Treatment methods for cystic fibrosis

Abstract

This application describes a method of treating cystic fibrosis or a CFTR-mediated disease, comprising administering Compound I or a pharmaceutically acceptable salt thereof. This application also describes pharmaceutical compositions comprising Compound I or a pharmaceutically acceptable salt thereof, and optionally one or more additional CFTR modulators. One aspect of the disclosure provides a pharmaceutical composition comprising 250 mg of the CFTR potentiator compound, N-(2-(tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (Compound I), or an equivalent amount of a pharmaceutically acceptable salt thereof. JPEG2023552828000095.jpg3161

Description

[Technical Field]

[0001] This application claims priority to U.S. Provisional Application No. 63 / 123,928 filed on 10 December 2020, U.S. Provisional Application No. 63 / 124,575 filed on 11 December 2020, and U.S. Provisional Application No. 63 / 150,434 filed on 17 February 2021, the contents of each of these applications being incorporated by reference in their entirety.

[0002] The present invention provides pharmaceutical compositions and methods for treating cystic fibrosis. [Background technology]

[0003] Cystic fibrosis (CF) is a recessive genetic disorder that affects approximately 83,000 children and adults worldwide. Despite advances in treatment, there is no cure for CF.

[0004] In patients with cystic fibrosis (CF), mutations in CFTR, which are endogenously expressed in the respiratory epithelium, lead to decreased apical anion secretion and imbalances in ion and fluid transport. The resulting decrease in anion transport contributes to increased mucus buildup in the lungs and associated microbial infections, which ultimately lead to death in CF patients. In addition to respiratory disease, CF patients typically suffer from gastrointestinal disorders and pancreatic insufficiency that, if left untreated, can be fatal. Furthermore, the majority of men with cystic fibrosis are infertile, and women with cystic fibrosis experience reduced fertilization rates.

[0005] Sequence analysis of the CFTR gene has revealed various disease-causing mutations (Cutting, G et al. (1990) Nature 346:366-369, Dean, M. et al. (1990) Cell 61:863:870, and Kerem, BS. et al. (1989) Science 245:1073-1080, Kerem, BS et al. (1990) Proc. Natl. Acad. Sci. USA 87:8447-8451). To date, more than 2000 mutations in the CF gene have been identified. CF mutations are listed in the "Cystic Fibrosis Mutation Database" at http: / / www.genet.sickkids.on.ca / app, and the entire database is incorporated herein by reference. The most common disease-causing mutation is the deletion of phenylalanine at position 508 of the CFTR amino acid sequence, commonly referred to as the F508del mutation. This mutation occurs in approximately 90% of cystic fibrosis cases and is associated with severe disease.

[0006] The deletion of residue 508 in CFTR prevents the nascent protein from folding correctly. This makes it impossible for the mutant protein to exit the endoplasmic reticulum (ER) and be transported to the plasma membrane. As a result, the number of CFTR channels for anion transport present in the membrane is far fewer than that observed in cells expressing wild-type CFTR, i.e., CFTR without the mutation. In addition to the transport impairment, this mutation leads to a defect in channel gating. The reduction in the number of channels in the membrane and the gating defect together lead to a decrease in anion and fluid transport across the epithelium. (Quinton, PM (1990), FASEB J.4:2709-2727). Channels defective due to the F508del mutation are less functional than wild-type CFTR channels, but are still functional. (Dalemans et al. (1991), Nature Lond. 354:526-528, Pasyk and Foskett (1995), J. Cell. Biochem. 270:12347-50). In addition to F508del, other disease-causing mutations in the CFTR that result in defective transport, synthesis, and / or channelgating may be upregulated or downregulated, altering anion secretion and modifying disease progression and / or severity.

[0007] CFTR is a cAMP / ATP-mediated anion channel expressed in various cell types, including absorptive and secretory epithelial cells, where CFTR regulates transmembrane anion flux and the activity of other ion channels and proteins. In epithelial cells, normal CFTR function is essential for maintaining systemic electrolyte transport, including in respiratory and digestive tissues. CFTR is composed of 1480 amino acids encoding a protein consisting of tandem repeats of transmembrane domains, each containing six transmembrane helices and a nucleotide-binding domain. Two transmembrane domains are linked to multiple phosphorylation sites that regulate channel activity and cell transport via a large polarity regulatory (R) domain.

[0008] Chloride transport occurs through the coordinated activity of ENaC (epithelial sodium channel) and CFTR present on the apical membrane, and Na + -K + -ATPase pump and Cl - channel. Secondary active transport of chloride from the lumen side results in the accumulation of intracellular chloride, which then passively leaves the cell via the Cl - channel and can result in vectorial transport. The Na + / 2Cl - / K + cotransporter, the Na + -K + -ATPase pump and the basolateral membrane K + channel, as well as the placement of CFTR on the lumen side, regulate chloride secretion. Since probably water itself is not actively transported, its flow across the epithelium depends on a small transepithelial osmotic gradient created by large fluxes of sodium and chloride.

[0009] Several CFTR modulating compounds have recently been identified. However, there is still a need for compounds that can treat or reduce the severity of cystic fibrosis and other CFTR-mediated diseases, particularly the more severe forms of these diseases.

Prior Art Documents

Non-Patent Documents

[0010]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

[0011] Accordingly, one aspect of the present disclosure provides a pharmaceutical composition comprising 250 mg of a CFTR enhancer compound, N-(2-(tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (Compound I) or a comparable amount of a pharmaceutically acceptable salt thereof. Compound I may be represented as having the following structure: [ka]

[0012] Compared to 150 mg of ibacaftol every 12 hours (i.e., twice daily at 12-hour intervals) and 150 mg of compound I once daily (i.e., once daily), administering 250 mg of compound I once daily was found to improve the therapeutic profile as measured by sweat chloride (SwCl).

[0013] In some embodiments, the Disclosure provides a pharmaceutical composition comprising 250 mg of Compound I and / or an equivalent amount thereof, wherein the composition may further comprise at least one additional active pharmaceutical ingredient and / or at least one carrier. Yet another embodiment of the Disclosure is a method for treating cystic fibrosis of a CFTR-mediated disease, comprising administering Compound I or an equivalent amount thereof, optionally, as part of a pharmaceutical composition comprising at least one additional ingredient, to a subject in need of treatment. In some embodiments, the pharmaceutical composition of the present invention comprises 250 mg of Compound I (or an equivalent amount thereof), 21.24 mg of Compound II, calcium salt hydrate form D, and 100 mg of Compound III (or an equivalent amount thereof). In some embodiments, the pharmaceutical composition of the present invention comprises 125 mg of compound I (or an equivalent amount of its pharmaceutically acceptable salt), 10.62 mg of compound II, calcium salt hydrate form D, and 50 mg of compound III (or an equivalent amount of its pharmaceutically acceptable salt).

[0014] One embodiment is a method for treating cystic fibrosis, a CFTR-mediated disease, comprising 250 mg of N-(2-(tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (compound I), alone, or 21.24 mg of (14S)-8-[3-(2-{dispiro[2.0.2.1]heptan-7-yl}ethoxy)-1H-pyrazole-1-yl]-12,12-dimethyl-2λ6-thia-3,9,11,18,23-pentazatetrasyl The present invention provides a method comprising administering chloro[17.3.1.111,14.05,10]tetracosa-1(22),5,7,9,19(23),20-hexaen-2,2,4-trione (compound II) calcium salt hydrate form D, and / or 50-100 mg of (R)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(1-(2,3-dihydroxypropyl)-6-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indole-5-yl)cyclopropanecarboxamide (compound III) in combination.

[0015] In some embodiments, 250 mg of Compound I (or an equivalent amount of its pharmaceutically acceptable salt) is administered in the same composition as 20 mg of Compound II calcium salt hydrate form D. In some embodiments, 250 mg of Compound I is administered in the same composition as 21.24 mg of Compound II calcium salt hydrate form D and 100 mg of Compound III (or an equivalent amount of its pharmaceutically acceptable salt). In some embodiments, a composition containing 250 mg of Compound I (or an equivalent amount of its pharmaceutically acceptable salt) is administered simultaneously with a separate composition containing 21.24 mg of Compound II calcium salt hydrate form D and / or 100 mg of Compound III (or an equivalent amount of its pharmaceutically acceptable salt). In some embodiments, 250 mg of Compound I is administered in the same composition as 21.24 mg of Compound II calcium salt hydrate form D and 100 mg of Compound III (or an equivalent amount of its pharmaceutically acceptable salt). In some embodiments, 250 mg of compound I, 21.24 mg of compound II calcium salt hydrate form D, and 100 mg of compound III (or an equivalent amount thereof of a pharmaceutically acceptable salt) are administered once daily in two equidose compositions. [Brief explanation of the drawing]

[0016] [Figure 1] The XRPD pattern of crystalline compound I (free form) morphology A is presented. [Figure 2] The 13C solid-state NMR spectrum of crystalline compound I (free form) form A is shown. [Figure 3] We present the XRPD pattern of crystalline compound I calcium salt hydrate form A. [Figure 4] The 13C solid-state NMR spectrum of compound I calcium salt hydrate form A is shown. [Figure 5] We present the XRPD pattern of crystalline compound I calcium salt hydrate morph D. [Figure 6] The 13C solid-state NMR spectrum of compound I calcium salt hydrate form D is shown. [Modes for carrying out the invention]

[0017] definition As used throughout this disclosure, “Compound I” is N-(2-(tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide, which may be shown as having the following structure: [ka] Compound I may be in the form of a pharmaceutically acceptable salt. Compound I and its pharmaceutically acceptable salts have been previously described in U.S. Patents 8,865,902, 9,181,192, and 9,512,079, and in International Patent Publications 2012 / 158885, 2014 / 078842, 2017 / 053455, and 2018 / 080591, each of which is incorporated herein by reference.

[0018] In some embodiments, the isotopic enrichment factors of each deuterium in compound I may vary. The term “isotopic enrichment factor” refers to the ratio between the isotopic abundance and the naturally occurring abundance of a particular isotope. In some embodiments, the isotopic enrichment factors of each designated deuterium atom in compound I are at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).

[0019] As used herein, "Compound II" refers to (14S)-8-[3-(2-{dispiro[2.0.2.1]heptan-7-yl}ethoxy)-1H-pyrazole-1-yl]-12,12-dimethyl-2λ6-thia-3,9,11,18,23-pentazatetracyclo[17.3.1.111,14.05,10]tetracosa-1(22),5,7,9,19(23),20-hexaene-2,2,4-trione, which may be represented by the following structure. [ka] Compound II, its deuterated derivatives, and pharmaceutically acceptable salts were first described in International Patent Publication 2019 / 161078 (incorporated herein by reference).

[0020] In some embodiments, compound II is in the form of calcium salt hydrate form D. 20 mg of compound II corresponds to 21.24 mg of compound II calcium salt hydrate form D. In some embodiments, compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta. In some embodiments, compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram having (a) signals at 6.1±0.2° 2-theta, 16.2±0.2° 2-theta, and 22.8±0.2° 2-theta, and (b) one or more signals at 5.5±0.2° 2-theta, 15.5±0.2° 2-theta, 19.7±0.2° 2-theta, 21.5±0.2° 2-theta, 22.1±0.2° 2-theta, 23.0±0.2° 2-theta, and 27.6±0.2° 2-theta.

[0021] In some embodiments, compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram having (a) signals at 6.1±0.2° 2-theta, 16.2±0.2° 2-theta, and 22.8±0.2° 2-theta, and (b) two or more signals from among 5.5±0.2° 2-theta, 15.5±0.2° 2-theta, 19.7±0.2° 2-theta, 21.5±0.2° 2-theta, 22.1±0.2° 2-theta, 23.0±0.2° 2-theta, and 27.6±0.2° 2-theta. In some embodiments, compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram having (a) signals at 6.1±0.2° 2-theta, 16.2±0.2° 2-theta, and 22.8±0.2° 2-theta, and (b) three or more signals from among 5.5±0.2° 2-theta, 15.5±0.2° 2-theta, 19.7±0.2° 2-theta, 21.5±0.2° 2-theta, 22.1±0.2° 2-theta, 23.0±0.2° 2-theta, and 27.6±0.2° 2-theta. In some embodiments, compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram having (a) signals at 6.1±0.2° 2-theta, 16.2±0.2° 2-theta, and 22.8±0.2° 2-theta, and (b) four or more signals from among 5.5±0.2° 2-theta, 15.5±0.2° 2-theta, 19.7±0.2° 2-theta, 21.5±0.2° 2-theta, 22.1±0.2° 2-theta, 23.0±0.2° 2-theta, and 27.6±0.2° 2-theta.

[0022] In some embodiments, Compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram having signals at 6.1±0.2° 2-theta, 16.2±0.2° 2-theta, 22.8±0.2° 2-theta, and 27.6±0.2° 2-theta. In some embodiments, Compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram having signals at 6.1±0.2° 2-theta, 15.5±0.2° 2-theta, 16.2±0.2° 2-theta, 19.7±0.2° 2-theta, 22.8±0.2° 2-theta, and 27.6±0.2° 2-theta. In some embodiments, Compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram substantially similar to that in Figure 5.

[0023] In some embodiments, compound II calcium salt hydrate form D has one or more peaks selected from 179.8±0.2 ppm, 130.2±0.2 ppm, 125.6±0.2 ppm, 120.9±0.2 ppm, 55.2±0.2 ppm, 44.3±0.2 ppm, 35.0±0.2 ppm, and 1.6±0.2 ppm. 13 C solid-state nuclear magnetic resonance ( 13 It is characterized as having a C ssNMR spectrum. In some embodiments, compound II calcium salt hydrate form D has two or more peaks selected from 179.8±0.2 ppm, 130.2±0.2 ppm, 125.6±0.2 ppm, 120.9±0.2 ppm, 55.2±0.2 ppm, 44.3±0.2 ppm, 35.0±0.2 ppm, and 1.6±0.2 ppm. 13 It is characterized as having a C ssNMR spectrum. In some embodiments, compound II calcium salt hydrate form D has three or more peaks selected from 179.8±0.2 ppm, 130.2±0.2 ppm, 125.6±0.2 ppm, 120.9±0.2 ppm, 55.2±0.2 ppm, 44.3±0.2 ppm, 35.0±0.2 ppm, and 1.6±0.2 ppm. 13It is characterized as having a C ssNMR spectrum. In some embodiments, compound II calcium salt hydrate form D has four or more peaks selected from 179.8±0.2 ppm, 130.2±0.2 ppm, 125.6±0.2 ppm, 120.9±0.2 ppm, 55.2±0.2 ppm, 44.3±0.2 ppm, 35.0±0.2 ppm, and 1.6±0.2 ppm. 13 It is characterized as having a C ssNMR spectrum. In some embodiments, compound II calcium salt hydrate form D has five or more peaks selected from 179.8±0.2 ppm, 130.2±0.2 ppm, 125.6±0.2 ppm, 120.9±0.2 ppm, 55.2±0.2 ppm, 44.3±0.2 ppm, 35.0±0.2 ppm, and 1.6±0.2 ppm. 13 It is characterized as having a C ssNMR spectrum. In some embodiments, compound II calcium salt hydrate form D has six or more peaks selected from 179.8±0.2 ppm, 130.2±0.2 ppm, 125.6±0.2 ppm, 120.9±0.2 ppm, 55.2±0.2 ppm, 44.3±0.2 ppm, 35.0±0.2 ppm, and 1.6±0.2 ppm. 13 It is characterized by having a C ssNMR spectrum.

[0024] In some embodiments, the compound II calcium salt hydrate form D has one or more peaks selected from 130.2±0.2 ppm, 125.6±0.2 ppm, and 35.0±0.2 ppm. 13 It is characterized as having a C ssNMR spectrum. In some embodiments, compound II calcium salt hydrate form D has two or more peaks selected from 130.2 ± 0.2 ppm, 125.6 ± 0.2 ppm, and 35.0 ± 0.2 ppm. 13 It is characterized as having a C ssNMR spectrum. In some embodiments, compound I calcium salt hydrate form D has peaks at 130.2±0.2 ppm, 125.6±0.2 ppm, and 35.0±0.2 ppm. 13It is characterized by having a C ssNMR spectrum.

[0025] In some embodiments, Compound II calcium salt hydrate form D has peaks at (a) 130.2±0.2 ppm, 125.6±0.2 ppm, and / or 35.0±0.2 ppm, and (b) peaks at 176.9±0.2 ppm, 160.9±0.2 ppm, 142.0±0.2 ppm, and / or 98.6±0.2 ppm. 13 It is characterized as having a C ssNMR spectrum. In some embodiments, compound II calcium salt hydrate form D has peaks at (a) 130.2±0.2 ppm, 125.6±0.2 ppm, and / or 35.0±0.2 ppm, and (b) peaks at 176.9±0.2 ppm, 160.9±0.2 ppm, 142.0±0.2 ppm, and 98.6±0.2 ppm. 13 It is characterized as having a C ssNMR spectrum. In some embodiments, compound II calcium salt hydrate form D has peaks at (a) 130.2±0.2 ppm, 125.6±0.2 ppm, and 35.0±0.2 ppm, and (b) peaks at 176.9±0.2 ppm, 160.9±0.2 ppm, 142.0±0.2 ppm, and / or 98.6±0.2 ppm. 13 It is characterized as having a C ssNMR spectrum. In some embodiments, compound II calcium salt hydrate form D has peaks at 130.2±0.2 ppm, 125.6±0.2 ppm, 35.0±0.2 ppm, 176.9±0.2 ppm, 160.9±0.2 ppm, 142.0±0.2 ppm, and 98.6±0.2 ppm. 13 It is characterized by having a C ssNMR spectrum.

[0026] In some embodiments, the compound II calcium salt hydrate form D is substantially similar to that shown in Figure 6. 13 Characterized by C ssNMR spectra.

[0027] In some embodiments, compound II calcium salt hydrate form D is Cu K α The crystal system is characterized by a triclinic crystal system, P1 space group, and the following unit cell dimensions, measured at 100 K using a Bruker diffractometer equipped with a linear (λ=1.5478 Å) and complementary metal-oxide-semiconductor (CMOS) detector. [Table 1]

[0028] As used throughout this disclosure, “Compound III” refers to (R)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(1-(2,3-dihydroxypropyl)-6-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indole-5-yl)cyclopropanecarboxamide. [ka] In some embodiments, compound III is in the form of a pharmaceutically acceptable salt. Compound III and its pharmaceutically acceptable salts are previously disclosed in International Patent Publication 2010 / 053471 (incorporated herein by reference).

[0029] As used herein, "CFTR" means cystic fibrosis membrane conductance regulator.

[0030] As used herein, “mutation” may refer to a mutation in the CFTR gene or the CFTR protein. “CFTR gene mutation” refers to a mutation in the CFTR gene, and “CFTR protein mutation” refers to a mutation in the CFTR protein. Generally, a genetic defect or mutation, or a change in nucleotides in a gene, results in a mutation or frameshift in the CFTR protein translated from that gene.

[0031] The term "F508del" refers to a mutant CFTR protein that lacks the amino acid phenylalanine at position 508.

[0032] As used herein, a patient who is "homozygous" for a particular gene mutation has the same mutation in each allele.

[0033] As used herein, a patient who is "heterozygous" for a particular gene mutation has a specific mutation in one allele and a different mutation in the other allele.

[0034] As used herein, the term “modulator” refers to a compound that increases the activity of a biological compound or molecule, such as a protein. For example, a CFTR modulator is a compound that increases the activity of CFTR. The increase in activity brought about by a CFTR modulator includes, but is not limited to, compounds that correct, enhance, stabilize, and / or amplify CFTR.

[0035] As used herein, the term "CFTR collector" refers to a compound that facilitates the processing and transport of CFTR, thereby increasing the amount of CFTR on the cell surface. Compounds II and III disclosed herein are CFTR collectors.

[0036] As used herein, the term "CFTR enhancer" refers to a compound that increases the channel activity of CFTR proteins located on the cell surface, resulting in enhanced ion transport. Compound I disclosed herein is a CFTR enhancer.

[0037] As used herein, the terms “active pharmaceutical ingredient” or “therapeutic agent” (“API”) refer to a biologically active compound.

[0038] As used herein, the term “pharmaceutically acceptable salt” refers to a salt form of the compound of the Disclosure in which the salt is nontoxic. Pharmaceutically acceptable salts of the compounds of the Disclosure include those derived from suitable inorganic and organic acids and bases. Pharmaceutically acceptable salts are well known in the art. For example, SMBerge, et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19.

[0039] As used herein, the term “amorphous” refers to a solid material that does not have long-range order in the position of its molecules. An amorphous solid is a generally supercooled liquid in which molecules are randomly arranged, such that there is no clearly defined arrangement, e.g., no molecular packing and no long-range order. Amorphous solids are generally isotropic, that is, they exhibit similar properties in all directions and do not have a distinct melting point. For example, an amorphous material is a solid material that does not have sharp, characteristic crystalline peaks in its X-ray powder diffraction (XRPD) pattern (i.e., is not crystalline as determined by XRPD). Instead, one or more broad peaks (e.g., halos) are observed in its XRPD pattern. Broad peaks are characteristic of amorphous solids. For a comparison of XRPDs of amorphous and crystalline materials, see US2004 / 0006237. In some embodiments, the solid material may contain amorphous compounds, for example, the solid material may be characterized by the absence of sharp, characteristic crystalline peaks in its XRPD spectrum (i.e., the solid material is not crystalline but amorphous as determined by XRPD). Instead, one or more broad peaks (e.g., halos) may be observed in the XRPD pattern of the solid material. For a comparison of XRPDs of amorphous and crystalline materials, see US2004 / 0006237. Solid materials containing amorphous compounds may be characterized by a broader temperature range of melting of the solid material compared to, for example, the melting range of a pure crystalline solid. Crystalline or amorphous forms can be characterized using other techniques, such as Raman spectroscopy, infrared spectroscopy, and solid-state NMR.

[0040] In some embodiments, the solid material may include a mixture of crystalline and amorphous solids. A solid material prepared to include an amorphous compound may, for example, contain up to 30% crystalline solid. In some embodiments, a solid material prepared to include an amorphous compound may also contain, for example, up to 25%, 20%, 15%, 10%, 5%, or 2% crystalline solid. In embodiments in which the solid material includes a mixture of crystalline and amorphous solids, characterization data such as XRPD may include indices for both crystalline and amorphous solids.

[0041] As used herein, the term “substantially amorphous” refers to a solid material that has little or no long-range order at the molecular positions. For example, a substantially amorphous material has a degree of crystallinity of less than 15% (e.g., less than 10%, or less than 5%, or less than 2%). Note that the term “substantially amorphous” also includes the descriptive term “amorphous,” which refers to a material that has no degree of crystallinity (0%).

[0042] As used herein, the term “substantially crystalline” refers to a solid material that contains little to no amorphous molecules. For example, a substantially crystalline material has less than 15% amorphous molecules (e.g., less than 10%, less than 5%, or less than 2%). Note that the term “substantially crystalline” also includes the descriptive term “crystalline,” which refers to a material that is 100% crystalline.

[0043] As used herein, the term "XRPD" refers to an analytical characterization method for X-ray powder diffraction. The XRPD patterns disclosed herein are recorded in transmission or reflection configuration using a diffractometer under ambient conditions.

[0044] As used herein, the term “ambient conditions” means room temperature, ambient conditions, and uncontrolled humidity conditions. The terms “room temperature” and “ambient temperature” mean 15°C to 30°C.

[0045] As used herein, the terms “X-ray powder diffractogram,” “X-ray powder diffraction pattern,” “XRPD pattern,” and “XRPD spectrum” are interchangeable to refer to experimentally obtained patterns plotting signal positions (on the x-axis) against signal intensities (on the y-axis). For amorphous materials, an X-ray powder diffractogram may contain one or more broad signals; for crystalline materials, an X-ray powder diffractogram may contain one or more signals, each identified by its angular value, measured as 2θ degrees (°2θ), shown on the x-axis of the X-ray powder diffractogram, which may be expressed as “...2-theta signal,” “[a]2-theta signal of…,” and / or “2-theta signal of at least… selected from….”

[0046] As used herein, “signal” or “peak” refers to a point in an XRPD pattern where the intensity measured by counting is maximum. Those skilled in the art will recognize that one or more signals (or peaks) in an XRPD pattern may overlap and, for example, may not be apparent to the naked eye. In fact, those skilled in the art will recognize that several methods recognized in the art, such as the Rietveld method, can and are suitable for determining whether a signal is present in a pattern.

[0047] As used herein, "a signal at 2° theta" refers to the X-ray reflection position measured and observed in an X-ray powder diffraction experiment (°2θ).

[0048] The repeatability of the measured angle values ​​is within ±0.2°2θ, meaning that the angle values ​​can be the listed angle values ​​+0.2°2theta, angle values ​​-0.2°2theta, or any value between the two endpoints (between angle values ​​+0.2°2theta and angle values ​​-0.2°2theta).

[0049] The terms "signal intensity" and "peak intensity" are interchangeable in referring to the relative signal intensity within a given X-ray powder diffractogram. Factors that may affect the relative signal intensity or peak intensity include the thickness of the sample and the preferred orientation (e.g., crystalline particles are not randomly distributed).

[0050] As used herein, an X-ray powder diffractogram is considered "substantially similar to that in a particular figure" if at least 90% of the signals in two diffractograms overlap, for example, at least 95%, at least 98%, or at least 99%. In determining "substantially similar," a person skilled in the art will understand that variations in intensity and / or signal position can exist in XRPD diffractograms even for the same crystalline form. Accordingly, a person skilled in the art will understand that the maximum signal value (at 2 degrees theta) in an XRPD diffractogram is generally identified as ±0.2 degrees 2 degrees theta of the reported value, which is a variation recognized in the art.

[0051] When used in this specification, 13 A C ssNMR spectrum is considered "substantially similar to that of [a particular] figure" if at least 90% of the signals in the two spectra overlap, such as at least 95%, at least 98%, or at least 99%. In determining "substantially similar," a person skilled in the art will understand that variations in intensity and / or signal position can exist in ssNMR spectra even for the same crystalline form. Accordingly, a person skilled in the art will understand that a chemical shift in an ssNMR spectrum (parts per million (ppm) as referred herein) generally means that the value is specified as ±0.2 ppm of the reported value, which is a variation recognized in the art.

[0052] As used herein, the term "X-ray powder diffractogram having a signal at 2-theta values" refers to an XRPD pattern that includes an X-ray reflection position (°2-theta) measured and observed in an X-ray powder diffraction experiment.

[0053] As used herein, the term "DSC" refers to the differential scanning calorimetry analytical method.

[0054] As used herein, the term “solvent” refers to any liquid in which the product is at least partially soluble (solubility of the product > 1 g / l).

[0055] As used herein, the term “dispersion” refers to a dispersion system in which a dispersed phase, which is a single substance, is distributed as distinct units throughout a second substance (a continuous phase or vehicle). The size of the dispersed phase can vary considerably (e.g., colloidal particles ranging from nanometers to several microns in size). Generally, the dispersed phase can be solid, liquid, or gas. In the case of a solid dispersion, both the dispersed phase and the continuous phase are solids. In pharmaceutical applications, a solid dispersion may contain a crystalline drug (dispersed phase) in an amorphous polymer (continuous phase), or alternatively, an amorphous drug (dispersed phase) in an amorphous polymer (continuous phase). In some embodiments, the solid dispersion contains the polymer constituting the dispersed phase and the drug constituting the continuous phase. Alternatively, the solid dispersion may contain the drug constituting the dispersed phase and the polymer constituting the continuous phase.

[0056] The terms "patient" and "subject" are used interchangeably and refer to animals, including humans.

[0057] As used herein, terms such as “treatment” and “to treat” generally mean improvement of CF or its symptoms in the subject, or reduction of the severity of one or more of the symptoms of CF. As used herein, “treatment” includes, but is not limited to, increased growth, improved weight gain, reduced mucous membranes in the lungs, improved pancreatic and / or hepatic function, reduction of lung infections, and / or reduction of cough or shortness of breath in the subject. Improvement of any of these symptoms or reduction of their severity can be readily assessed according to standard methods and techniques known in the art.

[0058] As used herein, the term “in combination with” means that, when referring to two or more compounds, drugs, or additional active pharmaceutical ingredients, the two or more compounds, drugs, or active pharmaceutical ingredients are administered to the patient before or after each other, or simultaneously with each other.

[0059] When used in relation to the dose, volume, or weight percentage of an ingredient in a composition or dosage form, the terms “about” and “approximately” include a specified dose, volume, or weight percentage value, or a range of doses, volumes, or weight percentages that are recognized by those skilled in the art as providing an equivalent pharmacological effect to that obtained from a specified dose, volume, or weight percentage. The terms “about” and “approximately” may also refer to an acceptable error to a particular value determined by those skilled in the art, which depends in part on how that value is measured or determined. In some embodiments, the terms “about” and “approximately” mean within 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, or 0.5% of a given value or range.

[0060] Those skilled in the art will recognize that when an amount of “a compound or a pharmaceutically acceptable salt thereof” is disclosed, the amount of the pharmaceutically acceptable salt form of the compound corresponds to an amount equivalent to the concentration of the free base of the compound. Note that the amounts of compounds or their pharmaceutically acceptable salts disclosed herein are based on their free base forms. For example, “100 mg of Compound I and at least one compound selected from its pharmaceutically acceptable salts” includes 100 mg of Compound I and a concentration of a pharmaceutically acceptable salt of Compound I equivalent to 100 mg of Compound I.

[0061] Suitable pharmaceutically acceptable salts are disclosed, for example, in SMBerge, et al. J. Pharmaceutical Sciences, 1977, 66, 1-19. For example, Table 1 of that paper provides the following pharmaceutically acceptable salts. [Table 2]

[0062] Non-limiting examples of pharmaceutically acceptable acid addition salts include salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, or perchloric acid; salts formed with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid; and salts formed using other methods used in the art, such as ion exchange. Non-limiting examples of pharmaceutically acceptable salts include adipine, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy- Examples include ethanesulfonates, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malons, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectins, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propions, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, and valersates. Pharmaceutically acceptable salts derived from suitable bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and N + (C 1-4This includes alkyl) tetra salts. This disclosure also assumes quaternization of any basic nitrogen-containing group of the compounds disclosed herein. Preferred non-limiting examples of alkali metal salts and alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium. Further non-limiting examples of pharmaceutically acceptable salts include ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfons, and aryl sulfons. Other preferred non-limiting examples of pharmaceutically acceptable salts include besylates and glucosamine salts.

[0063] Treatment method CFTR mutations can affect either the amount of CFTR, i.e., the number of CFTR channels on the cell surface, or CFTR function, i.e., the functional ability of each channel to open and transport ions. Mutations affecting CFTR amount include mutations that cause synthesis defects (Class I defects), mutations that cause processing and transport defects (Class II defects), mutations that cause decreased CFTR synthesis (Class V defects), and mutations that reduce CFTR surface stability (Class VI defects). Mutations affecting CFTR function include mutations that cause gating defects (Class III defects) and mutations that cause conductance defects (Class IV defects). Some CFTR mutations exhibit characteristics of multiple classes.

[0064] In some embodiments, a method for treating cystic fibrosis in a patient is disclosed herein, comprising administering an effective amount of the compound of the Disclosure, a pharmaceutically acceptable salt thereof, or any of the aforementioned deuterated analogs or pharmaceutical compositions to a patient, such as a human, the patient being a patient having cystic fibrosis. In some embodiments, the patient has the F508del / minimal function (MF) genotype, the F508del / F508del genotype (homozygous for the F508del mutation), the F508del / gating genotype, or the F508del / residual function (RF) genotype. In some embodiments, the patient is heterozygous and has one F508del mutation.

[0065] As used herein, “minimal function (MF) mutation” refers to a CFTR gene mutation associated with minimal CFTR function (a CFTR protein that is little to no function), and includes, for example, mutations associated with severe defects in the ability of CFTR channels to open and close, known as channel gating defects or “gating mutations,” mutations associated with severe defects in CFTR cellular processing and its delivery to the cell surface, mutations associated with no (or minimal) CFTR synthesis, and mutations associated with severe defects in channel conductance.

[0066] In some embodiments, the patient is heterozygous, having the F508del mutation in one allele and a mutation selected from Table 2 in the other allele. [Table 3-1] [Table 3-2]

[0067] In some embodiments, the patient is heterozygous, having the F508del mutation in one allele and a mutation selected from Table 3 in the other allele. [Table 4]

[0068] In some embodiments, the patient has at least one mutation selected from Table 4. In some embodiments, the patient does not have the F508del mutation but has at least one mutation selected from Table 4. [Table 5-1] [Table 5-2]

[0069] In some embodiments, the disclosure also relates to therapeutic methods using isotopically labeled derivatives of the aforementioned compounds. In some embodiments, isotopically labeled derivatives of the aforementioned compounds or pharmaceutically acceptable salts thereof are characterized in that one or more atoms are substituted (isotope-labeled) by atoms or groups of atoms having a different atomic mass or mass number than those of naturally occurring atoms. Examples of commercially available isotopes suitable for the disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, for example, 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F, and 36 Cl is one example.

[0070] Isotope-labeled compounds and salts can be used in several beneficial ways. They may be suitable for various types of assays, such as drug and / or substrate tissue distribution assays. For example, tritium ( 3 H) Labeling and / or carbon-14 ( 14C) Labeled compounds are particularly useful in various types of assays, such as substrate tissue distribution assays, due to their relatively simple preparation and excellent detection ability. For example, deuterium ( 2 H) Labeled compounds are therapeutically useful, non 2 It has potential therapeutic advantages over H-labeled compounds. Generally speaking, deuterium ( 2 H) Labeled compounds and salts may have higher metabolic stability compared to unlabeled compounds due to the kinetic isotope effects described below. Higher metabolic stability directly translates to a desired increase in in vivo half-life or lower dosage. Isotopically labeled compounds and salts can typically be prepared by following the procedures disclosed in the Synthesis Schemes and Related Descriptions, Examples, and Preparations sections of this specification, and by replacing unisotopically labeled reactants with readily available isotopically labeled reactants.

[0071] In some embodiments, the isotope-labeled compound and salt are deuterium ( 2 H) Labeled compounds and salts. In some specific embodiments, the isotope-labeled compounds and salts are deuterium ( 2 H) Labeled, in which one or more hydrogen atoms are replaced by deuterium. In the chemical structure, deuterium is represented as "D".

[0072] deuterium( 2 H) Labeled compounds and salts can manipulate the oxidative metabolism of compounds by first-order kinetic isotope effects. First-order kinetic isotope effects are changes in the rate of chemical reactions resulting from the exchange of isotopic nuclides, which are subsequently caused by a change in the ground state energy required for covalent bond formation after this isotope exchange. The exchange of heavier isotopes usually results in a decrease in the ground state energy of the chemical bond, and therefore a decrease in the rate-determining bond cleavage. If the bond cleavage occurs within or near the saddle point region along the coordination of a multi-product reaction, the product distribution ratio can change significantly. For illustrative purposes, if deuterium is bonded to a carbon atom in an inexchangeable position, k M / k DA typical rate difference is between 2 and 7. For further consideration, see S.L. Harbeson and R.D. Tung, Deuterium In Drug Discovery and Development, Ann. Rep. Med. Chem. 2011, 46, 403-417, which is incorporated herein by reference in its entirety.

[0073] The concentration of the isotope (e.g., deuterium) incorporated into the isotope-labeled compounds and salts of this disclosure may be defined by the isotope enrichment factor. As used herein, the term “isotope enrichment factor” means the ratio between the isotopic abundance and the natural abundance of a given isotope. In some embodiments, when the substituent in the compounds of the present disclosure is deuterium, such compounds have an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation in each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).

[0074] When discovering and developing therapeutic agents, those skilled in the art attempt to optimize pharmacokinetic parameters while maintaining desirable in vitro properties. It may be reasonable to assume that many compounds with insufficient pharmacokinetic profiles are susceptible to oxidative metabolism.

[0075] One embodiment disclosed herein provides a method for treating cystic fibrosis and other CFTR-mediated diseases, comprising administering 250 mg of compound I (or an equivalent amount of a pharmaceutically acceptable salt thereof). In some embodiments, 250 mg of compound I (or an equivalent amount of a pharmaceutically acceptable salt thereof) is administered once daily as a single dose alone or in combination with another CFTR modulator. In some embodiments, 250 mg of compound I (or an equivalent amount of a pharmaceutically acceptable salt thereof) is administered once daily as two 125 mg doses alone or in combination with another CFTR modulator.

[0076] One embodiment disclosed herein provides a method for treating cystic fibrosis and other CFTR-mediated diseases by daily administration of 250 mg of Compound I (or an equivalent amount of its pharmaceutically acceptable salt) in combination with 21.24 mg of Compound II calcium salt hydrate form D and 100 mg of Compound III (or an equivalent amount of its pharmaceutically acceptable salt). In some embodiments, 250 mg of Compound I (or an equivalent amount of its pharmaceutically acceptable salt), 21.24 mg of Compound II calcium salt hydrate form D, and 100 mg of Compound III (or an equivalent amount of its pharmaceutically acceptable salt) are administered daily in separate pharmaceutical compositions. In some embodiments, 250 mg of Compound I (or an equivalent amount of its pharmaceutically acceptable salt), 21.24 mg of Compound II calcium salt hydrate form D, and 100 mg of Compound III (or an equivalent amount of its pharmaceutically acceptable salt) are administered daily in a single pharmaceutical composition. In some embodiments, 250 mg of Compound I (or an equivalent amount of its pharmaceutically acceptable salt), 21.24 mg of Compound II calcium salt hydrate form D, and 100 mg of Compound III (or an equivalent amount of its pharmaceutically acceptable salt) are administered together once daily in two equivalent pharmaceutical compositions.

[0077] Pharmaceutical composition Another aspect of the present invention provides a pharmaceutical composition for use in the treatment of cystic fibrosis. In some embodiments, the pharmaceutical composition of the present invention comprises 250 mg of compound I (or an equivalent amount of a pharmaceutically acceptable salt thereof). In some embodiments, the pharmaceutical composition of the present invention comprises 125 mg of compound I (or an equivalent amount of a pharmaceutically acceptable salt thereof).

[0078] In some embodiments, the pharmaceutical composition of the present invention comprises 250 mg of Compound I, 21.24 mg of Compound II calcium salt hydrate form D, and 100 mg of Compound III. In some embodiments, the pharmaceutical composition of the present invention comprises 125 mg of Compound, 10.62 mg of Compound II calcium salt hydrate form D, and 50 mg of Compound III.

[0079] In some embodiments, the pharmaceutical compositions disclosed herein (e.g., tablets) comprise a first solid dispersion (e.g., a spray-dried dispersion) containing compound I, and a second solid dispersion (e.g., a spray-dried dispersion) containing compound III. Solid dispersions of non-deuterated analogs of compound I, and methods for preparing such dispersions, are disclosed in PCT Publication 2007 / 079139, incorporated herein by reference. These same solid dispersions are suitable for use with compound I. Solid dispersions of compound III and methods for preparing them are disclosed in PCT Publications 2011 / 119984 and 2015 / 160787, incorporated herein by reference.

[0080] In some embodiments, the pharmaceutical composition of the present invention comprises a solid dispersion containing about 39.9% by weight of Compound I relative to the weight of the composition (the solid dispersion contains 80% by weight of Compound I, 19.5% by weight of hypromellose acetate succinate, and 0.5% by weight of sodium lauryl sulfate), about 2.7% by weight of Compound II calcium salt hydrate form D relative to the weight of the composition, and a solid dispersion containing about 16.0% by weight of Compound III relative to the weight of the composition (the solid dispersion contains 80% by weight of Compound I and 20% by weight of hypromellose, relative to the weight of the solid dispersion).

[0081] Any suitable pharmaceutical composition known in the art can be used with Compound I, Compound II calcium salt hydrate form D, and Compound III. Several exemplary pharmaceutical compositions for Compound I and its pharmaceutically acceptable salts can be found in US8,865,902, US9,181,192, US9,512,079, WO2017 / 053455, and WO2018 / 080591, all of which are incorporated herein by reference. Exemplary pharmaceutical compositions containing Compound II and its pharmaceutically acceptable salts are disclosed in WO2019 / 161078 and WO2020 / 102346. Exemplary pharmaceutical compositions for Compound III and its pharmaceutically acceptable salts are disclosed in WO2011 / 119984 and WO2014 / 014841, which are incorporated herein by reference.

[0082] The pharmaceutical compositions disclosed herein may optionally further comprise at least one pharmaceutically acceptable carrier. The at least one pharmaceutically acceptable carrier may be selected from adjuvants and vehicles. As used herein, the at least one pharmaceutically acceptable carrier may include any solvent, diluent, other liquid vehicle, dispersant, suspension aid, surfactant, isotonic agent, thickener, emulsifier, preservative, solid binder, and lubricant suitable for the desired specific dosage form. Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. DB Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JCBoylan, 1988-1999, Marcel Dekker, New York disclose various carriers used in the formulation of pharmaceutical compositions and known techniques for their preparation. Unless any conventional carrier becomes incompatible with the compounds of this disclosure, for example, by producing any undesirable biological effect or by interacting with any other component of the pharmaceutical composition in a detrimental manner, its use is intended to be within the scope of this disclosure.Non-limiting examples of suitable pharmaceutically acceptable carriers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffers (e.g., phosphates, glycine, sorbic acid, and potassium sorbate), partial glyceride mixtures of saturated vegetable fatty acids, water, salts, and electrolytes (e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, lanolin fat, sugars (e.g., lactose, glucose, and sucrose), starches (e.g., corn starch and potato starch), cellulose and its derivatives (e.g., carboxymethylcellulose). Examples of ingredients include, but are not limited to, sodium sodium, ethylcellulose, and cellulose acetate, tragacanth powder, malt, gelatin, talc, excipients (e.g., cocoa butter and suppository wax), oils (e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil), glycols (e.g., propylene glycol and polyethylene glycol), esters (e.g., ethyl oleate and ethyl laurate), agar, buffers (e.g., magnesium hydroxide and aluminum hydroxide), alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, phosphate buffer solution, non-toxic compatible lubricants (e.g., sodium lauryl sulfate and magnesium stearate), colorants, release agents, coating agents, sweeteners, flavoring agents, fragrances, preservatives, and antioxidants.

[0083] In one embodiment, the pharmaceutical composition of the present disclosure comprises one or more fillers, disintegrants, and lubricants.

[0084] The fillers suitable for pharmaceutical compositions disclosed herein are compatible with the other components of the pharmaceutical composition, i.e., they do not substantially reduce the solubility, hardness, chemical stability, physical stability, or biological activity of the pharmaceutical composition. Exemplary fillers include cellulose, modified cellulose (e.g., sodium carboxymethylcellulose, ethylcellulose hydroxymethylcellulose, hydroxypropylcellulose), cellulose acetate, microcrystalline cellulose, calcium phosphate, dicalcium phosphate, starch (e.g., corn starch, potato starch), sugars (e.g., mannitol, lactose, sucrose, etc.), or any combination thereof. In some embodiments, the filler is microcrystalline cellulose.

[0085] In some embodiments, the pharmaceutical composition contains one or more fillers in an amount of at least 25% by weight (e.g., at least 27% by weight or at least 30% by weight) relative to the weight of the pharmaceutical composition. For example, the pharmaceutical composition contains 25% to 40% by weight (e.g., 25% to 35% by weight or 30% to 33% by weight) of fillers relative to the weight of the pharmaceutical composition. In another embodiment, the pharmaceutical composition contains at least 25% by weight (e.g., at least 27% by weight or at least 30% by weight) of microcrystalline cellulose, for example, Avicel PH102 or Avicel PH101, relative to the weight of the pharmaceutical composition. In yet another embodiment, the pharmaceutical composition contains 25% to 40% by weight (e.g., 25% to 35% by weight or 30% to 33% by weight) of microcrystalline cellulose relative to the weight of the pharmaceutical composition. In yet another embodiment, the pharmaceutical composition contains about 31.7% by weight of microcrystalline cellulose relative to the weight of the pharmaceutical composition. In another embodiment, the pharmaceutical composition contains approximately 32.6% by weight of microcrystalline cellulose relative to the weight of the pharmaceutical composition.

[0086] Disintegrants suitable for pharmaceutical compositions disclosed herein can enhance the dispersion of the pharmaceutical composition and are compatible with other components of the pharmaceutical composition, i.e., they do not substantially reduce the chemical stability, physical stability, hardness, or biological activity of the pharmaceutical composition. Exemplary disintegrants include croscarmellose sodium, sodium starch glycolate, crospovidone, or combinations thereof. In some embodiments, the disintegrant is croscarmellose sodium.

[0087] In some embodiments, the pharmaceutical compositions disclosed herein contain a disintegrant in an amount of 10% by weight or less (e.g., 8% by weight or 7% by weight or less) relative to the weight of the pharmaceutical composition. For example, the pharmaceutical composition contains 1% to 10% by weight (e.g., 2% to 8% by weight or 3% to 7% by weight) of a disintegrant relative to the weight of the pharmaceutical composition. In another embodiment, the pharmaceutical composition contains 10% by weight or less (e.g., 8% by weight or 7% by weight or less) of croscarmellose sodium relative to the weight of the pharmaceutical composition. In another embodiment, the pharmaceutical composition contains 1% to 10% by weight (e.g., 2% to 8% by weight or 3% to 7% by weight) of croscarmellose sodium relative to the weight of the pharmaceutical composition. In another embodiment, the pharmaceutical composition contains about 5.8% by weight of croscarmellose sodium relative to the weight of the pharmaceutical composition. In another embodiment, the pharmaceutical composition contains about 6.0% by weight of croscarmellose sodium relative to the weight of the pharmaceutical composition.

[0088] In some embodiments, the pharmaceutical compositions disclosed herein include a lubricant. The lubricant can prevent the adsorption of the mixed components to surfaces (e.g., mixing bowls, granulation rolls, compression dies, and / or punches). The lubricant can also reduce interparticle friction within the granules and improve the compression and discharge of the compressed pharmaceutical composition from the granulator and / or die press. Lubricants suitable for the pharmaceutical compositions disclosed herein are compatible with the other components of the pharmaceutical composition, i.e., they do not substantially reduce the solubility, hardness, or bioactivity of the pharmaceutical composition. Exemplary lubricants include magnesium stearate, sodium stearyl fumarate, calcium stearate, zinc stearate, sodium stearate, stearic acid, aluminum stearate, leucine, glyceryl behenylate, hydrogenated vegetable oils, or any combination thereof. In some embodiments, the lubricant is magnesium stearate.

[0089] In one embodiment, the pharmaceutical composition contains a lubricant in an amount of 5% by weight or less (e.g., 4% by weight or less, 3% by weight or less, or 2% by weight or less) relative to the weight of the pharmaceutical composition. For example, the pharmaceutical composition contains a lubricant in an amount of 0.10% to 5% by weight (e.g., 0.5% to 3% by weight or 0.75% to 2% by weight) relative to the weight of the pharmaceutical composition. In another embodiment, the pharmaceutical composition contains magnesium stearate in an amount of 5% by weight or less (e.g., 4% by weight or less, 3% by weight or less, or 2% by weight or less) relative to the weight of the pharmaceutical composition. In yet another embodiment, the pharmaceutical composition contains magnesium stearate in an amount of 0.10% to 5% by weight (e.g., 0.5% to 3% by weight or 0.75% to 2% by weight) relative to the weight of the pharmaceutical composition. In yet another embodiment, the pharmaceutical composition contains magnesium stearate in an amount of about 1.0% by weight relative to the weight of the pharmaceutical composition.

[0090] In some embodiments, the pharmaceutical composition disclosed herein is a tablet. In some embodiments, the tablet includes a film coating. In some embodiments, the film coating is Opadry20A100021.

[0091] In some embodiments, the tablets disclosed herein include the following: [Table 6]

[0092] In some embodiments, the tablets disclosed herein include the following: [Table 7]

[0093] In some embodiments, the tablets disclosed herein include the following: [Table 8]

[0094] In some embodiments, the tablets disclosed herein include the following: [Table 9]

[0095] In some embodiments, the tablets disclosed herein include the following: [Table 10]

[0096] In some embodiments, the tablets disclosed herein include the following: [Table 11]

[0097] In some embodiments, the tablets disclosed herein include the following: [Table 12]

[0098] In some embodiments, the tablets disclosed herein include the following: [Table 13]

[0099] In some embodiments, the tablets disclosed herein include the following: [Table 14]

[0100] In some embodiments, the tablets disclosed herein include the following: [Table 15]

[0101] In some embodiments, the tablets disclosed herein include the following: [Table 16]

[0102] In some embodiments, the tablets disclosed herein include the following: [Table 17]

[0103] General experimental procedure Unless otherwise specified, reagents and starting materials were obtained from commercial sources and used without purification. Proton and carbon NMR spectra were obtained at 400 MHz and 100 MHz, respectively. 1 H resonance frequency and 13The samples were acquired using either a Bruker Biospin DRX 400MHz FTNMR spectrometer or a 300MHz NMR spectrometer operating at the ¹¹C resonance frequency. One-dimensional proton and carbon spectra were acquired with a broadband observation (BBFO) probe at 20Hz sample rotation, with digital resolutions of 0.1834Hz / Pt and 0.9083Hz / Pt, respectively. All proton and carbon spectra were acquired under temperature control at 30°C using standard, previously published pulse sequences and routine processing parameters. The final purity of the compound was determined using a Waters Acquity UPLC BEH C 18 The purity was determined by reversed-phase ULC using a column (50 × 2.1 mm, 1.7 μm particle size) (product number: 186002350) and a dual gradient run of 1% to 99% mobile phase B over 3.0 minutes. Mobile phase A = H2O (0.05% CF3CO2H). Mobile phase B = CH3CN (0.035% CF3CO2H). Flow rate = 1.2 mL / min, injection volume = 1.5 μL, column temperature = 60°C. Final purity was calculated by averaging the area under the curve (AUC) of two UV traces (220 nm, 254 nm). Low-resolution mass spectra were obtained for hydrogen ions (H + The optical purity of (2S)-2,4-dimethyl-4-nitropentanoate methyl was determined by chiral gas chromatography (GC) analysis on an Agilent 7890A / MSD5975C instrument using a Restek Rt-βDEXcst column (30 m × 0.25 mm × 0.25 μm_df) at a flow rate of 2.0 mL / min (H2 carrier gas), injection temperature of 220 °C, oven temperature of 120 °C, and 15 minutes. The purity of compound I was determined by reversed-phase HPLC using a Poroshell 120 EC-C8 column (4.6 × 150 mm, 2.7 μm particle size) and a dual gradient of 30–95% mobile phase over 40 minutes. Mobile phase A = 5 mM ammonium acetate pH 4.50 and mobile phase B = acetonitrile. Flow rate = 1.0 mL / min, injection volume = 5 μL, 254 nm, column temperature = 30°C.

[0104] Compounds I, II, and III can be prepared by any suitable method in the art. A method for preparing compound I can be found in WO2019 / 109021 and U.S. Patent No. 9,512,079; a method for preparing compound II and its pharmaceutically acceptable salts is disclosed in WO2019 / 161078 and PCT / US2020 / 046116; and a method for preparing compound III and its pharmaceutically acceptable salts is disclosed in WO2011 / 119984 and WO2011 / 133751, all of which are incorporated herein by reference.

[0105] Example 1: Synthesis of N-(2-(tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-1,4-dihydroquinoline-3-carboxamide (Compound I) (2) The overall synthesis scheme for compound I is shown below, followed by the synthesis procedures for each intermediate. [ka]

[0106] A 5 L round-bottom flask equipped with an overhead stirrer was packed with 4- / er / -butylphenol (14, 503.2 g), K2CO3 (46.3 g), D2O (1949 g, 1761 mL, 3.5 vol), and MeOD (409 g, 503 mL, 1.0 vol). The mixture was heated and refluxed under a nitrogen atmosphere. The reaction mixture was aged under reflux for 16 hours. The reaction mixture was cooled to room temperature and sampled for conversion (D% inclusion). The reaction mixture was cooled to 5°C and 35% DC1 solution (90 g, 71.2 mL) was added. The reaction mixture was aged at 5°C for 2 hours. The resulting slurry was filtered, and the cake was washed with D2O (836 g, 755 mL, 1.5 vol). This process was repeated until the target D% inclusion was achieved (usually two exchanges are required). The moist cake is dried in a vacuum oven at 40°C with nitrogen bleed until a certain weight is obtained. 4-(tert-butyl)phen-2,6-d 2 The yield of -O-d(19) was 506 g of a white solid (98%) with a purity of 99.6% and 99.3% D% incorporated.

[0107] Synthesis procedure for 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phen-6-d-ol-d and 4-(tert-butyl)-2,6-bis(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenol-d(18) [ka] 4-(tert-butyl)phen-2,6-d 2 -O-d(19) (101.8 g, 0.66 mol, 1.0 equivalent) was dissolved in CH2Cl2 (400 mL) in a 2 L reactor.

number

[0108] Synthesis procedure for 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenol (17) [ka] 4-(tert-butyl)-2-(2-(methyl-d 3 )propane-2-yl-1,1,1,3,3,3-d 6 Dissolve phen-6-d-ol-d(18) (100 g, 0.462 mol, 1.0 equivalent) in CH2Cl2 (800 mL, 7 volumes) in a 2 L reactor and stir the solution. Cool the batch to 0 ± 3 °C. Add N-bromosuccinimide (84.4 g, 0.462 mol, 1.0 equivalent) to the batch portion-wise over 30 minutes. Stir the batch at 0 ± 2 °C for at least 30 minutes. Then heat the batch to 20 ± 2 °C over 2 hours and stir at 20 ± 2 °C for at least 12 hours. After complete conversion, add saturated NaHCCh aqueous solution (500 mL, 5 volumes) and stir the batch for at least 10 minutes. Stop stirring and allow the phases to separate for at least 5 minutes, drain the CH2Cl2 layer, and then remove the aqueous layer. Pack the CTLCh layer back into the container. A saturated NaHClE bicarbonate aqueous solution (500 mL, 5 vols) was added to the batch, and the batch was stirred for at least 10 minutes. Stirring was stopped, and the phases were separated for at least 5 minutes, the CH2Cl2 layer was drained, and then the aqueous layer was removed. The CH2Cl2 layer was packed back into the container and diluted with additional CH2Cl2 (300 mL, 3 vols). The batch was distilled (300 mL removed) and confirmed to be dry by kF. The resulting clear yellow solution was proceeded to the next step without further purification.

[0109] Synthesis procedure for 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (16) [ka] A clean reactor was filled with a CH2Cl2 solution of 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phen-6-d-ol-d(17) (136 g, 0.462 mol, 1.0 equivalent), followed by the addition of an additional CH2Cl2 (130 mL, 1 volume), and the solution was stirred. The batch was then filled with 4-(dimethylamino)pyridine (2.8 g, 0.023 mol, 0.05 equivalent) and triethylamine (70.1 g, 0.693 mol, 1.5 equivalent). The batch was cooled to 0 ± 3°C. While maintaining the batch temperature at <5°C, methyl chloroformate (48.0 g, 0.508 mol, 1.1 equivalent) was added dropwise to the batch over 40 minutes. The batch was stirred at 3±2°C for at least 30 minutes, then heated to 20±2°C over 1 hour. After complete conversion, 1N HCl (400 mL, 3 vols) was added. The batch was stirred for at least 10 minutes, then the layers were separated for at least 5 minutes. The lower organic layer was drained, followed by the aqueous layer (first aqueous layer). The organic layer was returned to the reactor with 1N HCl solution (400 mL, 3 vols). The batch was stirred for at least 10 minutes, then the layers were separated for at least 5 minutes. The lower organic layer was drained. The first aqueous layer was added to the reactor with CH2Cl2 (300 mL, 2.2 vols). The batch was stirred for at least 10 minutes, then the layers were separated for at least 5 minutes. The lower organic layer was drained and combined with the first organic layer, followed by the removal of the aqueous layer. The contents of both organic layers were packed into containers. The reactor was filled with water (500 mL, 3.7 vols). The batch was stirred for at least 10 minutes, then the layers were separated for at least 5 minutes. The lower organic layer was drained, followed by the aqueous layer. The organic layer was packed back into the reactor with CH2Cl2 (400 mL, 3 volumes). The batch was distilled to remove 800 mL, and thorough drying was confirmed by KF. The resulting 16 clear yellow solutions were incorporated into the next step without further purification.

[0110] Synthesis procedure for 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)-3-nitrophenylmethyl carbonate (15) [ka] The reactor was packed with 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (16), and the solution was then cooled to 0°C. While maintaining the temperature below 5°C, sulfuric acid (4.9 equivalents) and nitric acid (100%, 2.0 equivalents) were packed in. The reaction mixture was stirred at 0°C for 2 hours until complete conversion was achieved. The reaction mixture was then quenched with water (8.8 vol) and diluted with CH2Cl2 (1.7 vol). The layers were separated, and the upper aqueous layer was extracted with CH2Cl2 (2.8 vol). After separating the layers, the organic layers were combined and returned to the reactor, and washed with sodium bicarbonate (7.4% w / w, 6.8 vol). After separating the layers, the organic layers were returned to the reactor, and washed with sodium chloride (23% w / w, 3.8 vol). After separating the layers, the organic layer was returned to the reactor and concentrated to the minimum volume. 1.2 volumes of methanol were added, and the mixture was then concentrated to the minimum volume. 1.7 volumes of methanol were added, and the slurry was heated under reflux for 30 minutes, then slowly cooled to 5°C over 4 hours. The solid product (15) was filtered, and the cake was washed with cold methanol (1.0 volume). The solid 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)-3-nitrophenylmethyl carbonate (15) was dried under vacuum at 40°C to 50°C to obtain an off-white solid with 99.9% purity and 99% D content.

[0111] Synthesis procedure for 5-amino-4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (7) [ka] Pack the reactor with 5% by weight of 5% Pd / C (50-65% by weight wet, JM type 37). Pack with (4.0 vol) methanol. Close the system. Inflate with N2 at 2.0 Bar.(g >Purge. H2 at 2.0 Bar (g) It is activated by H2 in a container at 25℃±5℃. (g Fill to 2.0 Bar. Stir for at least 2 hours while maintaining a temperature of 25°C ± 5°C. Evacuate and inject N2 at 2.0 Bar. (g) Purge. Pack compound 15 (1.0 equivalent) into the reactor together with Na2HPO4 (2.3 equivalents). Pack with (11.0 volume) methanol. Close the system. N2 at 2.0 Bar (g >Purge. H2 at 2.0 Bar (g) It is activated by H2 in a container at 25℃±5℃. (g Fill the reactor to 2.0 Bar. Stir for approximately 24 hours while maintaining a reaction temperature of 25°C ± 5°C. Once the conversion is complete, add 7.7 volumes of MeOH to dilute the reaction mixture. Heat the reaction mixture to 35.0°C ± 5°C. Filter off the catalyst and Na2HPO4. Wash the reactor and filter cake with methanol (4.0 volumes) and filter together with the initial filtrate. Check the Pd content and perform resin treatment if necessary (for resin treatment, fill with SPM-32 resin (5 wt%)). Stir the resin treatment solution at 35.0°C ± 5°C for at least 3 hours. Filter off the resin.

[0112] Wash the reactor and filter cake with methanol (2.0 vol), filter, and combine with the first filtrate. Pack with Norit CASP activated carbon (3 wt%). Stir at 35.0°C ± 5°C for at least 3 hours. Filter off the activated carbon. Wash the reactor and filter cake with methanol (2.0 vol), filter, and combine with the first filtrate. Distill under vacuum at below 50°C to 8.0 vol. Pack with water (2.0 vol) while maintaining the temperature at 45°C ± 5°C. Cool the resulting slurry to 0°C ± 5°C over 2 hours. Hold the slurry at 0°C ± 5°C for at least 1 hour and stir. Filter, and wash the cake with 2.0 vol methanol / water (8:2) at 0°C ± 5°C. 5-amino-4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (7) is dried under vacuum at 40°C or below to obtain a white solid with a purity of >99.5%.

[0113] Synthesis procedure for 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)-5-(4-oxo-l,4-dihydroquinoline-3-carboxamide)phenylmethyl carbonate (8) [ka] The procedure for converting compound 7 to compound 8 can be carried out using a similar procedure to that for compound 5.

[0114] Synthesis procedure for N-(2-(tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-l,4-dihydroquinoline-3-carboxamide (2) (Compound I) [ka] The procedure for converting compound 8 to compound 2 can be carried out using a similar procedure to that for the synthesis of compound 1.

[0115] Example 2: Synthesis of 5-amino-4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (7) Another overall scheme for the synthesis of compound 7 is shown below, followed by the synthesis steps for each synthetic intermediate. [ka]

[0116] Synthesis procedure for 4-(tert-butyl)phen-2,6-d2-ol-d(19) [ka] A clean, dry 500 mL reactor was filled with 4- / c / 7-butylphenol(14) (24.6 g, 0.162 mmol, 1.00 equivalent), CH2Cl2 (64 mL, 2.6 vol), and heptane (64 mL, 2.6 vol). The mixture was heated to 25°C and stirred until all solids dissolved. Deuterium chloride (35% w / w in deuterium oxide, 25 mL, 1.0 vol) was added to the solution, and the mixture was stirred for at least 3.5 hours. Stirring was stopped, the phases were separated, and the aqueous layer (bottom) was then discharged from the reactor. Deuterium chloride (35% w / w in deuterium oxide, 25 mL, 1.0 vol) was added to the reactor, and the mixture was stirred for at least 3.5 hours. Stirring was stopped, the phases were separated, and the aqueous layer (bottom) was then discharged from the reactor. The reactor was filled with deuterium chloride (35% w / w in deuterium oxide, 25 mL, 1.0 vol), and the mixture was stirred for at least 3.5 hours. Stirring was stopped, the phases were separated, and the aqueous layer (bottom) was then discharged from the reactor. The resulting solution was sampled and confirmed to be at least 99% of the desired deuterium-integrated product 4-(tert-butyl)phen-2,6-d2-ol-d(19) relative to the starting material 4- / er / -butylphenol. The solution in the reactor was then continued to the next steps described below.

[0117] Synthesis procedure for 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phen-6-d-ol (18) [ka] A methylene chloride solution containing the reaction mixture of 4-(tert-butyl)phen-2,6-d2-ol-d(19) was packed with CH2Cl2 (125 mL, 5 vols). Using a distillation head, the reactor was heated to 60°C, and approximately 125 mL of the reaction solution was distilled from the reactor. CH2Cl2 (125 mL, 5 vols) was then packed into the reactor. Next, approximately 100 mL of the reaction solution was distilled from the reactor, and at this point, the solution was sampled to confirm that the water content (KF) was less than 300 ppm, and the CH2Cl2 and heptane content was measured. After measuring the batch volume, CH2Cl2 (8 mL, 0.24 vols) was packed in to adjust the total CH2Cl2 content to 3 vols, and heptane (68 mL, 2.8 vols) was packed in to adjust the heptane content to 4.5 vols. The solution was filled with / cvv-butylacetoate e-dg (30.2 g, 1.46 equivalents), and the resulting solution was cooled to 0°C. Sulfuric acid-J2 (8.12 g, 0.49 equivalents) was filled into the solution over at least 15 minutes, and the solution was stirred for 2 hours while maintaining a temperature of 0-5°C. After this time, the temperature was set to rise to 20°C over 2 hours, and the solution was stirred for a further 14 hours. The solution was sampled to confirm the presence of 4- / c77-butylphenol (14) or 4-(tert-butyl)phen-2,6-d2-ol-d (19) at a concentration of less than 3%. The reactor was filled with CH2Cl2 (58 mL, 2.4 vols) and heptane (90 mL, 3.7 vols), and after the solution was cooled to 0-5°C, water (125 mL, 5 vols) was added. The mixture was stirred for 15 minutes, then stirring was stopped, and the phases were separated. After draining the aqueous phase (bottom) from the reactor, 125 mL of 0.5 N NaOH aqueous solution (5 volumes) was added, and the temperature was adjusted to 20°C. The mixture was stirred for 20 minutes, then stirring was stopped, and the phases were separated. The organic phase (top) was sampled, and it was confirmed that 4-tert-butylphenol (14) or 4-(tert-butyl)phen-2,6-d2-ol-d (18) was present at a concentration of less than 0.5%. The aqueous phase (bottom) was drained from the reactor. The solution in the reactor was allowed to continue to the next steps described below.

[0118] Synthesis procedure for 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenol (17) [ka] After stirring the alkylation reaction solution to produce 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phen-6-d-ol-d(18) to 0-5°C, bromine (38.4 g, 1.45 equivalents) was added for at least 1 hour while maintaining the temperature below 5°C. The solution was sampled to confirm the presence of 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phen-6-d-ol at a concentration of less than 1%. Sodium disulfite (20% w / w aqueous solution, 147 g, 0.95 equivalents) was added to the solution for at least 1 hour while maintaining the temperature below 10°C, and after adjusting the temperature to 20°C, the mixture was stirred for a further 1 hour. Stirring was stopped and the phases were separated. The aqueous phase (bottom) was drained from the reactor, and water (125 mL, 5 volumes) was added to the reactor. The mixture was stirred for 15 minutes, then the stirring was stopped, and the phases were separated. The aqueous phase (bottom) was drained from the reactor. The 17 solutions in the reactor were continued to the next steps described below.

[0119] Surprisingly, this bromination reaction significantly improved the selectivity of the nitration reaction. Another unexpected benefit of this process was that bromination converted the mixture of compound 18 and 4-(tert-butyl)-2,6-bis(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenol to the same desired product (17). This resulted in a significant improvement in the overall yield.

[0120] Synthesis procedure for 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (16) [ka] The bromination reaction solution producing 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenol (17) was packed with CH2Cl2 (125 mL, 5 volumes). Using a distillation head, the reactor was heated to 60°C and approximately 125 mL of the reaction solution was distilled from the reactor. CH2Cl2 (125 mL, 5 volumes) was packed into the reactor. Approximately 125 mL of the reaction solution was distilled from the reactor. CH2Cl2 (125 mL, 5 volumes) was packed into the reactor. Next, approximately 125 mL of the reaction solution was distilled from the reactor, and at this point, the solution was sampled to confirm that the water content (KF) was less than 300 ppm, and the CH2Cl2 and heptane content was measured. After measuring the batch volume, the solution was filled with CFLCh to adjust the total CH2Cl2 content to 5.3 volumes, and then filled with heptane to adjust the heptane content to 8 volumes. Triethylamine (31.7 g, 1.91 equivalents) was added to the solution, and the solution was cooled to 0-5°C. Methyl chloroformate (24.1 g, 1.56 equivalents) was added to the solution over at least 1 hour while maintaining the temperature below 10°C. The solution was stirred for 1 hour, and a sample of the solution was taken to confirm the presence of 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenol (17) at a concentration of less than 1%. 1N hydrochloric acid aqueous solution (125 mL, 0.76 equivalents) was added to the solution over at least 30 minutes while maintaining the temperature below 10°C. Next, the temperature was adjusted to 20°C, stirring was stopped, and the phases were separated. After the aqueous phase (bottom) was drained from the reactor, water (125 mL, 5 volumes) was added to the reactor. The mixture was stirred for 15 minutes, then stirring was stopped, and the phases were separated. After the aqueous phase (bottom) was drained from the reactor, water (125 mL, 5 volumes) was added to the reactor. The mixture was stirred for 15 minutes, then stirring was stopped, and the phases were separated. The aqueous phase (bottom) was drained from the reactor. The solution of (16) in the reactor was continued to the next steps described below.

[0121] Synthesis procedure for 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)-3-nitrophenylmethyl carbonate (15) [ka] CH2Cl2 (125 mL, 5 volumes) was packed into the protective reaction solution that produces 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (16). Using a distillation head, the reactor was heated to 60°C and approximately 125 mL of the reaction solution was distilled from the reactor. CH2Cl2 chloride (125 mL, 5 volumes) was packed into the reactor. Approximately 125 mL of the reaction solution was distilled from the reactor. CH2Cl2 (125 mL, 5 volumes) was packed into the reactor. CH2Cl2 (125 mL, 5 volumes) was packed into the reactor. Approximately 125 mL of the reaction solution was distilled from the reactor. Next, approximately 125 mL of the reaction mixture was distilled from the reactor. At this point, the solution was sampled to confirm that the water content (KF) was less than 300 ppm, and the CH2Cl2 and heptane content was measured. After measuring the batch volume, CH2Cl2 was added to adjust the total CH2Cl2 content to 6 volumes, and heptane was added to adjust the heptane content to 9 volumes. After cooling the solution to 0-5°C, sulfuric acid (172 g, 10.3 equivalents) was added for at least 30 minutes while maintaining the temperature below 5°C. Nitric acid (70% w / w, 19.1 g, 1.31 equivalents) was added to the mixture for at least 30 minutes while maintaining the temperature below 10°C. After stirring the mixture for 1 hour, a sample was taken and analyzed to confirm the presence of 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (16) at a concentration of less than 1%. Water (100 mL, 4 volumes) was added to the mixture over at least 1 hour while maintaining the temperature below 10°C. Stirring was stopped, the phases were separated, and the aqueous phase (bottom) was discharged from the reactor. After restarting stirring, sodium bicarbonate (8% w / w aqueous solution, 100 mL, 4 volumes, 0.62 equivalents) was added over at least 10 minutes while maintaining the temperature below 10°C. The temperature was adjusted to 20°C, stirring was stopped, and the phases were separated. After draining the aqueous phase (bottom) from the reactor, water (100 mL, 4 volumes) was added to the reactor, and the mixture was stirred for 15 minutes. Stirring was stopped, the phases were separated, and the aqueous phase (bottom) was drained from the reactor.The mixture was filled with water (100 mL, 4 vols) and stirred for 15 minutes. Stirring was stopped, the phases were separated, and the aqueous phase (bottom) was discharged from the reactor. After marking the solvent level in the reactor, a distillation head was attached and the temperature was set to 80°C. Methanol (570 mL, 23 vols) was filled into the solution and distilled simultaneously, matching the addition rate to the distillation rate by maintaining the solvent level at the mark. Distillation was continued until the batch volume reached approximately 264 mL (11 vols) and approximately 1.10 kg of distillate was removed. The mixture was sampled and analyzed to confirm the presence of heptane at less than 1% v / v. The temperature was adjusted to 0°C for 4 hours. The mother liquor was collected and analyzed to determine the concentration of 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)-3-nitrophenylmethyl carbonate (15), and the mixture was filtered. Methanol (51.1 mL, 2 volumes) was packed into the reactor and stirred until the temperature reached 0-5°C. The filter cake was washed with this solution and then dried by vacuum for at least 1 hour. The solid was then vacuum dried to produce 41.5 g of 2-bromo-4-(tert-butyl)-6-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)-3-nitrophenylmethyl carbonate (15) as an off-white solid (purity 98.4% w / w, yield 63% after purity correction).

[0122] Synthesis procedure for 5-amino-4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (7) [ka] Pack the reactor with 5% by weight of 5% Pd / C (50-65% by weight wet, JM type 37). Pack with (4.0 vol) methanol. Close the system. Inflate with N2 at 2.0 Bar. (g >Purge. H2 at 2.0 Bar (g) It is activated by H2 in a container at 25℃±5℃. (gFill to 2.0 Bar. Stir for at least 2 hours while maintaining a temperature of 25°C ± 5°C. Evacuate and inject N2 at 2.0 Bar. (g) Purge. Pack compound 15 (1.0 equivalent) into the reactor together with Na2HPO4 (2.3 equivalents). Pack with (11.0 volume) methanol. Close the system. N2 at 2.0 Bar (g >Purge. H2 at 2.0 Bar (g) It is activated by H2 in a container at 25℃±5℃. (g Fill the reactor to 2.0 Bar. Stir for approximately 24 hours while maintaining a reaction temperature of 25°C ± 5°C. Once the conversion is complete, add 7.7 volumes of MeOH to dilute the reaction mixture. Heat the reaction mixture to 35.0°C ± 5°C. Filter off the catalyst and Na2HPO4. Wash the reactor and filter cake with methanol (4.0 volumes) and filter together with the initial filtrate. Check the Pd content and perform resin treatment if necessary (for resin treatment, fill with SPM-32 resin (5 wt%). Stir the resin treatment solution at 35.0°C ± 5°C for at least 3 hours. Filter off the resin.

[0123] Wash the reactor and filter cake with methanol (2.0 vol), filter, and combine with the first filtrate. Pack with Norit CASP activated carbon (3 wt%). Stir at 35.0°C ± 5°C for at least 3 hours. Filter off the activated carbon. Wash the reactor and filter cake with methanol (2.0 vol), filter, and combine with the first filtrate. Distill under vacuum at below 50°C to 8.0 vol. Pack with water (2.0 vol) while maintaining the temperature at 45°C ± 5°C. Cool the resulting slurry to 0°C ± 5°C over 2 hours. Hold the slurry at 0°C ± 5°C for at least 1 hour and stir. Filter, and wash the cake with 2.0 vol methanol / water (8:2) at 0°C ± 5°C. 5-amino-4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (7) is dried under vacuum at 40°C or below to obtain a white solid with a purity of >99.5%.

[0124] Synthesis procedure for 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)-5-(4-oxo-l,4-dihydroquinoline-3-carboxamide)phenylmethyl carbonate (8) [ka] The procedure for converting compound 7 to compound 8 can be carried out using a similar procedure to that for compound 5.

[0125] Synthesis procedure for N-(2-(tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-l,4-dihydroquinoline-3-carboxamide (2) (Compound I) [ka] The procedure for converting compound 8 to compound 2 can be carried out using a similar procedure to that for the synthesis of compound 1.

[0126] Example 3: Synthesis of 5-amino-4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (7) An alternative scheme for the synthesis of compound 7 is shown below, followed by the synthesis procedures for each synthetic intermediate. [ka]

[0127] Synthesis procedure for 5-(tert-butyl)-2-hydroxybenzoic acid (15) [ka] 3.49 g of 1.6 M nBuLi in hexane was added to a round-bottom flask equipped with a magnetic stirring bar, thermocouple, and N2 bubbler. The round-bottom flask was cooled to -20°C and stirring was started. A solution of 2-bromo-4-tert-butylphenol (26) (5.00 g) in MTBE (12.5 mL) was prepared, cooled to -20°C, and added dropwise to the round-bottom flask while maintaining the temperature at -20°C ± 5°C. The reaction mixture was stirred at -20°C ± 5°C for 15 minutes, and then warmed to 23°C. After 15 minutes at room temperature, the completeness of lithiation was measured by ¹H NMR (200 pL of the reaction mixture diluted in 0.75 mL of d₄-MeOH). The reaction was considered complete when less than 1% of 2-bromo-4-tert-butylphenol was observed. The reaction mixture was cooled to 0°C, dry ice (solid CO2) was added, and the reaction was stirred at room temperature for 45 minutes. Water (50.0 mL) was added to quench the reaction. The mixture was transferred to a separatory funnel, the phases were separated, and the organic phase was discarded. The aqueous phase was acidified to approximately pH 2 with 1 M HCl (15.0 mL) and then extracted three times with MTBE (25.0 mL). The combined organic extracts were concentrated under reduced pressure to obtain 5-(tert-butyl)-2-hydroxybenzoic acid (25) as a yellow solid (2.25 g, yield 53.15%). 3 / 4 NMR (400 MHz, d4-MeOH): 7.86 (1H, d, J = 2.6 Hz), 7.54 (1H, dd, J = 8.7, 2.6 Hz), 6.85 (1H, d, J = 2.7 Hz), 1.30 (9H, s).

[0128] Synthesis procedure for methyl 5-(tert-butyl)-2-hydroxybenzoate (24) [ka] This reaction can be carried out according to the procedure described in Bioorganic and Medicinal Chemistry Letters, 2005, vol.15(21), pp.4752-4756.

[0129] Synthesis procedure for 2-((tert-butoxycarbonyl)oxy)-5-(tert-butyl)methyl benzoate (23) [ka] Di- / e / 7-butyl carbonate (230.55 g) and CH2Cl2 (400 mL) were packed into a 1 L reactor, and the mixture was stirred until the solid was completely dissolved. Dimethylaminopyridine (0.587 g) was packed into the stirred solution together with methyl 5-( / er / -butyl)-2-hydroxybenzoate (24) (200 g). The reaction mixture was stirred at 15-30°C, and after 60 minutes, the integrity was measured by HPLC using sample aliquots. The reaction was considered complete when the peak area of ​​5-tert-butyl-2-hydroxybenzoate (24) was less than 1%. A semi-saturated solution of ammonium chloride was prepared in a separate flask by diluting saturated ammonium chloride solution (200 mL) with water (200 mL). The reaction mixture was washed twice with saturated aqueous ammonium chloride solution (200 mL each wash). During each wash, the mixture was stirred for 15 minutes and held for 15 minutes. Next, the organic solution was washed twice with water (100 mL each time). During each wash, the mixture was stirred for 15 minutes and held for 15 minutes. The organic solution was transferred to a 1 L round-bottom flask and concentrated under vacuum at 35°C or below to obtain a white solid (275.51 g and HPLC analysis (method) showed a purity of 99.46% and a yield of 93.0% for methyl 2(( / er / -butoxycarbonyl)oxy)-5-(ter / -butyl)benzoate (23)). 3 / 4 NMR (400 MHz, CDCb): 8.01 (m, 1H); 7.57 (m, 1H); 7.11 (m, 1H); 3.89 (s, 3H); 1.58 (s, 9H); 1.33 (s, 9H).

[0130] Synthesis procedure for 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenol (22) [ka] THF (176 mL) was packed into a 500 mL jacketed reactor and cooled to 5°C. A solution of (methyl-d3) magnesium iodide (60.5 g) in dibutyl ether (145 mL) was slowly added to the stirring solvent at 0-35°C. The obtained slurry was maintained at 20-30°C, and a solution of 2-((tert-butoxycarbonyl)oxy)-5-( / er / 7-butyl(benzoic acid (23) (22 g)) in THF (44 mL) was added over 4-6 hours. The reaction mixture was stirred at 20-30°C, and after 60 minutes, the integrity was measured by HPLC using sample aliquots. The reaction was considered complete when the peak area of ​​2-(( / er / -butoxycarbonyl)oxy)-5-( / er / -butyl)benzoic acid (23) fell to less than 1%. A second reactor was filled with 6N aqueous hydrochloric acid (110 mL), and the stirred solution was cooled to 0-10°C. The reaction slurry was slowly transferred to an acidic solution at 0-35°C. The phase was stirred for 15 minutes, held for 15 minutes, and then separated. The aqueous phase was extracted with dibutyl ether (132 mL). During the process, the phases were stirred for 15 minutes, held for 15 minutes, and then separated. The combined organic phases were sequentially washed with water (2 × 77 mL), 5% sodium thiosulfate aqueous solution (77 mL), and water (77 mL). During each wash, the mixture was stirred for 15 minutes and held for 15 minutes. The organic solution was transferred to a round-bottom flask and concentrated under vacuum at a temperature below 80°C to obtain 4-( / er / -butyl)-2-(2-(methyl-d3)propane-2- Il-1,1,1,3,3,3-d6)phenol (22) was obtained as a crude oily product (5.94 g and measured by HPLC analysis (method) with a purity of 83.8%, LC / MS analysis showed a D9 isotope purity of 99.3%, and HPLC analysis yielded 84.9% methyl-4-( / er / -butyl)-2-(2-(methyl-i3 / 4)propan-2-yl-1,1,1,3,3,3-^phenol (23)). 3 / 4 NMR (400 MHz, CD3OD): 7.22 (m, 1H); 7.00 (m, 1H); 6.65 (m, 1H); 1.26 (s, 9H).

[0131] The Grignard reaction in (23) resulted in some deuterium incorporation in (22). The mixture was subjected to a series of HCl washes to perform an H / D exchange: [ka]

[0132] HDD replacement procedure Fill the reactor with a deuterated analog of compound 22 (1.00 equivalent). Fill with DCM (5 volumes). Set the jacket to 20°C. Stir to dissolve the solid. Fill with 35% hydrochloric acid (5 volumes). Stir for at least 6 hours to mix the layers. Stop stirring and let the layers stand for at least 30 minutes. Discharge the bottom layer (organic layer) from the reactor. Discharge the aqueous layer from the reactor. Refill the reactor with the organic portion. Repeat the HCl washing procedure twice. Charge pre-mixed water (2.5 volumes) and saturated NaCl aqueous solution (2.5 volumes). Stir for 30 minutes to mix the layers. Stop stirring and let the layers stand for at least 30 minutes. Discharge the bottom layer (organic layer) from the reactor. Discharge the aqueous portion from the reactor. Refill the reactor with the organic portion. Fill with water (5 volumes). Mix the layers by stirring for 30 minutes. Stop stirring and let the layers stand for at least 30 minutes. Discharge the bottom layer (organic layer) from the reactor. Discharge the aqueous components from the reactor. Refill the reactor with the organic components. Distill the solvent to the minimum volume under reduced pressure (using a rotovap with a bath temperature of 35°C). Refill with DCM (5 volumes). Distill the solvent to the minimum volume under reduced pressure (using a rotovap with a bath temperature of 35°C). Refill with DCM (5 volumes). Sample the solution and measure the water content with KF. Repeat until the water content is less than 300 ppm. Note: Since this solution was used directly in the next reaction, the final amount of DCM should be the amount required for the alkoxyformylation reaction of compound 22.

[0133] Synthesis procedure for 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (21) [ka] The procedure for converting compound 22 to compound 21 can be carried out using a similar procedure to that for compound 12.

[0134] Synthesis procedure for 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)-5-nitrophenylmethyl carbonate (20) [ka] The procedure for converting compound 21 to compound 20 can be carried out using a similar procedure to that for compound 11A.

[0135] Synthesis procedure for 5-amino-4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenylmethyl carbonate (7) [ka] The procedure for converting compound 20 to compound 7 can be carried out using a similar procedure to that for compound 4.

[0136] Synthesis procedure for 4-(tert-butyl)-2-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)-5-(4-oxo-l,4-dihydroquinoline-3-carboxamide)phenylmethyl carbonate (8) [ka] The procedure for converting compound 7 to compound 8 can be carried out using a similar procedure to that for compound 5.

[0137] Synthesis procedure for N-(2-(tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-l,4-dihydroquinoline-3-carboxamide (2) (Compound I) [ka] The procedure for converting from (8) to (2) can be carried out using a similar procedure to that for the synthesis of compound I.

[0138] Example 4: Synthesis of (14S)-8-[3-(2-{dispiro[2.0.2.1]heptan-7-yl}ethoxy)-1H-pyrazole-1-yl]-12,12-dimethyl-2λ6-thia-3,9,11,18,23-pentazatetracyclo[17.3.1.111,14.05,10]tetracosa-1(22),5,7,9,19(23),20-hexaen-2,2,4-trione (compound II) Unless otherwise specified, reagents and starting materials were obtained from commercial sources and used without purification.

[0139] Proton and carbon NMR spectra were obtained at 400 MHz and 100 MHz, respectively. 1 H resonance frequency and 13 The spectra were acquired using either a Bruker Biospin DRX 400MHz FTNMR spectrometer or a 300MHz NMR spectrometer operating at the ¹¹¹ C resonance frequency. One-dimensional proton and carbon spectra were acquired with a broadband observation (BBFO) probe at 20Hz sample rotation, with digital resolutions of 0.1834Hz / Pt and 0.9083Hz / Pt, respectively. All proton and carbon spectra were acquired under temperature control at 30°C using standard, previously published pulse sequences and routine processing parameters.

[0140] Part A: Synthesis of 2-chloro-6-[3-(2-dispiro[2.0.2.1]heptan-7-ylethoxy)pyrazole-1-yl]pyridine-3-carboxylic acid [ka] Step 1: 7-(bromomethyl)dispiro[2.0.2.1]heptane [ka] A 1000 mL three-necked round-bottom flask was fitted with a mechanical stirrer, a cooling bath, an addition funnel, a J-Kem temperature probe, and a nitrogen inlet / outlet. Under a nitrogen atmosphere, triphenylphosphine (102.7 mL, 443.2 mmol) and dichloromethane (1 L) were added to this container, yielding a clear, colorless solution. Stirring was started, and acetone was added to the cooling bath. Dry ice was added to the cooling bath in small amounts until a pot temperature of -15°C was achieved. A solution of bromine (22.82 mL, 443.0 mmol) in dichloromethane (220 mL, 10 mL / g) was added to the addition funnel, and then added dropwise over 1 hour. Dry ice was added to the cooling bath in small amounts during the addition process to maintain the pot temperature at -15°C. After the addition of bromine was complete, the pale yellow suspension was stirred at -15°C for 15 minutes, at which point the suspension was cooled to -30°C. A funnel was filled with a solution of dispiro[2.0.2.1]heptane-7-ylmethanol (50 g, 402.6 mmol), pyridine (35.82 mL, 442.9 mmol), and dichloromethane (250 mL, 5 mL / g). The clear, pale yellow solution was then added dropwise over 1.5 hours while maintaining the pot temperature at -30°C. The resulting clear, pale yellow reaction mixture was gradually warmed to a pot temperature of -5°C and then stirred at -5°C for 1 hour. The reaction mixture was then poured into hexane (2000 mL), which caused a precipitate to form. The suspension was stirred at room temperature for 30 minutes and then filtered through a glass fritbuchner funnel with a 20 mm Celite layer. The clear filtrate was concentrated under reduced pressure (water bath temperature 20°C) to obtain a yellow oil, although some precipitate was present. The oil was diluted with some hexane and allowed to stand at room temperature for 15 minutes, then filtered through a glass fritbuchner funnel with a 20 mm Celite layer. The clear filtrate was concentrated under reduced pressure (water bath temperature 20°C) to obtain 7-(bromomethyl)dispiro[2.0.2.1]heptane (70 g, 93%) as a clear yellow oil. 11H NMR (400 MHz, chloroform-d) δ 3.49 (d, J = 7.5 Hz, 2H), 1.90 (t, J = 7.5 Hz, 1H), 1.06 - 0.84 (m, 4H), 0.71 (ddd, J = 9.1, 5.1, 4.0 Hz, 2H), 0.54 (dddd, J = 8.6, 4.8, 3.8, 1.0 Hz, 2H).

[0141] Step 2: 2-Dispiro[2.0.2.1]heptan-7-ylacetonitrile

Chemical Structure

[0142] Step 3: 2-Dispiro[2.0.2.1]heptan-7-ylacetic acid

Chem.

[0143] Step 4: 2-Dispiro[2.0.2.1]heptan-7-ylethanol

Chem.

[0144] Step 5: 3-(2-dispiro[2.0.2.1]heptane-7-ylethoxy)pyrazole-1-carboxylate tert-butyl [ka] Triphenylphosphine (4.399 g, 16.77 mmol) was added to a solution of tert-butyl 5-oxo-1H-pyrazole-2-carboxylate (2.942 g, 15.97 mmol) and 2-dispiro[2.0.2.1]heptan-7-ylethanol (2.318 g, 16.77 mmol) in tetrahydrofuran (36.78 mL). Diisopropyl azodicarboxylate (3.391 g, 3.302 mL, 16.77 mmol) was slowly added dropwise to the mixture over 10 minutes (slight exothermic reaction was observed). The reaction mixture was stirred at room temperature for 30 minutes, then at 50°C for 30 minutes. Tetrahydrofuran was removed under vacuum. Toluene (23.54 mL) was added to the crude residue, and the mixture was stirred overnight, causing the precipitate to gradually crystallize. The mixture was slurryed on Celite, then the precipitate was filtered off, washed with toluene (8.705 mL), and washed again with toluene (8.705 mL). The filtrate was concentrated under vacuum. The crude product was purified by silica gel chromatography using a gentle gradient from 100% hexane to 100% ethyl acetate to obtain 3-(2-dispiro[2.0.2.1]heptan-7-ylethoxy)pyrazole-1-carboxylate tert-butyl (3.449 g, 71%). ESI-MS m / z calculation: 304.17868, measured value: 305.1 (M+1). + ; Retention time: 0.82 minutes (LC method A).

[0145] Step 6: 3-(2-dispiro[2.0.2.1]heptane-7-ylethoxy)-1H-pyrazole [ka] 3-(2-dispiro[2.0.2.1]heptane-7-ylethoxy)pyrazole-1-carboxylate tert-butyl (5.304 g, 17.43 mmol) was dissolved in dichloromethane (53.04 mL) with trifluoroacetic acid (29.81 g, 20.14 mL, 261.4 mmol), and the reaction mixture was stirred at room temperature for 120 minutes. The reaction mixture was evaporated, and the resulting oil was partitioned between ethyl acetate and a saturated sodium bicarbonate solution to separate the layers. The aqueous portion was extracted twice more with ethyl acetate, and then the organic layers were combined, washed with brine, dried over sodium sulfate, filtered, and evaporated to obtain 3-(2-dispiro[2.0.2.1]heptane-7-ylethoxy)-1H-pyrazole (3.56 g, 100%) as oil. ESI-MS m / z calculated value 204.12627, measured value 205.1 (M+1) + ; Retention time: 0.59 minutes (LC method A).

[0146] Step 7: 2-Chloro-6-[3-(2-dispiro[2.0.2.1]heptan-7-ylethoxy)pyrazole-1-yl]pyridine-3-carboxylate tert-butyl [ka] 2,6-Dichloropyridine-3-carboxylate tert-butyl (4.322 g, 17.42 mmol), 3-(2-dispiro[2.0.2.1]heptane-7-ylethoxy)-1H-pyrazole (3.559 g, 17.42 mmol), and potassium carbonate (2.891 g, 20.92 mmol) were combined in anhydrous dimethyl sulfoxide (71.18 mL). 1,4-Diazabicyclo[2.2.2]octane (391.1 mg, 3.487 mmol) was added, and the mixture was stirred under nitrogen at room temperature for 16 hours. The reaction mixture was diluted with water (136.9 mL) and stirred for 15 minutes. The resulting white solid was filtered and washed with water. The solid was dissolved in dichloromethane and dried over magnesium sulfate. The mixture was filtered and evaporated to obtain 2-chloro-6-[3-(2-dispiro[2.0.2.1]heptan-7-ylethoxy)pyrazole-1-yl]pyridine-3-carboxylate tert-butyl (5.69 g, 79%) as a white solid. 1 H NMR (400MHz, chloroform-d) δ8.35(d,J=2.9Hz,1H), 8.18(d,J=8.4Hz,1H), 7.69(d,J=8.4Hz,1H), 5.94(d,J=2.9Hz,1H), 4.25(s,2H) , 1.90(d,J=6.8Hz,2H), 1.62(s,9H), 1.49(t,J=6.6Hz,1H), 0.85(d,J=1.5Hz,4H), 0.65(d,J=1.5Hz,2H), 0.52(d,J=1.1Hz,2H). ESI-MSm / z calculated value: 415.16626, measured value: 360.0 (M-tBu)+; holding time: 2.09 minutes (LC method B).

[0147] Step 8: 2-Chloro-6-[3-(2-dispiro[2.0.2.1]heptan-7-ylethoxy)pyrazole-1-yl]pyridine-3-carboxylic acid [ka] tert-Butyl 2-chloro-6-[3-(2-dispiro[2.0.2.1]heptan-7-yl ethoxy)pyrazol-1-yl]pyridine-3-carboxylate (5.85 g, 14.07 mmol) was dissolved in dichloromethane (58.5 mL) together with trifluoroacetic acid (16.26 mL, 211.1 mmol), and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was evaporated, ether was added to the resulting solid, and then the ether was removed under reduced pressure. Evaporation from this ether was repeated two more times to afford 2-chloro-6-[3-(2-dispiro[2.0.2.1]heptan-7-yl ethoxy)pyrazol-1-yl]pyridine-3-carboxylic acid as a white solid (5.06 g, 100%). 1 H NMR (400 MHz, chloroform-d) δ 8.41 (d, J = 8.5 Hz, 1H), 8.37 (d, J = 2.9 Hz, 1H), 7.75 (d, J = 8.5 Hz, 1H), 5.97 (d, J = 2.9 Hz, 1H), 4.27 (s, 2H), 1.91 (d, J = 6.7 Hz, 2H), 1.50 (s, 1H), 0.85 (d, J = 1.5 Hz, 4H), 0.71 - 0.62 (m, 2H), 0.52 (d, J = 1.1 Hz, 2H). ESI-MS m / z calc. 359.10367, found 360.2 (M+1) + ; Retention time: 2.16 min (LC method B).

[0148] Part B: Synthesis of tert-Butyl (4S)-2,2-Dimethyl-4-[3-[(6-Sulfamoyl-2-pyridyl)amino]propyl]pyrrolidine-1-carboxylate

Chem.

Chem.

[0149] Step 2: 3-Methylenetetrahydropyran-2-one [ka] A 5 L three-necked round-bottom flask was fitted with a mechanical stirrer, heating mantle, addition funnel, J-Kem temperature probe / controller, and nitrogen inlet / outlet. Under a nitrogen atmosphere, (E)-(2-oxotetrahydropyran-3-ylidene)methylone (sodium salt) (205 g, 1.366 mol) and tetrahydrofuran (1640 mL) were packed into the flask, yielding a white suspension. Stirring was initiated, and the pot temperature was recorded at 19°C. Paraformaldehyde (136.6 g, 4.549 mol) was then added to the flask as a solid in a single addition. The resulting suspension was heated to 63°C and maintained at this temperature for 15 hours. Upon heating, the reaction mixture became slightly gelatinous. The white gelatinous mixture was concentrated under reduced pressure to remove most of the tetrahydrofuran. The remaining residue was partitioned in a separatory funnel with ethyl acetate (1000 mL), saturated sodium chloride (500 mL), and saturated sodium bicarbonate (500 mL). Organic matter was removed, and the residual aqueous solution was extracted with ethyl acetate (5 × 300 mL). The combined organic matter was dried over sodium sulfate (500 g) and then vacuum filtered through a glass fritbuchner funnel equipped with a 20 mm Celite layer. The filtration cake was washed by displacement with ethyl acetate (250 mL). The clear filtrate was concentrated under reduced pressure to obtain a clear, pale yellow oil (135 g) as the desired crude product. This substance was purified by silica gel column flash chromatography (liquid load) eluting with a gradient from 100% hexane to 60% ethyl acetate in hexane over 1 hour, and a 450 mL fraction was collected. The product was detected by TLC analysis on silica gel eluting with 3:1 hexane / ethyl acetate and visualized under UV light. The product fractions were combined and concentrated under reduced pressure to obtain the desired product, 3-methylenetetrahydropyran-2-one, as a clear, colorless oil (132 g, 1.18 mol, 72% yield, containing 16 wt% residual ethyl acetate as determined by NMR). ¹H NMR (400 MHz, dimethyl sulfoxide-d6) showed δ6.18 (q, J=1.9 Hz, ¹H), 5.60 (q, J=1.9 Hz, ¹H), 4.40-4.26 (m, ²H), 2.61 (ddt, J=7.0, 6.3, 2.0 Hz, ²H), 1.90-1.75 (m, ²H).

[0150] Step 3: 3-(2-methyl-2-nitro-propyl)tetrahydropyran-2-one [ka] A 5000 mL three-necked round-bottom flask was fitted with a mechanical stirrer, a cooling bath used as a secondary containment vessel, a J-Kem temperature probe, an addition funnel, and nitrogen inlet / outlet. The flask was filled with 2-nitropropane (104.9 g, 1.177 mol) under a nitrogen atmosphere. Stirring was started, and the pot temperature was recorded at 19°C. Next, 1,8-diazabicyclo[5.4.0]undeca-7-ene (22.41 g, 147.2 mmol) was added to the flask in one lump sum, filling it with a clear, pale yellow solution. No exothermic reaction was observed. The addition funnel was filled with a solution of 3-methylenetetrahydropyran-2-one (110 g, 981.0 mmol) in acetonitrile (1100 mL), and this was added dropwise over 1 hour, yielding a clear, pale yellow solution that gradually became exothermic, reaching 24°C. The reaction mixture was stirred at room temperature for 3.5 hours and then concentrated under reduced pressure. The remaining residue was dissolved in dichloromethane (1000 mL) and partitioned with 500 mL of a 3:2 mixture of 1 mole of citric acid solution / saturated sodium chloride solution. The resulting organic phase was a clear, pale blue solution, and the aqueous phase was a slightly cloudy, very pale blue solution. The organic layer was removed, and the remaining aqueous layer was extracted with dichloromethane (300 mL). The combined organic layers were washed with saturated sodium chloride solution (300 mL), dried over sodium sulfate (250 g), and then filtered through a glass frit-Buchner funnel. The filtrate was concentrated under reduced pressure to a volume of approximately 200 mL. The clear, pale blue dichloromethane solution was diluted with methyl tert-butyl ether (1500 mL), and the cloudy solution was concentrated under reduced pressure to a volume of approximately 200 mL to obtain a suspension. The mixture was again diluted with methyl tert-butyl ether (1500 mL) and concentrated under reduced pressure to a volume of approximately 250 mL. The resulting suspension was allowed to stand overnight (approximately 12 hours) at room temperature. The solid was collected by vacuum filtration in a glass frit-Buchner funnel, and the filtration cake was washed by displacement with cold methyl tert-butyl ether (2 × 150 mL), followed by aspiration for 30 minutes. The substance was further dried in a vacuum oven at 45°C for 5 hours to obtain the desired product, 3-(2-methyl-2-nitro-propyl)tetrahydropyran-2-one, as a white solid (160 g, 0.795 mol, yield 81%). 1¹H NMR (400MHz, dimethyl sulfoxide-d6) δ 4.34 (ddd, J=11.1, 9.3, 4.3Hz, 1H), 4.20 (dt, J=11.1, 5.1Hz, 1H), 2.75-2.62 (m, 1H), 2.56 (dd, J=14.9, 5.2Hz, 1H), 2.01-1.89 (m, 2H), 1.89-1.67 (m, 2H), 1.55 (d, J=6.0Hz, 6H), 1.44 (dddd, J=12.8, 11.5, 8.1, 6.6Hz, 1H).

[0151] Step 4: 3-(3-hydroxypropyl)-5,5-dimethylpyrrolidine-2-one [ka] A 1000 mL three-necked round-bottom flask was fitted with a Teflon stirring rod, heating mantle, J-Kem temperature probe / controller, and rubber septum. The flask was filled with 3-(2-methyl-2-nitro-propyl)tetrahydropyran-2-one (25 g, 124.2 mmol) and ethyl alcohol (375 mL) to obtain a white suspension. Stirring was started, and the suspension was heated to 40°C for 10 minutes to obtain a clear, colorless solution. A gas dispersion tube was then fitted to the flask, and the solution was degassed with nitrogen for 15 minutes. The flask was then filled with Raney nickel (8.019 g, 50% w / w, 68.31 mmol), and the septum was fitted. The flask was evacuated and placed under a hydrogen atmosphere. This process was repeated for three cycles. The flask was then placed under a hydrogen atmosphere at 1 atm, and the reaction mixture was gradually heated to 60°C. The reaction mixture was stirred at 60°C for 24 hours. After cooling to room temperature, a gas dispersion tube was fitted to the container, and the reaction mixture was degassed with nitrogen for 15 minutes. The mixture was vacuum filtered through a glass fritbuchner funnel with a 20 mm Celite layer. The filter cake was washed by displacement with ethanol (2 × 100 mL), aspirated until only slight moisture from the ethyl alcohol remained, then moistened with water, and the used Raney nickel catalyst was discarded in water. The clear, pale amber filtrate was concentrated under reduced pressure to obtain a clear, viscous, pale amber oil. The oil was diluted with methyl tert-butyl ether (1500 mL), and the turbid solution was concentrated under reduced pressure to a volume of approximately 150 mL to obtain a suspension. The mixture was again diluted with methyl tert-butyl ether (1500 mL) and concentrated under reduced pressure to a volume of approximately 150 mL. The resulting suspension was allowed to stand overnight (approximately 12 hours) at room temperature. The solid was collected by vacuum filtration in a glass frit-Buchner funnel, and the filtration cake was washed by displacement with cold methyl tert-butyl ether (2 × 50 mL) and then aspirated for 30 minutes. The substance was further dried in a vacuum oven at 45°C for 3 hours to obtain the product, 3-(3-hydroxypropyl)-5,5-dimethylpyrrolidine-2-one, as a white solid (19 g, 0.111 mol, 89% yield). 1¹H NMR (400MHz, dimethyl sulfoxide-d6) δ 7.63 (s, 1H), 3.38 (t, J=6.5Hz, 2H), 2.37 (tdd, J=9.8, 8.5, 4.4Hz, 1H), 2.02 (dd, J=12.3, 8.6Hz, 1H), 1.72 (tdd, J=9.6, 7.5, 4.4Hz, 1H), 1.52-1.32 (m, 3H), 1.28-1.03 (m, 7H).

[0152] Step 5: 3-(5,5-dimethylpyrrolidine-3-yl)propan-1-ol [ka] A 5 L three-necked round-bottom flask was fitted with a mechanical stirrer, heating mantle, addition funnel, J-Kem temperature probe / controller, and nitrogen inlet / outlet. The flask was filled with lithium aluminum hydride pellets (19.39 g, 510.9 mmol) under a nitrogen atmosphere. Next, tetrahydrofuran (500 mL, 20 mL / g) was added to the flask. Stirring was initiated, and the pot temperature reached 20°C. The mixture was stirred at room temperature for 0.5 hours to dissolve the pellets. The resulting grayish suspension reached a pot temperature of 24°C. A tetrahydrofuran (500 mL) solution of 3-(3-hydroxypropyl)-5,5-dimethylpyrrolidine-2-one (25 g, 146.0 mmol) was added to the addition funnel, and the clear, pale yellow solution was added dropwise over 90 minutes. Slight heating was necessary to achieve homogeneity. After completion of the addition, the resulting grayish suspension reached a pot temperature of 24°C. Next, the mixture was heated to a pot temperature of 65°C and maintained at that temperature for 72 hours. Analysis of the reaction mixture at this point showed that some residual starting material still remained and there was no change in product formation. Subsequently, the reaction stopped at this point. The heating mantle was removed and a cooling bath was attached to the vessel. The suspension was cooled to 0°C in a crushed ice / water cooling bath and then quenched by very slow dropwise addition of water (19.93 mL), followed by a 15 wt% sodium hydroxide solution (19.93 mL), and finally water (59.79 mL). The pot temperature of the resulting white suspension was recorded at 5°C. The cooling bath was removed and the heating mantle was again attached to the vessel. The suspension was heated to 60°C and maintained at that temperature for 30 minutes. The warm suspension was vacuum filtered through a glass frit-Buchner funnel with a 20 mm Celite layer. Next, the filtrate was washed by displacement with tetrahydrofuran (2 × 250 mL) at 60°C, followed by aspiration for 30 minutes. The clear filtrate was concentrated under reduced pressure to obtain the desired product, 3-(5,5-dimethylpyrrolidine-3-yl)propan-1-ol, a clear, pale yellow viscous oil (23.5 g, 0.149 mol, 99% yield). 1¹H NMR (400MHz, dimethyl sulfoxide-d6) δ 3.37 (dt, J=8.3, 6.4Hz, 3H), 2.95 (dd, J=10.6, 7.6Hz, 1H), 2.40 (dd, J=10.7, 7.7Hz, 1H), 2.04 (dt, J=16.1, 8.1Hz, 1H), 1.69 (dd, J=12.2, 8.2Hz, 1H), 1.50-1.24 (m, 5H), 1.11-0.94 (m, 7H).

[0153] Step 6: 4-(3-hydroxypropyl)-2,2-dimethylpyrrolidine-1-carboxylate tert-butyl [ka] A 1 L three-neck round-bottom flask was equipped with a mechanical stirrer, a cooling bath, an addition funnel, a J-Kem temperature probe, and a nitrogen inlet / outlet. The vessel was charged with 3-(5,5-dimethylpyrrolidin-3-yl)propan-1-ol (15 g, 95.39 mmol) and dichloromethane (225 mL, 15 mL / g) under a nitrogen atmosphere to obtain a clear pale yellow solution. Stirring was initiated and the pot temperature was recorded at 19 °C. The cooling bath was filled with crushed ice / water to lower the pot temperature to 0 °C. The addition funnel was charged with triethylamine (12.55 g, 124.0 mmol) and then added dropwise as is over 5 minutes. No exotherm was observed. Next, the addition funnel was charged with di-tert-butyl dicarbonate (22.89 g, 104.9 mmol) dissolved in dichloromethane (225 mL). The clear pale yellow solution was then added dropwise over 30 minutes, resulting in gentle gas evolution. No exotherm was observed. The cooling bath was removed and the resulting clear pale yellow solution was warmed to room temperature and stirred at room temperature for 3 hours. The reaction mixture was transferred to a separatory funnel and partitioned with water (75 mL). The organic layer was removed, washed with saturated sodium chloride solution (75 mL), dried over sodium sulfate (150 g), and then filtered through a glass frit Buchner funnel. The filtrate was concentrated under reduced pressure to afford a clear pale yellow oil (30 g) as the desired crude product. This material was purified by silica gel column flash chromatography (liquid injection with dichloromethane) eluting over 60 minutes with a gradient from 100% dichloromethane in dichloromethane to 10% methyl alcohol in dichloromethane, collecting 50 mL fractions. The desired product fractions were combined and concentrated under reduced pressure to afford tert-butyl 4-(3-hydroxypropyl)-2,2-dimethyl-pyrrolidine-1-carboxylate (22 g, 0.0855 mol, 90% yield) as a clear pale yellow viscous oil. 1H NMR(400MHz,DMSO-d6)δ4.38(td,J=5.2,1.4Hz,1H), 3.54(dt,J=10.3,6.7Hz,1H), 3.38(td,J=6.6,3.5Hz,2H), 2.76(q,J=1 0.3Hz,1H), 2.07(td,J=11.6,5.7Hz,1H), 1.87(ddd,J=16.7,12.1,6.0Hz,1H), 1.37(dd,J=14.2,10.4Hz,17H), 1.24(s,3H).

[0154] Step 7: 2,2-dimethyl-4-(3-methylsulfonyloxypropyl)pyrrolidine-1-carboxylate tert-butyl [ka] 4-(3-hydroxypropyl)-2,2-dimethyl-pyrrolidine-1-carboxylate tert-butyl (50.5 g, 196.22 mmol) and triethylamine (39.711 g, 54.698 mL, 392.44 mmol) were dissolved in dichloromethane (500 mL), and the resulting solution was cooled in an ice bath for 30 minutes. Mesyl chloride (24.725 g, 16.706 mL, 215.84 mmol) was added dropwise over 30 minutes, then the ice bath was removed, and the mixture was stirred at room temperature for 1 hour. The reaction product was then quenched with saturated sodium bicarbonate solution (200 mL). The phases were separated, and the organic phase was extracted with saturated sodium bicarbonate (200 mL) and water (2 × 100 mL). The aqueous phase was discarded, the organic phase was dried over sodium sulfate, filtered, and concentrated under vacuum to obtain 2,2-dimethyl-4-(3-methylsulfonyloxypropyl)pyrrolidine-1-carboxylate tert-butyl (64.2 g, 93%) as a pale yellow oil. ESI-MS m / z calculated value: 335.1766, measured value: 336.4 (M+1). + ; Retention time: 5.54 minutes (LC method Q).

[0155] Step 8: 4-(3-aminopropyl)-2,2-dimethylpyrrolidine-1-carboxylate tert-butyl [ka] 2,2-dimethyl-4-(3-methylsulfonyloxypropyl)pyrrolidine-1-carboxylate tert-butyl (64.2 g, 191.38 mmol) was dissolved in dioxane (650 mL), then ammonium hydroxide (650 mL) was added, and the resulting mixture was heated at 45°C for 18 hours. After 18 hours, the reaction mixture was cooled to room temperature. The solution was diluted with 1 M sodium hydroxide (200 mL) and then extracted with diethyl ether (3 × 650 mL). The aqueous phase was discarded, and the combined organic phase was extracted with water (2 × 200 mL). The aqueous phase was discarded, the organic phase was dried over sodium sulfate, filtered, and concentrated under vacuum to obtain 4-(3-aminopropyl)-2,2-dimethyl-pyrrolidine-1-carboxylate tert-butyl (48.9 g, 95%) as a pale yellow oil. ESI-MS m / z calculated value: 256.2151, measured value: 257.3 (M+1) + ; Retention time: 3.70 minutes (LC method Q).

[0156] Step 9: 2,2-dimethyl-4-[3-[(6-sulfamoyl-2-pyridyl)amino]propyl]pyrrolidine-1-carboxylate tert-butyl [ka] 4-(3-aminopropyl)-2,2-dimethylpyrrolidine-1-carboxylate tert-butyl (8.91 g, 34.8 mmol) and 6-fluoropyridine-2-sulfonamide (6.13 g, 34.8 mmol) were added to dimethyl sulfoxide (75 mL), and potassium carbonate (4.91 g, 35.5 mmol) was added. The mixture was stirred at 100 °C for 12 hours, then cooled to ambient temperature and stirred for a further 4 hours (total 16 hours). The reaction mixture was slowly poured into hydrochloric acid (35 mL of 1 M, 35.00 mmol) in water (200 mL) (some foaming) and diluted with ethyl acetate (250 mL). The organic phase was separated and washed with 100 mL of brine. The organic phase was dried on magnesium sulfate, filtered on Celite, and concentrated under vacuum to obtain a dark yellow oil. The crude product was purified by silica gel chromatography eluting with 0% to 100% ethyl acetate in hexane. Both a pure (9.0g) fraction and a non-pure (3g) fraction were collected. The non-pure fraction was purified by silica gel chromatography eluting with 0% to 100% ethyl acetate in hexane to obtain a total of 2,2-dimethyl-4-[3-[(6-sulfamoyl-2-pyridyl)amino]propyl]pyrrolidine-1-carboxylate tert-butyl (10.0g, 69%). 1 H NMR (400MHz, dimethyl sulfoxide-d6) δ7.52(dd,J=8.5,7.2Hz,1H), 7.07(s,2H), 6.95(dd,J=7.2,0.7Hz,2H), 6.61(d,J=8.5Hz,1H), 3.55(q,J=9.1Hz,1H), 3.32-3 .24(m,2H), 2.79(q,J=10.0Hz,1H), 2.13(d,J=16.1Hz,1H), 1.96-1.82(m,1H ), 1.51(dt,J=18.0,9.3Hz,2H), 1.37(dd,J=12.9,10.6Hz,15H), 1.24(s,3H). ESI-MS m / z calc. 412.21442, measured value 413.1 (M+1) + ; Retention time: 2.34 minutes (LC method D).

[0157] Step 10: (4S)-2,2-dimethyl-4-[3-[(6-sulfamoyl-2-pyridyl)amino]propyl]pyrrolidine-1-carboxylate tert-butyl [ka] Racemic 2,2-dimethyl-4-[3-[(6-sulfamoyl-2-pyridyl)amino]propyl]pyrrolidine-1-carboxylate tert-butyl (7 g, 16.97 mmol) was subjected to chiral separation by SFC chromatography using a ChiralPak IG (250 × 21.2 mm column, 5 μm particle size) with a 40% methanol / 60% carbon dioxide mobile phase at 70 mL / min for 11.0 minutes (injection volume = 500 μL of 32 mg / mL solution in methanol). The first peak to elute was (4S)-2,2-dimethyl-4-[3-[(6-sulfamoyl-2-pyridyl)amino]propyl]pyrrolidine-1-carboxylate tert-butyl (3.4481 g, 99%). ESI-MS m / z calculated value: 412.21442, measured value: 413.2 (M+1). + ; Retention time: 0.63 minutes (LC method A).

[0158] Part C: Synthesis of (14S)-8-[3-(2-{dispiro[2.0.2.1]heptan-7-yl}ethoxy)-1H-pyrazole-1-yl]-12,12-dimethyl-2λ6-thia-3,9,11,18,23-pentazatetracyclo[17.3.1.111,14.05,10]tetracosa-1(22),5,7,9,19(23),20-hexaene-2,2,4-trione (compound II) [ka] Step 1: (4S)-4-[3-[[6-[[2-chloro-6-[3-(2-dispiro[2.0.2.1]heptan-7-ylethoxy)pyrazole-1-yl]pyridine-3-carbonyl]sulfamoyl]-2-pyridyl]amino]propyl]-2,2-dimethylpyrrolidine-1-carboxylate tert-butyl [Chemistry] To a solution of 2-chloro-6-[3-(2-dispiro[2.0.2.1]heptan-7-yl ethoxy)pyrazol-1-yl]pyridine-3-carboxylic acid (5.2 g, 14.45 mmol) in tetrahydrofuran (THF) (100 mL) was added carbonyldiimidazole (2.8 g, 16.51 mmol), and the mixture was stirred at ambient temperature for 1 hour. To this mixture was added (4S)-2,2-dimethyl-4-[3-[(6-sulfamoyl-2-pyridyl)amino]propyl]pyrrolidine-1-carboxylic acid tert-butyl (6.0 g, 14.54 mmol) in THF (15 mL), followed by 1,8-diazabicyclo[5.4.0]undec-7-ene (6.5 mL, 43.47 mmol), and the mixture was stirred at ambient temperature for 16 hours. The reaction was diluted with water (150 mL), and the mixture was acidified with aqueous hydrochloric acid (6 M, 15 mL, 90.00 mmol). The mixture was extracted with ethyl acetate (300 mL), and the organic phase was separated. The organic phase was washed with brine, dried over magnesium sulfate, filtered through celite, and concentrated in vacuo to give a white precipitate. The precipitate was slurried in acetonitrile, and the solid was collected by filtration using a medium glass frit and washed with acetonitrile. The filtrate was concentrated in vacuo to give a yellow oil. The crude oil was diluted with acetonitrile and some N-methyl-2-pyrrolidone and chromatographed on a 415 g reverse phase C 18 column eluting with 50% - 100% acetonitrile in water to give tert-butyl (4S)-4-[3-[[6-[[2-chloro-6-[3-(2-dispiro[2.0.2.1]heptan-7-yl ethoxy)pyrazol-1-yl]pyridine-3-carbonyl]sulfamoyl]-2-pyridyl]amino]propyl]-2,2-dimethyl-pyrrolidine-1-carboxylate (4.5 g, 41%). ESI-MS m / z calculated 753.30756, found 754.4 (M + 1) + ; Retention time: 3.79 minutes (LC method D).

[0159] Step 2: 2-Chloro-N-[[6-[3-[(3S)-5,5-dimethylpyrrolidine-3-yl]propylamino]-2-pyridyl]sulfonyl]-6-[3-(2-dispiro[2.0.2.1]heptan-7-ylethoxy)pyrazole-1-yl]pyridine-3-carboxamide (trifluoroacetate) [ka] (4S)-4-[3-[[6-[[2-chloro-6-[3-(2-dispiro[2.0.2.1]heptan-7-ylethoxy)pyrazole-1-yl]pyridine-3-carbonyl]sulfamoyl]-2-pyridyl]amino]propyl]-2,2-dimethyl-pyrrolidine-1-carboxylate tert-butyl (5.9 g, 7.821 mmol) was dissolved in dichloromethane (30 mL) and toluene (15 mL), to which trifluoroacetic acid (6.0 mL, 77.88 mmol) was added, and the mixture was stirred at ambient temperature for 18 hours. The solvent was removed under vacuum with the bath temperature set to 45°C to obtain a dark yellow oil. The oil was diluted with toluene (125 mL), and the solvent was removed under vacuum with the bath temperature set to 45°C. The oil was diluted with toluene, and the solvent was removed under vacuum to obtain a thick, viscous yellow oil, 2-chloro-N-[[6-[3-[(3S)-5,5-dimethylpyrrolidine-3-yl]propylamino]-2-pyridyl]sulfonyl]-6-[3-(2-dispiro[2.0.2.1]heptan-7-ylethoxy)pyrazole-1-yl]pyridine-3-carboxamide (trifluoroacetate) (6.0 g, 100%), which was used in the next step without further purification. ESI-MS m / z calculated value: 653.2551, measured value: 654.3 (M+1) + ; Retention time: 2.6 minutes (LC method B).

[0160] Step 3: (14S)-8-[3-(2-{dispiro[2.0.2.1]heptan-7-yl}ethoxy)-1H-pyrazole-1-yl]-12,12-dimethyl-2λ6-thia-3,9,11,18,23-pentazatetracyclo[17.3.1.111,14.05,10]tetracosa-1(22),5,7,9,19(23),20-hexaene-2,2,4-trione (compound II) [ka] Potassium carbonate (5.3 g, 38.35 mmol) was added to a solution of 2-chloro-N-[[6-[3-[(3S)-5,5-dimethylpyrrolidine-3-yl]propylamino]-2-pyridyl]sulfonyl]-6-[3-(2-dispiro[2.0.2.1]heptan-7-ylethoxy)pyrazole-1-yl]pyridine-3-carboxamide (trifluoroacetate) (6.0 g, 7.810 mmol) in NMP (140 mL). The mixture was purged with nitrogen for 5 minutes. The mixture was then heated at 150 °C for 22 hours. The reaction mixture was cooled to room temperature and added to water (300 mL) to obtain an off-white solid precipitate. The mixture was carefully acidified with aqueous hydrochloric acid (6 M 12 mL, 72.00 mmol) to obtain a foamy slurry. The solid was collected by filtration using a medium-sized glass frit. The wet filter cake was dissolved in ethyl acetate (500 mL) and washed with 200 mL of brine. The aqueous phase was slightly turbid, so it was acidified with a small amount of 6N hydrochloric acid and returned to the organic phase. The aqueous phase was separated, the organic phase was dried over magnesium sulfate, filtered, and concentrated under vacuum to obtain a pale yellow oil. This crude product was diluted with acetonitrile and eluted with 415 g of C in 50% to 100% acetonitrile in water. 18The product was chromatographically treated using a reverse-phase column. The product was isolated as a creamy foam. The foam was dried under vacuum at 45°C for 48 hours to obtain (14S)-8-[3-(2-{dispiro[2.0.2.1]heptan-7-yl}ethoxy)-1H-pyrazole-1-yl]-12,12-dimethyl-2λ6-thia-3,9,11,18,23-pentazatetracyclo[17.3.1.111,14.05,10]tetracosa-1(22),5,7,9,19(23),20-hexaen-2,2,4-trione (compound II) (3.32 g, 68%). 1 H NMR (400MHz, dimethyl sulfoxide-d6) δ12.48(s,1H), 8.20(d,J=2.8Hz,1H), 7.81(d,J=8.2Hz,1H), 7.57(dd,J=8.5,7.2Hz,1H), 7.05( d,J=7.1Hz,1H), 6.97(d,J=8.5Hz,1H), 6.91(d,J=8.2Hz,1H), 6.71(d,J=8.5Hz,1H), 6.08(d,J=2.7Hz,1H), 4.21(td,J=6.7,1. 3Hz,2H), 3.92(d,J=12.0Hz,1H), 3.16(s,1H), 2.95(d,J=13.3Hz,1H), 2.78-2.66(m,1H), 2.07(s,1H), 1.92-1.72(m,4H), 1.6 0(s,6H), 1.51(s,3H), 1.47(t,J=6.5Hz,1H), 1.31(q,J=12.2Hz,1H), 0.89-0.77(m,4H), 0.69-0.61(m,2H), 0.53-0.45(m,2H). ESI-MS m / z calculated value 617.27844, measured value 618.4(M+1) + , retention time: 10.29 min (LC method F).

[0161] Example 5: Compound II (free form) Form A The reactor was equipped with an overhead stirrer, reflux condenser, N2 bubble line and outlet, and a temperature probe. (14S)-8-bromo-12,12-dimethyl-2λ 6 -Tia-3,9,11,18,23-Pentazatetracyclo[17.3.1.1 11 , 14 .0 5, 10 ]Tetracosa-1(23),5,7,9,19,21-hexaene-2,2,4-trione (86% w / w IPAc 120g [103.2g (14S)-8-bromo-12,12-dimethyl-2λ 6 -Tia-3,9,11,18,23-Pentazatetracyclo[17.3.1.1 11 , 14 .0 5 , 10 ]Tetracosa-1(23),5,7,9,19,21-hexaene-2,2,4-trione], 0.21 mol, 1 equivalent), 3-(2-(dispiro[2.0.2 4 .1 3 A mixture of heptane-7-yl)ethoxy)-1H-pyrazole (42.6 g, 0.21 mol, 1 equivalent), 325 mesh K2CO3 (63.4 g, 0.46 mol, 2.2 equivalents), CuI (3.3 g, 17.2 mmol, 0.083 equivalents), and BuOAc (740 mL) was packed into the reactor. The mixture was stirred at ambient temperature. Then DMF (300 mL, 2.9 volumes) and N,N'-dimethylcyclohexane-1,2-diamine (14.6 g or 16.2 ml, 0.1 mol, 0.49 equivalents) was packed into the reactor, and the mixture was purged with three N2 / vacuum / N2 cycles. The mixture was then heated to 120°C for 4 hours and then cooled to ambient temperature. 10% w / v aqueous oxalic acid solution (860 mL, 0.96 mol, 4.6 equivalents) was added dropwise, and the mixture was stirred for at least 1 hour. The mixture was then filtered to remove the suspended solid. The removed solid was washed with (2 × 120 mL). The layers were separated from the filtrate. The organic layer was washed with 8% w / v trisodium citrate solution (600 mL). Brine was added as needed to aid phase separation. The organic layer was washed with 1:1 v / v water / brine (400 mL). The organic layer was filtered through a Celite pad. The filter pad was washed with IPAc (150 mL). The filtrate was concentrated, and then 800 mL of 1-PrOH (7.8 vol) was added to concentrate the mixture. This step was repeated once more. Toluene (800 mL) was added, and the mixture was concentrated. This step was repeated once more to obtain a concentrated slurry. The crude mixture was concentrated to the volume of 300 mL (2.9 vol) of toluene. After stirring the slurry overnight, the solid was collected by filtration and washed with toluene (2 × 100 mL, 0.97 vol). The solid was dried under vacuum at 50°C with nitrogen bleeding until the loss on drying was less than 1.0%, yielding compound II (107.0 g, 83%, 94.5% (AUC) HPLC purity) as a white / off-white solid.

[0162] Recrystallization: Compound II form A [22.2 g, 94.6% (AUC)] was suspended in toluene (440 mL, 20 vols based on Compound II), and the mixture was heated under reflux. After maintaining reflux for up to 2 hours, the mixture was cooled to ambient temperature over 8 hours. After stirring the slurry overnight at ambient temperature, the solid was collected by filtration and washed with toluene (40 mL, 1.8 vol). The solid was dried under vacuum at 50°C with nitrogen bleeding until the loss on drying was less than 1.0%, yielding Compound II form A (18.8 g, 84%, 96.8% (AUC) HPLC purity) as a white / off-white solid.

[0163] Second recrystallization: Compound II form A [17.5 g, 97.0% (AUC) Compound II form A] was suspended in toluene (350 mL, 20 vols based on Compound II), and the mixture was heated under reflux. After holding under reflux for more than 2 hours, the mixture was cooled to ambient temperature over 8 hours. After stirring the slurry overnight at ambient temperature, the solid was collected by filtration and washed with toluene (40 mL, 1.8 vols). The solid was dried under vacuum at 50°C with nitrogen bleeding until the loss on drying was less than 1.0%, yielding Compound II form A (free form) (15.7 g, 89%, 98.4% (AUC) HPLC purity) as a white / off-white solid.

[0164] Free form A of compound II is the most stable polymorph, with a water activity of ≤0.95 at ambient temperature.

[0165] AX-ray powder diffraction XRPD patterns were acquired at room temperature in reflection mode using a Bruker Advance with a Vantec-1 detector. Samples were analyzed in a silicon sample holder in continuous mode with a step size of 0.0144531° and a step time of 0.25 seconds, at 3–40° 2-theta. The sample was rotated at 15 rpm. The XRPD diffractogram of compound II (free form) morph A is provided in Figure 1, and the XRPD data are summarized in Table 5 below. [Table 18]

[0166] B. Elucidation of Single Crystals A single crystal with the structure of compound II (free form) form A was grown from acetone / heptane. X-ray diffraction data was obtained from Mo K α The diffractometer was equipped with a line (λ=0.71073Å) and a CCD detector, and acquired at 298K. The structure was elucidated and refined using the SHELX program (Sheldrick, GM, Acta Cryst., (2008) A64, 112-122), and the results are summarized in Table 6 below. [Table 19]

[0167] C. Solid-state NMR 1. Solid-state NMR experiment (applicable to all crystalline forms of compound II): A Bruker-Biospin 400MHz wide-bore spectrometer equipped with a Bruker-Biospin 4mm HFX probe was used. The sample was packed into a 4mm ZrO2 rotor and rotated under magic angle rotation (MAS) conditions at a rotation speed typically set to 12.5kHz. 13 In order to set an appropriate recycling delay in C cross-polarization (CP) MAS experiments, the proton relaxation time 1 Measurements were performed using the H MAS T1 saturation recovery relaxation experiment. The CP contact time for the carbon CPMAS experiment was set to 2 milliseconds. A CP proton pulse with a linear ramp (50%~100%) was used. The carbon Hartmann-Hahn match was optimized with an external reference sample (glycine). The carbon spectrum was recorded by proton decoupling using a TPPM15 decoupling array at an electric field strength of approximately 100 kHz.

[0168] 2. Solid-state NMR of Compound II (free form) in form A Compound II (free form): Solid form A 13 The 13C NMR data is shown in Figure 2 and summarized in Table 7 below. [Table 20]

[0169] D. Differential Scanning Calorimetry Analysis DSC was performed using a TA Discovery differential scanning calorimeter (TA Instruments, New Castle, DE). The instrument was calibrated with indium. Approximately 1–10 mg of sample was weighed into a sealed pan and crimped using a lid with a single hole. The DSC sample was scanned at a heating rate of 10°C / min from 25°C to 300°C. Data was collected and analyzed using Trios Analysis software (TA Instruments, New Castle, DE). The thermogram showed a single fusion endothermic peak at approximately 227°C.

[0170] Example 6: Compound II calcium salt hydrate form A Form A of the calcium salt hydrate of compound II is the kinetically most preferred form of calcium salt hydrate, resulting in higher solubility, solubility, and exposure compared to other forms of calcium salt hydrate.

[0171] The calcium salt hydrate form A of compound II was prepared by packing 0.2 mmol of compound II (free form) form A and 0.1 mmol of Ca(OMe)2 dry powder with approximately 45 mg / mL of IPA, spiking with approximately 10% water, and heating to 70°C. Initially, all solids dissolved. After 5 minutes, a white solid precipitated. The resulting slurry was stirred at room temperature for 4 days. The solid was isolated as the calcium salt hydrate form A of compound II by vacuum filtration and dried overnight at 40°C under vacuum (isolation yield approximately 78%).

[0172] Another method for preparing Compound II calcium salt hydrate form A involved using 10 g of Compound I (free form A) packed with 63 mL of IPA and 7 mL of water. The slurry was heated to 55-65°C. 1.1 equivalents of NaOH were added to the mixture. The mixture was stirred until the solution was homogeneous. The solution was then cooled to 25°C, and 0.1 g of Compound II sodium salt hydrate form A was seeded in. The slurry was stirred for 18 hours. The solution was then heated to 45°C. 0.1 g of Compound II calcium salt hydrate form A was seeded into the slurry. A solution of 0.55 equivalents of CaCl2, 9 mL of IPA, and 1 mL of water was added over 5 hours. The resulting slurry was stirred for 2 hours. The slurry was cooled to 20°C over 5 hours. The resulting solid was collected by vacuum filtration, and the resulting wet cake was washed with 50 mL of water. The washed wet cake was air-dried for 1 hour. The air-dried, moist cake was transferred to a vacuum oven at 45°C for 20 hours with slight nitrogen bleeding to obtain crystalline compound II calcium salt hydrate form A (8.5 g, isolation yield 82%).

[0173] AX-ray powder diffraction: XRPD patterns were acquired in reflection mode at room temperature using a Bruker Advance with a Vantec-1 detector. Samples were analyzed in a silicon sample holder in continuous mode with a step size of 0.0144531° and a step time of 0.25 seconds, at 3–40° 2-theta. The sample was rotated at 15 rpm. The XRPD diffractogram of compound II calcium salt hydrate form A is shown in Figure 3 and summarized in Table 8. [Table 21]

[0174] B. Elucidation of Single Crystals Crystals having the structure of compound II calcium salt hydrate form A were grown by dissolving 1 mg of compound II calcium salt hydrate form A in 350 μL of a 90 / 10 mixture of dichloroethane / ethanol, and then diffusing the vapor with pentane for several days. X-ray diffraction data were obtained from Cu K αThe structures were acquired at both 100K and 298K using a Bruker diffractometer equipped with a line (λ=1.5478Å) and a CCD detector. The structures were elucidated and refined using the SHELX program (Sheldrick, GM, Acta Cryst., (2008) A64, 112-122), and the results are summarized in Table 9 below. [Table 22]

[0175] C. Solid-state NMR Solid form of compound II calcium salt hydrate A 13 The 13C NMR spectra are shown in Figure 4 and summarized in Table 10. [Table 23]

[0176] D. Differential scanning calorimetry analysis: DSC thermograms were obtained using a TA Instruments DSC Q2000. The sample was heated at 10°C / min from 30°C to 350°C. The thermogram showed an endothermic peak at approximately 223°C.

[0177] Example 7: Compound II calcium salt hydrate form D Compound II calcium salt hydrate form D is the most stable form of calcium salt hydrate under specific conditions, such as a mixture of ethanol and water.

[0178] Approximately 25 mg of Compound II calcium salt hydrate form A was added to 0.5 mL of EtOH:water 67:33 w / w. The slurry was heated at 65°C for 8 days. The resulting solid, collected by vacuum filtration, was Compound II calcium salt hydrate form D.

[0179] Alternatively, the calcium salt hydrate form D of compound II was prepared from 89 g of sodium hydrate form A of compound II, mixed with 1080 mL of IPA and 120 mL of water. The slurry was heated to 55-65°C. 18 g of compound II calcium salt hydrate form D seeds were added to the slurry. This slurry was wet-milled as a solution of 0.55 equivalents of CaCl2, 81 mL of IPA, and 9 mL of water, added over 5 hours. The wet mill was operated until it was confirmed by X-ray powder diffraction that the slurry consisted entirely of compound I calcium salt hydrate form D. The obtained solid was collected by vacuum filtration, and the wet cake was washed with 350 mL of water. The washed wet cake was air-dried for 1 hour. The air-dried wet cake was transferred to a vacuum oven at 45°C for 20 hours with slight nitrogen bleeding to obtain crystalline compound II calcium salt hydrate form D (83.15 g, isolation yield 90.6%).

[0180] Compound II calcium salt hydrate form D has a water activity of 0.1–0.95 from ambient temperature to 60°C and is the most stable polymorph in IPA / water.

[0181] AX-ray powder diffraction: X-ray powder diffraction (XRPD) spectra were recorded in reflection mode at room temperature using a PANalytical Empyrean system equipped with a sealed tube source and a PIXcel 1D Medipix-2 detector (Malvern PANalytical Inc., Westborough, Massachusetts). The X-ray generator was operated with a voltage of 45 kV and a current of 40 mA using a copper wire (1.54060 Å). The powder sample was placed in a back-filled sample holder and loaded into the instrument. The sample was scanned over a range of approximately 3° to approximately 40°²θ with a step size of 0.0131303° and 49.725 seconds per step. The XRPD diffractogram of compound I calcium salt hydrate form D is shown in Figure 5 and summarized in Table 11. [Table 24-1] [Table 24-2]

[0182] B. Elucidation of Single Crystals Crystals are selected from a compound II calcium salt hydrate form D seeding process in ethanol / water. X-ray diffraction data are provided for Cu K provided with a Rigaku MM007HF rotating anode. a The images were acquired at 100K using a Bruker diffractometer equipped with a line (l=1.5478) and a CMOS detector. The structure was elucidated and refined using the SHELX program (Sheldrick, GM, Acta Cryst., (2008) A64, 112-122), and the results are summarized in Table 12. [Table 25]

[0183] C. Solid-state NMR: Solid form of compound I calcium salt hydrate C 13 The 13C NMR spectra are shown in Figure 6 and summarized in Table 13. [Table 26-1] [Table 26-2]

[0184] D. Differential Scanning Calorimetry Analysis DSC was performed using a TA Discovery differential scanning calorimeter (TA Instruments, New Castle, DE). The instrument was calibrated with indium. Approximately 1–10 mg of sample was weighed into a sealed pan and crimped using a lid with one hole. The DSC sample was scanned at a heating rate of 10°C / min from 25°C to 300°C. Data was collected and analyzed using Trios Analysis software (TA Instruments, New Castle, DE). The thermogram showed multiple endothermic peaks at approximately 182°C and 208°C.

[0185] Example 8: Efficacy data of 250 mg of compound I In the clinical trial, a comparative study was conducted on the absolute change in SwCl at 12 weeks in subjects with gating mutations who were receiving stable treatment with ibakhtol. Compound I was generally considered safe and well-tolerated at 150 mg and 250 mg doses for 12 weeks. The 250 mg dose showed improvement in SwCl at 12 weeks compared to ibakhtol baseline, while the 150 mg dose of compound I showed a decrease in SwCl compared to ibakhtol baseline. [Table 27]

[0186] Example 9: Preparation of Exemplary Tablet Formulation The granular components listed in Table 15 (spray-dried dispersion (SDD) of Compound I, calcium salt hydrate form D of Compound II, SDD of Compound III, microcrystalline cellulose, and croscarmellose sodium) were weighed, sieved through a screen, and placed in a bin blender. These components were blended to prepare the granular powder blend. The granular powder blend was dry-granulated using a roller compactor and then crushed into granules. The extragranular microcrystalline cellulose was weighed, passed through a screen, and mixed with the crushed granules in the bin blender. Magnesium stearate was weighed, sieved, and then added to the bin blender and mixed. The blended components were compressed using an electric assisted rotary tablet press to prepare a table with the required core weight and hardness. The tablets were then placed in a coater and a non-functional coating was applied. [Table 28]

[0187] Other Embodiments The foregoing description discloses and describes only exemplary embodiments of the present disclosure. Those skilled in the art will readily recognize from such description, as well as the accompanying drawings and claims, that various changes, modifications, and variations may be made without departing from the spirit and scope of the present disclosure as defined in the following claims. In one embodiment, for example, the following items are provided. (Item 1) A method for treating cystic fibrosis, (a) 250 mg of compound I or an equivalent amount of a pharmaceutically acceptable salt thereof, (b) 21.24 mg of Compound II calcium salt hydrate in form D [ka] (c) Compound III 100 mg [ka] A method comprising administering a comparable amount of the pharmaceutically acceptable salt daily. (Item 2) A method for treating cystic fibrosis, for patients who require treatment. (a) 250 mg of compound I. (b) 21.240 mg of Compound II calcium salt hydrate form D, and (c) 100 mg of compound III, A method that includes administering the medication daily. (Item 3) The method according to item 1 or 2, wherein compounds I, II, and III are administered in separate compositions. (Item 4) The method according to item 1 or 2, wherein compounds I, II, and III are administered in a single composition. (Item 5) The method according to item 1 or 2, wherein compounds I, II, and III are administered once daily as two compositions, each composition comprising 125 mg of compound I, 10.62 mg of compound II calcium salt hydrate form D, and 50 mg of compound III. (Item 6) A method for treating cystic fibrosis, comprising 250 mg of compound I [ka] A method comprising administering, or a comparable amount of the pharmaceutically acceptable salt thereof, once daily to a patient in need of treatment. (Item 7) The method according to any one of items 1 to 6, wherein the patient is homozygous for the F508del mutation or has the F508del / minimal-function genotype, F508del / gating genotype, or F508del / residual-function genotype. (Item 8) The method according to any one of items 1 to 6, wherein the patient has a heterozygous genotype and one F508del mutation. (Item 9) The method described in item 8, wherein the patient has one mutation selected from Table 3. (Item 10) The method described in item 8, wherein the patient has one mutation selected from Table 4. (Item 11) The method according to any one of items 1 to 6, wherein the patient has at least one mutation selected from Table 4. (Item 12) (a) 250 mg of compound I or an equivalent amount of a pharmaceutically acceptable salt thereof (b) 21.24 mg of Compound II calcium salt hydrate form D, and (c) A pharmaceutical composition comprising 100 mg of compound III or an equivalent amount thereof of a pharmaceutically acceptable salt. (Item 13) (a) 250 mg of compound I, (b) 21.24 mg of Compound II calcium salt hydrate form D, and (c) A pharmaceutical composition containing 100 mg of compound III. (Item 14) (a) 125 mg of compound I or an equivalent amount of a pharmaceutically acceptable salt thereof (b) 10.62 mg of Compound II calcium salt hydrate form D, and (c) A pharmaceutical composition comprising 50 mg of compound III or an equivalent amount thereof of a pharmaceutically acceptable salt. (Item 15) (a) 125 mg of compound I, (b) 10.62 mg of Compound II calcium salt hydrate form D, and (c) A pharmaceutical composition containing 50 mg of compound III. (Item 16) (a) A solid dispersion containing compound I in a quantity of 156.3 mg, wherein the solid dispersion is (i) Compound I, in an amount of 80% by weight relative to the weight of the solid dispersion. (ii) 19.5% by weight of the solid dispersion, and (iii) A solid dispersion comprising 0.5% by weight of sodium lauryl sulfate relative to the weight of the solid dispersion, (b) 10.6 mg of Compound II calcium salt hydrate form D, (c) A solid dispersion containing compound III in a quantity of 62.5 mg. The solid dispersion is (i) 80% by weight of compound III and (ii) A solid dispersion comprising 20% ​​by weight of hypromellose relative to the weight of the solid dispersion, (d) 124.0 mg of microcrystalline cellulose and (e) 22.8 mg of croscarmellose sodium, (f) A tablet containing 3.8 mg of magnesium stearate. (Item 17) (a) A solid dispersion containing compound I in a quantity of 156.3 mg, wherein the solid dispersion is (i) Compound I, in an amount of 80% by weight relative to the weight of the solid dispersion. (ii) 19.5% by weight of the solid dispersion, and (iii) A solid dispersion comprising 0.5% by weight of sodium lauryl sulfate relative to the weight of the solid dispersion, (b) 10.6 mg of Compound II calcium salt hydrate form D, (c) A solid dispersion containing compound III in a quantity of 62.5 mg. The solid dispersion is (i) 80% by weight of compound III and (ii) A solid dispersion comprising 20% ​​by weight of hypromellose relative to the weight of the solid dispersion, (d) 124.0 mg of microcrystalline cellulose and (e) 22.8 mg of croscarmellose sodium, (f) 3.8 mg of magnesium stearate and (g) A tablet containing 11.4 mg of film-coated material. (Item 18) (a) A solid dispersion containing compound I in a quantity of 156.3 mg, wherein the solid dispersion is (i) Compound I, in an amount of 80% by weight relative to the weight of the solid dispersion. (ii) 19.5% by weight of the solid dispersion, and (iii) A solid dispersion comprising 0.5% by weight of sodium lauryl sulfate relative to the weight of the solid dispersion, (b) 10.6 mg of Compound II calcium salt hydrate form D, (c) A solid dispersion containing compound III in a quantity of 62.5 mg. The solid dispersion is (i) 80% by weight of compound III and (ii) A tablet comprising a solid dispersion containing 20% ​​by weight of hypromellose relative to the weight of the solid dispersion. (Item 19) The tablet according to item 18, wherein the tablet further comprises 70 to 170 mg of microcrystalline cellulose. (Item 20) The tablet according to item 18, wherein the tablet further comprises 10 to 40 mg of croscarmellose sodium. (Item 21) The tablet according to item 18, wherein the tablet further comprises 70 to 170 mg of microcrystalline cellulose and 10 to 40 mg of croscarmellose sodium. (Item 22) (a) A solid dispersion containing compound I in a quantity of 156.3 mg, wherein the solid dispersion is (i) Compound I, in an amount of 80% by weight relative to the weight of the solid dispersion. (ii) 19.5% by weight of the solid dispersion, and (iii) A solid dispersion comprising 0.5% by weight of sodium lauryl sulfate relative to the weight of the solid dispersion, (b) 10.6 mg of Compound II calcium salt hydrate form D, (c) A solid dispersion containing compound III in a quantity of 62.5 mg. The solid dispersion is (i) 80% by weight of compound III and (ii) A solid dispersion comprising 20% ​​by weight of hypromellose relative to the weight of the solid dispersion, (d) 125.0 mg of microcrystalline cellulose and (e) 22.8 mg of croscarmellose sodium, (f) A tablet containing 2.9 mg of magnesium stearate. (Item 23) (a) A solid dispersion containing compound I in a quantity of 156.3 mg, wherein the solid dispersion is (i) Compound I, in an amount of 80% by weight relative to the weight of the solid dispersion. (ii) 19.5% by weight of the solid dispersion, and (iii) A solid dispersion comprising 0.5% by weight of sodium lauryl sulfate relative to the weight of the solid dispersion, (b) 10.6 mg of Compound II calcium salt hydrate form D, (c) A solid dispersion containing compound III in a quantity of 62.5 mg. The solid dispersion is (i) 80% by weight of compound III and (ii) A solid dispersion comprising 20% ​​by weight of hypromellose relative to the weight of the solid dispersion, (d) 124.5 mg of microcrystalline cellulose and (e) 22.8 mg of croscarmellose sodium, (f) 3.8 mg of magnesium stearate and optionally A tablet containing (g) 15.9 mg of film-coated material. (Item 24) 250 mg of compound I

change

Claims

1. A combination for use in the treatment of cystic fibrosis, the following: (a) Compound I: 【Chemistry 57】 or a pharmaceutically acceptable salt thereof, (b) Compound II calcium salt hydrate form D: 【Chemistry 1】 (c) Compound III: 【Chemistry 58】 or a pharmaceutically acceptable salt thereof Includes, The aforementioned treatments are as follows: (a) 250 mg of compound I, or an equivalent amount of a pharmaceutically acceptable salt thereof, and (b) 21.24 mg of Compound II calcium salt hydrate form D, and (c) 100 mg of compound III, or an equivalent amount of its pharmaceutically acceptable salt. This includes daily administration to patients requiring treatment, A combination of the aforementioned compound II calcium salt hydrate form D, characterized by X-ray powder diffractograms (XRPDs) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

2. A combination for use in the treatment of cystic fibrosis, (a) Compound I: 【Transformation 3】 (b) Compound II calcium salt hydrate form D: 【Chemistry 1】 (c) Compound III: 【Chemistry 59】 Includes, The aforementioned treatments are as follows: (a) 250 mg of compound I, (b) 21.24 mg of Compound II calcium salt hydrate form D, and (c) 100 mg of compound III This includes daily administration to patients requiring treatment, A combination of the aforementioned compound II calcium salt hydrate form D, characterized by X-ray powder diffractograms (XRPDs) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

3. The combination according to claim 1, wherein the treatment comprises administration of separate compositions of compound I, II, and III.

4. The combination according to claim 1, wherein the treatment comprises administration of a single composition of compound I, II, and III.

5. The combination according to claim 1, wherein the treatment comprises once-daily administration of two compositions of compound I, II, and III, each composition comprising 125 mg of compound I, 10.62 mg of compound II calcium salt hydrate form D, and 50 mg of compound III.

6. The combination according to claim 1, wherein the patient is homozygous for the F508del cystic fibrosis membrane conductance regulator (CFTR) mutation, or has the F508del / minimal-function CFTR genotype, the F508del / gating CFTR genotype, or the F508del / residual-function CFTR genotype.

7. The combination according to claim 1, wherein the patient has a heterozygous genotype and one F508del cystic fibrosis membrane conductance regulatory factor (CFTR) mutation.

8. The aforementioned patient: Table 29 The combination according to claim 7, having one CFTR mutation selected from.

9. The aforementioned patient, as follows: Table 30-1 Table 30-2 Table 30-3 Having one CFTR mutation selected from, Herein, D443Y;G576A;R668C, R74W;D1270N, R74W;V201M, R74W;V201M;D1270N, F508C;S1251N, and G576A;R668C are compound mutations, wherein a single allele of the CFTR gene has multiple mutations, and the multiple mutations exist independently of the presence of mutations on other alleles, as described in claim 7.

10. The aforementioned patient, as follows: Table 31-1 Table 31-2 Table 31-3 Having at least one cystic fibrosis membrane conductance regulator (CFTR) mutation selected from, Herein, D443Y;G576A;R668C, R74W;D1270N, R74W;V201M, R74W;V201M;D1270N, F508C;S1251N, and G576A;R668C are compound mutations, wherein a single allele of the CFTR gene has multiple mutations, and the multiple mutations exist independently of the presence of mutations on other alleles, as described in claim 1.

11. (a) 250 mg of compound I: 【Chem.99】 Or a comparable amount of its pharmaceutically acceptable salt, (b) 21.24 mg of Compound II calcium salt hydrate in form D: 【Chemistry 100】 , and (c) Compound III in 100 mg: 【Chemistry 101】 or containing a substantial amount of its pharmaceutically acceptable salt, A pharmaceutical composition for use in the treatment of cystic fibrosis, wherein the compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

12. (a) 250 mg of compound I: 【Chemical Engineering 102】 、 (b) 21.24 mg of Compound II calcium salt hydrate in form D: 【Chemistry 103】 , and (c) Compound III in 100 mg: 【Chemical 104】 Includes, A pharmaceutical composition for use in the treatment of cystic fibrosis, wherein the compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

13. (a) 125 mg of compound I: 【Chemistry 61】 Or a comparable amount of its pharmaceutically acceptable salt, (b) 10.62 mg of Compound II calcium salt hydrate in form D: 【Chemistry 1】 (c) Compound III at 50 mg: 【Transformation 62】 or containing a substantial amount of its pharmaceutically acceptable salt, A pharmaceutical composition for use in the treatment of cystic fibrosis, wherein the compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

14. (a) 125 mg of compound I: 【Transformation 3】 (b) 10.62 mg of Compound II calcium salt hydrate in form D: 【Chemistry 1】 (c) Compound III at 50 mg: 【Chemistry 105】 Includes, A pharmaceutical composition for use in the treatment of cystic fibrosis, wherein the compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

15. (a) Compound I: 【Transformation 63】 A solid dispersion containing 156.3 mg, wherein the solid dispersion is (i) 80% by weight of compound I, (ii) 19.5% by weight of the solid dispersion, and (iii) A solid dispersion comprising 0.5% by weight of sodium lauryl sulfate relative to the weight of the solid dispersion, (b) 10.6 mg of Compound II calcium salt hydrate in form D: 【Chemistry 64】 and, (c) A 62.5 mg solid dispersion containing compound III, wherein the solid dispersion is (i) Compound III in an amount of 80% by weight relative to the weight of the solid dispersion: 【Transformation 65】 , and (ii) A solid dispersion comprising 20% ​​by weight of hypromellose relative to the weight of the solid dispersion, (d) 124.0 mg of microcrystalline cellulose and (e) 22.8 mg of croscarmellose sodium, (f) Contains 3.8 mg of magnesium stearate, A tablet for use in the treatment of cystic fibrosis, wherein the compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

16. (a) Compound I: 【Chemical Formula 66】 A solid dispersion containing 156.3 mg, wherein the solid dispersion is (i) 80% by weight of compound I, (ii) 19.5% by weight of the solid dispersion, and (iii) A solid dispersion comprising 0.5% by weight of sodium lauryl sulfate relative to the weight of the solid dispersion, (b) 10.6 mg of Compound II calcium salt hydrate in form D: 【Transformation 67】 and, (c) A 62.5 mg solid dispersion containing compound III, wherein the solid dispersion is (i) Compound III in an amount of 80% by weight relative to the weight of the solid dispersion: 【Transformation 68】 , and (ii) A solid dispersion comprising 20% ​​by weight of hypromellose relative to the weight of the solid dispersion, (d) 124.0 mg of microcrystalline cellulose and (e) 22.8 mg of croscarmellose sodium, (f) 3.8 mg of magnesium stearate and (g) Contains 11.4 mg of film coating, A tablet for use in the treatment of cystic fibrosis, wherein the compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

17. (a) Compound I: 【Transformation 69】 A solid dispersion containing 156.3 mg, wherein the solid dispersion is (i) 80% by weight of compound I, (ii) 19.5% by weight of the solid dispersion, and (iii) A solid dispersion comprising 0.5% by weight of sodium lauryl sulfate relative to the weight of the solid dispersion, (b) 10.6 mg of Compound II calcium salt hydrate in form D: 【Transformation 70】 and, (c) A 62.5 mg solid dispersion containing compound III, wherein the solid dispersion is (i) Compound III in an amount of 80% by weight relative to the weight of the solid dispersion: 【Chemistry 71】 , and (ii) A solid dispersion comprising 20% ​​by weight of hypromellose relative to the weight of the solid dispersion, A tablet for use in the treatment of cystic fibrosis, wherein the compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

18. The tablet according to claim 17, wherein the tablet further comprises 70 to 170 mg of microcrystalline cellulose.

19. The tablet according to claim 17, wherein the tablet further comprises 10 to 40 mg of croscarmellose sodium.

20. The tablet according to claim 17, wherein the tablet further comprises 70 to 170 mg of microcrystalline cellulose and 10 to 40 mg of croscarmellose sodium.

21. (a) Compound I: 【Chemistry 72】 A solid dispersion containing 156.3 mg, wherein the solid dispersion is (i) 80% by weight of compound I, (ii) 19.5% by weight of the solid dispersion, and (iii) A solid dispersion comprising 0.5% by weight of sodium lauryl sulfate relative to the weight of the solid dispersion, (b) 10.6 mg of Compound II calcium salt hydrate in form D: 【Transformation 73】 and, (c) Compound III: 【Chemistry 74】 A 62.5 mg solid dispersion containing, the solid dispersion is (i) 80% by weight of compound III and (ii) A solid dispersion comprising 20% ​​by weight of hypromellose relative to the weight of the solid dispersion, (d) 125.0 mg of microcrystalline cellulose and (e) 22.8 mg of croscarmellose sodium, (f) Contains 2.9 mg of magnesium stearate, A tablet for use in the treatment of cystic fibrosis, wherein the compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

22. (a) Compound I: 【Chemistry 75】 A solid dispersion containing 156.3 mg, wherein the solid dispersion is (i) 80% by weight of compound I, (ii) 19.5% by weight of the solid dispersion, and (iii) A solid dispersion comprising 0.5% by weight of sodium lauryl sulfate relative to the weight of the solid dispersion, (b) 10.6 mg of Compound II calcium salt hydrate in form D: 【Transformation 76】 and, (c) A 62.5 mg solid dispersion containing compound III, wherein the solid dispersion is (i) Compound III in an amount of 80% by weight relative to the weight of the solid dispersion: 【Chemical 77】 , and (ii) A solid dispersion comprising 20% ​​by weight of hypromellose relative to the weight of the solid dispersion, (d) 124.5 mg of microcrystalline cellulose and (e) 22.8 mg of croscarmellose sodium, (f) 3.8 mg of magnesium stearate and optionally (g) Contains 15.9 mg of film coating, A tablet for use in the treatment of cystic fibrosis, wherein the compound II calcium salt hydrate form D is characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

23. The combination according to any one of claims 1 to 10, the pharmaceutical composition according to any one of claims 11 to 14, or the tablet according to any one of claims 15 to 22, wherein the compound II calcium salt hydrate form D is characterized by an XRPD having (a) signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta, and (b) one or more signals selected from 5.5 ± 0.2 degrees 2-theta, 15.5 ± 0.2 degrees 2-theta, 19.7 ± 0.2 degrees 2-theta, 21.5 ± 0.2 degrees 2-theta, 22.1 ± 0.2 degrees 2-theta, 23.0 ± 0.2 degrees 2-theta, and 27.6 ± 0.2 degrees 2-theta.

24. The combination, pharmaceutical composition, or tablet according to claim 23, wherein the compound II calcium salt hydrate form D is characterized by an XRPD having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, 22.8 ± 0.2 degrees 2-theta, and 27.6 ± 0.2 degrees 2-theta.

25. The combination, pharmaceutical composition, or tablet according to claim 23, wherein the compound II calcium salt hydrate form D is characterized by an XRPD having signals of 6.1±0.2 degrees 2-theta, 15.5±0.2 degrees 2-theta, 16.2±0.2 degrees 2-theta, 19.7±0.2 degrees 2-theta, 22.8±0.2 degrees 2-theta, and 27.6±0.2 degrees 2-theta.

26. The aforementioned compound II calcium salt hydrate form D is 【Transformation 78】 A combination according to any one of claims 1 to 10, a pharmaceutical composition according to any one of claims 11 to 14, or a tablet according to any one of claims 15 to 22, characterized by an X-ray powder diffractogram substantially similar to that of the combination according to any one of claims 1 to 10.

27. The aforementioned compound II calcium salt hydrate form D is Cu K α The following were measured at 100 K using a Bruker diffractometer equipped with a line (λ = 1.5478 Å): Triclinic system, P1 space group, and below. 【Transformation 79】 A combination according to any one of claims 1 to 10, a pharmaceutical composition according to any one of claims 11 to 14, or a tablet according to any one of claims 15 to 22, characterized by the unit cell dimensions.

28. The compound II calcium salt hydrate form D has one or more peaks selected from 130.2 ± 0.2 ppm, 125.6 ± 0.2 ppm, and 35.0 ± 0.2 ppm. １３ C solid-state nuclear magnetic resonance ( １３ A combination according to any one of claims 1 to 10, a pharmaceutical composition according to any one of claims 11 to 14, or a tablet according to any one of claims 15 to 22, characterized by a C ssNMR spectrum.

29. The compound II calcium salt hydrate form D has one or more peaks selected from 179.8±0.2 ppm, 130.2±0.2 ppm, 125.6±0.2 ppm, 120.9±0.2 ppm, 55.2±0.2 ppm, 44.3±0.2 ppm, 35.0±0.2 ppm, and 1.6±0.2 ppm. １３ The combination, pharmaceutical composition, or tablet according to claim 28, characterized by a C ssNMR spectrum.

30. The compound II calcium salt hydrate form D has (a) one or more peaks selected from 130.2 ± 0.2 ppm, 125.6 ± 0.2 ppm, and 35.0 ± 0.2 ppm, and (b) one or more peaks selected from 176.9 ± 0.2 ppm, 160.9 ± 0.2 ppm, 142.0 ± 0.2 ppm, and 98.6 ± 0.2 ppm. １３ The combination, pharmaceutical composition, or tablet according to claim 28, characterized by a C ssNMR spectrum.

31. The aforementioned compound II calcium salt hydrate form D is 【Chemistry 80】 It is substantially similar to １３ A combination according to any one of claims 1 to 10, a pharmaceutical composition according to any one of claims 11 to 14, or a tablet according to any one of claims 15 to 22, characterized by a solid-state nuclear magnetic resonance spectrum.

32. A pharmaceutical composition for use in the treatment of cystic fibrosis, Compound I: 【Chemistry 81】 or containing a pharmaceutically acceptable salt thereof, The aforementioned pharmaceutical composition, Compound II calcium salt hydrate form D: 【Chemistry 82】 , and Compound III: 【Chemistry 83】 or a pharmaceutically acceptable salt thereof Administered in combination, The aforementioned treatments are as follows: (a) 250 mg of compound I, or an equivalent amount of a pharmaceutically acceptable salt thereof, and (b) 21.24 mg of Compound II calcium salt hydrate form D, and (c) 100 mg of compound III, or an equivalent amount of its pharmaceutically acceptable salt. It is characterized by including daily administration of the drug, A pharmaceutical composition characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

33. A pharmaceutical composition for use in the treatment of cystic fibrosis, Compound II calcium salt hydrate form D: 【Chemical 84】 Includes, The aforementioned pharmaceutical composition, Compound I: 【Chemical 85】 or a pharmaceutically acceptable salt thereof, Compound III: 【Chemical 86】 or a pharmaceutically acceptable salt thereof Administered in combination, The aforementioned treatments are as follows: (a) 250 mg of compound I, or an equivalent amount of a pharmaceutically acceptable salt thereof, and (b) 21.24 mg of Compound II calcium salt hydrate form D, and (c) 100 mg of compound III, or an equivalent amount of its pharmaceutically acceptable salt. It is characterized by including daily administration of the drug, A pharmaceutical composition characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

34. A pharmaceutical composition for use in the treatment of cystic fibrosis, Compound III: 【Chemistry 87】 or a pharmaceutically acceptable salt thereof Includes, The aforementioned pharmaceutical composition, Compound II calcium salt hydrate form D: 【Chemical 88】 , and Compound I: 【Chemistry 89】 or a pharmaceutically acceptable salt thereof Administered in combination, The aforementioned treatments are as follows: (a) 250 mg of compound I, or an equivalent amount of a pharmaceutically acceptable salt thereof, and (b) 21.24 mg of Compound II calcium salt hydrate form D, and (c) 100 mg of compound III, or an equivalent amount of its pharmaceutically acceptable salt. It is characterized by including daily administration of the drug, A pharmaceutical composition characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

35. A pharmaceutical composition for use in the treatment of cystic fibrosis, Compound I: [Chemical 90] Includes, The aforementioned pharmaceutical composition, Compound II calcium salt hydrate form D: 【Chemistry 91】 , and Compound III: 【Chemistry 92】 Administered in combination, The aforementioned treatments are as follows: (a) 250 mg of compound I, (b) 21.24 mg of Compound II calcium salt hydrate form D, and (c) 100 mg of compound III It is characterized by including daily administration to patients requiring treatment, A pharmaceutical composition characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

36. A pharmaceutical composition for use in the treatment of cystic fibrosis, Compound II calcium salt hydrate form D: 【Chemistry 93】 Includes, The aforementioned pharmaceutical composition, Compound I: 【Chemical 94】 , and Compound III: 【Chemical 95】 Administered in combination, The aforementioned treatments are as follows: (a) 250 mg of compound I, (b) 21.24 mg of Compound II calcium salt hydrate form D, and (c) 100 mg of compound III It is characterized by including daily administration to patients requiring treatment, A pharmaceutical composition characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

37. A pharmaceutical composition for use in the treatment of cystic fibrosis, Compound III: 【Chemistry 96】 Includes, The aforementioned pharmaceutical composition, Compound II calcium salt hydrate form D: 【Chemistry 97】 , and Compound I: 【Chem.98】 Administered in combination, The aforementioned treatments are as follows: (a) 250 mg of compound I, (b) 21.24 mg of Compound II calcium salt hydrate form D, and (c) 100 mg of compound III It is characterized by including daily administration to patients requiring treatment, A pharmaceutical composition characterized by an X-ray powder diffractogram (XRPD) having signals of 6.1 ± 0.2 degrees 2-theta, 16.2 ± 0.2 degrees 2-theta, and 22.8 ± 0.2 degrees 2-theta.

Images