289–1329). It should be understood that the use of any conventional excipient in any therapeutic or pharmaceutical composition is assumed by this application, provided that the conventional excipient is not incompatible with the active ingredient.

[0175] Pharmaceutical compositions can be formulated for specific routes of administration, such as oral, parenteral, and rectal administration. In addition, pharmaceutical compositions described herein can be prepared in solid form (including, without limitation, capsules, tablets, pills, granules, powders, or suppositories) or in liquid form (including, without limitation, liquids, suspensions, or emulsions). Pharmaceutical compositions can be subjected to conventional pharmaceutical procedures such as sterilization and / or may contain conventional inert diluents, smoothing agents, carriers, or buffers, as well as auxiliary agents such as solvents, preservatives, stabilizers, wetting agents, emulsifiers, and bulking agents.

[0176] Typically, a pharmaceutical composition is a tablet or capsule containing an active ingredient together with at least one excipient, such as: a) Diluents, such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine; b) Lubricants, such as silica, talc, stearic acid, its magnesium or calcium salt, sodium stearyl fumarate and / or polyethylene glycol; for tablets, further c) Binders, e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone and / or polyvinylpyrrolidone-vinyl acetate copolymer; if desired; d) Carriers such as aqueous vehicles containing cosolvating materials such as captisol, PEG, glycerin, cyclodextrin, or other similar materials; e) Tablet decomposition substances, such as starch, agar, alginic acid or its sodium salt, croscarmellose sodium, crospovidone, sodium starch glycolate or effervescent mixtures; and / or f) Absorbents, colorants, flavorings, and sweeteners.

[0177] The tablets may be film-coated or enteric-coated according to methods known in the art. In one embodiment, the compound or composition is prepared for oral administration, for example, as a tablet or capsule, and optionally packaged as a multi-dose type suitable for storing and / or dispensing unit doses of the pharmaceutical product. Examples of suitable packaging include, but are not limited to, airtight foil, unit dose containers (e.g., vials), blister packaging and strip packaging.

[0178] Tablets may contain an active ingredient mixed with non-toxic, pharmaceutically acceptable excipients suitable for tablet manufacture. These excipients include, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulators and disintegrants such as corn starch or alginic acid; binders such as starch, gelatin, or gum arabic; and smoothing agents such as magnesium stearate, stearic acid, or talc. Tablets may be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained effect over a longer period. For example, time-delaying materials such as glyceryl monostearate or glyceryl distearate may be used. Formulations for oral use may be provided as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oily medium, such as peanut oil, liquid paraffin, or olive oil.

[0179] Since water may promote the breakdown of certain compounds, the present invention further includes anhydrous pharmaceutical compositions and dosage forms comprising remibrutinib as an active ingredient, which is prepared or can be prepared by the methods described herein.

[0180] Anhydrous pharmaceutical compositions and dosage forms can be prepared using anhydrous or low-moisture-content components and low-moisture or low-humidity conditions. Anhydrous pharmaceutical compositions may be prepared and stored in such a way that their anhydrous properties are maintained. Therefore, anhydrous compositions can be packaged using materials known to prevent exposure to water so that they can be included in suitable standard kits. Examples of suitable packaging include, but are not limited to, airtight foil, plastic, unit dose containers (e.g., vials), blister packaging, and strip packaging.

[0181] The present invention further includes pharmaceutical compositions and dosage forms comprising one or more agents that reduce the rate at which the compounds described herein as active ingredients degrade. Such agents, referred to herein as “stabilizers,” include, but are not limited to, antioxidants such as ascorbic acid, pH buffers, or salt buffers.

[0182] In some embodiments, remibrutinib substantially free of nitrosamine impurities can be provided in pharmaceutical compositions and dosage forms described in International Publication No. 2022 / 162513 (Agent Case No. PAT059011-WO-PCT), which is thereby incorporated in whole by reference.

[0183] The pharmaceutical composition may be further formulated into a final dosage form. Examples of dosage forms include capsules or tablets. In one example, the dosage form is a film-coated tablet, as disclosed, for example, in Example 8 of International Publication No. 2022 / 162513.

[0184] In one embodiment, the pharmaceutical composition or combination of the present invention may be administered in a unit dose of about 1 to 1000 mg of the active ingredient or about 1 to 500 mg, about 1 to 250 mg, about 1 to 150 mg, about 0.5 to 100 mg, or about 10 to 50 mg of the active ingredient to a subject weighing about 50 to 70 kg. In one embodiment, the pharmaceutical composition or combination of the present invention may be administered in a unit dose of about 10 mg, about 25 mg, or about 50 mg. The therapeutically effective dose or amount of a compound, pharmaceutical composition, or combination thereof depends on the species, weight, age, and individual condition of the subject, the disorder or disease being treated, or its severity. A physician, clinician, or veterinarian of ordinary skill can easily determine the effective amount of each active ingredient necessary to prevent, treat, or inhibit the progression of the disorder or disease.

[0185] The active pharmaceutical ingredient (i.e., remibrutinib active pharmaceutical ingredient substantially free of nitrosamine impurities) may be present in the pharmaceutical composition (e.g., a film-coated tablet) in amounts of approximately 10 mg, approximately 15 mg, approximately 20 mg, approximately 25 mg, approximately 50 mg, or approximately 100 mg.

[0186] In one embodiment, the present invention relates to a pharmaceutical composition comprising LOU064 active pharmaceutical ingredient substantially free of nitrosamines, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide described in any one of the preceding claims, and one or more pharmaceutically acceptable excipients.

[0187] In another embodiment, the present invention relates to a pharmaceutical composition comprising LOU064 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition is substantially free of nitrosamines, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. The content of nitrosamine impurities in the pharmaceutical composition is as defined above.

[0188] In another embodiment, the present invention relates to a final dosage form comprising the pharmaceutical composition described above and one or more pharmaceutically acceptable excipients. In one embodiment, the final dosage form is a film-coated tablet. In another embodiment, the final dosage form is a film-coated tablet in which LOU064 is present in amounts of about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, or about 100 mg, for example, about 25 mg or about 100 mg.

[0189] In one embodiment, the remibrutinib active pharmaceutical ingredient (i.e., remibrutinib active pharmaceutical ingredient that is substantially free of nitrosamine impurities (e.g., impurities N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) is present in the pharmaceutical composition (e.g., a film-coated tablet) in an amount of 25 mg. In one embodiment of this design, the content of nitrosamines (e.g., nitrosamine impurities N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) in the remibrutinib drug substance or composition is less than about 550 ppb, e.g. less than about 530 ppb; less than about 400 ppb, e.g. less than about 360 ppb; less than about 300 ppb; less than about 200 ppb; less than about 150 ppb, e.g. less than about 130 ppb; less than about 100 ppb, e.g. less than about 90 ppb; less than about 50 ppb or less than about 25 ppb, all based on the total amount of LOU064 in free or salt form. In another embodiment of this embodiment, the level of nitrosamines in or in the remibrutinib drug substance, for example N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is between about 25 ppb and about 550 ppb, for example between about 25 ppb and about 530 ppb; or between about 25 ppb and about 400 ppb, for example between about 25 ppb and about 360 ppb; or between about 25 ppb and about 300 ppb; or between about 25 ppb and about 200 ppb, for example between about 25 ppb and about 90 ppb; or between about 25 ppb and about 100 ppb, for example between about 25 ppb and about 90 ppb, all based on the total amount of LOU064 in free or salt form.In yet another embodiment, the levels of nitrosamines in or within the remibrutinib active pharmaceutical ingredient, such as N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, are between about 100 ppb and about 550 ppb, for example between about 100 ppb and about 530 ppb; or between about 100 ppb and about 400 ppb, for example between about 100 ppb and about 360 ppb; or between about 100 ppb and about 350 ppb, for example between about 100 ppb and about 320 ppb; or between about 100 ppb and about 250 ppb; or between about 100 ppb and about 150 ppb, for example between about 100 ppb and about 130 ppb, all based on the total amount of LOU064 in free or salt form.

[0190] In another embodiment, the active pharmaceutical ingredient (i.e., remibrutinib active pharmaceutical ingredient substantially free of nitrosamine impurities) is present in a pharmaceutical composition (e.g., a film-coated tablet) in an amount of 100 mg. In one aspect of this embodiment, the content of nitrosamines (e.g., nitrosamine impurities N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) in the remibrutinib active pharmaceutical ingredient or composition is less than about 550 ppb, e.g. less than about 130 ppb; less than about 100 ppb, e.g. less than about 90 ppb; less than about 50 ppb or less than about 25 ppb, all based on the total amount of LOU064 in free or salt form. In another embodiment of this embodiment, the level of nitrosamines in or in the remibrutinib drug substance, for example N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is between about 25 ppb and about 130 ppb or between about 35 ppb and about 90 ppb; or between about 25 ppb and about 100 ppb, for example between about 25 ppb and about 90 ppb or between about 100 ppb and about 130 ppb, all based on the total amount of LOU064 in free or salt form.

[0191] The efficacy of the above-mentioned dosages can be demonstrated in in vitro and in vivo studies using mammals, such as mice, rats, dogs, monkeys, or their excised organs, tissues, and preparations. The compounds described herein can be administered in vitro in the form of liquids, such as aqueous solutions, and enterally, parenterally, and preferably intravenously in vivo, for example, as suspensions or in aqueous solutions. The in vitro dosage is approximately 10 -3 Molar concentration ~10 -9The molar concentration may range between two levels. The effective in vivo dose may range between approximately 0.1 and 500 mg / kg or between approximately 1 and 100 mg / kg, depending on the route of administration. In one embodiment, the effective in vivo dose ranges between approximately 10 mg and 200 mg daily, for example, approximately 10 mg, 20 mg, 25 mg, 35 mg, 50 mg, 100 mg, or 200 mg daily. In one embodiment, the effective in vivo dose is selected from approximately 10 mg, 35 mg, 50 mg, or 100 mg once daily. In one embodiment, the effective in vivo dose is selected from approximately 10 mg, 25 mg, 50 mg, or 100 mg twice daily.

[0192] Manufacturing of pharmaceutical compositions (formulations) The present invention further provides a method for preparing a pharmaceutical composition comprising (i) remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition is substantially free of nitrosamines, in particular N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. Methods for preparing such pharmaceutical compositions are described herein, but it will be readily apparent to those skilled in the art that two or more of these methods may also be used in combination.

[0193] Accordingly, the present invention also provides combinations of methods for preparing a pharmaceutical composition comprising (i) remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition is substantially free of nitrosamines, in particular N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. The pharmaceutical compositions produced by these methods may be used in pharmaceutical products described herein.

[0194] Thus, an aspect of the present invention is a method for preparing a pharmaceutical composition, comprising mixing ruxolitinib or a pharmaceutically acceptable salt thereof with one or more pharmaceutically acceptable excipients, wherein each of the one or more pharmaceutically acceptable excipients and optionally each of the one or more pharmaceutically acceptable excipients has a nitrous acid content of less than 1.5 ppm, less than 1 ppm or less than about 0.5 ppm. Each of the one or more pharmaceutically acceptable excipients, for example each of the one or more pharmaceutically acceptable excipients, may have a nitrous acid content of less than about 0.4 ppm, less than about 0.3 ppm, less than about 0.2 ppm or less than about 0.1 ppm.

[0195] Thus, the present invention provides a method for preparing a pharmaceutical composition, comprising mixing ruxolitinib or a pharmaceutically acceptable salt thereof with one or more pharmaceutically acceptable excipients, wherein each of the one or more pharmaceutically acceptable excipients has a nitrous acid content of less than 1.5 ppm, less than 1 ppm or less than about 0.5 ppm based on the amount of each excipient. Each of the one or more pharmaceutically acceptable excipients may have a nitrous acid content of less than about 0.4 ppm, less than about 0.3 ppm, less than about 0.2 ppm or less than about 0.1 ppm based on the amount of each excipient.

[0196] Another aspect of the present invention is a method for preparing a pharmaceutical composition, comprising mixing ruxolitinib or a pharmaceutically acceptable salt thereof with more than one pharmaceutically acceptable excipients, wherein the combination of excipients has a total nitrous acid content of less than 1.5 ppm, less than 1 ppm or less than about 0.5 ppm based on the combined amount of the excipients. The combination of excipients may have a total nitrous acid content of less than about 0.4 ppm, less than about 0.3 ppm, less than about 0.2 ppm or less than about 0.1 ppm based on the combined amount of the excipients.

[0197] A method for preparing a pharmaceutical composition comprising remibrutinib or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients may include the step of mixing remibrutinib or the pharmaceutically acceptable salt thereof with one or more pharmaceutically acceptable excipients. The method may further include the step of drying the resulting mixture. Accordingly, aspects of the present invention relate to a method for preparing a pharmaceutical composition comprising (i) remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, the method comprising (a) mixing remibrutinib or the pharmaceutically acceptable salt thereof with one or more excipients and (b) drying the resulting mixture. The drying step may be carried out until the water activity value is less than 0.12, less than 0.10, less than 0.09, or less than 0.08. The method may further include, before or after the drying step, a step of processing the composition into a solid oral dosage form. For example, the method may include a further step of compressing the composition into tablets. Alternatively, the method may include a further step of filling the composition into capsules. After the drying step, the method may include a further step of storing the composition in the presence of a desiccant, for example, storing the composition in a sealed container that also contains a desiccant, or storing the composition in a sealed pharmaceutical package that also contains a desiccant in an individual container.

[0198] A method for preparing a pharmaceutical composition comprising remibrutinib or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients may include one or more steps described in International Publication 2022 / 162513 (Agent Case No. PAT059011-WO-PCT), which is incorporated in whole by reference thereby.

[0199] In one embodiment, the one or more pharmaceutically acceptable excipients may be selected from the group consisting of lactose, crystalline cellulose, mannitol, sucrose, starch, granular hydrophilic fumed silica, polyvinyl pyrrolidone-vinyl acetate copolymer, polyvinyl pyrrolidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hypromellose, carboxymethyl cellulose, methylcellulose, hydroxyethyl cellulose, carboxyethyl cellulose, carboxymethyl hydroxyethyl cellulose, polyethylene glycol, polyvinyl alcohol, shellac, polyvinyl alcohol-polyethylene glycol copolymer, polyethylene-propylene glycol copolymer, sodium lauryl sulfate, potassium lauryl sulfate, ammonium lauryl sulfate, sodium lauryl ether sulfate, polysorbate, perfluorobutane sulfonate, dioctyl sulfosuccinate, magnesium carbonate, kaolin, cellulose (e.g., crystalline cellulose, powdered cellulose), calcium phosphate or sodium phosphate, croscarmellose sodium, crospovidone, sodium starch glycolate, corn starch or alginic acid, magnesium stearate, sodium stearyl fumarate, stearic acid and talc. In one aspect of the above embodiment, the one or more pharmaceutically acceptable excipients may be selected from the group consisting of magnesium stearate, sodium stearyl fumarate, crystalline cellulose, crospovidone, croscarmellose sodium, lactose, mannitol, sodium lauryl sulfate and polyvinyl pyrrolidone-vinyl acetate copolymer (copovidone).

[0200] The one or more pharmaceutically acceptable excipients may be or may contain magnesium stearate or sodium stearyl fumarate. The content of nitrous acid in magnesium stearate or sodium stearyl fumarate may be less than about 1.5 ppm, optionally less than about 1 ppm, less than about 0.5 ppm, less than about 0.2 ppm.

[0201] One or more pharmaceutically acceptable excipients may be or may contain crystalline cellulose. The nitrite content in the crystalline cellulose may be less than about 500 ppb, less than about 400 ppb, less than about 300 ppb, less than about 200 ppb, less than about 100 ppb, less than about 100 ppb, less than about 90 ppb, less than about 80 ppb, less than about 70 ppb, less than about 60 ppb, less than about 50 ppb, less than about 40 ppb, less than about 30 ppb, less than about 20 ppb, or less than about 10 ppb. Preferably, the nitrite content in the crystalline cellulose may be less than about 100 ppb.

[0202] Suitable crystalline cellulose excipients are known to those skilled in the art and include types MCC PH102 and MCC PH105. The crystalline cellulose may be MCC PH102. One or more pharmaceutically acceptable excipients may be sodium lauryl sulfate (SLS) or contain it. The nitrite content in SLS is less than about 1.5 ppm, less than about 1 ppm, less than about 0.5 ppm, optionally less than or equal to about 500 ppb, less than or equal to about 200 ppb, or less than or equal to about 100 ppb.

[0203] One or more pharmaceutically acceptable excipients may be or may contain polyvinylpyrrolidone-vinyl acetate copolymer (copovidone). The nitrite content in copovidone may be less than about 500 ppb, less than about 400 ppb, less than about 300 ppb, less than about 200 ppb, or less than about 100 ppb.

[0204] As disclosed elsewhere in this specification, the amount of nitrite in a composition, for example, the amount of nitrite in a pharmaceutically acceptable excipient, may be determined using the Griess test (e.g., Example 16). Accordingly, the present invention provides a method for preparing a composition comprising (i) remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, the method comprising the use of the Griess test to determine the amount of nitrite in one, more or more excipients, or each of one or more excipients, optionally, one or more excipients in which the amount of nitrite is determined, selected from the group consisting of magnesium stearate, sodium stearyl fumarate, crystalline cellulose, crospovidone, croscamellose sodium, lactose, mannitol, sodium lauryl sulfate, and polyvinylpyrrolidone-vinyl acetate copolymer (copovidone).

[0205] Remibrutinib or a pharmaceutically acceptable salt thereof used in a method for preparing a pharmaceutical composition may be prepared by a method for preparing remibrutinib or a pharmaceutically acceptable salt thereof as described elsewhere in this specification.

[0206] Testing of active pharmaceutical ingredients and pharmaceutical compositions (formulations) The present invention provides a method for testing remibrutinib or a pharmaceutically acceptable salt thereof for the presence and / or amount of a nitrosamine, particularly N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, and further provides a method for testing a pharmaceutical composition comprising (i) remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients. Accordingly, aspects of the present invention relate to a method for evaluating a composition comprising remibrutinib or a pharmaceutically acceptable salt thereof, comprising testing the composition for the presence and / or amount of a nitrosamine, particularly N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

[0207] The method may be used to verify a process for producing a composition comprising remibrutinib or a pharmaceutically acceptable salt thereof. Accordingly, aspects of the present invention relate to a method for verifying a process for producing a composition comprising remibrutinib or a pharmaceutically acceptable salt thereof, comprising testing the composition produced by the process for the presence and / or amount of a nitrosamine, in particular N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

[0208] Another aspect of the present invention relates to a method for obtaining regulatory approval for a pharmaceutical composition comprising or including remibrutinib or a pharmaceutically acceptable salt thereof, the method comprising (i) testing the composition for the presence and / or amount of nitrosamines, particularly N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, and (ii) submitting the results of the said test to a regulatory authority. Suitable regulatory authorities to whom the results may be submitted are, for example, the FDA, EMA, MHRA, Swissmedic, or PMDA.

[0209] In some embodiments of the method of the present invention, a batch of composition is tested to determine the presence and / or amount of nitrosamines in the batch, in particular the presence and / or total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. Specifically, a sample of the batch is tested. The batch test may be used to determine whether a pharmaceutical product should be prepared from the batch. For example, a pharmaceutical product may be prepared from a batch only if it is determined that the batch has a total amount of nitrosamines less than approximately 1000 ppb (e.g., less than approximately 550 ppb, e.g., less than approximately 530 ppb; less than approximately 400 ppb, e.g., less than approximately 360 ppb; less than approximately 150 ppb, e.g., less than approximately 130 ppb; less than approximately 100 ppb, e.g., less than approximately 90 ppb); less than approximately 50 ppb; or less than approximately 25 ppb, based on remibrutinib in free or salt form.

[0210] Accordingly, aspects of the present invention are a process for preparing a pharmaceutical product comprising (i) a composition comprising remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, a. Obtain a batch of remibrutinib or a pharmaceutically acceptable salt thereof. b. Determining the total amount of nitrosamines in the batch, particularly by testing a sample of the batch, and c. A process is provided for preparing a pharmaceutical product from a batch only if it is determined that the batch has a total amount of nitrosamines less than approximately 1000 ppb (e.g., less than approximately 550 ppb, e.g., less than approximately 530 ppb; less than approximately 400 ppb, e.g., less than approximately 360 ppb; less than approximately 150 ppb, e.g., less than approximately 130 ppb; less than approximately 100 ppb, e.g., less than approximately 90 ppb); less than approximately 50 ppb; or less than approximately 25 ppb, based on the amount of remibrutinib in free or salt form.

[0211] In some embodiments, a pharmaceutical product may be prepared from a batch only if it is determined that the batch has a total amount of nitrosamine (e.g., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) which corresponds to an amount of 100 ng / day or less of free nitrosamine base (e.g., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) which will consequently be administered to the patient when the composition is administered according to the approved dosage and administration for remibrutinib or a pharmaceutically acceptable salt thereof. Accordingly, aspects of the present invention are a process for preparing a pharmaceutical product comprising (i) a composition comprising remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, a. Obtain a batch of remibrutinib or a pharmaceutically acceptable salt thereof. b. Determining the total amount of nitrosamines (e.g., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) in the batch, particularly by testing a sample of the batch, and c. When the composition is administered according to the approved dosage and administration for remibrutinib or a pharmaceutically acceptable salt thereof, the amount of nitrosamine (e.g., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide or less than 100 ng / day corresponds to the amount of the nitrosamine (e.g., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl) when administered according to the approved dosage and administration for remibrutinib or a pharmaceutically acceptable salt thereof. The present invention provides a process for preparing a pharmaceutical product from a batch only if it is determined that the batch contains the total amount of nitrosamines (e.g., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) which will consequently be administered to the patient.

[0212] In some embodiments, a batch of the composition is tested to determine the total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide in the batch. In particular, a sample of the batch is tested. This batch test may be used to determine whether a pharmaceutical product should be prepared from the batch. For example, a pharmaceutical product may be prepared from a batch only if it is determined that the batch has a total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide less than about 1000 ppb based on the total amount of remibrutinib in free or salt form; (e.g., less than about 550 ppb, e.g., less than about 530 ppb; less than about 400 ppb, e.g., less than about 360 ppb; less than about 150 ppb, e.g., less than about 130 ppb; less than about 100 ppb, e.g., less than about 90 ppb); less than about 50 ppb; or less than about 25 ppb.

[0213] Accordingly, aspects of the present invention are a process for preparing a pharmaceutical product comprising (i) a composition comprising remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, a. Obtain a batch of remibrutinib or a pharmaceutically acceptable salt thereof. b. Determining the total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide in the batch, particularly by testing a sample of the batch. c. A process is provided for preparing a pharmaceutical product from a batch only if it is determined that the batch has a total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide less than about 1000 ppb (e.g., less than about 550 ppb, e.g., less than about 530 ppb; less than about 400 ppb, e.g., less than about 360 ppb; less than about 150 ppb, e.g., less than about 130 ppb; less than about 100 ppb, e.g., less than about 90 ppb); less than about 50 ppb; or less than about 25 ppb, based on the total amount of remibrutinib in free or salt form.

[0214] In some embodiments, a batch of the composition is tested to determine the amount of total nitrosamines in the batch, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines. In particular, a sample of the batch is tested. This batch test may be used to determine whether a pharmaceutical product should be prepared from the batch. For example, a pharmaceutical product may be prepared from a batch only if it is determined that the batch has a total amount of nitrosamines, including N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines, in an amount less than 1000 ppb (e.g., less than 550 ppb, e.g., less than 530 ppb; less than 400 ppb, e.g., less than 360 ppb; less than 150 ppb, e.g., less than 130 ppb; less than 100 ppb, e.g., less than 90 ppb); less than 50 ppb; or less than 25 ppb, based on the total amount of remibrutinib in free or salt form. Preferably, the pharmaceutical product may be prepared from a batch only if it is determined that the batch has a total amount of nitrosamines, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines, based on the total amount of remibrutinib in free or salt form, less than 360 ppb; less than 150 ppb, for example less than 130 ppb; and less than 100 ppb, for example less than 90 ppb. Accordingly, aspects of the present invention are processes for preparing a pharmaceutical product comprising (i) a composition comprising remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, a. Obtain a batch of remibrutinib or a pharmaceutically acceptable salt thereof. b. Determining the total amount of nitrosamines in the batch, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines, particularly by testing a sample of the batch. c. A process is provided for preparing a pharmaceutical product from a batch only if it is determined that the batch has a total amount of nitrosamines, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines, in an amount less than 1000 ppb (e.g., less than 550 ppb, e.g., less than 530 ppb; less than 400 ppb, e.g., less than 360 ppb; less than 150 ppb, e.g., less than 130 ppb; less than 100 ppb, e.g., less than 90 ppb); less than 50 ppb or less than 25 ppb, based on the total amount of remibrutinib in free or salt form.

[0215] In some embodiments, a pharmaceutical product may be prepared from a batch only if it is determined that the batch has a total amount of nitrosamines, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines, which would result in the amount of total nitrosamines administered to a patient being 100 ng / day or less of free nitrosamine base when the composition is administered according to the approved dosage and administration for remibrutinib or a pharmaceutically acceptable salt thereof. Accordingly, embodiments of the present invention are processes for preparing a pharmaceutical product comprising a composition comprising (i) remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, a. Obtain a batch of remibrutinib or a pharmaceutically acceptable salt thereof. b. Determining the total amount of nitrosoamines, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosoamines in the batch, particularly by testing samples of the batch, and c. Providing a process that includes preparing a pharmaceutical product from a batch only if it is determined that the batch has a total amount of nitrosoamines that would result in an amount of nitrosoamine free base of 100 ng / day or less being administered to a patient when the composition is administered according to the approved use and dosage for lenvatinib or a pharmaceutically acceptable salt thereof, said total amount of nitrosoamines including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosoamines.

[0216] In some embodiments, stability testing may be performed using a sample of a batch of the composition. After this stability test, the batch sample may be tested for the total amount of nitrosamine (N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, etc. This test on the batch sample may be used to determine whether the batch is suitable for sale and / or administration to patients. For example, a batch may be determined to be suitable for sale only if a sample of the batch after stability testing is determined to have a total amount of nitrosamines (such as N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide less than approximately 1000 ppb (e.g., less than approximately 550 ppb, e.g., less than approximately 530 ppb; less than approximately 400 ppb, e.g., less than approximately 360 ppb; less than approximately 150 ppb, e.g., less than approximately 130 ppb; less than approximately 100 ppb, e.g., less than approximately 90 ppb); less than approximately 50 ppb or less than approximately 25 ppb.

[0217] Accordingly, aspects of the present invention relate to a process for marketing a verified batch of pharmaceutical products comprising (i) remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, a. To generate batches of pharmaceutical products. b. Perform stability testing on the sample from the aforementioned batch. c. Determine the total amount of nitrosamines (N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, etc. in the batch sample after the stability test. d. Provide a process for validating a batch for sale only if it is determined that a batch sample after stability testing has a total amount of nitrosamines (such as N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide less than approximately 1000 ppb (e.g., less than approximately 550 ppb, e.g., less than approximately 530 ppb; less than approximately 400 ppb, e.g., less than approximately 360 ppb; less than approximately 150 ppb, e.g., less than approximately 130 ppb; less than approximately 100 ppb, e.g., less than approximately 90 ppb); less than approximately 50 ppb; or less than approximately 25 ppb.

[0218] In some embodiments, stability testing may be performed using a sample of a batch of the composition, and the batch sample may be tested for the total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide in the batch after stability testing. This test on the batch sample may be used to determine whether the batch is suitable for sale and / or administration to patients. For example, a batch may be determined to be suitable for sale only if it is determined that a sample of the batch after stability testing has a total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide less than about 1000 ppb (e.g., less than about 550 ppb, e.g., less than about 530 ppb; less than about 400 ppb, e.g., less than about 360 ppb; less than about 150 ppb, e.g., less than about 130 ppb; less than about 100 ppb, e.g., less than about 90 ppb); less than about 50 ppb; or less than about 25 ppb, based on the total amount of remibrutinib in free or salt form. Accordingly, an aspect of the present invention is a process for selling a verified batch of a pharmaceutical product comprising (i) a composition comprising remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients. a. To generate batches of pharmaceutical products. b. Perform stability testing on the sample from the aforementioned batch. c. After the stability test, determine the total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide in the sample of the batch and d. Provide a process for validating a batch for sale only if it is determined that a batch sample after stability testing has a total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide less than approximately 1000 ppb (e.g., less than approximately 550 ppb, e.g., less than approximately 530 ppb; less than approximately 400 ppb, e.g., less than approximately 360 ppb; less than approximately 150 ppb, e.g., less than approximately 130 ppb; less than approximately 100 ppb, e.g., less than approximately 90 ppb); less than approximately 50 ppb; or less than approximately 25 ppb, based on the total amount of remibrutinib in free or salt form.

[0219] In some embodiments, stability testing may be performed using a sample of a batch of the composition, which may be tested after stability testing for the amount of total nitrosamines in the batch, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines. This test on a sample of a batch may be used to determine whether the batch is suitable for sale and / or administration to patients. For example, a batch may be determined to be suitable for sale only if it is determined that a sample of the batch after stability testing has a total amount of nitrosamines, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines, of less than approximately 1000 ppb (e.g., less than approximately 550 ppb, e.g., less than approximately 530 ppb; less than approximately 400 ppb, e.g., less than approximately 360 ppb; less than approximately 150 ppb, e.g., less than approximately 130 ppb; less than approximately 100 ppb, e.g., less than approximately 90 ppb); less than approximately 50 ppb; or less than approximately 25 ppb, based on the total amount of remibrutinib in free or salt form. Accordingly, aspects of the present invention relate to a process for marketing a verified batch of pharmaceutical products comprising (i) remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients, a. To generate batches of pharmaceutical products. b. Perform stability testing on the sample from the aforementioned batch. c. After stability testing, determine the total amount of nitrosamines in the batch, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines, and d. Provide a process for validating a batch for sale only if it is determined that a batch sample after stability testing has a total amount of nitrosamines, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines, in an amount less than approximately 1000 ppb (e.g., less than approximately 550 ppb, e.g., less than approximately 530 ppb; less than approximately 400 ppb, e.g., less than approximately 360 ppb; less than approximately 150 ppb, e.g., less than approximately 130 ppb; less than approximately 100 ppb, e.g., less than approximately 90 ppb); less than approximately 50 ppb; or less than approximately 25 ppb.

[0220] In some embodiments, a batch may be determined to be suitable for sale only if it is determined that a batch sample after stability testing has a total amount of nitrosamines, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines, which would result in the amount of free nitrosamines administered to a patient if the composition were administered according to the approved dosage and administration for remibrutinib or a pharmaceutically acceptable salt thereof. Accordingly, embodiments of the present invention are processes for selling verified batches of pharmaceutical products comprising (i) remibrutinib or a pharmaceutically acceptable salt thereof and (ii) one or more pharmaceutically acceptable excipients. a. To generate batches of pharmaceutical products. b. Perform stability testing on the sample from the aforementioned batch. c. After stability testing, determine the total amount of nitrosamines in the batch sample, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines, and d. Provide a process for validating a batch for sale only if it is determined that a batch sample after stability testing has a total amount of nitrosamines equivalent to an amount of 100 ng / day or less of free nitrosamine base when the composition is administered according to the approved dosage and administration for remibrutinib or a pharmaceutically acceptable salt thereof, which would result in the total amount of nitrosamines, including both N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and other nitrosamines being administered to a patient.

[0221] In any of these methods, the step of testing for the presence and / or amount of nitrosamines may be performed using high-performance liquid chromatography (HPLC) and / or gas chromatography (GC)-mass spectrometry. Similarly, the step of determining the total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide may be performed using high-performance liquid chromatography (HPLC) and / or gas chromatography (GC)-mass spectrometry. For example, the HPLC-MS method performed may be the method provided in Example 15.

[0222] The present invention also provides the use of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide as a standard sample for detecting impurities in compositions containing remibrutinib or a pharmaceutically acceptable salt thereof. The composition may contain remibrutinib or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. The impurity may be N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. Detection of the impurity may be performed using high-performance liquid chromatography (HPLC)-and / or gas chromatography (GC)-mass spectrometry.

[0223] Methods for treatment The present invention also provides a method for treating a disorder mediated by or restored by inhibition of BTK, comprising administering a therapeutically effective dose of remibrutinib API (a remibrutinib API substantially free of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide as described herein) prepared or prepareable by the method described herein to a patient in need of such treatment.

[0224] The present invention also includes a method for treating a disorder mediated by or restored by inhibition of BTK, comprising administering to a patient requiring such treatment a pharmaceutical composition substantially free of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide as described herein.

[0225] In another embodiment, the present invention also includes the use of remibrutinib prepared or prepareable by the methods described herein (remibrutinib substantially free of the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide as described herein) for the preparation of pharmaceuticals for the treatment of disorders mediated by or restored by inhibition of BTK.

[0226] In another embodiment, the present invention also includes a remibrutinib API prepared or prepareable by the method described herein for use in the treatment of disorders mediated by or restored by BTK (a remibrutinib API substantially free of the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide as described herein). Pharmaceutical compositions substantially free of the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide as described herein for use in the treatment of disorders mediated by or restored by BTK

[0227] Remibrutinib APIs prepared or prepareable by the methods described herein (e.g., remibrutinib APIs substantially free of nitrosamines (e.g., nitrosamine impurities N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) and nitrosamines (e.g., nitrosamine impurities N-(3-(6-amino-5-(2(methyl)( Pharmaceutical compositions substantially free of nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide are useful in the treatment of the following diseases or disorders mediated by or restored by inhibition of BTK: autoimmune disorders, inflammatory diseases, allergic diseases, respiratory diseases such as asthma and chronic obstructive pulmonary disease (COPD), transplant rejection; diseases in which antibody production, antigen presentation, cytokine production or lymphoid organ formation is abnormal or adverse; rheumatoid arthritis, systemic juvenile idiopathic arthritis (SO4). JIA), gout, pemphigus vulgaris, idiopathic thrombocytopenic purpura, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, Sjögren's syndrome, sweat gland abscess, IgE-driven allergies, e.g., drug, poison, food allergies; autoimmune hemolytic anemia, anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis, cryoglobulinemia, thrombotic thrombocytopenic purpura, chronic urticaria (chronic idiopathic urticaria, induced urticaria), chronic allergies (atopic dermatitis, contact dermatitis, allergic rhinitis), atherosclerosis, type 1 diabetes, type 2 diabetes, inflammatory bowel disease, ulcerative colitis , including Crohn's disease, pancreatitis, glomerulonephritis, Goodpasture syndrome, Hashimoto's thyroiditis, Graves' disease, antibody-mediated transplant rejection (AMR), graft-versus-host disease, B-cell-mediated hyperacute, acute and chronic transplant rejection; thromboembolic disorders, myocardial infarction, angina pectoris, stroke, ischemic disorders, pulmonary embolism; hematopoietic cancers, including but not limited to multiple myeloma; leukemia; acute myeloid leukemia; chronic myeloid leukemia; lymphocytic leukemia; myeloid leukemia; non-Hodgkin lymphoma; lymphoma; polycythemia vera; essential thrombocythemia; myelofibrosis with myeloid metaplasia; and Waldenström disease.

[0228] Remibrutinib active pharmaceutical ingredient prepared or prepareable by the methods described herein (remibrutinib substantially free of the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide as described herein) is particularly useful in the treatment of chronic urticaria, such as chronic idiopathic urticaria or chronic induced urticaria; Sjögren's syndrome, multiple sclerosis, sweat gland abscesses and food allergies. [Examples]

[0229] Example 1: Method for the quantitative determination of nitrosamine (N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) Example 1a: Method for determining nitrosamines in intermediate F8 Limit test by LC-MS (SIM mode), AJS-ES, positive mode, SIM ion: 483.2[M+H]+ Column: YMC-Triart C18, 100×3.0mm, particle size 1.9μm, 12nm Mobile phase A: Dissolve 0.315 g of ammonium formate in 1000 mL of water and add 0.5 mL of formic acid. Mobile phase B: methanol Sample solvent: methanol + 0.2% (v / v) formic acid Sample solution: Approximately 100 mg of sample in 20 mL Flow rate: 0.6mL / min Injection volume: 3μL Autosampler cooling, 8℃ Column temperature: 0.80℃ gradient:

[0230] [Table 4]

[0231] Nitrosamines at 1 ppb in F8 are isolated. 9 This refers to nitrosamines in a proportion (by mass) of 1 part F8 (i.e., F8 may contain other impurities).

[0232] Example 1b: Method for determining nitrosamines in the active pharmaceutical ingredient Limit test by LC-MS (SIM mode), AJS-ES, positive mode, SIM ion: 483.2[M+H]+ Column: YMC-Triart C18, 100×3.0mm, particle size 1.9μm, 12nm Mobile phase A: Dissolve 0.315 g of ammonium formate in 1000 mL of water and add 0.5 mL of formic acid. Mobile phase B: methanol Sample solvent: methanol + 0.2% (v / v) formic acid Sample solution: Approximately 100 mg of sample in 20 mL Flow rate: 0.6mL / min Injection volume: 3μL Autosampler cooling, 8℃ Column temperature: 0.80℃ gradient:

[0233] [Table 5]

[0234] Example 2: Improved Ames Test (EAT) for N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide was evaluated in an EAT performed under GLP conditions in accordance with EMA and FDA guidance (2023) for N-nitrosamines. In this study, mutations of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide were evaluated in 30% rat and hamster metabolic activation (β-naphthoflavone / phenobarbital-induced rat and hamster liver post-mitochondrial S-9 fraction) in Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537, as well as Escherichia coli strain WP2 uvrA pKM101, both in the absence and in the presence of β-naphthoflavone / phenobarbital-induced metabolic activation (β-naphthoflavone / phenobarbital-induced rat and hamster liver post-mitochondrial S-9 fraction). The tests were conducted using the pre-incubation method (30 minutes). N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide was mixed with dimethyl sulfoxide (DMSO) at concentrations of 5, 16, 50, 160, 500, 1600, and 5000 μg / plate.

[0235] The results of the formulation analysis demonstrated the stability and homogeneity of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide in the formulation (at room temperature for 6 hours), achieving a concentration within 100% + / - 10% of the nominal concentration.

[0236] Precipitation was observed in all tester strains at 1600 μg / plate and / or 5000 μg / plate, both in the absence and in the presence of S-9.

[0237] Bacterial toxicity was observed in strain TA1537 only at ≥1600 μg / plate in the presence of hamster S-9.

[0238] Following EAT recommendations, in addition to parallel strain-specific positive controls, two nitrosamine positive controls known to be mutagenic in the presence of S-9, namely N-nitrosodiethylamine (NDEA) and N-methyl-N-nitroso-(2-phenylethyl)amine (MNPA), were used. In the presence of 30% rat S-9, NDEA induced >2x mutations in strain TA100. In the presence of hamster S-9, ≥2x or ≥3x increases were observed in all tester strains except TA1537. MNPA was mutagenic in Salmonella strains TA100, TA1535 and E. coli strain WP2 uvrA pKM101 in the presence of rat and hamster S-9 (≥2x or ≥3x increases). An increase of up to 1.9 was observed in strain TA98 in the presence of hamster S-9. After treatment with N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, in the presence of hamster S-9, there was an increase in the number of revertant mutants of ≥2 and ≥3 in strains TA100 and TA1535, respectively. N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide was not mutagenic without metabolic activation or in the presence of rat S-9.

[0239] In conclusion, N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide induced mutations in Salmonella strains TA100 and TA1535 in the presence of hamster S-9 under EAT conditions. Therefore, it is considered mutagenic under susceptible metabolic conditions.

[0240] Example 3: Stability of intermediate F8: Nitrosamine content over time 7 g of F7 and 84 ml of iPrOAc were packed into a 250 ml reactor under argon. 5.12 g of 37% HCl was added to the suspension over 2 hours. The reaction mixture was stirred overnight and quenched with 55 ml of distilled water. The two-phase suspension was warmed to 30°C. 0.35 g of charcoal was added and the suspension was stirred for 2 hours. The solid was filtered and rinsed with 2 ml of distilled water. The clear aqueous layer was transferred to a clean reactor under argon. The pH was adjusted to pH 6 with 30% NaOH (approximately 10 ml). 28.3 ml of EtOH was added and the suspension was warmed to 60°C. At this temperature, the pH was gradually adjusted to pH 10 with 1N NaOH. The white suspension was cooled to RT and filtered under a protective atmosphere. The filtered cake was rinsed with an ethanol / water mixture and ethanol. The F8 aqueous product was dried.

[0241] The nitrosamine content in intermediate F8 was determined using the method described in Example 1a.

[0242] [Table 6]

[0243] Further stability data showed that nitrosamine levels in the active pharmaceutical ingredient increased during storage. The effects of storage under light exposure versus dark conditions, with a protective gas (nitrogen) versus an unprotected atmosphere, and the effect of temperature were investigated. In addition, stability samples were taken after 12M storage at various temperatures, and humidity was included in the assessment.

[0244] [Table 7]

[0245] The following observations were made: No increase in nitrosamine content was detected after 12 months of storage in a frozen state (-20°C), and only a slight increase was observed under low-temperature conditions (5°C). However, a rapid growth of >100% was observed during storage at 30°C / 65%RH and 40°C / 75%RH. These results were supported by other stress tests. When stored under inert gas, a slight but consistent formation of nitrosamines was observed at room temperature in the laboratory over several days, whereas when stored in an open vial, a rapid increase in nitrosamine levels was observed over the same period. Exposure to light was shown to slightly accelerate the conversion of F8 to nitrosamines. The nitrosamine content in intermediate F8 gradually increased upon exposure to air or during storage, for example, due to exposure to air, suggesting that intermediate F8 should be handled with care.

[0246] Example 4: Conversion of F8 to LOU064 API with a lower nitrosamine content (i.e., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) [ka]

[0247] Example 4a: 0.5% sulfuric acid solution: Purified water (284.0 g) was packed into the reaction vessel, sulfuric acid (1.4 g) was added, and the reaction mixture was stirred for 30 minutes to prepare for later use.

[0248] Solution of acrylic anhydride: Ethyl acetate (36.0 g) was packed into the reaction vessel, acrylic anhydride (5.8 g, 1.05 equivalents) was added, and the mixture was stirred for 30 minutes and set aside for later use.

[0249] A reactor under nitrogen was filled with (548.0 g) ethyl acetate, (100 g) purified water, 1.2 equivalents of base (e.g., sodium carbonate with low nitrite content [Na2CO3.10H2O: 5.6 g or anhydrous Na2CO3]) and F8 (20.0 g). The reaction mixture was stirred and heated to an internal temperature IT = 52-62°C.

[0250] A freshly prepared solution of acrylic anhydride was gradually added over 2 hours. The dropping funnel was rinsed with ethyl acetate (11.4 g), and the reaction mixture was stirred for 30 minutes at an internal temperature of 52-62°C.

[0251] The phases were separated and the aqueous layer was discarded. The organic layer (internal temperature = 52-62°C) was washed with the 0.5% aqueous solution of sulfuric acid prepared above (285.4 g), and then with purified water (200.0 g). The internal temperature (IT) was adjusted to 60-66°C. Filtration was performed. The filter was rinsed with aqueous ethyl acetate (28.4 g and 1.5 g of purified water). The filtrates were combined and the IT was adjusted to 58-64°C. The organic phase was concentrated until the remaining material in the flask was 622.0 g / 688 ml, and at the same time, 144.0 g of ethyl acetate was added dropwise. After concentration, the mixture of crystalline species in ethyl acetate (0.52 g) [0.060 g of anhydrous crystalline form A species disclosed in Example 1 of International Publication No. 2020 / 234779] was packed into the flask and stirred at IT = 55-61°C for 15 minutes. After constant temperature stirring, the reaction mixture in the flask was checked to see if it was a suspension. If it was not a suspension, an additional mixture of 0.060 g of LOU064 crystal species in 0.52 g of ethyl acetate was added to the flask. If it was a suspension, the organic phase was concentrated at IT=55-61°C until the remaining material in the flask was 622.0 g / 688 ml, while simultaneously gradually adding 72.0 g of ethyl acetate dropwise. After distillation, the mixture was cooled to IT=22-38°C, but the cooling time should be longer than 200 minutes. The organic phase was concentrated at IT=22-38°C until the remaining material in the flask was 622.0 g / 688 ml, and controlled. Simultaneously, 556.0 g of ethyl acetate was gradually added dropwise. After concentration, the mixture was cooled to -3-3°C, but the cooling time should be longer than 3 hours. The mixture was stirred for 4 hours, the cake was filtered, and the filtered cake was rinsed in two parts with ethyl acetate (80.0 g) pre-cooled to -3-3°C. Remibrutinib was subsequently obtained in crystalline form (anhydrous variety A, disclosed in Example 1 of International Publication No. 2020 / 234779).

[0252] The nitrosamine content in the active pharmaceutical ingredient was determined according to Example 1b.

[0253] [Table 8]

[0254] [Table 9]

[0255] Example 4b: Larger Scale Preparation of H2SO4 0.5%: 66.6 g of sulfuric acid was diluted in 15 L of purified water at room temperature.

[0256] Preparation of acrylic anhydride solution (immediately before use): 292 g of F9 was diluted in 1.8 kg of ethyl acetate.

[0257] Na2CO3 (281 g, 1.2 equivalents) was added to a suitable container and dissolved by adding purified water (6 kg, 150 equivalents compared to F8). Ethyl acetate (15 kg) was filled into the container and stirred at RT for 10 minutes. F8 (1 kg - nitrosamine content is 48 ppb) was filled into the container and the container was rinsed with ethyl acetate (13 kg). The total amount of ethyl acetate was 28 kg.

[0258] The reaction mixture was then heated to 60°C. As soon as the temperature was reached, a freshly prepared solution of F9 in ethyl acetate was added over 2 hours. After the addition was complete, the reaction mixture was stirred at 60°C for 30 minutes. Once the mixture was almost in solution (i.e., no longer a suspension), the reaction mixture was quenched with purified water (9 kg) and stirred for 10 minutes. Stirring was stopped and the phases were separated over 30 minutes. The aqueous layer was discarded and an aqueous solution of 40.5% H2SO4 was added. The reaction mixture was stirred for 15 minutes. Stirring was stopped and the phases were separated over 15 minutes. The aqueous layer was discarded and purified water (10 kg) was added. The reaction mixture was stirred at 60°C for 30 minutes. Stirring was stopped and the phases were separated over 30 minutes. The aqueous layer was discarded and the organic phase was subjected to the same clear filtration / crystallization procedure as in Example 4a. The obtained active pharmaceutical ingredient (F11) was isolated, and the content of nitrosamine (N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) was 55 ppb, as measured according to Example 1b.

[0259] Therefore, it can be seen that adding water before adding acrylic anhydride has the effect of reducing the amount of nitrosamine in the active pharmaceutical ingredient (F11). Adding water before adding acrylic anhydride can reduce the amount of nitrosamine impurities.

[0260] For example, adding 12.5 molar equivalents of water before adding acrylic anhydride reduced the amount of nitrosamine impurities by three times. A further twofold reduction can be achieved by using 125 molar equivalents of water per mole of F8 before adding acrylic anhydride.

[0261] Example 5: Synthesis of F8 having varying amounts of nitrosamine (i.e., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) Example 5a: [ka] 7 g of F7 and 84 ml of iPrOAc were packed into a 250 ml reactor under argon. 5.12 g of 37% HCl was added to this suspension over 2 hours. The reaction mixture was stirred overnight and quenched with 55 ml of tap water. The two-phase suspension was warmed to 30°C. 0.35 g of charcoal was added and the suspension was stirred for 2 hours. The solid was filtered and rinsed with 2 ml of tap water. The clear aqueous layer was transferred to a clean reactor under argon. The pH was adjusted to pH 6 with 30% NaOH (approximately 10 ml). 28.3 ml of EtOH was added and the suspension was warmed to 60°C. At this temperature, the pH was gradually adjusted to pH 10 with 1N NaOH. The white suspension was cooled to RT and filtered under a protective atmosphere. The filtered cake was rinsed with a mixture of ethanol and water and with ethanol. The F8 aqueous product was dried. The nitrosamine (N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) content in F8 is 840 ppb.

[0262] Example 5b: Optimization conditions for reducing the nitrosamine content in F8: 7 g of F7 and 84 ml of iPrOAc were packed into a 250 ml reactor under argon. 5.12 g of 37% HCl was added to this suspension over 2 hours. The reaction mixture was stirred overnight and quenched with 55 ml of distilled water. The two-phase suspension was warmed to 30°C. 0.35 g of charcoal was added and the suspension was stirred for 2 hours. The solid was filtered and rinsed with 2 ml of distilled water. The clear aqueous layer was transferred to a clean reactor under Ar. The pH was adjusted to 6 with a freshly opened bottle of 30% NaOH (approximately 10 ml). The freshly opened bottle of NaOH contributes to a low level of nitrite content (the nitrite content can be determined using the Griess test as disclosed in Example 16). 28.3 ml of EtOH was added and the suspension was warmed to 60°C. At this temperature, the pH was gradually adjusted to 10 with 1N NaOH. The white suspension was cooled to RT and filtered under a protective atmosphere. The filtered cake was rinsed with an ethanol-water mixture and ethanol. The F8 aqueous product was dried. The content of nitrosamine (N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) in F8 was 31 ppb.

[0263] In conclusion, the use of purification (e.g., distilled water) and low-nitrite content NaOH (e.g., by using a newly opened bottle of NaOH) in the deprotection step of F7 to obtain F8 has a significant effect on the nitrosamine content in F8.

[0264] [Table 10]

[0265] Example 6: Converting F7 to F11 without drying intermediate F8 7 g of F7 and 84 ml of iPrOAc were packed into a 250 ml reactor under argon. 5.12 g of 37% HCl was added to this suspension over 2 hours. The reaction mixture was stirred overnight and quenched with 55 ml of distilled water. The two-phase suspension was warmed to 30°C. 0.35 g of charcoal was added and the suspension was stirred for 2 hours. The solid was filtered and rinsed with 2 ml of distilled water. The clear aqueous layer was transferred to a clean reactor under argon. The pH was adjusted to 6 with a freshly opened bottle of 30% NaOH (approximately 10 ml). 28.3 ml of EtOH was added and the suspension was warmed to 60°C. At this temperature, the pH was gradually adjusted to 10 with 1N NaOH. The white suspension was cooled to RT and filtered under a protective atmosphere. The filtered cake was rinsed with a mixture of ethanol and water and with ethanol. A hydrated cake (containing 7 ppb of nitrosamines) was then directly added to the following reaction. To evaluate the F8 analyte, the sample was dried and the loss on drying was measured (LOD approximately 40%).

[0266] A 250 ml reactor under argon was filled with 1.4 g of Na2CO3 and 30 g of purified water. 8.63 g of hydrated F8 (5.05 g calculated based on LOD) was suspended in 140 g of ethyl acetate and added to the reactor. The white suspension was heated to 60°C, and a solution of 1.46 g of acrylic anhydride in 10 ml of ethyl acetate was added over 2 hours. The reaction mixture was stirred at 60°C for a further 30 minutes, and 73 ml of purified water was added. After stirring at 60°C for 10 minutes, the phases were separated, and the organic layer was washed with a 0.5% H2SO4 solution in purified water and with purified water. Azeotropic distillation of ethyl acetate allowed for the crystallization of LOU064, which was isolated by filtration. The nitrosamine content in the active pharmaceutical ingredient was 25 ppb.

[0267] Examples 7-11: All synthesis steps for producing F7 as disclosed in PCT / IB2023 / 059664 Preparation of Example 7-F2 [ka] To a suspension of AlCl3 in xylene at 5°C, a xylene solution of F1 was added for over 40 minutes. The mixture was heated to over 30°C for over 60 minutes and stirred at this temperature overnight. Depositphotos was added, and the resulting solution was quenched with a 0.5N aqueous solution of HCl at 0°C for 1 hour. The mixture was heated to 25°C, and the phases were separated. The aqueous layer was discarded, and the organic layer was concentrated. The resulting dilute suspension was cooled to 20°C at 0.3 K / min. The solid was filtered, and the filter cake was washed with a 1:1 solution of xylene and heptane, dried, and F2 was obtained as a white solid in approximately 83% yield.

[0268] Preparation of Example 8-F6 [ka] Preparation of F3 solution: 13.0 g of water, 2.4 g of 30% sodium hydroxide solution, 68.0 g of toluene, and 13.0 g of 2-methylaminoethanol were packed into a reaction flask. The internal temperature was adjusted to 10-30°C. The reaction mixture was stirred for 25-35 minutes. Anhydrous Boc (37.8 g, 1.00 equivalent) was added dropwise, and the reaction mixture was stirred for a further 6-12 hours at 10-30°C. The reaction was quenched with water (13.0 g), and the resulting two-phase mixture was stirred for 25-35 minutes. The lower aqueous layer was removed, and the organic layer was washed with another 13.0 g of water. The organic layer was used directly in the next step.

[0269] Mitsunobu reaction to F4: A solution of F3 in toluene (1.4 eq) was dried by Dean-Stark distillation to a water content of 0.07 wt% NMT. ​​Triphenylphosphine (42 g, 1.32 eq) was added to the dried solution of F3 at 20-30°C, and the reaction mixture was stirred at room temperature until a clear solution was observed. The reactor was inertized and cooled to approximately -30°C. F2 (20 g, 1.0 eq) was then added, followed by DIAD (31.8 g, 1.30 eq) over 4-8 hours while maintaining the internal temperature between -25°C. The slightly cloudy solution was warmed to 10°C within 4 hours and stirred for a further 15-20 hours between 5-15°C. After the completion of the reaction, toluene was distilled at 55°C to produce a slightly viscous yellowish-brown suspension. The mixture was cooled to 10°C, and n-heptane (140 g) was added. The mixture was stirred for 2 hours to obtain a pale brown, well-stirrable suspension. The suspension was filtered, and the filtered cake was washed with cooled n-heptane. The filtered cake containing triphenylphosphine oxide and H2-DIAD was discarded. The combination of mother liquor and washing solution was concentrated at JT 55°C and 150 mg bar to approximately 1 / 3 of the original volume to produce a clear yellow solution of F4.

[0270] Amination to F6: The solvent in the F4 solution was then switched to iPrOH via distillation and the addition of iPrOH. To the yellow solution of F4 in iPrOH, H2O (3.5 w / w wrt F2) and a 25 wt% NH3 solution (3.5 w / w F2) were added. The resulting yellow solution was stirred at 70°C for 16 hours. Immediately upon heating to 70°C, a small gas release (NH3) was observed. After the completion of the reaction, the resulting yellow solution was cooled to over 45°C for 40 minutes, and F6 seed crystals were added as a suspension in iPrOH. The suspension was aged for approximately 20 minutes. The dilute suspension was then cooled to 10-20°C at 10°C / hour and aged for a further 30 minutes. The suspension was filtered, and the filter cake was washed with a mixture of H2O and iPrOH (1:1) (40 g). The wet product was dried at 50°C under full vacuum (for approximately 20 hours) to obtain F6 as a white crystalline solid in approximately 70% yield.

[0271] Preparation of Example 9-X6b [ka] The synthesis of X6b was a highly convergent process, starting with the preparation of an N6a solution, the preparation of an acyl chloride X6c solution, and the combination of the two solutions to form X6b.

[0272] Autoclave: Preparation of N6a solution: N6b (20 g, 1.0 equivalent) was packed into an autoclave under N2 conditions and diluted with isopropyl acetate (105 g). Then, approximately 1 wt% of wet Pt(V) / C (0.126 g dry weight) was added, and the atmosphere was changed from N2 to H2. Hydrogenation was carried out under 3 bar H2 for 12 hours at an internal temperature below 30°C. At the end of the reaction, the suspension was filtered to remove the catalyst. The reactor and filter cake were rinsed with isopropyl acetate. The N6a solution can be used as is or water can be removed by azeotropic distillation.

[0273] Reactor A: Preparation of X6c solution: Under an N2 atmosphere, X6d (17 g, 1.1 equivalents) was suspended in toluene (56 g). A catalytic amount of pyridine was added, and the reaction mixture was heated to 50°C. Thionyl chloride was then added dropwise for over 2 hours, and the resulting mixture was stirred at 50°C for 1 hour. The turbid solution was then distilled to half its volume, the reactor was replenished with toluene to its original volume, and the process was repeated to remove excess thionyl chloride. The X6c mixture was then cooled to RT.

[0274] Reactor A: Formation of X6b: To a solution of X6c (1.1 equivalents) in toluene, the previously prepared solution of N6a (1.0 equivalent) in iPrOAc was added for over 1 hour. At the end of the addition, DIPEA (13.4 g, 1.2 equivalents) was carefully added for over 2 hours. The reaction mixture was stirred for 3 hours after the completion of DIPEA addition, and the reaction was quenched with iPrOH (26.4 g). The reaction was stirred overnight at RT (room temperature), and the suspension was filtered. The moist cake was rinsed with iPrOH and iPrOH / water. The cake was drained and dried under reduced pressure. X6b was typically isolated in 87–93% yield.

[0275] Example 10a: Optimization of Suzuki conditions for converting X6a to F7 Previously, a coupling reaction between F6 and X6a was reported to have been carried out at 75°C for 8 hours using 1 eq of F6, 1.15 eq of X6a, 5 mol% Pd(PPh3)2Cl2, 3 eq of Na2CO3, 12 vol DME, and 10 vol water, with an isolation yield of 74% (DOI:10.1021 / acs.jmedchem.9b01916).

[0276] The cross-coupling reaction was optimized to replace the DME solvent with a Class 3 solvent suitable for commercial processes, while also reducing Pd usage and production costs. [ka]

[0277] Details of the design and experiment 1) Suzuki's 12 catalyst precursors and 6 solvent systems (80°C: tertoamyl alcohol, CPME, and toluene; 60°C: THF, Me-THF, and MeCN, each combined with water) were screened at a level of 2.0 mol% Pd using 1.15 eq. X6a in the presence of 3.0 eq. K3PO4. After 16 hours, a series of catalyst precursor / solvent combinations were found in which the reaction could be promoted, fully converted, and devolonate was the main byproduct; it was decided to perform screening of all ligands in both toluene (80°C) and Me-THF (60°C). 2) Forty-eight ligands were screened in 10.0 vol. Me-THF / 3.0 vol. water at 60°C or in 10.0 vol. toluene / 3.0 vol. water at 80°C using 2.0 mol% Pd(OAc)2, 1.1 eq. boronate, and 3.0 eq. K3PO4. After 16 hours, it was found that five ligands (RuPhos, dppf, S-Phos, Cy3P·HBF4, and Ph2P(t-Bu)) could promote the reaction, be completely converted, yield Prod / IS in Me-THF / water at 60°C, and allow for control of deboronate byproducts at levels of 3% to 8%.

[0278] [Table 11]

[0279] 3) Maintaining a P:Pd ratio of 2:1, six Pd precursors (Pd(OAc)2, [Pd(C3H5)Cl]2, Pd(TFA)2, Pd(MeCN)2Cl2, Pd2(dba)3, and PdBr2) were screened at a 1.0 mol% Pd level in combination with RuPhos, dppf, S-Phos, Cy3P·HBF4, and Ph2P(t-Bu), respectively, in the presence of 3.0 eq. K3PO4 and 1.05 eq. X6a in 10.0 vol. Me-THF / 3.0 vol. water at 60°C. After 16 hours, Cy3P·HBF4 and Ph2P(t-Bu) remained as optimal ligand candidates, while Pd(TFA)2, Pd(MeCN)2Cl2, and PdBr2 continued to be excellent Pd precursors.

[0280] [Table 12]

[0281] 4) Maintaining a P:Pd ratio of 2:1, and using Cy3P·HBF4 and / or Ph2P(t-Bu) as ligands, the following combinations were screened in 10.0 vol. Me-THF / 3.0 vol. water at 60°C with a Pd usage of 0.1-2.0 mol% in the presence of 3.0 eq. K3PO4 and 1.05 eq. X6a. After 16 hours, it was found that Pd(MeCN)2Cl2 / Ph2P(t-Bu) was the best and most optimal catalyst precursor combination, allowing the Pd usage to be reduced to 0.3-0.5 mol%, and enabling control of the devoronate / Prod ratio to approximately 1%.

[0282] [Table 13]

[0283] 5) Using Pd(MeCN)2Cl2 / Ph2P(t-Bu) and 1.05eq.X6a as the optimal catalyst precursor combination, we screened 0.1-0.5 mol% Pd usage in the presence of K2CO3, Cs2CO3, K3PO4, and KF, respectively. We found that K3PO4 was the optimal base, and that 0.3-0.5 mol% Pd(MeCN)2Cl2 / Ph2P(t-Bu) catalyst precursor is recommended for scale-up reactions.

[0284] Most efficient conditions 1) The reaction of 1.0 eq. F6, 1.05 eq. X6a, 0.5 mol% Pd(MeCN)2Cl2, 1.0 mol% Ph2P(t-Bu), and 3.0 eq. K3PO4 in 10.0 vol. Me-THF / 3.0 vol. water at 60°C for 16 hours achieved complete conversion, with an IPC purity of 90.6% by HPLC and a devoronate / Prod ratio of 1%. 2) The reaction of 1.0 eq. F6, 1.05 eq. X6a, 0.3 mol% Pd(MeCN)2Cl2, 0.6 mol% Ph2P(t-Bu), and 3.0 eq. K3PO4 in 10.0 vol. Me-THF / 3.0 vol. water at 60°C for 16 hours achieved 99% conversion, with an IPC purity of 88.5% by HPLC and a devoronate / Prod ratio of 1%.

[0285] [ka]

[0286] Next, the optimal conditions 1) The reaction of 1.0 eq. F6, 1.05 eq. X6a, 0.8 mol% Pd(TFA)2, 1.6 mol% Ph2P(t-Bu), and 3.0 eq. K3PO4 in 10.0 vol. Me-THF / 3.0 vol. water at 60°C for 16 hours achieved complete conversion, with an IPC purity of 90.9% by HPLC and a devolonate / Prod ratio of 2%. 2) The reaction of 1.0 eq. F6, 1.05 eq. X6a, 0.8 mol% Pd(MeCN)2Cl2, 1.6 mol% Ph2P(t-Bu), and 3.0 eq. K3PO4 in 10.0 vol. Me-THF / 3.0 vol. water at 60°C for 16 hours achieved complete conversion, with an IPC purity of 91.2% by HPLC and a devoronate / Prod ratio of 2%.

[0287] Preparation of F7 from X6b by one-pot boration-Suzuki cross-coupling using the optimized conditions of Example 10b-10a [ka] Miyaura Boration: X6b (1.0 eq), B2pin2 (1.06 equivalents), and KOAc (2.5 equivalents) were packed into a reactor under an N2 atmosphere containing defasted Me-THF. The water content of the reaction mixture was measured and adjusted to between 1000 and 2500 ppm. After inertization of the vessel, a solution of Pd(MeCN)2Cl2 (0.5 mol%) in defasted MeTHF and a solution of PPh2tBu (1.0 mol%) in defasted MeTHF were successively added. The reaction mixture was then heated to 70°C for 16 hours.

[0288] Suzuki coupling: Once the complete conversion of X6b was achieved (X6b < 0.25%, conversion is approximately 98%), the reaction mixture was cooled to RT and quenched with an aqueous solution of KOH (21% wt / wt). The aqueous layer was separated and discarded, and a fresh aqueous solution of KOH (21% wt / wt) was added. F6 (0.96 equivalents compared to X6b) was added as a solid, followed by another PPh2tBu (2 mol%) in defasted MeTHF and another Pd(MeCN)2Cl2 (1 mol%) in defasted MeTHF after proper degassing. The reaction mixture was then heated to 60°C for approximately 24 hours. After the completion of the reaction, an aqueous solution of N-acetylcysteine ​​was added to the reaction mixture at 60°C. After stirring for 2 hours, the aqueous layer was discarded. Another aqueous solution of N-acetylcysteine ​​was added, and the pH was adjusted to ≥9.5 by adding an aqueous solution of KOH. After stirring for 2 hours, the aqueous layer was discarded. The organic layer was then washed with water for 30 minutes, and the aqueous layer was discarded. The solution was filtered through activated carbon at 60°C, and the solution was concentrated to half its volume by vacuum distillation. n-heptane was gradually added, and the resulting suspension was cooled to 20°C, stirred for 2 hours, and filtered. The filtered cake was washed with a 1:5 mixture of Me-THF and n-heptane. If the purity was not satisfactory, the wet cake could be slurryed again with Me-THF and n-heptane (1:5). The cake was drained and dried under reduced pressure. F7 is typically isolated in 92% yield.

[0289] Development of a one-pot borylation / Suzuki cross-coupling using tetrahydroxydiborone for use in the preparation of Example 10c-F7 A one-pot boration / Suzuki cross-coupling process using tetrahydroxydiborone was developed for the synthesis of F7 from X6b using BBA as the boration reagent. This process was characterized by the use of significantly reduced amounts of Pd-catalyst, avoidance of pinacol hydrate precipitation in the final product, and the use of methanol as the green alcohol solvent throughout both steps. This process addresses some of the previous problems associated with the use of bis(pinacolate)diborone as the boration reagent, thus resulting in a more atomically efficient and cost-effective approach. The results below demonstrate the feasibility of this one-pot process on a 2.2g scale using a FlexyALR reactor. [ka]

[0290] Results and Discussion Miyaura Boration: To develop optimal reaction conditions for Miyaura boration using BBA, critical reaction parameters such as catalyst system, base, solvent, and temperature were screened. This boration was limited to the use of a Pd(II)-catalyst precursor that promotes rapid Pd(0) formation. In fact, the use of a second-generation Buchwald catalyst precursor combined with two equivalents of an additional ligand proved to be the most efficient catalyst system in the inventors' reaction (Table 1, entries 1-6). Of all the screened catalyst precursors, only Pd-XPhos-2G obtained complete conversion of the starting material while providing the highest yield and selectivity toward the formation of X6a (entry 2). Similarly, the use of ethylene glycol as an additive also proved to be very beneficial, as complete conversion could not be achieved without ethylene glycol (entries 1 vs 2). BBA can be stabilized at insights through the formation of the corresponding boronic acid ester derivative, reducing the amount of borylation reagent and Pd while increasing the rate of boration. Further attempts were made to reduce the amount of catalyst used (entries 8-10). Surprisingly, by reducing the amount of catalyst used, smaller amounts of reduction and dimerization products IMP1 and IMP2 were obtained while still achieving a nearly complete conversion of X6b (entry 8). Moreover, higher conversion was observed by increasing the reaction time, thus suggesting that BBA was still present in the reaction mixture (entry 9). These results could be shown that the formed boronic acid may undergo a decomposition pathway catalyzed by Pd(II) and that a larger supply of Pd in ​​the presence of trace amounts of oxygen may facilitate this pathway. Finally, by simply increasing the reaction temperature to 50°C, complete conversion to the final product was observed with high selectivity and yield (entry 10).

[0291] [Table 14]

[0292] The reaction was also evaluated by replacing ethylene glycol with amine bases, DIPEA, and other Buchwald catalyst precursors to determine whether the results of the Miyaura borylation could be further improved and whether the amount of catalyst working could be increased (Table 2, entries 1-5). Most catalysts did not perform well under these conditions, but improvement was seen by using Pd-cataCXium 3G (entry 5). Slightly larger amounts of IMP1 and IMP2 were formed compared to already optimized conditions, but these results were promising considering that cataCXium is superior to XPhos when used in combination with DIPEA (entry 5 vs 1). In addition to these results, the inventors screened other decisive reaction parameters to determine whether these results could be further improved (entries 6-9). Considering the inventors' previous results, the reduction in catalyst usage was investigated first (entry 6). Importantly, the inventors found that 0.05 mol% Pd was sufficient to complete the reaction, suggesting that the catalytic activity of Pd-cataCxium-3G under these conditions was much higher than that of Pd-XPhos-2G. Importantly, it was found that heating to 50°C was optimal, as decreasing the temperature resulted in an incomplete reaction (Entry 7). Surprisingly, the inventors found that the addition of ethylene glycol was detrimental to the conversion of the reaction, thus suggesting that cyclic diborone species may be less reactive under these conditions (Entry 8). Although remarkably high catalytic activity was observed under these newly optimized conditions, the relative amounts of byproducts IMP1 and IMP2 could not be further reduced, and the conditions based on the use of Pd-Xphos-2G, KOAc, and ethylene glycol remained superior.

[0293] [Table 15]

[0294] Suzuki Cross-Coupling: Having determined two sets of optimization conditions for the synthesis of boronic acid X6a using BBA as the boronating reagent, the feasibility of subsequent Suzuki coupling was then investigated, with the ultimate goal of developing a one-pot process for the synthesis of F7. For this purpose, Suzuki coupling of X6a and F6 was attempted at 60°C under reaction conditions previously developed by Molanders (Gurung, SR, et al., Org. Process Res. Dev. 2017, 21, 65-74) (Table 3, entry 1). Contrary to expectations, heterogeneous and incomplete conversion of X6a and F6 was observed after heating to 60°C for 17 hours. Furthermore, F6 was converted to EtOH and S N It was found that the reaction partially occurred via the Ar pathway, forming the corresponding ether. At this point, we considered whether the use of milder organic bases, such as amines, could help reduce this side reaction. Indeed, the use of Et3N resulted in minimal formation of the CO coupling product and led to a homogeneous and nearly perfect conversion of X6a and F6 (Entry 2). Surprisingly, MeOH proved superior to EtOH, providing complete conversion of X6a and F6 as well as a higher yield of the coupling product (Entry 3). Moreover, F7 precipitated directly from the reaction mixture, thus considerably simplifying the final work-up purification. The formation of reduction and dimerization products IMP1 and IMP2 demonstrated the presence of trace amounts of oxygen in the reaction solvent, but we anticipated that scaling up the process would effectively eliminate this problem (Entries 1-3).

[0295] [Table 16]

[0296] One-pot borylation and coupling: Since Pd-XPhos 2G and Pd-cataCXium 3G proved to be excellent catalyst precursors in the Miyaura borylation using BBA, we decided to compare the efficiency of these two catalysts in a one-pot process using our optimized conditions (Table 4). As shown in entry 1, Pd-XPhos 2G proved superior to Pd-cataCXium 3G in the one-pot procedure, demonstrating that it yielded a 79% isolation yield of F7 with a purity of 78% starting from X6b. As expected, work-up and purification of F7 could be carried out by direct filtration and washing of the formed precipitate with a MeOH / H2O mixture.

[0297] [Table 17]

[0298] Scale-up: Having developed the conditions for both steps in MeOH using the same catalyst precursor under mild conditions, we attempted the one-pot reaction on a larger scale (2.2 g of X6b) using a Flexy ALR-1 300 ml reactor (Table 5).

[0299] X6b (2.20 g, 1.0 equivalent), potassium acetate (1.76 g, 3.0 equivalent), ethylene glycol (1.0 ml, 3.0 equivalent), and MeOH (100 ml) were packed into a 300 ml FlexyALR reactor. The reaction mixture was degassed through a continuous vacuum / N2 cycle, and a solid mixture of BBA (807 mg, 1.5 equivalent), Pd XPhos 2G (12 mg, 0.25 mol%), and XPhos (14 mg, 0.50 mol%) was added under N2. After a second degassing, the reaction was heated to 50°C and stirred overnight. The mixture containing boronic acid was then cooled to 20°C, and F6 (1.73 g, 0.95 equivalent), Pd XPhos 2G (24 mg, 0.5 mol%), Et3N (2.5 ml), and degassed water (30 ml) were added under N2. The reaction was degassed for the third time and stirred overnight at 60°C. The reaction was then cooled to 40°C and concentrated under reduced pressure (removing approximately 40 ml of MeOH). The reaction mixture was then cooled to 20°C and stirred for 3 hours. The pale brown suspension was filtered, washed with a cold solution of MeOH / H2O 4 / 1 (40 ml), and dried to obtain F7 (1.87 g, 56%) as a brown solid.

[0300] [Table 18]

[0301] The Miyaura borylation of X6b resulted in the formation of the desired intermediate X6a with excellent yield and selectivity. Interestingly, as described by Molanders for the use of Pd-XPhos 2G, the completion of the borylation was demonstrated by a sudden color change of the reaction mixture from white to pale orange-yellow. Subsequently, Suzuki coupling was carried out by adding F6, a fresh batch of catalyst, Et3N, and H2O to the reaction mixture. Filtration and washing of the final product yielded F7 with an isolation yield of 56% across both steps and an IPC purity of 87%. Importantly, as the inventors predicted, the formation of by-products IMP1 and IMP2 was minimized by removing all trace amounts of oxygen by performing both steps in the reactor.

[0302] Example 12: Stability of LOU064 active pharmaceutical ingredient To determine the stability behavior of the nitrosamine impurity (N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) in the LOU064 active pharmaceutical ingredient, an accelerated predictive stability (APS) test was performed.

[0303] Method explanation: A statistical-based approach using the Arrhenius equation with humidity modifications, the Accelerated Stability Prediction Test (APS) [Waterman et al., International Journal of Pharmaceutics 293(1-2), 101-125 (2005)], is applied to understand stability behavior and predict retest periods. The design of the predictive study is based on literature-reported studies demonstrating the modeling of observed degradation for solid oral dosage forms (Waterman et al., Pharmaceutical Research 24(4), 780-790 (2007) and Waterman et al., Journal of Pharmaceutical Sciences, 99(11), 4437-4452 (2010)). Short-term studies are performed on open petri dish samples of representative batches of the active pharmaceutical ingredient under accelerated conditions using a wide range of temperatures and humidity, with the goal of reaching specification limits within a specified retest period that limits the properties under each of the conditions detailed below. Humidity determines the water activity in the active pharmaceutical ingredient (API) and can have a significant effect on the reaction rate in solid APIs, even for reactions that do not contain water themselves. The humidity-corrected Arrhenius equation reflects the influence of both temperature and moisture on the dynamics of degradation product formation. The resulting open petri dish data is shown using the humidity-corrected Arrhenius equation with ASAPprime® (current version 6.0). 1 To fit: formula 1 In k = In A - Ea / RT + B(%RH) Arrhenius equation modified for humidity [1]; k is the decomposition rate, A is the Arrhenius frequency factor (collision frequency), E a is the activation energy for the chemical reaction, R is the gas constant, T is the temperature in Kelvin, and B is the humidity sensitivity constant, with relative humidity percentage denoted as %RH.

[0304] Experimental conditions: Three batches of the LOU064 drug substance were placed in open petri dishes under the protocol summarized below.

[0305]

Table 19

[0306] Samples of the LOU064 drug substance batch were prepared in two ways in a closed container, one sample was purged with nitrogen and the other was not.

[0307]

Table 20

[0308] The withdrawn samples were analyzed by HPLC-MS according to the method described in Example 1 and reported below in Tables 8 and 9.

[0309]

Table 21

[0310] Nitrosamine was found to be stable and did not increase or decrease beyond the expected analytical variation under all APS open petri dish conditions.

[0311]

Table 22

[0312] When comparing LOU064 active pharmaceutical ingredient stored in a sealed container under nitrogen with that stored in a sealed container without nitrogen, no difference was observed beyond the variability observed in the analysis.

[0313] Conclusion: Based on APS data, an increase in nitrosamine impurities is not expected in the LOU064 API prepared with low levels of nitrosamine as described in the present invention. Nitrogen purging had no measurable effect on nitrosamine formation.

[0314] Example 13: Evaluation of the formulation manufacturing process for increased nitrosamine impurities (N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) The formulations (film-coated tablets) were prepared using LOU064 active pharmaceutical ingredient, which is substantially free of nitrosamines, as disclosed in International Publication No. 2022 / 162513, Example 8. The manufacturing process includes the steps of grinding the hydrated drug, spray granulation and granulation, final compounding, and tableting.

[0315] The levels of nitrosamine impurities (N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) were measured at each step of the process and summarized in Table 10 below.

[0316] [Table 23]

[0317] Conclusion: The active pharmaceutical ingredient (API) process (impurity content) is the primary factor in the presence of nitrosamine impurities in the formulation. The level of nitrosamine impurities is minimally affected by the formulation manufacturing process, particularly when the nitrosamine level in the API is less than 200 ppb, as disclosed in International Publication No. 2022 / 162513.

[0318] Example 14: Stability of LOU064 formulation (film-coated tablet) A statistical-based approach using the Arrhenius equation with a modification to humidity as described in Example 12, and an accelerated stability prediction test (APS), were used to understand the stability behavior and predict the shelf life.

[0319] Several batches of LOU064 formulations (film-coated tablets) prepared according to Example 8 of International Publication No. 2022 / 162513 using a substantially nitrosamine-free active pharmaceutical ingredient were analyzed for stability under various storage conditions. The nitrosamine (i.e., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) content was analyzed by HPLC-MS as described in Example 1.

[0320] [Table 24]

[0321] On day 28, the batch was further analyzed using XRPD.

[0322] [Table 25]

[0323] Conclusion: Even under extreme conditions, no trend of nitrosamine impurities was observed in the APS study (i.e., the level of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide did not increase). Furthermore, no changes in polymorphism were observed.

[0324] Example 15: Method for verifying the nitrosamine content in a pharmaceutical formulation (e.g., film-coated tablets) The method can also be applied to verify the nitrosamine content in the active pharmaceutical ingredient.

[0325] In this explanation, the remibrutinib nitrosamine impurity (i.e., N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide) will be referred to as RNI.

[0326] RNI-d7, used as a reference material below, has the following structure. [ka]

[0327] HPLC-MS Principle (U) Detection and internal standard method using HPLC and HRAM (High Resolution Accurate Mass) mass spectrometry reagent Methanol HPLC gradient grade or equivalent, e.g., JT Baker 8402 Ammonia 25% (w / V) LC-MS grade or equivalent, e.g., Merck 533003 Purified water Reference material RNI RNI-d7) device (U)HPLC connected to an HRAM-MS instrument (e.g., Thermo Orbitrap Exploris 120 or equivalent) Waters UPLC BEH C18 column, 100mm length, 2.1mm inner diameter, and 1.7μm particle size, or equivalent column. Filter: 0.2 μm PVDF filter or equivalent. Solvent: Water:MeOH = 20:80 (V / V) Chromatography conditions Mobile phase A: 0.025% (w / V) ammonia in water. For example, add 1000 μL of 25% ammonia to 1000 mL of water and mix thoroughly. Mobile phase B MeOH

[0328] [Table 26]

[0329] Flow rate 0.4mL / min Detection by high-resolution mass spectrometry detectors Column temperature 50℃ Condenser after the column: 40°C Autosampler temperature: 22.5°C Injection volume 5μL The approximate retention time for RNI and RNI-d7 is about 7 minutes (for reference only). Note: It is recommended to switch the LC flow to wastewater before and after the elution of the RNI and RNI-d7 peaks.

[0330] MS detector settings Ionized H-ESI, cation Spray voltage 3500V Sheath gas 50AU Auxiliary gas 5AU Sweep gas 0AU Ion transfer tube temperature: 300℃ Vaporizer temperature: 350℃ Scan type tMS 2 Precursor ion (m / z) RNI 483.1951 [M+H] + Precursor ion (m / z) RNI-d7 490.2390[M+H] + RNI: Product ion (m / z) 87.0553±10ppm (quantitative) 397.1476±10ppm (confirmed) RNI-d7: Product ion (m / z) (93.0929 + 94.0992) ± 10 ppm (quantification) 398.1539 ± 10 ppm (confirmation) Resolution 60000 Isolation window (m / z) 0.4 HCD collision energy (V) 18 RF lens (%) 70 Maximum injection time (ms) Automatic Expected LC peak width (s): 20 Note: All MS detector settings reported above are guidelines only and may be adapted to various MS detectors and optimized as long as the SST requirements are met. MS conditions can be adjusted to optimize detection.

[0331] System Conformity Testing (SST) There are no peaks interfering with the peaks of the blank RNI and RNI-d7. Reproducibility (STD) S rel ≤20% (n=6) (RNI peak area in standard solution) Reporting threshold: Signal-to-noise ratio of ≥10 for the peak of RNI in LOQ solution. Note: If there is no noise in the blank, the signal-to-noise ratio (S / N) does not need to be calculated. In this case, the SST (Service Level Test) is suitable. The proportionality constant (PF) is the proportional relationship between the peak ratio of RNI to IS between the standard solution and the LOQ solution: 0.7 ≤ PF ≤ 1.3 (start of sequence). Drift of standard solution: drift of ±25% or less Note: Ph.Eur.2.2.46 "Chromatographic Separation Techniques" or USP <621> Calculations are performed according to the "Chromatography, System Suitability Test".

[0332] procedure Internal standard solution (IS) (c RNI-d7 ≈ 5 ng / mL): Dissolve RNI-d7 in methanol and dilute with solvent (approximately 5 ng RNI-d7 / mL). For example, accurately weigh approximately 2.5 mg of RNI-d7 reference material into a 25 mL volumetric flask, dissolve it in methanol, and dilute with methanol (c RNI-d7 ≈ 100 μg / mL). Further dilute 100 μL of the above solution with solvent to 10 mL (c RNI-d7 ≈ 1 μg / mL). Further dilute 1000 μL of the above solution with solvent to 200 mL. The test solution IS contains approximately 2.5 mg LOU064 / mL (approximately 2.5 mg LOU064 / mL and approximately 5 ng RNI-d7 / mL). For example, crush and homogenize 10 tablets (an IKA mill can be used, for example). Accurately weigh approximately 136 mg of tablet powder (equivalent to approximately 25 mg of LOU064) into a 10 mL volumetric flask. Add approximately 2 / 3 of the flask volume of IS and shake in a shaker for at least 15 minutes. Fill the flask with IS and shake thoroughly. Filter into an HPLC vial through a 0.2 μm PVDF filter (or other suitable filter) and discard the first 0.5 mL. Note: Alternative test solution preparations may be used if validated. RNI stock solution (stock SS) (c RNI ≈ 100 μg / mL) Approximately 100 μg RNI / mL in methanol For example, accurately weigh approximately 5 mg of the RNI reference substance into a 50 mL volumetric flask, dissolve it in methanol, and dilute it. Reference standard solution (REF) (c RNI=1μg / mL) (c RNI-d7≈5ng / mL) 1μg RNI / mL in IS (1μg RNI / mL and approximately 5ng RNI-d7 / mL For example, add the exact volume (500 μL) of stock SS to a 50 mL volumetric flask using a pipette, taking into account the exact weight and purity of the RNI reference substance, according to the following formula. Dilute with IS.

number

number

number

[0333] Example 16: Method for verifying the nitrite content Chromatography conditions: GRIESS reaction and nitrite analysis by HPLC Method HPLC UV: Mobile phase A: 0.1M formic acid in water Mobile phase B: Acetonitrile Analytical column: Water XBridge BEH C18, 100 x 3.0 mm, 2.5 μm Injection volume: 20uL Column temperature: 40℃ Flow rate: 0.8mL / min Needle cleaning: Wash with acetonitrile / water in a 1:1 v / v ratio. Detection: 548nm gradient:

[0334] [Table 27]

[0335] Sample preparation for the determination of nitrite content in sodium carbonate: Sodium carbonate sample preparation: Solvent: 85% o-H3PO4:Water = 37.5:62.5 Neutralization reagent: 8 mL solvent + 1 mL GRIESS reagent

[0336] Sample solution: Weigh approximately 600 mg of sodium carbonate sample into a 10 mL volumetric flask and add the neutralizing reagent (9 mL total) in three separate additions while in a water bath. (Since carbon dioxide is generated during the addition process, the rate of carbon dioxide generation should be controlled by the addition rate to ensure that bubbles do not overflow from a full bottle, and the solution should be shaken while adding.) After adding the neutralizing reagent, sonicate the solution until the bubbles disappear (approximately 1-2 minutes), and then dilute with water.

[0337] Sample preparation for comparison standards: Water and 1 mg / mL of 1 mg / mL nitrite Std solution in 85% o-H3PO4 Comparison stock solution: 0.1 mL of nitrite Std solution in 100 mL of water = SSS (0.001 mg / mL) Comparison solution: Transfer 60 μL of the comparison stock solution to a 10 mL volumetric flask, add 8 mL of solvent and 1 mL of GRIESS reagent, dilute with water, and mix thoroughly (100 ppb).

[0338] Sample preparation for the quantification of nitrite content in sodium hydroxide: Sodium hydroxide sample preparation: Solvent: 85% o-H3PO4:Water = 37.5:62.5

[0339] Sample stock solution: Weigh 8000 mg NaOH into a 10 mL volumetric flask, dilute with MQ water, and mix thoroughly. (Avoid prolonged ultrasonic exposure; experimental data shows that prolonged ultrasonic exposure of glassware increases NO2 levels).

[0340] Sample solution: Transfer 1.5 mL of 85% o-H3PO4 and 0.5 mL of GRIESS reagent to a 5 mL volumetric flask. Place the volumetric flask in an ice bath, transfer 2.5 mL of the stock sample solution, and gradually add it dropwise to the 5 mL volumetric flask. Shake the volumetric flask as much as possible. After the addition is complete, mix well, allow to return to room temperature, and dilute with 85% o-H3PO4. Mix thoroughly.

[0341] Sample preparation for comparison standards: Water and 1 mg / mL of 1 mg / mL nitrite Std solution in 85% o-H3PO4 Water and 1 mg / mL of 1 mg / mL nitrite Std solution in 85% o-H3PO4 Comparison stock solution: 100 mL of water with 100 μL of nitrite Std solution = SSS (0.001 mg / mL) Comparison solution: Transfer 50 μL of the comparison stock solution to a 5 mL volumetric flask, add 2 mL of water and 0.5 mL of GRIESS reagent, dilute with 85% o-H3PO4, and mix thoroughly (25 ppb).

Claims

1. Substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide. 【Chemistry 1】 drug substance.

2. The LOU064 active pharmaceutical ingredient according to claim 1, which is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, wherein the content of the aforementioned impurities is less than about 550 ppb, for example less than about 530 ppb.

3. The LOU064 active pharmaceutical ingredient according to claim 1, which is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, wherein the content of the aforementioned impurities is less than about 400 ppb, for example less than about 360 ppb.

4. LOU064 active pharmaceutical ingredient substantially free of the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide according to claim 1, wherein the content of the aforementioned impurities is less than about 150 ppb, for example less than about 130 ppb.

5. The LOU064 active pharmaceutical ingredient according to claim 1, which is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, wherein the content of the aforementioned impurities is about 100 ppb or less, for example, less than about 90 ppb.

6. The content of the aforementioned impurities is determined by HPLC-MS, for example, MS by selective ion monitoring, for example, HPLC-MS using the conditions described in Example 1 or Example 15, for example, LOU064 active pharmaceutical ingredient substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, according to any one of claims 2 to 5.

7. When measured at a temperature of approximately 25°C and an X-ray wavelength of 1.5405 Å, LOU064 showed the following values: 7.8±0.2°2θ, 9.2±0.2°2θ, 12.0±0.2°2θ, 13.6±0.2°2θ, 15.6±0.2°2θ, 16.0±0.2°2θ, 17.8±0.2°2θ, 18.3±0.2°2θ, 18.7±0.2°2θ, 19.2±0.2°2θ, 19.9±0.2°2θ, 22.1±0.2°2θ, 23.4±0.2°2θ, 23.9±0.2°2θ, 24.8±0.2°2θ, 25.2±0.2°2θ, and 25.5±0.2°2θ. LOU064 active pharmaceutical ingredient according to any one of claims 1 to 6, having a crystalline form characterized by an X-ray powder diffraction pattern including one or more representative peaks with respect to 2θ selected from the group consisting of θ, 27.2 ± 0.2° 2θ and 29.6 ± 0.2° 2θ, substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

8. LOU064 is a crystalline substance characterized by an X-ray powder diffraction pattern that includes representative peaks at 7.8±0.2°2θ, 9.2±0.2°2θ, and 12.0±0.2°2θ when measured at a temperature of approximately 25°C and an X-ray wavelength of 1.5405 Å, λ, and is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, according to any one of claims 1 to 7.

9. LOU064 is a crystalline form A in which the phase is substantially pure, and is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, as described in claim 7 or 8.

10. LOU064 is a substantially chemically pure LOU064 active pharmaceutical ingredient that is substantially free of nitrosamine impurities according to any one of claims 1 to 9, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

11. A pharmaceutical composition comprising the LOU064 active pharmaceutical ingredient described in any one of claims 1 to 10 and one or more pharmaceutically acceptable excipients.

12. A pharmaceutical composition comprising LOU064 or a pharmaceutically acceptable salt thereof, wherein the composition is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

13. A pharmaceutical composition comprising LOU064 or a pharmaceutically acceptable salt thereof, wherein the total amount of nitrosamine impurities in the composition, for example, the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is less than or equal to the maximum amount of nitrosamine impurities permitted in the composition by the regulatory authority at the time the composition is prepared and / or administered.

14. The pharmaceutical composition according to claim 12, wherein the total amount of nitrosamine impurities in the composition, for example, the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is less than about 550 ppb, for example less than about 530 ppb, based on the total amount of LOU064 in free or salt form.

15. The pharmaceutical composition according to claim 12, wherein the total amount of nitrosamine impurities in the composition, for example, the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is less than about 400 ppb, for example less than about 360 ppb, based on the total amount of LOU064 in free or salt form.

16. The pharmaceutical composition according to claim 12, wherein the total amount of nitrosamine impurities in the composition, for example, the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is less than about 150 ppb, for example less than about 130 ppb, based on the total amount of LOU064 in free or salt form.

17. The pharmaceutical composition according to claim 12, wherein the total amount of nitrosamine impurities, for example, the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is about 100 ppb or less, for example less than about 90 ppb, based on the total amount of LOU064 in free or salt form.

18. A pharmaceutical composition comprising LOU064 or a pharmaceutically acceptable salt thereof, which has been tested and found to contain a total amount of nitrosamine impurities less than about 550 ppb, for example less than about 400 ppb, for example less than about 150 ppb, for example less than about 100 ppb, based on the total amount of LOU064 in free or salt form, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

19. a) a pharmaceutical composition comprising LOU064 or a pharmaceutically acceptable salt thereof, and b) a pharmaceutical product comprising documentation, either directly or via a link to an electronic database, assuring that the total amount of nitrosamine impurities in the composition, for example, the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, is less than about 550 ppb, for example less than about 400 ppb, for example less than about 150 ppb, for example less than about 100 ppb, based on the total amount of LOU064 in free or salt form.

20. a) a pharmaceutical composition comprising LOU064 or a pharmaceutically acceptable salt thereof, and b) a pharmaceutical product comprising documentation, either directly or via a link to an electronic database, assuring that the total amount of nitrosamine impurities in the composition, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, at the time the composition is prepared and / or administered, is less than or equal to the maximum amount of nitrosamine impurities permitted in the composition by the regulatory authority.

21. The pharmaceutical product according to claim 19 or 20, wherein the total amount of LOU064 or a pharmaceutically acceptable salt thereof in the composition is the same amount of LOU064 as about 5 mg, or about 10 mg, about 25 mg, about 50 mg, or about 100 mg of free base.

22. A final dosage form comprising the pharmaceutical composition according to any one of claims 11 to 18.

23. The final dosage form according to claim 22, wherein the dosage form is a film-coated tablet.

24. The final dosage form according to any one of claims 22 or 23, wherein the free form of LOU064 is present in an amount of about 5 mg, about 10 mg, about 25 mg, or about 100 mg, for example, about 25 mg or about 100 mg.

25. LOU064 active pharmaceutical ingredient according to any one of claims 1 to 10, or a pharmaceutical composition according to any one of claims 11 to 18, or a final dosage form according to claim 22, 23, or 24, for use in the treatment of diseases or disorders mediated by or restored by inhibition of BTK.

26. A method for treating a disease or disorder mediated by or restored by inhibition of BTK, comprising administering a therapeutically effective amount of the LOU064 active pharmaceutical ingredient described in any one of claims 1 to 10, the pharmaceutical composition described in any one of claims 11 to 18, or the final dosage form described in claim 22, 23, or 24 to a subject in need thereof.

27. The diseases or disorders mediated by or restored by BTK include autoimmune disorders, inflammatory diseases, allergic diseases, airway diseases such as asthma and chronic obstructive pulmonary disease (COPD), transplant rejection; diseases in which antibody production, antigen presentation, cytokine production, or lymphoid organ formation is abnormal or adverse; rheumatoid arthritis, systemic juvenile idiopathic arthritis (SOJIA), gout, pemphigus vulgaris, and idiopathic thrombocytopenic purpura. Systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, Sjögren's syndrome, sweat gland abscess, IgE-driven allergies, such as drug, poison, and food allergies; autoimmune hemolytic anemia, anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis, cryoglobulinemia, thrombotic thrombocytopenic purpura, chronic urticaria (chronic idiopathic urticaria, induced urticaria), chronic allergies (atopic dermatitis, contact dermatitis, allergic rhinitis) LOU064 active pharmaceutical ingredient for use according to claim 25 or a pharmaceutical composition for use according to claim 25 or a method for treatment according to claim 26, selected from: inflammation), atherosclerosis, type 1 diabetes, type 2 diabetes, inflammatory bowel disease, ulcerative colitis, Crohn's disease, pancreatitis, glomerulonephritis, Goodpasture syndrome, Hashimoto's thyroiditis, Graves' disease, antibody-mediated transplant rejection (AMR), graft-versus-host disease, B-cell-mediated hyperacute, acute and chronic transplant rejection; thromboembolic disorders, myocardial infarction, angina pectoris, stroke, ischemic disorders, pulmonary embolism; hematopoietic cancers including but not limited to multiple myeloma; leukemia; acute myeloid leukemia; chronic myeloid leukemia; lymphocytic leukemia; myeloid leukemia; non-Hodgkin lymphoma; lymphoma; polycythemia vera; essential thrombocythemia; myelofibrosis with myeloid metaplasia; and Waldenström disease.

28. The disease or disorder mediated by or restored by inhibition of BTK is selected from chronic urticaria, for example, chronic idiopathic urticaria or chronic induced urticaria; Sjögren's syndrome, multiple sclerosis, sweat gland abscess and food allergy, and is the LOU064 active pharmaceutical ingredient for use according to claim 25, the pharmaceutical composition for use according to claim 25, or the method for treatment according to claim 26.

29. A process for preparing an LOU064 drug substance that is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide, a. 【Chemistry 2】 To provide a suspension containing a base, water, and a solvent; b. A process comprising reacting the suspension with acrylic anhydride to provide a LOU064 active pharmaceutical ingredient that is substantially free of nitrosamine impurities, such as the nitrosamine impurity N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

30. The aforementioned base is an inorganic base (e.g., Na ２ CO ３ _K ２ CO ３ , NaOH, KOH, Mg(OH) ２ Ca(OH) ２ For example, Na ２ CO ３ The process according to claim 29, selected from ).

31. The process according to claim 29, wherein the base is a base with a low nitrite content, for example, less than about 250 ppb or less than about 120 ppb.

32. Sodium carbonate with a low nitrite content is Na ２ CO ３ .10H ２ O, K ２ CO ３ .7H ２ O, Na with a low nitrite content ２ CO ３ or K with a low nitrite content ２ CO ３ , for example, with a nitrite content of less than about 250 ppb, the process according to claim 31.

33. The process according to any one of claims 29 to 32, wherein the base is used in an amount between about 1.1 molar equivalents and about 1.2 molar equivalents.

34. The process according to any one of claims 29 to 33, wherein the solvent is selected from MeTHF, THF, alcohol (isopropanol), dichloromethane, toluene, ethyl acetate, isopropyl acetate, acetonitrile, acetone, tert-butyl methyl ether (TBME), for example ethyl acetate, and optionally the solvent has a low nitrite content (e.g., less than 1.5 ppm, less than 1 ppm, less than 0.5 ppm, less than 0.2 ppm).

35. The process according to any one of claims 29 to 34, wherein the suspension in a) comprises at least about 25 molar equivalents of water, optionally purified water.

36. The process according to claim 35, wherein the suspension or solution in a) comprises at least about 35 molar equivalents of water.

37. The process according to claim 36, wherein the suspension in a) comprises at least about 125 molar equivalents of water (for example, between about 125 and about 150 molar equivalents of water).

38. The process according to any one of claims 29 to 37, wherein the suspension in a) is heated to an internal temperature of about 50 to about 65 degrees Celsius.

39. The process according to any one of claims 29 to 38, wherein the acrylic anhydride is added to the suspension of a) as a solution in a solvent (for example, in ethyl acetate).

40. The step further includes deprotecting F7 and providing F8: 【Transformation 3】 The process according to any one of claims 29 to 39, wherein P is an amino protecting group, for example, tert-butyloxycarbonyl (BOC).

41. The process according to claim 40, wherein F7 is deprotected in the presence of an acid (e.g., HCl), and F8 is isolated after a neutralization step, e.g., with a base, e.g., with a low nitrite-containing NaOH (e.g., a solution of low nitrite-containing NaOH in purified water).

42. The process according to claim 40 or 41, wherein F8 is added to step a) without any drying step.

43. The method further includes reacting compound X6b and compound F6 to provide compound F7: 【Chemistry 4】 The process according to any one of claims 29 to 42, wherein X and Y are each independently Cl, Br, or I, and P is an amine protecting group.

44. A LOU064 active pharmaceutical ingredient prepared or prepareable by the process described in any one of claims 29 to 43.

45. A process for preparing a pharmaceutical composition, comprising mixing remibrutinib with one or more pharmaceutically acceptable excipients, wherein the one or more excipients contain less than 1.5 ppm, less than 1 ppm, less than 0.5 ppm, more preferably less than 0.2 ppm of nitrite, based on the amount of each of the excipients.

46. The process according to claim 45, wherein the excipient is sodium lauryl sulfate (SLS), and optionally the nitrite content in the SLS excipient is less than 1.5 ppm, less than 1.0 ppm, less than 0.5 ppm, or less than 0.2 ppm.

47. The process according to claim 45, wherein the excipient is magnesium stearate or magnesium stearate and sodium stearyl fumarate.

48. The process according to claim 45, wherein the excipient is crystalline cellulose, and optionally the nitrite content in the crystalline cellulose is less than 200 ppb or less than 100 ppb.

49. The process according to claim 45, wherein the excipient is a polyvinylpyrrolidone-vinyl acetate copolymer (copovidone), and optionally the nitrite content in the copovidone is less than 200 ppb or less than 100 ppb.

50. A method for evaluating a pharmaceutical composition comprising remibrutinib or a pharmaceutically acceptable salt thereof, comprising testing the composition for the presence or amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

51. A method for verifying a process for producing a pharmaceutical composition comprising remibrutinib or a pharmaceutically acceptable salt thereof, comprising testing the composition produced by the process for the presence or amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide.

52. A method for obtaining regulatory approval for a pharmaceutical composition comprising remibrutinib or a pharmaceutically acceptable salt thereof, comprising testing a sample of the composition for the presence or amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide and submitting the results of the test to the regulatory authority.

53. A batch of the composition is tested according to any one of claims 50 to 52.

54. A process for preparing a pharmaceutical product comprising remibrutinib or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients, a. Obtain a batch of remibrutinib or a pharmaceutically acceptable salt thereof; b. Determining the total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide in the batch; and c. A process comprising preparing the pharmaceutical product from the batch only if it is determined that the batch has a total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide less than about 1000 ppb (e.g., less than about 550 ppb, e.g., less than about 530 ppb; less than about 400 ppb, e.g., less than about 360 ppb; less than about 150 ppb, e.g., less than about 130 ppb; less than about 100 ppb, e.g., less than 90 ppb); less than about 50 ppb; or less than about 25 ppb.

55. A process for marketing a verified batch of a pharmaceutical product comprising remibrutinib or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients, a. To generate batches of the aforementioned pharmaceutical products; b. Determining the total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide in the batch; and d. A process of verifying the batch for sale only if it is determined that the sample of the batch has a total amount of N-(3-(6-amino-5-(2(methyl)(nitroso)amino)ethoxy)pyrimidine-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide less than about 1000 ppb (e.g., less than about 550 ppb, e.g., less than about 530 ppb; less than about 400 ppb, e.g., less than about 360 ppb; less than about 150 ppb, e.g., less than about 130 ppb; less than about 100 ppb, e.g., less than about 90 ppb); less than about 50 ppb; or less than about 25 ppb.

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