Crystallographic form and formulation of KRAS inhibitors

Abstract

This provides a novel soluble form of the compound. [Solution] The present invention relates to the crystalline form of 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile, and to pharmaceutical formulations, methods of manufacture, and use for the treatment of diseases comprising the compound or a pharmaceutically acceptable salt thereof.

Description

[Technical Field]

[0001] This disclosure relates to the crystalline form of 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile, and pharmaceutical compositions / formulations comprising the compound or pharmaceutically acceptable salts thereof, and their use for the treatment of diseases. [Background technology]

[0002] Therapeutic agents can be administered to patients via different routes, including orally, intravenously, and subcutaneously. Oral administration of therapeutic agents offers several advantages over other routes of administration. For example, orally delivered drugs are easily self-administered, thereby increasing patient compliance and eliminating the need for injectable or inhaled therapies or dedicated delivery devices in the therapeutic setting. Furthermore, because oral administration does not require complex devices or punctures of the body surface or membranes, it is typically the safest route for introducing drugs into the body.

[0003] Despite the advantages offered by oral administration, achieving a consistent and adequate circulating concentration of a drug by oral administration can be difficult, particularly due to its low water solubility, slow dissolution rate in biological fluids, poor stability of the drug at physiological pH, low permeability to biological membranes, high pre-circulating metabolism, and inadequate or inconsistent systemic absorption between individuals or within specific regions of the gastrointestinal system. From a safety perspective, minimizing the total dose requirement for efficacy and reducing absorption variability should allow for a further reduction of undesirable side effects such as diarrhea and emesis, commonly referred to as vomiting.

[0004] Oncogenic KRAS mutations have been identified in approximately 30% of human cancers and have been shown to activate multiple downstream signaling pathways.

[0005] Despite the prevalence of KRAS mutations, it remains a challenging therapeutic target. (Cox, ADDrugging the Undruggable RAS: Mission Possible? Nat. Rev. Drug Disc. 2014, 13, 828-851; Pylayeva-Gupta, y et al. RAS Oncogenes: Weaving a Tumorigenic Web. Nat. Rev. Cancer 2011, 11, 761-774).

[0006] To date, research has focused on KRAS G12C variant inhibitors (for example, International Publications 2020 / 081282, 2020 / 101736, 2020 / 146613, and 2021 / 118877 disclose KRas G12C inhibitors), while Publication 2021 / 041671 discloses KRas G12D small molecule inhibitors, Publication 2017 / 011920 discloses KRas G12C, G12D, and G12V small molecule inhibitors, and Publication 2023 / 183585 discloses KRAS G12D inhibitors.

[0007] An example of KRAS G12D is 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrili, which has the following structure:

[0008] [ka] It has the following properties. This compound is disclosed in International Publication No. 2023 / 183585 as an amorphous free base compound (hereinafter referred to as "Formula II").

[0009] There is still a need to provide a more soluble form of Formula II. Specifically, there is a need for an alternative method for preparing the compound. Furthermore, there is a need to provide a pharmaceutical composition containing the compound or a pharmaceutically acceptable salt thereof that provides an improved pharmacokinetic profile in which harmful or undesirable effects such as vomiting or diarrhea are reduced or minimized. The present invention addresses one or more of these needs by providing a novel crystalline form of the compound and / or a pharmaceutical composition containing the compound or a pharmaceutically acceptable salt thereof. [Overview of the project]

[0010] In one embodiment, the following saccharinate compound:

[0011] [ka] A solvent is provided, and the presence of a solvent is optional. In some embodiments, the solvent is selected from water, tetrahydrofuran (THF), acetone, methanol, ethanol, and benzyl alcohol.

[0012] In another embodiment, this disclosure relates to formula I:

[0013] [ka] The target compound is one in which the solvent may be optionally present.

[0014] The compound of formula I is a disaccharinate, and the chemical name of formula I may be 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharinate. In some embodiments, the compound of formula I comprises one or more solvents such as water, tetrahydrofuran (THF), acetone, methanol, ethanol, and benzyl alcohol.

[0015] In another embodiment, the disclosure relates to a method for preparing a compound of formula I, the method comprising suspending 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (i.e., formula II) in a solvent, adding saccharin, and isolating the compound of formula I.

[0016] In another embodiment, the disclosure relates to a solid oral pharmaceutical dosage form comprising: a) a core composition comprising a compound disclosed herein (e.g., formulas I and II); b) a capsule containing the core composition, the capsule having a body and a cap; c) a polymer seal covering the transition between the capsule cap and the body; and d) an enteric coating coating the polymer seal and the capsule.

[0017] In another embodiment, the present disclosure is directed to a process for preparing a dosage form of a compound disclosed herein (e.g., Formula I, Formula II), comprising: a) blending all components contained in the core composition in a suitable blender; b) metering an amount of the blend formed in step (a) to obtain the core composition; c) compressing the blend metered in step (b) using a capsule slug mold; d) placing the slug formed in step (c) into a capsule; e) covering the transition between the capsule cap and the capsule body with a polymer seal; and f) coating the capsule with an enteric coating.

[0018] In another embodiment, the present disclosure is directed to a solid pharmaceutical composition comprising: (1) a compound disclosed herein, which constitutes about 30% to 67% of the composition; (2) microcrystalline cellulose, which constitutes about 20% to 55% of the composition; (3) mannitol, which constitutes about 0% to 10% of the composition; (4) crospovidone, which constitutes about 2% to about 5% of the composition; (5) colloidal silicon dioxide, which constitutes up to about 0.5% to 2% of the composition; (6) magnesium stearate, which constitutes about 1% to 3% of the composition; and (7) an enteric coating, wherein all percentages are percentages by weight and the total weight is 100%. In some embodiments, the solid pharmaceutical composition is in the form of a tablet.

[0019] In another embodiment, there is provided a therapy for treating a disease such as cancer, comprising the composition disclosed herein, or a pharmaceutically acceptable salt thereof.

[0020] In another embodiment, there is provided a therapy for treating a disease such as cancer, comprising the pharmaceutical composition disclosed herein. Brief Description of the Drawings

[0021] [Figure 1]This is the XRPD chromatogram of 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile 1.25 acetate 0.75 hydrate. [Figure 2] This is the XRPD chromatogram of 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile monocinnamate. [Figure 3] This is the XRPD chromatogram for Equation I. [Figure 4] The following data are from in vivo pharmacokinetic studies of enteric-coated and non-enteric-coated capsules containing formula I, administered orally to monkeys (60 mg / kg). The free plasma concentration of formula I versus time (average of 3 animals) is shown. [Figure 5] The following data are from an in vivo pharmacokinetic study of non-enteric-coated capsules containing formula I, administered orally to monkeys (60 mg / kg). The free plasma concentration of formula I versus time (per animal) is shown. [Figure 6] The following data are from an in vivo pharmacokinetic study of enteric-coated capsules containing formula I, administered orally to monkeys (60 mg / kg). The free plasma concentration of formula I versus time (per animal) is shown. [Modes for carrying out the invention]

[0022] In this specification,

[0023] [ka] The crystalline form of the compound and alternative methods for preparing the crystalline form of the compound are disclosed. Furthermore, this disclosure relates to pharmaceutical compositions comprising the compounds disclosed herein or pharmaceutically acceptable salts thereof (e.g., Formula I or Formula II) that provide an improved pharmacokinetic profile in which harmful or undesirable effects such as vomiting or diarrhea are reduced or minimized.

[0024] In some embodiments, this disclosure provides pharmaceutical compositions (e.g., enteric-coated capsules, enteric-coated tablets) comprising the compounds disclosed herein (e.g., formulas I, II). In some embodiments, the pharmaceutical compositions disclosed herein can reduce the frequency of vomiting without significantly affecting exposure by delivering the compounds disclosed herein (e.g., formulas I, II) to the upper gastrointestinal tract. In some embodiments, the pharmaceutical compositions comprising the compounds disclosed herein exhibit improved pharmacokinetics (e.g., lower compound variability and higher exposure of the compound in the subject). In some embodiments, vomiting is not observed (or the frequency of vomiting is reduced) in subjects administered with the pharmaceutical composition comprising the compounds disclosed herein (e.g., formulas I, II).

[0025] As used herein, “compounds disclosed herein” or “compounds(s) disclosed herein” means any compound disclosed herein, including formula I, formula II (free base), and pharmaceutically acceptable salts of formula II. Exemplary salts of compounds disclosed herein include saccharates, such as disaccharates. In some embodiments, the compounds disclosed herein optionally contain one or more solvents. In some embodiments, one or more solvents are selected from water, tetrahydrofuran (THF), acetone, methanol, ethanol, and benzyl alcohol.

[0026] In some embodiments, the compounds disclosed herein (e.g., Formula I) can be used to treat cancers such as KRAS G12D-associated cancer.

[0027] KRAS G12D-related cancer is a cancer in which one or more cancer cells express a KRAS G12D mutant protein or contain a KRAS G12D mutation. Examples of cancers that can be treated with the compounds disclosed herein include, but are not limited to, lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer. More specific but non-limiting examples of KRAS G12D-related cancers that can be treated with the compounds disclosed herein (e.g., Formula I) include pancreatic adenocarcinoma, colon adenocarcinoma, lung adenocarcinoma, colorectal adenocarcinoma, and rectal adenocarcinoma. In one embodiment, the cancer is non-small cell lung cancer. In another embodiment, the cancer is colorectal cancer. In yet another embodiment, the cancer is pancreatic cancer. In yet another embodiment, the cancer is gastric cancer.

[0028] In one embodiment, KRAS G12D-associated cancer is a solid tumor containing one or more cancer cells that express the KRAS G12D mutant protein or contain the KRAS G12D mutation.

[0029] In one embodiment, a method is disclosed for treating a solid tumor containing one or more cancer cells that express a KRAS G12D mutant protein or contain a KRAS G12D mutation, the method comprising administering a compound disclosed herein (e.g., Formula I) to a patient in need of such treatment.

[0030] In one embodiment, the patient is also administered an effective dose of one or more of the following: PD-1 inhibitors, PD-L1 inhibitors, CDK4 / CDK6 inhibitors, EGFR inhibitors, ERK inhibitors, Aurora A inhibitors, SHP2 inhibitors, platinum agents, and pemetrexed, or pharmaceutically acceptable salts thereof. An example of a PD-1 inhibitor is pembrolizumab. An example of a CDK4 / CDK6 inhibitor is abemaciclib. An example of an EGFR inhibitor is cetuximab. In one embodiment, the compounds disclosed herein (e.g., Formula I) are administered simultaneously, separately, or sequentially in combination with an effective dose of a PD-1 inhibitor. In one embodiment, the compound (e.g., Formula I) is administered simultaneously, separately, or sequentially in combination with an effective dose of a PD-1 inhibitor containing pembrolizumab. In one embodiment, the compound (e.g., Formula I) is administered simultaneously, separately, or sequentially in combination with an effective dose of a PD-L1 inhibitor. In one embodiment, the compound (e.g., formula I) is administered simultaneously with an effective amount of a CDK4 / CDK6 inhibitor, either separately or sequentially. In one embodiment, the compound (e.g., formula I) is administered simultaneously with an effective amount of a CDK4 / CDK6 inhibitor containing abemaciclib, either separately or sequentially. In one embodiment, the compound (e.g., formula I) is administered simultaneously with an effective amount of an EGFR inhibitor, either separately or sequentially. In one embodiment, the compound (e.g., formula I) is administered simultaneously with an effective amount of an EGFR inhibitor containing cetuximab, either separately or sequentially. In one embodiment, the compound (e.g., formula I) is administered simultaneously with an effective amount of an ERK inhibitor, either separately or sequentially. In one embodiment, the compound (e.g., formula I) is administered simultaneously with an effective amount of an Aurora A inhibitor, either separately or sequentially. In one embodiment, the compound (e.g., formula I) is administered simultaneously with an effective amount of an SHP2 inhibitor, either separately or sequentially. In one embodiment, the compound (e.g., formula I) is administered simultaneously, separately, or sequentially in combination with an effective amount of platinum. In another embodiment, the compound (e.g., formula I) is administered simultaneously, separately, or sequentially in combination with an effective amount of pemetrexed.

[0031] This specification discloses a method for treating gastric cancer, comprising administering a compound disclosed herein (e.g., Formula I) to a patient in need of such treatment.

[0032] This specification discloses a method for treating G12D-related cancer, comprising administering a compound disclosed herein (e.g., Formula I) to a patient in need of such treatment.

[0033] This specification discloses a method for treating G12D-associated cancers, the cancers including lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, or colorectal cancer, the method comprising administering a compound disclosed herein (e.g., Formula I) to a patient in need of such treatment.

[0034] This specification discloses a method for treating gastric cancer, comprising administering a compound disclosed herein (e.g., Formula I) and a PD-1 inhibitor to a patient in need of such treatment. In embodiments, the PD-1 inhibitor includes pembrolizumab.

[0035] This specification discloses a method for treating G12D-associated cancer, comprising administering a compound disclosed herein (e.g., Formula I) and a PD-1 inhibitor to a patient in need of such treatment. In embodiments, the PD-1 inhibitor includes pembrolizumab.

[0036] This specification discloses a method for treating G12D-associated cancers, wherein the cancer includes lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, or colorectal cancer, and the method comprises administering a compound disclosed herein (e.g., Formula I) and a PD-1 inhibitor to a patient in need of such treatment. In embodiments, the PD-1 inhibitor includes pembrolizumab.

[0037] This specification discloses a method for treating gastric cancer, comprising administering a compound disclosed herein (e.g., Formula I) and an EGFR inhibitor to a patient in need of such treatment. In embodiments, the EGFR inhibitor includes cetuximab.

[0038] This specification discloses a method for treating G12D-associated cancer, comprising administering a compound disclosed herein (e.g., Formula I) and an EGFR inhibitor to a patient in need of such treatment. In embodiments, the EGFR inhibitor includes cetuximab.

[0039] This specification discloses a method for treating G12D-associated cancers, the cancers including lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, or colorectal cancer, the method comprising administering a compound disclosed herein (e.g., Formula I) and an EGFR inhibitor to a patient in need of such treatment. In embodiments, the EGFR inhibitor includes cetuximab.

[0040] This specification discloses a method for treating gastric cancer, comprising administering a compound disclosed herein (e.g., Formula I) together with a CDK4 / CDK6 inhibitor to a patient in need of such treatment. In embodiments, the CDK4 / CDK6 inhibitor includes abemaciclib.

[0041] This specification discloses a method for treating G12D-associated cancer, comprising administering a compound disclosed herein (e.g., Formula I) together with a CDK4 / CDK6 inhibitor to a patient in need of such treatment. In embodiments, the CDK4 / CDK6 inhibitor includes abemaciclib.

[0042] This specification discloses a method for treating G12D-associated cancers, the cancers including lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, or colorectal cancer, the method comprising administering a compound disclosed herein (e.g., Formula I) together with a CDK4 / CDK6 inhibitor to a patient in need of such treatment. In embodiments, the CDK4 / CDK6 inhibitor includes abemaciclib.

[0043] This specification discloses a method for treating gastric cancer, comprising administering a compound disclosed herein (e.g., Formula I) together with a PD-1 inhibitor and an EGFR inhibitor to a patient in need of such treatment. In embodiments, the PD-1 inhibitor comprises pembrolizumab, and the EGFR inhibitor comprises cetuximab.

[0044] This specification discloses a method for treating G12D-associated cancer, comprising administering a compound disclosed herein (e.g., Formula I) together with a PD-1 inhibitor and an EGFR inhibitor to a patient in need of such treatment. In embodiments, the PD-1 inhibitor comprises pembrolizumab, and the EGFR inhibitor comprises cetuximab.

[0045] This specification discloses a method for treating G12D-associated cancers, the cancers including lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, or colorectal cancer, the method comprising administering a compound disclosed herein (e.g., Formula I) together with a PD-1 inhibitor and an EGFR inhibitor to a patient in need of such treatment. In embodiments, the PD-1 inhibitor comprises pembrolizumab, and the EGFR inhibitor comprises cetuximab.

[0046] This specification discloses a method for treating gastric cancer, comprising administering a compound disclosed herein (e.g., Formula I) to a patient in need of such treatment. In embodiments, the gastric cancer is KRAS G12D gastric cancer.

[0047] This specification discloses a method for treating KRAS G12D-related cancers, the method comprising administering a compound disclosed herein (e.g., Formula I) to a patient in need of such treatment, where KRAS G12D-related cancers include colorectal cancer, pancreatic cancer, and non-small cell lung cancer.

[0048] In all methods and uses described herein in which two or more therapeutic agents are administered, the two or more therapeutic agents may be administered simultaneously, separately, or sequentially.

[0049] Pharmaceutical compositions containing the compounds disclosed herein (e.g., Formula I) may be prepared by combining the disclosed compounds with pharmaceutically acceptable excipients. The term "excipients" refers to diluents or fillers, binders, granulators, adhesives, polymers and copolymers, disintegrants, stabilizers, lubricants, anti-adhesion agents, flow enhancers, surfactants, dispersants or wetting agents, dissolution retarders or enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, and sweeteners, or combinations thereof, that are not harmful to the patient. Examples of pharmaceutical compositions and processes for their preparation are found in "Remington: The Science and Practice of Pharmacy," Loyd, V., et al. Eds., 22. nd This can be found in Ed., Mack Publishing Co., 2012.

[0050] In some embodiments of the solid formulation, the excipient includes a diluent or filler. In some embodiments, the diluent or filler is selected from the group consisting of dibasic calcium phosphate, kaolin, lactose, dextrose, magnesium carbonate, sucrose, mannitol, glucose or other monosaccharides, dextrin or other polysaccharides, microcrystalline cellulose, powdered cellulose, cellulose derivatives—for example, HPMC, precipitated calcium carbonate, calcium sulfate, sorbitol, inositol, and starch, or combinations of two or more thereof. In some embodiments, the diluent or filler is present in an amount of about 5% to about 95% by weight, or about 10% to about 90% by weight, or about 20% to about 85% by weight, or about 25% to about 70% by weight.

[0051] This specification discloses orally administered pharmaceutical compositions comprising a compound disclosed herein (e.g., Formula I). ​​In one embodiment, the orally administered pharmaceutical composition is enterically coated. The enterically coated composition provides at least one of higher bioavailability or reduced stomach upset compared to an orally administered, non-enterically coated pharmaceutical composition. The compositions disclosed herein may be administered to a patient in either a fasted or fed state. Accordingly, in some embodiments, the compound of any embodiment is administered to the subject before a meal. In some such embodiments, the compound is administered 3 hours, 2 hours, 1 hour, 30 minutes, or 15 minutes before a meal. In other embodiments, the compound of any embodiment described herein is administered to the subject during a meal. In other embodiments, the compound of any embodiment described herein is administered to the subject within 3 hours, 2 hours, 1 hour, 30 minutes, or 15 minutes before a meal.

[0052] This specification discloses 1) enteric-coated capsules, 2) enteric-coated tablets, and 3) enteric-coated pellets, each of which comprises a compound disclosed herein. In one embodiment, the compound is of formula I.

[0053] In one embodiment, the pharmaceutical preparation is an enteric-coated capsule containing formula I without any excipients. In another embodiment, the pharmaceutical preparation is an enteric-coated tablet containing formula I.

[0054] In some embodiments, pharmaceutical compositions containing a pure active ingredient (API) such as Formula I may be filled into capsules made of acid-resistant materials such as Eudracaps® enteric-coated capsules and Capsugel® Enprotect® capsules. In some embodiments, the pharmaceutical formulation (e.g., enteric-coated capsules) may be administered under fasting or non-fasting conditions.

[0055] In some embodiments, enteric-coated capsules containing the compound disclosed herein (e.g., Formula I) can significantly reduce vomiting in fasted and non-fasted subjects compared to HPMC capsules containing the compound disclosed herein. In some embodiments, enteric-coated capsules containing the compound disclosed herein (e.g., Formula I) provide the compound disclosed herein (e.g., Formula I) with a higher area under the curve (AUC) exposure and much lower variability compared to HPMC capsules containing the same compound disclosed herein. In some embodiments, no vomiting is observed in subjects administered with enteric-coated capsules containing the compound disclosed herein (e.g., Formula I). ​​In some embodiments, subjects administered with enteric-coated capsules containing the compound disclosed herein (e.g., Formula I) exhibit similar PK profiles (e.g., lower drug variability between subjects when the same dose is administered) and reduced and / or no vomiting, regardless of whether the subject is administered in a non-fasted or fasted state.

[0056] In some embodiments, administering enteric-coated capsules with food may help reduce nausea / vomiting.

[0057] In some embodiments, administering enteric-coated tablets with food may help reduce nausea / vomiting.

[0058] In some embodiments, this disclosure relates to a method for treating cancer in subjects requiring cancer treatment, comprising administering to the subjects a therapeutically effective amount of a solid pharmaceutical composition disclosed herein (e.g., enteric-coated tablets, enteric-coated capsules). In some embodiments, the cancer is KRAS G12D-associated cancer. In some embodiments, the incidence of diarrhea in subjects is less than the incidence of vomiting associated with administering the same amount of a non-enteric-coated capsule composition of formula I. In some embodiments, the incidence of diarrhea in subjects is less than the incidence of diarrhea associated with administering the same amount of a capsule composition of formula I. In some embodiments, vomiting or diarrhea occurs in less than 5% of the entire subject population. In some embodiments, vomiting or diarrhea occurs in less than 10% of the entire subject population. In some embodiments, vomiting or diarrhea occurs in less than 15% of the entire subject population. In some embodiments, vomiting or diarrhea occurs in less than 20% of the entire subject population. In some embodiments, vomiting or diarrhea as a side effect occurs in less than 25% of the entire population of subjects.

[0059] Enteric-coated pharmaceutical compositions Enteric-coated pharmaceutical formulations are manufactured so that the product passes through the patient's stomach without alteration, dissolves upon leaving the stomach and entering the small intestine, and rapidly releases the active ingredient. Such formulations have been used for many years and can be in the form of capsules, tablets, or pellets. In the case of tablets or pellets, the active ingredient is located inside the tablet or pellet and is encapsulated in a film or envelope, i.e., an "enteric coating," which is insoluble in acidic environments such as the stomach but soluble in near-neutral environments such as the small intestine.

[0060] This specification discloses orally administered pharmaceutical compositions comprising the compounds disclosed herein. In one embodiment, the orally administered pharmaceutical composition is enterically coated. The enterically coated composition offers at least one of higher bioavailability or reduced stomach upset compared to an orally administered, unenterically coated pharmaceutical composition. The compositions disclosed herein may be administered to a patient in either a fasted or fed state.

[0061] This specification discloses 1) enteric-coated capsules, 2) enteric-coated tablets, and 3) enteric-coated pellets, each of which comprises a compound disclosed herein. In one embodiment, the compound is of formula I.

[0062] Enteric-coated capsules Dosage forms for oral administration are disclosed herein. The dosage forms described herein consist of a core composition comprising the compounds disclosed herein (e.g., Formula I, Formula II), wherein the compounds are contained in a sealed capsule coated with an enteric coating.

[0063] In one embodiment, an enteric-coated, orally administered pharmaceutical composition is an enteric-coated capsule, the capsule comprising: a) a core composition comprising a compound disclosed herein; b) a capsule containing the core composition, having a body and a cap; c) a polymer seal covering the transition between the capsule cap and the body; and d) an enteric coating coating the polymer seal and the capsule.

[0064] Enteric-coated capsules and tablets release little to no of the compounds disclosed herein in fluids with pH < 4.5 (i.e., the stomach), and release most of the compounds in fluids with pH > 6.0 (i.e., the intestines). In one embodiment, the dosage form releases no of the compounds disclosed herein (e.g., formulas I and II) for up to 2 hours in fluids with pH 4.5 to 6.0, and releases most of the compounds in fluids with pH > 6.0.

[0065] In one embodiment, the enteric-coated capsule may be Evonik (Eudracaps® (size 0)), Capsugel (Enprotect® (size 0)), or CanadaCaps. In one embodiment, the enteric-coated capsule is Eudracaps®. In one embodiment, the enteric-coated capsule is Enprotect®. In one embodiment, the enteric-coated capsule is CanadaCaps.

[0066] Enteric coatings are made from at least one polymer. In one embodiment, the enteric coating comprises a copolymer. The copolymer may contain at least one of methyl acrylate, methyl methacrylate, methacrylic acid, or ethyl acrylate.

[0067] According to another embodiment, the enteric coating comprises poly[methacrylic acid, ethyl acrylate], where methacrylic acid and ethyl acrylate are present in a ratio of approximately 1:1.

[0068] According to another embodiment, the enteric coating comprises methyl polyacrylate, methyl methacrylate, and methacrylic acid in a ratio of approximately 7:3:1.

[0069] According to another embodiment, the core composition further comprises a permeation enhancer selected from the group consisting of sodium N-[8-(2-hydroxybenzoyl)amino]caprylate (SNAC), sodium salcaprozate, sodium caprate (CIO), or 8-(N-2-hydroxy-5-chlorobenzoyl)-amino-caprylic acid (5-CNAC).

[0070] Another embodiment of the present disclosure provides a solid oral pharmaceutical dosage form comprising: a) a core composition comprising a compound disclosed herein (e.g., formulas I and II) and CIO; b) a capsule containing the core composition, the capsule having a body and a cap; c) a polymer seal covering the transition between the capsule cap and the body; and d) an enteric coating coating the polymer seal and the capsule, wherein the coating comprises one or more copolymers selected from the group consisting of i) poly[methacrylic acid and ethyl acrylate] in which methacrylic acid and ethyl acrylate are present in a ratio of about 1:1 and ii) poly[methyl acrylate, methyl methacrylate, methacrylic acid] in which methyl acrylate, methyl methacrylate and methacrylic acid are present in a ratio of about 7:3:1.

[0071] In yet another embodiment of this disclosure, a method is provided for treating a disease or condition in a patient who requires treatment of a disease or condition, the method comprising administering a dosage form described herein.

[0072] In yet another embodiment, a dosage form of the present disclosure is provided for use in the treatment of a disease or condition in a patient.

[0073] Another embodiment of the present disclosure provides a process for preparing the dosage forms disclosed herein, the process comprising: a) blending all components to be contained in a core composition in a suitable blender; b) weighing the amount of the blend formed in step (a) to become the core composition; c) compressing the blend weighed in step (b) using a capsule slag mold; d) placing the slag formed in step (c) inside a capsule; e) covering the transition between the capsule cap and the body with a polymer seal; and f) coating the capsule with an enteric coating.

[0074] Oral dosage forms that release the disclosed compounds (e.g., Formulas I and II) in the central or distal region of the intestine offer at least one of the following compared to oral dosage forms that release earlier in the gastrointestinal tract, such as the stomach, duodenum, or proximal region of the small intestine: higher bioavailability of the therapeutic agent, or reduced gastric upset compared to immediate-release formulations. Such targeted release can be achieved through the use of the dosage forms described herein.

[0075] The dosage forms described herein can achieve such targeted release whether they are administered to patients in a fed or fasted state. The potential benefits of the availability of such flexible timing and scheduling of administration include improved adherence and efficacy.

[0076] Dosage forms designed to target different regions of the intestine have been described to date. For example, Maroni, Alessandra, et al. In vitro and in vivo evaluation of an oral multiple-unit formulation for colonic delivery of insulin.EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS 108 (2016):76-82, Schellekens, RCA, et al. Pulsatile drug delivery to ileo-colonic segments by structured incorporation of disintegrants in pH-responsive polymer coatings.JOURNAL OF CONTROLLED RELEASE 132.2(2008):91-98, Liu,Fang,et al.Evolution of a physiological pH 6.8 bicarbonate buffer system:application to the dissolution testing of enteric coated products.European Journal of Pharmaceutics and Biopharmaceutics 78.1 (2011):151-157, Maroni, Alessandra, et al. In vitro and in vivo evaluation of an oral multiple-unit formulation for colonic delivery of insulin.EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS 108(2016):76-82, Schellekens, RCA, et al.Pulsatile drug delivery to ileo-colonic segments by structured incorporation of disintegrants in pH-responsive polymer coatings.JOURNAL OF CONTROLLED RELEASE 132.2 (2008):91-98, Liu, Fang, et al.Evolution of a physiological pH 6.8 bicarbonate buffer system:application to the dissolution testing of enteric coated products.EUROPEAN JOURNAL OF See PHARMACEUTICS AND BIOPHARMACEUTICS 78.1(2011):151-157. .

[0077] Enteric coatings providing the targeted release properties described herein include coatings composed of one or more polymers or copolymers. Non-limiting examples of such polymers or copolymers are specified in Table 1 below, where the trade names listed are often commercial products containing the polymers or copolymers, typically shown as aqueous dispersions.

[0078] [Table 1]

[0079] In certain embodiments, the coatings that may be used in the dosage forms of the present disclosure consist of one or more of the following: poly[methacrylic acid, ethyl acrylate] (1:1 ratio) (e.g., Eudragit® L30D-55), poly[methyl acrylate, methyl methacrylate, methacrylic acid] (7:3:1 ratio) (e.g., Eudragit® S30D), poly[methacrylic acid, methyl methacrylate] (1:2 ratio) (e.g., Eudragit® S), poly[methacrylic acid, methyl methacrylate] (1:1 ratio) (e.g., Eudragit® L), and HPMCAS(H). Examples of specific copolymers include poly[methacrylic acid, ethyl acrylate] (1:1 ratio) (e.g., Eudragit® L30D-55), poly[methyl acrylate, methyl methacrylate, methacrylic acid] (7:3:1 ratio) (e.g., Eudragit® FS30D), and mixtures thereof.

[0080] In some embodiments, the coating contains 10-70% poly[methyl acrylate, methyl methacrylate, methacrylic acid] (in a 7:3:1 ratio) (e.g., Eudragit® FS30D). In certain embodiments, the coating contains about 51.6% poly[methyl acrylate, methyl methacrylate, methacrylic acid] (in a 7:3:1 ratio) (e.g., Eudragit® FS30D). In certain embodiments, the coating contains about 53.3% poly[methyl acrylate, methyl methacrylate, methacrylic acid] (in a 7:3:1 ratio) (e.g., Eudragit® FS30D).

[0081] In certain embodiments, the coating includes a combination of poly[methacrylic acid, ethyl acrylate] (1:1 ratio) (e.g., Eudragit® L30D-55) and poly[methyl acrylate, methyl methacrylate, methacrylic acid] (7:3:1 ratio) (e.g., Eudragit® FS30D). In certain embodiments, the coating includes a combination of about 13.3 percent poly[methacrylic acid, ethyl acrylate] (1:1 ratio) (e.g., Eudragit® L30D-55) and about 53.3 percent poly[methyl acrylate, methyl methacrylate, methacrylic acid] (7:3:1 ratio) (e.g., Eudragit® FS30D). In certain embodiments, the coating includes a combination of approximately 12.9 percent poly[methacrylic acid, ethyl acrylate] (1:1 ratio) (e.g., Eudragit® L30D-55) and approximately 51.6 percent poly[methyl acrylate, methyl methacrylate, methacrylic acid] (7:3:1 ratio) (e.g., Eudragit® S30D).

[0082] In addition to the coating composition, the release profile of a dosage form is also influenced by the amount of coating applied to the sealed capsule, which is referred to herein as the "coating level." This dimension is expressed herein as the mass of coating per unit area of ​​the capsule, most typically in mg / cm² in relation to the dosage forms described herein. 2 In certain embodiments, the coating level is 5-20 mg / cm². 2 In certain embodiments, the coating level is 6-16 mg / cm². 2 In certain embodiments, the coating level is 7-10 mg / cm². 2 In certain embodiments, the coating level is approximately 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 mg / cm². 2In certain embodiments, the coating level is approximately 6.5, 7.5, 8.5, 9.5, 10.2, 10.5, 10.9, 11.9, 12.9, 13.0, or 13.7 mg / cm³. 2 That is the case.

[0083] The polymers and copolymers described above are typically provided in the form of aqueous dispersions. Therefore, a key component of the enteric coatings described herein consists of purified water. In certain embodiments, the water content is 20–40 percent. In certain embodiments, the water content is approximately 33 percent.

[0084] In some embodiments, the coating includes other excipients that improve the manufacturability and / or functionality of the coating. Examples of such excipients that may be included are plasticizers. Commonly used plasticizers include propylene glycol, glycerol, polyethylene glycol (e.g., PEG-400), glyceryl triacetate (triacetin), triethyl citrate (TEC), acetyl triethyl citrate, phthalates, diethyl phthalates, acetylated phthalates, castor oil, and mineral oil. In certain embodiments, the coating contains TEC. In certain embodiments, the concentration of TEC is 1 to 10 percent. In certain embodiments, the TEC content is about 3 percent.

[0085] Another possible excipient is a pH adjuster, which can be used to adjust the pH at which the polymer or copolymer dissolves. Examples of such pH adjusters are known to those skilled in the art and include sodium carbonate, sodium bicarbonate, potassium dihydrogen phosphate, and ammonium hydroxide.

[0086] Another component that may be added as an excipient to the coating composition and / or to the outside of the coating is a material used to prevent stickiness during storage. An example of such a material is talc.

[0087] In addition to the effects of the coating described above, the release profiles of the dosage forms described herein also result from the use of sealed capsules. The capsules themselves are hard-shell capsules known in the art and are generally made from natural materials such as gelatin, polysaccharide derivatives such as carrageenan, or cellulose such as methylcellulose or hydroxypropyl methylcellulose (HPMC). The capsule consists of two parts: a capsule body filled with the core composition and a cap that fits onto the body. The selection of a particular capsule for use in the dosage forms described herein is within the scope of the skill of those skilled in the art and depends in part on the volume of the core composition contained therein. In certain embodiments, the capsule is made of HPMC.

[0088] Capsules are typically soluble in gastrointestinal fluids, and therefore, to avoid release upstream of the desired area of ​​the intestine, the capsules must be protected from contact with gastrointestinal fluids before reaching that point. Such protection is provided by using the preferred enteric coatings described above, but for the enteric coating to provide such protection, the entire capsule must be coated, i.e., the coating must be applied so that there are no gaps into which fluids can enter.

[0089] It has been found that adding a polymer seal over the edge of the capsule cap where it transitions to the capsule body, prior to the application of an enteric coating, helps prevent release in the proximal small intestine and contributes to improved bioavailability. The amount of capsule surface area covered by the seal is not critical, as long as it covers the transition from cap to body. Therefore, the seal may partially or completely cover the capsule. For example, in certain embodiments, the seal may be a thin band around the capsule, and in other embodiments, it may be a coat or subcoat that covers the entire capsule.

[0090] In some embodiments, polymer seals are provided by a process known as banding. Banding is typically used in liquid-filled capsule dosage forms to prevent leakage of liquid from the inside, but in relation to this disclosure, banding serves to prevent the external liquid from reaching the core composition of the capsule until the enteric coating dissolves as described above.

[0091] The seal for use in the dosage form of the composition including the band may be made from a material having the same solubility properties as the enteric coating itself, or it may not have enteric properties. Therefore, the seal including the band can be made from the same material as described above for the enteric coating, or from the same material as the capsule itself. In certain embodiments, the band is composed of the same composition as the enteric coating. In certain embodiments, the band is composed of HPMC. In certain embodiments, the band contains HPMC, ethanol, and water. In certain embodiments, the band contains about 17% HPMC, about 58% ethanol, and 25% water.

[0092] Permeability enhancers increase local permeability (Twarog et al. 2019), resulting in increased oral bioavailability for therapeutic use. Examples of permeability enhancers that may be used in formulations disclosed herein include sodium N-[8-(2-hydroxybenzoyl)amino]caprylate (SNAC), sodium caprylate (C8), sodium caprate or sodium decanoate (CIO), or 8-(N-2-hydroxy-5-chlorobenzoyl)-amino-caprylic acid (5-CNAC). Sodium salcaprozate is generally considered safe (GRAS) and is included in FDA-approved medical foods (Eligen® - Vitamin B12, Emisphere, Roseland, NJ, USA). Rybelsus® tablets, approved by the FDA, European Medicines Agency, and Japan Pharmaceuticals and Medical Devices Agency, contain 300 mg of SNAC. CIO has the status of a food additive with no daily consumption limit.

[0093] The core compositions of this disclosure may contain a capsule filler to improve at least one of the fluidity, compressibility, tackiness, and density of the blend in the core. An example of such material is microcrystalline cellulose (MCC). In certain embodiments, the dosage forms of this disclosure include MCC. The core compositions disclosed herein may contain up to about 80 mg of MCC. In certain embodiments, the core compositions of this disclosure contain about 60 mg or less of MCC.

[0094] The composition may contain additional functional excipients, such as common tableting / encapsulating excipients. Examples include lubricants (e.g., magnesium stearate, sodium stearyl fumarate (SFF)), disintegrants (sodium starch glycolate, crospovidone, croscarmellose sodium, starch, etc.), and / or flow enhancers (colloidal silica, starch, silicone oil, talc, etc.).

[0095] Another advantage of the dosage forms described herein is that they can achieve acceptable bioavailability whether administered while the patient is in a fed or fasting state. As used herein, the term “fed state,” also known as the absorbed state, refers to the state of a person’s body after they have eaten food and their body has digested the food and absorbed the nutrients. This state begins when the food is eaten and can last up to about four hours, depending on the type and amount of nutrients ingested. The term “fasting state,” in contrast, refers to the state when a person’s body has not digested food and has not absorbed nutrients. Because the duration of the fed state varies, drugs that require administration during a fasting state generally need to be taken in the morning before breaking the overnight fast. In certain embodiments, when the dosage forms of this disclosure are administered concurrently with food, no negative food effect on bioavailability is observed. The dosage forms of this disclosure can be used to treat a variety of diseases or disorders.

[0096] The dosage forms described herein may further (or alternatively) reduce the stomach discomfort that patients may experience compared to when they take non-enteric coated dosage forms.

[0097] When used to treat diseases or conditions such as cancer, more specifically, cancers that are sensitive to treatment with KRAS G12D inhibitors. Examples of such cancers include pancreatic cancer, colorectal cancer, and non-small cell lung cancer (NSCLC). The dosage forms disclosed herein may be administered once daily, twice daily, every other day, every three days, every four days, every five days, every six days, or once a week. Preferably, the dosage form is administered once daily.

[0098] As used herein, terms such as “treatment,” “to treat,” and “to treat” include delaying or reducing the progression of a disease or disorder. These terms also include alleviating, relieving, reducing, eliminating, or reducing one or more symptoms of a disorder or condition, even if the disorder or condition is not actually eliminated, and even if the progression of the disorder or condition itself is not delayed or reversed.

[0099] As used herein, the term “approximately” is intended to refer to the degree of error of an amount or quantity, taking into account the nature or precision of the measurement. For example, the degree of error may be indicated by the number of significant figures provided for the measurement, as understood in the art, and may include, but is not limited to, a variation of ±1 of the most accurate significant figures reported for the amount or quantity. Typically, an exemplary degree of error is within 20 percent (%) of a given value or range of values, preferably within 10%, and more preferably within 5%. For example, “approximately” may mean that a number may be changed by only ±5%, ±4%, ±3%, ±2%, ±1%, ±0.5%, ±0.4%, ±0.3%, ±0.2%, ±0.1%, or ±0.05%. The term “approximately” includes a specific identifier. For example, “approximately 5.0” includes 5.0.

[0100] Preparation of enteric-coated capsules Enteric-coated capsules may be prepared by blending compounds disclosed herein, such as Formula I, and CIO in a suitable blender (on a small scale, weighing all components, transferring them to a jar, and blending them in a resonant acoustic mixer at 25G for 5 minutes), weighing the target blend, compressing the blend into a slag using a suitable capsule slag mold, and arranging it to fit inside the enteric-coated capsule body.

[0101] Next, fill the capsules using bread and measure mg / cm³. 2 The capsules are coated to different coating levels represented by [specific coating levels]. After coating, each capsule is manually sealed around the cap and body transition area using 10 μL of coating solution, dried, and then refrigerated. To achieve optimal manufacturability and the required capsule attributes, the blend state and encapsulation / coating / sealing process are further modified during scale-up, depending on the scale and manufacturing equipment used.

[0102] Enteric-coated tablets The compounds disclosed herein (e.g., Formula I) can be formulated as enteric-coated tablets. In one embodiment, the tablet comprises a compound disclosed herein, such as Formula I, microcrystalline cellulose, croscarmellose sodium, silicon dioxide, hydroxypropyl methylcellulose, and magnesium stearate. In one embodiment, the enteric-coated tablet contains about 30% to about 50% by weight of the compound disclosed herein. In one embodiment, the tablet further comprises titanium dioxide, talc, glyceryl monocaprylate and glyceryl dicaprate, polyethylene glycol-polyvinyl alcohol graft copolymer, polyvinyl alcohol, and optionally a coloring agent. The enteric layer is described below.

[0103] Enteric-coated pellets The compounds disclosed herein may be formulated as enteric-coated pellets, which are then optionally encapsulated. The capsules may or may not be enteric-coated. In one embodiment, the enteric-coated pellets are contained within an unenteric-coated capsule, such as a gelatin capsule.

[0104] The enteric-coated pellet comprises a) a core consisting of the compound disclosed herein and one or more pharmaceutically acceptable excipients; b) an optional separation layer; c) an enteric layer containing hydroxypropyl methylcellulose acetate succinate (HPMCAS) and a pharmaceutically acceptable excipient; and d) an optional finishing layer. In one embodiment, the compound is of formula I.

[0105] Furthermore, this specification discloses a method for producing enteric-coated pellets, comprising: a) providing a core comprising a compound disclosed herein and one or more pharmaceutically acceptable excipients; b) optionally applying a separation layer comprising one or more pharmaceutically acceptable excipients to the core; c) applying an enteric layer comprising HPMCAS and one or more pharmaceutically acceptable excipients, wherein the HPMCAS is applied as an aqueous solution or suspension, and the application is carried out in a fluidized bed apparatus; and d) optionally applying a finishing layer.

[0106] Pellet core The pellet core is prepared by applying a layer containing the compound disclosed herein (e.g., Formula I) to an inert core. Such inert cores have been conventionally used in pharmaceuticals and are readily available in all industrialized countries. In some embodiments, the core is prepared from starch and sucrose for use in confectionery and pharmaceutical manufacturing. However, cores of any pharmaceutically acceptable excipient can be used, including, for example, microcrystalline cellulose, plant gum, wax, etc. The main characteristic of the inert core is that it is inert with respect to both the compound disclosed herein and other excipients in the pellet, and ultimately with respect to the patient who will ingest the pellet.

[0107] The core size depends, for example, on the desired size of the pellets to be manufactured. Generally, pellets can be as small as 0.1 mm or as large as 2 mm. The core is approximately 0.3 to 0.8 mm in size to provide finished pellets in the desired size range of approximately 0.5 to 1.5 mm in diameter.

[0108] In some embodiments, the core may be specified as having a particle size range such as 18–20 U.S. mesh, 20–25 U.S. mesh, 25–30 U.S. mesh, or 30–35 U.S. mesh in order to obtain an acceptable size distribution of various absolute sizes.

[0109] The amount of core used depends on the weight and thickness of the added layer, and generally, the core constitutes about 10 to 70% of the product. More preferably, the amount of core filling accounts for about 15 to 45% of the product.

[0110] When pellet production is initiated with an inert core, the compound is typically coated onto the core to produce a final drug concentration of approximately 10-25% of the product. The amount of compound naturally depends on the desired dose of the drug and the amount of pellet to be administered.

[0111] A convenient method for coating cores with compounds is the "powder coating" process, in which the core is moistened with a viscous liquid or binder, the compound is added in powder form, and the mixture is dried. Such methods are routinely performed in the practice of industrial pharmaceuticals, and appropriate equipment is used on a daily basis. Such equipment is actually used in several steps of the process of the present invention and is therefore described herein. Historically, this method has been carried out in conventional coating pans similar to those used in sugar coating. While this process can be used to prepare pellets, the equipment has inefficient airflow and drying capacity, thereby limiting the rate of application and potentially resulting in longer processing times to minimize aggregation.

[0112] Alternatively, the products of the present invention can be manufactured using a fluidized bed apparatus (using a rotary processor) or a rotary plate apparatus, such as the Freund CF-Granulator (Vector Corporation, Marion, Iowa). A rotary plate apparatus typically consists of a cylinder with a rotatable plate at its bottom. The movement of the clump of particles to be coated is provided by friction between the clump and the stationary wall of the cylinder and its rotating bottom. Means can be provided to apply hot air to dry the clump, spray a liquid onto the clump, and balance the drying rate as in the case of a fluidized bed.

[0113] When applying a powder coating, the pellet clumps are kept in an adhesive state, and the powder, in this case the compound, that adheres to them is added continuously or periodically to adhere to the adhesive pellets. Once all of the compound has been applied, the spraying is stopped and the clumps are dried in an airflow. Adding an inert powder to the compound may be appropriate or convenient.

[0114] Additional solids may be added to the layer containing the compound. These solids may be added to facilitate the coating process as needed to aid flow, reduce static charge, aid bulk accumulation, and form a smooth surface. Inert substances such as talc, kaolin, and titanium dioxide, lubricants such as magnesium stearate, finely powdered silicon dioxide, crospovidone, and non-reducing sugars, such as sucrose, can be used. The amount of such substances ranges from a few tenths of the product to about 20% of the product. Such solids should have a fine particle size of less than 50 μm to produce a smooth surface.

[0115] The compound is adhered to the core by spraying a pharmaceutical excipient that is sticky and adhesive when wet, and then dried to form a strongly cohesive film. Pharmaceutical scientists are familiar with and have conventionally used many such substances, most of which are polymers. Examples of such polymers include hydroxypropyl methylcellulose, hydroxypropylcellulose, and polyvinylpyrrolidone. Further examples of such substances include, for example, methylcellulose, carboxymethylcellulose, acacia, and gelatin. The amount of adhesive excipient ranges from about 4% to about 12% of the product and largely depends on the amount of compound to be attached to the core.

[0116] Compounds can also be accumulated on the core by spraying a slurry containing compounds either dissolved in enough water to make the slurry sprayable, or suspended in a solution of excipients in a suspended compound layer. Such slurries can be ground through a machine adapted to grind suspensions to reduce the particle size of the compound. Grinding in suspension form is desirable because it avoids the dust generation and containment problems that occur when grinding dry powdered pharmaceuticals. A method for applying this suspension is a classic pharmaceutical fluidized bed coating apparatus such as a Wurster column, which simply consists of a vertical cylinder with an air-permeable bottom and an upward-facing spray nozzle just above the bottom, or a downward-facing spray nozzle mounted on the mass of product. The cylinder is filled with the particles to be coated, a sufficient amount of air is drawn in through the bottom of the cylinder to suspend the mass of particles, and the liquid to be applied is sprayed onto the mass. The temperature of the fluidized air is balanced with respect to the spraying rate to maintain the pellet or tablet mass at the desired moisture level and viscosity while the coating is being formed.

[0117] On the other hand, the core may contain monolithic particles incorporating the compound. Such cores can be prepared by granulation techniques, which are widely used in pharmaceutical science, particularly in the preparation of granular materials for compressed tablets. Although the particle size of the core is too small for preparation by compression techniques, the core can be prepared by mixing the compound with a mass of pharmaceutical excipients, wetting the mass with water or a solvent, drying it, and grinding the mass into particles within the same particle size range as described above for inert cores. This can be achieved through the processes of extrusion and marmelization.

[0118] The pellet cores can also be prepared by mixing the compound with conventional pharmaceutical ingredients to obtain a desired concentration, and then forming the mixture into cores of a desired size by conventional procedures or processes.

[0119] separation layer A separation layer between the compound-containing core and the enteric coating is not essential, but it is a characteristic feature of the formulation. If necessary, the separation layer provides a smooth base for applying the enteric coating, extends the pellet's resistance to acidic conditions, and improves stability by inhibiting any interactions between the drug and the enteric polymer in the enteric coating.

[0120] The smoothing function of the separation layer is purely mechanical, and its purpose is to improve the coating of the enteric layer and avoid thin spots within it caused by ridges and irregularities on the core. Therefore, the smoother and less irregular the core can be, the less material is required for the separation layer, and if the compound has an extremely fine particle size and the core is fabricated to be as close to a perfect sphere as possible, the need for smooth properties in the separation layer can be completely avoided.

[0121] Generally, the separation layer consists of a cohesive or polymeric material and a finely powdered solid excipient that constitutes the packing material. When sugar is used in the separation layer, the sugar is applied in the form of an aqueous solution and constitutes part or all of the cohesive material that binds the separation layers together. In addition to or instead of sugar, polymeric materials can be used in the separation layer. For example, small amounts of substances such as hydroxypropyl methylcellulose, polyvinylpyrrolidone, and hydroxypropylcellulose can be used to increase the adhesion and cohesiveness of the separation layer.

[0122] Furthermore, it is desirable to use packing and excipients in the separation layer to increase the smoothness and solidity of the layer. Substances such as fine talc and silicon dioxide are widely accepted as pharmaceutical excipients and can be conveniently added as needed to pack and smooth the separation layer.

[0123] Generally, the amount of sugar in the separation layer may be in the range of about 2% to about 10% of the product when sugar is used, and the amount of polymer or other adhesive material may be in the range of about 0.1% to about 5%. The amount of filler such as talc should be in the range of about 5% to about 15% based on the weight of the final product.

[0124] The separation layer can be applied by spraying an aqueous solution of sugar or polymer material onto the filler as described in the preparation of the compound layer. However, the smoothness and homogeneity of the separation layer can be improved by completely dispersing the filler as a suspension in a solution of sugar and / or polymer material, and then spraying this suspension onto the core using the apparatus as described above in the preparation of the core having the compound layer, and then drying it.

[0125] enteric layer The enteric coating layer consists of an enteric polymer, which must be selected for its compatibility with the compound as described above. The enteric polymer must be a polymer having only a small number of carboxylic acid groups per unit weight or repeating unit of the polymer. In one embodiment, the enteric polymer is hydroxypropyl methylcellulose acetate succinate (HPMCAS), and this product is defined as containing 4% to 28% succinoyl groups, which are the only free carboxyl groups in the compound. See Japanese Standards of Pharmaceutical Ingredients 1991, pages 1216-21, Standard No. 19026. HPMCAS is a trademark of AQOAT and is available from Shin-Etsu Chemical Co., Ltd., Tokyo, Japan. It is available in two particle size grades and three molecular weight ranges. In this embodiment, we use the L grade with a number average molecular weight of 93,000, but other grades are also expected to be usable.

[0126] Enteric-coated polymers can be applied as coatings from aqueous suspensions, from aqueous solutions, from organic solvent solutions, or as powders. Application from organic solvents is currently not preferred in the pharmaceutical industry due to the cost of the solvent and the difficulty of either disposing of solvent vapors or recovering evaporated solvents. Therefore, a detailed discussion of the application of enteric coatings from organic solvents will not be given here, however pharmaceutical scientists will recognize that such applications are entirely possible if circumstances favor them.

[0127] Enteric-coated polymers can also be applied according to the method described by Shin-Etsu Chemical Co. Ltd. (Obara, et al., Poster PT6115, AAPS Annual Meeting, Seattle, Wash., Oct. 27-31, 1996). When applying enteric-coated polymers in powder form, the enteric-coated polymer is added directly to the tablet or pellet in a solid state, and a plasticizer is sprayed onto the tablet or pellet at the same time. The deposit of solid enteric-coated particles is then converted into a film by curing. Curing is performed by spraying a small amount of water onto the coated tablet or pellet, and then heating the tablet or pellet for a short time. This method of applying enteric coating can be carried out using the same type of apparatus as described above for the preparation of cores having a compound layer.

[0128] When enteric-coated polymers are applied as aqueous suspensions, problems often arise in obtaining a uniform and cohesive film. Therefore, it is highly desirable to purchase fine-particle grade polymers or to grind the polymer particles to an extremely small size before application. This can be done by grinding the dry polymer in an air-shock mill or by preparing a suspension and grinding the polymer in slurry form. Slurry grinding is generally preferred, especially since it can also be used to grind the filler portion of the enteric layer in the same process. It is desirable to reduce the average particle size of the enteric-coated polymer to a range of about 1 μm to about 5 μm, preferably 3 μm or less.

[0129] When applying enteric-coated polymers in suspension form, it is important to ensure that the suspension remains homogeneous and that conditions favorable to polymer aggregation do not arise. Such care includes maintaining the suspension in a gently agitated state, but not agitating it so vigorously that it produces bubbles, and ensuring that the suspension does not swirl and remain stationary, for example, in the nozzle body or in a delivery tube that is too large. Often, polymers in suspension form will aggregate if the suspension becomes too warm, and the critical temperature can be as low as 30°C in individual cases. Since the spray nozzle and tube are exposed to hot air in typical fluidized bed systems, care must be taken to ensure that the suspension continues to move actively through the system to cool the tube and nozzle. Particularly when using HPMCAS, it is desirable to cool the suspension to below 20°C before application, to cool the tube and nozzle by pumping a small amount of cold water before initiating pumping of the suspension, and to use a delivery tube with a diameter small enough to allow for a spray rate that allows the suspension to continue moving rapidly through the tube.

[0130] However, in this disclosure, enteric polymers are applied as aqueous solutions whenever possible. In the case of HPMCAS, dissolution of the polymer can preferably be achieved by neutralizing the polymer with ammonia. Neutralization of the polymer can be achieved simply by adding ammonia, preferably in the form of an aqueous ammonium hydroxide solution, to the aqueous suspension of the polymer. Complete neutralization results in complete dissolution of the polymer at approximately pH 5.7–5.9. Good results can also be obtained by partially neutralizing the polymer by adding less than an equivalent amount of ammonia. In such cases, the unneutralized polymer remains in a suspended state suspended in the solution of the neutralized polymer. As mentioned above, when using such methods, it is clearly important to control the particle size of the polymer. Using a neutralized polymer more easily provides a smooth, cohesive enteric layer than using a suspended polymer, while using a partially neutralized polymer provides moderate smoothness and cohesiveness. In particular, when the enteric layer is applied on a very smooth separation layer, excellent results can be obtained from a partially neutralized enteric polymer.

[0131] The degree of neutralization can be varied within a range that does not adversely affect the results or ease of operation. For example, operations involving neutralization of about 25% to about 100% are described in this disclosure. Another condition is neutralization of about 45% to about 100%, and another condition is neutralization of about 65% to about 100%. Yet another neutralization method results in neutralization of about 25% to about 65%. However, it has been found that the enteric polymer in the product obtained after drying is neutralized to a lesser extent than when applied. When neutralized or partially neutralized HPMCAS is applied, the HPMCAS in the final product is neutralized by about 0% to about 25%, more preferably by about 0% to about 15%.

[0132] Most enteric-coated polymers require the addition of plasticizers for best results. In the case of HPMCAS, the plasticizer is triethyl citrate, used in amounts of up to approximately 15% to 30% of the enteric-coated polymer in aqueous suspension applications. When neutralized HPMCAS is used, lower levels of plasticizer may be required, or no plasticizer may be required at all.

[0133] Small amounts of components such as defoamers, suspending agents when the polymer is in suspension form, and surfactants that help smooth the film are also commonly used. For example, silicone defoamers, surfactants (e.g., polysorbate 80, sodium lauryl sulfate, etc.) and suspending agents (e.g., carboxymethylcellulose, plant gum, etc.) can generally be used in amounts ranging from up to 1% of the product.

[0134] Typically, the enteric layer is filled with powder excipients such as talc, glyceryl monostearate, or silicon dioxide hydrate to increase the layer thickness, strengthen it, reduce static charge, and decrease particle aggregation. Such solids can be added to the enteric polymer mixture in amounts ranging from about 1% to about 10% of the final product, while the amount of the enteric polymer itself is usually in the range of about 5% to about 25%, more preferably about 10% to about 20%.

[0135] The application of enteric-coated polymers to pellets follows the same general procedure as described above, using a fluidized bed apparatus to simultaneously spray the enteric polymer solution or suspension and dry it with hot air. The temperature of the dry air and the temperature of the circulating pellet mass should be maintained within the range recommended by the enteric polymer manufacturer.

[0136] Finishing layer While a finishing layer on the enteric coating is not essential, it can often improve the aesthetics, handling, storage, and machinability of the product, and can also provide additional benefits. The simplest finishing layer is simply a small amount (less than about 1%) of an antistatic component, such as talc or silicon dioxide, sprinkled on the surface of the pellets. Another simple finishing layer is a small amount (about 1%) of wax, such as beeswax, melted onto the circulating mass of pellets to further smooth the pellets, reduce static charge, prevent the pellets from sticking together, and increase the hydrophobicity of the surface.

[0137] More complex finishing layers can constitute the final spray layer of the components. For example, thin layers of polymer materials such as hydroxypropyl methylcellulose and polyvinylpyrrolidone can be applied in amounts ranging from about 2% to about 10%. The polymer material can also support a suspension of opaque microparticles such as opaque agents, fillers such as talc, or coloring materials, particularly red or yellow iron oxides. Such layers dissolve rapidly in the stomach, protecting the compound while leaving an enteric coating, but providing further means of pharmaceutically elegantness and protection from mechanical damage to the product.

[0138] The finishing layer applied to the product of the present invention is essentially the same type commonly used in pharmaceuticals to smooth, seal, and color enteric-coated products, and can be formulated and applied in the usual manner.

[0139] Embodiment In one embodiment, the compound of formula I is intended for therapeutic use.

[0140] In one embodiment, the compound of formula I is intended for use in the treatment of cancer.

[0141] In one embodiment, the compound of formula I is intended for use in the treatment of KRAS G12D-related cancers.

[0142] In one embodiment, examples of cancers that can be treated with the compounds disclosed herein (e.g., Formula I) include, but are not limited to, lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, gastric cancer, and colorectal cancer. In some embodiments, the cancer includes lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, or colorectal cancer.

[0143] In one embodiment, the compounds disclosed herein (e.g., Formula I) are intended for use in combination with, separately or sequentially with, one or more of the following pharmaceutically acceptable salts in cancer treatment: PD-1 or PD-L1 inhibitors, CDK4 / CDK6 inhibitors, EGFR inhibitors, ERK inhibitors, Aurora A inhibitors, SHP2 inhibitors, platinum agents, and pemetrexed. An example of a PD-1 inhibitor is pembrolizumab. An example of a CDK4 / CDK6 inhibitor is abemaciclib. An example of an EGFR inhibitor is cetuximab. In one embodiment, the compound of Formula I is administered simultaneously, separately or sequentially with an effective amount of a PD-1 inhibitor. In one embodiment, the compound of Formula I is administered simultaneously, separately or sequentially with an effective amount of a PD-1 inhibitor containing pembrolizumab. In one embodiment, the compound of Formula I is administered simultaneously, separately or sequentially with an effective amount of a PD-L1 inhibitor. In one embodiment, the compound of formula I is administered simultaneously with an effective amount of a CDK4 / CDK6 inhibitor, separately or in combination with it. In one embodiment, the compound of formula I is administered simultaneously with an effective amount of a CDK4 / CDK6 inhibitor containing abemaciclib, separately or in combination with it. In one embodiment, the compound of formula I is administered simultaneously with an effective amount of an EGFR inhibitor, separately or in combination with it. In one embodiment, the compound of formula I is administered simultaneously with an effective amount of an EGFR inhibitor containing cetuximab, separately or in combination with it. In one embodiment, the compound of formula I is administered simultaneously with an effective amount of an ERK inhibitor, separately or in combination with it. In one embodiment, the compound of formula I is administered simultaneously with an effective amount of an Aurora A inhibitor, separately or in combination with it. In one embodiment, the compound of formula I is administered simultaneously with an effective amount of an SHP2 inhibitor, separately or in combination with it. In one embodiment, the compound of formula I is administered simultaneously with an effective amount of a platinum agent, separately or in combination with it. In one embodiment, the compound of formula I is administered simultaneously with, separately from, or in combination with, an effective amount of pemetrexed.

[0144] In embodiments according to any one of the preceding “Use” embodiments, the crystalline form of the compound disclosed herein is used. In some embodiments, the compound is of formula I. In further embodiments, the crystalline form of the compound of formula I is characterized by having an X-ray powder diffraction (XRPD) pattern with peaks at 2θ values ​​of 4.4±0.2 and 11.3±0.2. The crystalline form may be further characterized by having peaks at 2θ values ​​of 12.9±0.2 and 16.2±0.2. The crystalline morphology can be further characterized by the presence of peaks at 4.4±0.2, and 11.3±0.2, 17.5±0.2, 20.1±0.2, and 20.8±0.2 or at 2θ values ​​of 4.4±0.2, 11.3±0.2, 12.9±0.2, 16.2±0.2, 17.5±0.2, 20.1±0.2, and 20.8±0.2.

[0145] Further Embodiments The following embodiments are further embodiments of the present disclosure. 1. The following saccharinate compounds:

[0146] [ka] The solvent may optionally be present, preferably a saccharinate compound selected from water, tetrahydrofuran (THF), acetone, methanol, ethanol, and benzyl alcohol. 2.Formula:

[0147] [ka] A compound wherein a solvent may optionally be present, preferably the solvent being selected from water, tetrahydrofuran (THF), acetone, methanol, ethanol, and benzyl alcohol. 3. The crystalline form of the compound according to Embodiment 1 or 2, characterized by having an X-ray powder diffraction (XRPD) pattern with peaks at 2θ values ​​of 4.4±0.2 and 11.3±0.2. 4. The crystal morphology according to Embodiment 3, characterized by having an X-ray powder diffraction (XRPD) pattern that further includes peaks at 2θ values ​​of 12.9±0.2 and 16.2±0.2. 5. The crystal morphology according to Embodiment 3 or 4, characterized by having an X-ray powder diffraction (XRPD) pattern that further includes peaks at 2θ values ​​of 17.5±0.2, 20.1±0.2, and 20.8±0.2. 6. A pharmaceutical composition comprising a compound or crystalline form described in any one of Embodiments 1 to 5, and at least one excipient. 7. A method for treating a patient with cancer, comprising administering an effective amount of the pharmaceutical composition described in Embodiment 6 to a patient in need of treatment. 8. The method according to Embodiment 7, wherein the cancer is a KRAS G12D-related cancer. 9. The method according to Embodiment 7 or 8, wherein the cancer includes lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, or colorectal cancer. 10. A compound or crystalline form described in any one of Embodiments 1 to 5 for use in therapy. 11. A compound or crystalline form according to any one of Embodiments 1 to 5, for use in the treatment of cancer. 12. The cancer is a KRAS G12D-associated cancer, and the compound or crystalline form for use as described in Embodiment 11. 13. The cancer is selected from lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer, and is the compound or crystalline form for use according to Embodiment 11 or 12. 14. The cancer is selected from colorectal cancer, pancreatic cancer, and gastric cancer, and is a compound or crystalline form for use as described in Embodiment 12. 15. A compound or crystalline form according to any one of Embodiments 1 to 5 for use in combination with, simultaneously with, separately from, or sequentially with, a PD-1 inhibitor in the treatment of cancer. 16. The cancer is a KRAS G12D-associated cancer, and the compound or crystalline form for use as described in Embodiment 15. 17. The cancer is selected from lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer, and is the compound or crystalline form for use as described in Embodiment 15 or 16. 18. The cancer is selected from colorectal cancer, pancreatic cancer, and non-small cell lung cancer, and is a compound or crystalline form for use according to Embodiment 16 or 17. 19. A compound or crystalline form according to any one of Embodiments 1 to 5 for use simultaneously, separately, or sequentially in combination with a PD-1 inhibitor in the treatment of cancer, wherein the cancer is selected from colorectal cancer, pancreatic cancer, and gastric cancer. 20. The PD-1 inhibitor is a compound or crystalline form for use according to any one of embodiments 15 to 19, comprising pembrolizumab. 21. A compound or crystalline form according to any one of Embodiments 1 to 5 for use in combination with, simultaneously with, separately from, or sequentially with, an EGFR inhibitor in the treatment of cancer. 22. The compound or crystal form according to Embodiment 21, wherein the cancer is a KRAS G12D-associated cancer. 23. The cancer is selected from lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer, and is the compound or crystalline form according to Embodiment 21 or 22. 24. The cancer is selected from colorectal cancer, pancreatic cancer, and non-small cell lung cancer, and is the compound or crystalline form described in Embodiment 23. 25. The cancer is selected from colorectal cancer, pancreatic cancer, and gastric cancer, and is the compound or crystalline form described in Embodiment 21. 26. The EGFR inhibitor is a compound or crystalline form for use according to any one of embodiments 21 to 25, comprising cetuximab. 27. A method for preparing a compound or crystalline form described in any one of Embodiments 1 to 5, comprising suspending 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrili in a solvent, adding saccharin, and isolating a disaccharinate. 28. The method according to Embodiment 27, wherein saccharin is dissolved in a solvent and then added to 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile in the solvent. 29. The method according to Embodiment 27 or 28, wherein the solvent is an alcohol. 30. The method according to Embodiment 29, wherein the alcohol is a C1-C4 alcohol. 31. The method according to any one of embodiments 28 to 30, wherein the solvent comprises at least one of ethanol or methanol. 32. The method according to any one of Embodiments 27 to 31, wherein saccharin is dissolved in a solvent, and the resulting solution is gradually added to 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile in the solvent. 33. The method according to any one of Embodiments 27 to 32, wherein the solvent is ethanol, and 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile in ethanol is heated to about 47±5°C, after which saccharin in ethanol is added. 34. The method according to any one of Embodiments 27 to 33, wherein saccharin is used in molar excess with respect to 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile. 35. Equation I:

[0148] [ka] A compound wherein a solvent may be optionally present, and which can be obtained by treating 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrili with saccharin. 36.a) a core composition comprising a compound disclosed herein (e.g., formula I, formula II); b) a capsule containing the core composition, the capsule having a body and a cap; c) a polymer seal covering the transition between the capsule cap and the body; and d) an enteric coating coating the polymer seal and the capsule. 37. The compound is

[0149] [ka] The dosage form according to Embodiment 36, which is a saccharin salt. 38. Compounds are

[0150] [ka] The dosage form described in Embodiment 36. 39. The dosage form according to Embodiment 36, wherein the compound is not released at all into a fluid having pH < 4.5, and the majority of the compound is released into a fluid having pH > 6.0. 40. The dosage form according to Embodiment 36 or 39, wherein no of the compounds disclosed herein are released into a fluid having a pH of 4.5 to 6.0 for up to 2 hours, and the majority of the compounds are released into a fluid having a pH of >6.0. 41. The dosage form is the dosage form according to Embodiment 36 or 39, wherein no of the compounds disclosed herein are released into a fluid having a pH of 4.5 to 6.0 for up to 2 hours, and the majority of the compounds are released into a fluid having a pH of >6.8. 42. A polymer seal comprising the same composition as the enteric coating, according to any one of embodiments 36 to 41. 43. The dosage form according to any one of embodiments 36 to 42, wherein the polymer seal comprises HPMC. 44. The polymer seal is a band, according to any one of embodiments 36 to 43. 45. The dosage form according to any one of Embodiments 36 to 44, wherein the enteric coating comprises a copolymer comprising at least one polymer selected from the group consisting of methyl acrylate, methyl methacrylate, methacrylic acid and ethyl acrylate. 46. The dosage form according to any one of Embodiments 36 to 45, wherein the enteric coating comprises a copolymer comprising at least one polymer selected from polymethacrylic acid and ethyl acrylate. 47. The dosage form according to Embodiment 45, wherein methacrylic acid and ethyl acrylate are present in a ratio of about 1:1. 48. The dosage form according to Embodiment 46, wherein the enteric coating comprises polymethacrylic acid and ethyl acrylate at a concentration of 10 to 70%. 49. The dosage form according to Embodiment 46, wherein the enteric coating comprises polymethacrylic acid and ethyl acrylate at a concentration of about 65%. 50. The dosage form according to any one of Embodiments 36 to 38, wherein the coating further comprises methyl polyacrylate, methyl methacrylate, and methacrylic acid. 51. The dosage form according to Embodiment 45, wherein methyl acrylate, methyl methacrylate, and methacrylic acid are present in a ratio of about 7:3:1. 52. The dosage form according to Embodiment 50, wherein the enteric coating comprises methyl polyacrylate, methyl methacrylate, and methacrylic acid at a concentration of 40 to 60%. 53. The dosage form according to Embodiment 50, wherein the enteric coating comprises methyl polyacrylate, methyl methacrylate, and methacrylic acid at a concentration of about 52%. 54. The dosage form according to any one of Embodiments 36 to 53, wherein the enteric coating comprises triethyl citrate (TEC). 55. The dosage form according to Embodiment 54, wherein the enteric coating comprises TEC at a concentration of about 3%. 56. The dosage form according to Embodiment 54, wherein the enteric coating comprises TEC at a concentration of about 2%. 57. The coating level is 5 to 20 mg / cm 2 , the dosage form according to any one of Embodiments 36 to 55. 58. The dosage form according to any one of embodiments 36 to 57, wherein the core composition further comprises a permeation enhancer. 59. The dosage form according to Embodiment 58, wherein the permeation enhancer is selected from the group consisting of sodium N-[8-(2-hydroxybenzoyl)amino]caprylate (SNAC), sodium caprate (CIO), sodium caprylate (C8), and 8-(N-2-hydroxy-5-chlorobenzoyl)-amino-caprylic acid (5-CNAC). 60. The dosage form according to Embodiment 58 or 59, wherein the permeation enhancer is CIO. 61. The dosage form according to Embodiment 60, wherein the core composition contains 1 to 100 mg of CIO. 62. The dosage form according to any one of embodiments 36 to 61, wherein the core composition further comprises microcrystalline cellulose (MCC). 63. The dosage form according to Embodiment 62, wherein the core composition contains approximately 66 mg of MCC. 64.a) A solid oral pharmaceutical dosage form comprising: a) a core composition comprising a compound disclosed herein (e.g., formulas I and II) and CIO; b) a capsule containing the core composition, having a body and a cap; c) a polymer seal covering the transition between the capsule cap and the body; and d) an enteric coating coating the polymer seal and the capsule, wherein the coating comprises one or more copolymers selected from the group consisting of i) polymethacrylic acid, ethyl acrylate in a ratio of about 1:1, and ii) poly[methyl acrylate, methyl methacrylate, methacrylic acid] in a ratio of about 7:3:1. 65. The core composition comprises (i) the compounds disclosed herein and (ii) about 280 mg of CIO; the polymer seal comprises (i) about 17% HPMC, (ii) about 58% ethanol, and (iii) about 25% water; the enteric coating comprises (i) about 12.9% poly[methacrylic acid, ethyl acrylate] with methacrylic acid and ethyl acrylate in a ratio of about 1:1, (ii) about 51.6% polymethyl acrylate, methyl methacrylate, methacrylic acid] with methyl acrylate, methyl methacrylate, and methacrylic acid in a ratio of about 7:3:1, (iii) about 2.9% TEC, and (iv) about 32.6% water, with a coating level of about 10 mg / cm² 2 The dosage form described in Embodiment 64. 66. The core composition comprises (i) the compounds disclosed herein, (ii) about 280 mg of CIO, and (iii) about 66 mg of MCC; the polymer seal comprises (i) about 17% HPMC, (ii) about 58% ethanol, and (iii) about 25% water; the enteric coating comprises (i) about 12.9% poly[methacrylic acid, ethyl acrylate] with methacrylic acid and ethyl acrylate in a ratio of about 1:1, (ii) about 51.6% polymethyl acrylate, methyl methacrylate, methacrylic acid in a ratio of about 7:3:1, (iii) about 2.9% TEC, and (iv) about 32.6% water, with a coating level of about 10 mg / cm² 2 The dosage form described in Embodiment 64. 67. The core composition comprises (i) the compounds disclosed herein, (ii) about 280 mg of CIO, and (iii) about 66 mg of MCC; the polymer seal comprises (i) about 17% HPMC, (ii) about 58% ethanol, and (iii) about 25% water; the enteric coating comprises (i) about 64.5% poly[methacrylic acid, ethyl acrylate] with methacrylic acid and ethyl acrylate in a ratio of about 1:1, (ii) about 2.9% TEC, and (iii) about 32.6% water, with a coating level of about 7 mg / cm². 2 The dosage form described in Embodiment 64. 68. A dosage form according to any one of embodiments 36 to 67, wherein the bioavailability of the compound is not adversely affected when the dosage form is administered orally with food. 69. A process for preparing a dosage form according to any one of embodiments 36 to 68, comprising: a) blending all components to be contained in the core composition in a suitable blender; b) weighing the amount of the blend formed in step (a) to become the core composition; c) compressing the blend weighed in step (b) using a capsule slag mold; d) placing the slag formed in step (c) inside a capsule; e) covering the transition between the capsule cap and the body with a polymer seal; and f) coating the capsule with an enteric coating. 70. an enteric pellet comprising: a) a core comprising a compound disclosed herein (e.g., formula I, formula II) and one or more pharmaceutically acceptable excipients; b) an optional separation layer comprising a non-reducing sugar and one or more pharmaceutically acceptable excipients; c) an enteric layer comprising hydroxypropyl methylcellulose acetate succinate (HPMCAS) and one or more pharmaceutically acceptable excipients; and d) an optional finishing layer. 71. The pellet according to Embodiment 70, wherein HPMCAS is partially neutralized with ammonium ions to such an extent that approximately 0% to approximately 25% of the succinic acid groups are neutralized. 72. The pellet according to Embodiment 70, wherein HPMCAS is partially neutralized to the extent that approximately 0% to approximately 15% of the succinic acid groups are neutralized. 73. The pellet according to embodiment 70, wherein a separation layer is present. 74. The pellet according to Embodiment 70, wherein the average particle size of the compound is approximately 50 μm or less. 75. The pellet according to Embodiment 70, wherein the core comprises an inert core, on which the compound is deposited as a layer further comprising a pharmaceutically acceptable excipient. 76. The pellet according to embodiment 70, wherein a separation layer is present. 77. The pellet according to Embodiment 70, wherein HPMCAS is partially neutralized with ammonium ions to such an extent that approximately 0% to approximately 25% of the succinate groups are neutralized. 78. The pellet according to Embodiment 70, wherein the separation layer contains a pharmaceutically acceptable sugar. 79. The pellet according to embodiment 78, wherein the sugar is sucrose. 80. A process for preparing enteric-coated pellets, comprising: a) providing a core comprising a compound disclosed herein (e.g., formula I, formula II) and one or more pharmaceutically acceptable excipients; b) optionally applying a separation layer comprising a non-reducing sugar and one or more pharmaceutically acceptable excipients to the core; c) applying an enteric layer comprising HPMCAS and one or more pharmaceutically acceptable excipients, wherein the HPMCAS is supplied as an aqueous solution or suspension, and the application is carried out in a fluidized bed apparatus; and d) optionally applying a finishing layer. 81. The process according to Embodiment 80, wherein the compound is of formula I. 82. The process according to Embodiment 80, wherein HPMCAS is completely or partially neutralized with ammonium ions. 83. The process according to Embodiment 82, wherein HPMCAS is neutralized to such an extent that approximately 25% to approximately 100% of the succinic acid groups are neutralized. 84. The process according to embodiment 80, wherein a separation layer is applied. 85. The process according to Embodiment 84, wherein the separation layer contains a pharmaceutically acceptable sugar. 86. The process according to embodiment 85, wherein the sugar is sucrose. 87. The process according to Embodiment 80, wherein the core is prepared by applying fluoxetine and one or more pharmaceutically acceptable excipients to an inactive core. 88. The process according to Embodiment 80, wherein the separation layer is applied and contains a pharmaceutically acceptable sugar. 89. Gelatin capsules containing the pellets of Embodiments 70-79. 90. A solid pharmaceutical composition comprising: (1) a compound disclosed herein (e.g., Formula I, Formula II) comprising about 30 to 67% of the composition; (2) microcrystalline cellulose comprising about 20 to 55% of the composition; (3) mannitol comprising about 0 to 10% of the composition; (4) crospovidone comprising about 2 to about 5% of the composition; (5) colloidal silicon dioxide comprising up to about 0.5 to 2% of the composition; (6) magnesium stearate comprising about 1 to 3% of the composition; and (7) an enteric coating, all percentages being weight percentages and the total weight being 100%. 91. A solid pharmaceutical composition of Embodiment 90, wherein the compound is of formula I. 92. The solid pharmaceutical composition according to Embodiment 90, wherein the solid pharmaceutical composition is in the form of a tablet. 93. The solid pharmaceutical composition according to Embodiment 90, wherein the solid pharmaceutical composition further comprises a film coating. 94. (1) Compounds disclosed herein (e.g., Formula I, Formula II) constitute about 33.3 percent of the composition; (2) Microcrystalline cellulose constitutes about 51.2 percent of the composition; (3) Mannitol constitutes about 10 percent of the composition; (4) Crospovidone constitutes about 3 percent of the composition; (5) Colloidal silicon dioxide constitutes about 1 percent of the composition; (6) Magnesium stearate constitutes about 1.5 percent of the composition; (7) Enteric coating. All percentages are by weight, and the total weight is 100 percent. This is the solid pharmaceutical composition according to Embodiment 90. 95. The solid pharmaceutical composition according to Embodiment 94, wherein the solid pharmaceutical composition is in the form of a tablet. 96. The solid pharmaceutical composition according to Embodiment 95, wherein the solid pharmaceutical composition further comprises a film coating. 97. An oral solid pharmaceutical composition in tablet form, comprising: (1) a compound disclosed herein (e.g., Formula I, Formula II) constituting about 33.3% of the composition; (2) microcrystalline cellulose constituting about 51.2% of the composition; (3) mannitol constituting about 10.0% of the composition; (4) crospovidone constituting about 3.0% of the composition; (5) colloidal silicon dioxide constituting about 1.0% of the composition; (6) magnesium stearate constituting about 1.5% of the composition; and (7) an enteric coating, all percentages being weight percentages, with a total weight of 100.0%, wherein the solid pharmaceutical composition further comprises a film coating. 98. The solid pharmaceutical composition according to Embodiment 97, wherein the compound is of formula I. 99. A method for treating cancer in a patient requiring cancer treatment, comprising administering to the patient a compound, crystalline form, pharmaceutical composition, dosage form, pellet, or solid pharmaceutical composition described in any one of Embodiments 1 to 98. 100. The method according to Embodiment 99, wherein the cancer is a G12D-mediated cancer. 101. The method according to Embodiment 99 or 100, wherein the cancer is selected from the group consisting of lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer. 102. The method according to Embodiment 101, wherein the cancer is selected from the group consisting of pancreatic cancer, colorectal cancer, and non-small cell lung cancer. 103. The method according to embodiment 102, wherein the cancer is pancreatic cancer. 104. The method according to embodiment 102, wherein the cancer is colorectal cancer. 105. The method according to Embodiment 102, wherein the cancer is non-small cell lung cancer. 106. The dosage form according to any one of Embodiments 99 to 102, administered once daily, twice daily, every other day, every three days, every four days, every five days, every six days, or once a week. 107. The method according to any one of embodiments 99 to 106, wherein the dosage form is administered once daily. 108. The dosage form according to any one of Embodiments 99 to 107, which may be administered with or without food. 109. Compounds, crystalline forms, pharmaceutical compositions, dosage forms, pellets, or solid pharmaceutical compositions described in any one of Embodiments 1 to 108 for use in therapy. 110. A compound, crystalline form, pharmaceutical composition, dosage form, pellet or solid pharmaceutical composition according to any one of Embodiments 1 to 109 for use in the treatment of cancer. 111. The use described in Embodiment 110, wherein the cancer is a KRAS G12D-associated cancer. 112. The use according to Embodiment 111, wherein cancer includes lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, or colorectal cancer. 113. The use according to Embodiment 111 or 112, wherein the cancer is pancreatic cancer. 114. The use according to Embodiment 111 or 112, wherein the cancer is colorectal cancer. 115. The use according to Embodiment 111 or 112, wherein the cancer is non-small cell lung cancer. 116. The dosage form is the dosage form for use described in Embodiment 110, administered once daily, twice daily, every other day, every three days, every four days, every five days, every six days, or once a week. 117. The dosage form is administered once daily, as described in Embodiment 116. 118. The dosage form is a dosage form for use as described in Embodiment 116 or 117, which can be administered with or without food. 119. A method for preparing a solid pharmaceutical composition comprising a compound disclosed herein, comprising: (a) pre-blending, blending together a diluent, a flow accelerator, and the compound disclosed herein; (b) de-lumping the component from step (a) using a screening mill; (c) blending the component from step (b) with a disintegrant and a flow accelerator; (d) lubricating the component from step (c) by adding a lubricant; and (e) dry-forming the component from step (d) using a roller compactor. A method comprising: (f) granulating to produce a granular blend; (g) blending a diluent, a disintegrant, and a flow promoter with the granular blend of step (e); (h) lubricating the components of step (f) by adding a lubricant to produce a lubricated blend; (g) compressing the lubricated blend of step (g) by filling a rotary tablet press and compressing the lubricated blend into a tablet core; and (i) filling the core tablets of step (h) into a pan coater and performing enteric film coating by adding an enteric film coating agent. 120. The method according to Embodiment 119, wherein the compound is of formula I. 121. Use in a treatment or therapy as described above, wherein nausea in a patient receiving treatment or therapy in an enteric-coated dosage form containing the compound of formula I is less than nausea associated with administering the same amount of a non-enteric-coated dosage form of the compound of formula I. 122. Use in a treatment or therapy as described above, wherein vomiting in a patient receiving treatment or therapy with an enteric-coated dosage form containing the compound of formula I is less than vomiting associated with the administration of the same amount of a non-enteric-coated dosage form of the compound of formula I. one two three.

[0151] [ka] A compound that is 124.

[0152] [ka] It is a compound. 125. Equation I:

[0153] [ka] A compound wherein a solvent may be optionally present, and which can be obtained by treating 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrili with saccharin. 126. The compound according to Embodiment 125, wherein saccharin is used in excess. 127. A method for treating gastric cancer, comprising administering a compound of formula I to a patient in need of such treatment. 128. The method according to embodiment 127, wherein the gastric cancer is KRAS G12D gastric cancer. 129. A method for treating KRAS G12D-related cancer, comprising administering a compound of formula I to a patient in need of such treatment, wherein the KRAS G12D-related cancer includes colorectal cancer, pancreatic cancer, or non-small cell lung cancer. 130. A method for treating KRAS G12D-associated cancer, comprising administering a compound of formula I to a patient in need of such treatment, wherein the KRAS G12D-associated cancer includes colorectal cancer, pancreatic cancer, or non-small cell lung cancer. 131. A method for treating a solid tumor containing one or more cancer cells that express a KRAS G12D mutant protein or contain a KRAS G12D mutation, comprising administering a compound of formula I to a patient in need of such treatment.

[0154] Abbreviation: "ACN" refers to acetonitrile, "Alloc" refers to the allyloxycarbonyl group, "APC" refers to allylpalladium(II) chloride dimer, "aq." refers to aqueous solution, "atm" refers to atmospheric pressure, "B2pin2" refers to bis(pinacolato)diborone, "Boc" refers to tert-butoxycarbonyl, "Cbz" refers to the benzyloxycarbonyl group, "CV" refers to column volume, "DBDMH" refers to 1,3-dibromo-5,5-dimethylimidazolidined-2,4-dione, "DCM" refers to dichloromethane, "DCE" refers to 1,2-dichloroethane, "DEA" refers to diethanolamine, and "DIBAL-H" refers to diisobutylaluminum hydride. "Hydride" refers to dimethylacetamide, "DIEA" and "DIPEA" refer to N,N-diisopropylethylamine, "DMA" or "DMAC" refers to dimethylacetamide, "DMAP" refers to 4-dimethylaminopyridine, "DMF" refers to N,N-dimethylformamide, "DMSO" refers to dimethyl sulfoxide, "DTT" refers to dithiothreitol, "ERK" refers to extracellular signal-regulated kinase, "EtI" refers to ethyl iodide, "Â" refers to ethyl acetate, "EtOH" refers to ethanol, "F" refers to bioavailability, "F a " refers to the absorbed fraction, "FaSIF" refers to the fasting simulated intestinal fluid, "FeSIF" refers to the feeding simulated intestinal fluid, and "F g" refers to the fraction that survives metabolism in the intestines, "Fmoc" refers to the fluorenylmethyloxycarbonyl group, "GDP" refers to guanosine diphosphate, "GTP" refers to guanosine triphosphate, "h" refers to time, "Hex" or "hex" refers to hexane or hexanes, "HPLC" refers to high-performance liquid chromatography, "IPA" refers to isopropyl alcohol, "KOAc" refers to potassium acetate, "LiHMDS" refers to lithium bis(trimethylsilyl)amide, "MAPK" refers to mitogen-activated protein kinase, "mCPBA" refers to 3-chloroperoxybenzoic acid, "Me" refers to the methyl group, "MeOH" refers to methanol, "min" refers to minutes, and "MTBE" refers to methyl tert-butyl ether. ether) refers to, "NaBH(OAc)3" refers to sodium triacetoxyborohydride, "NaOMe" refers to sodium methoxide, "NBS" refers to N-bromosuccinimide, "NCS" refers to N-chlorosuccinimide, "NMP" refers to 1-methylpyrrolidine-2-one, and "Pd-117" refers to dichloro[bis(2-(diphenylphosphino)phenyl)ether]palladium(II), CAS 205319-06-8 is referred to as "Pd-170," which refers to chloro(clotyl)(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)palladium(II), (1,2,3-η)-2-buten-1-yl]chloro[dicyclohexyl[2',4',6'-tris(1-methylethyl)[1,1'-biphenyl]-2-yl]phosphine]palladium, CAS 1798782-02-1, "Pd(OAc)2" refers to palladium(II) acetate, "RT" refers to room temperature, and "sat."" refers to saturated, "SCX" refers to strong cation exchange, "SDD" refers to spray-dried dispersion, "STAB" refers to sodium triacetoxyborohydride, and ". t "Bu" refers to the tert-butyl group, "TEA" refers to triethylamine, "TFA" refers to trifluoroacetic acid, "THF" refers to tetrahydrofuran, "XantPhos" refers to 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, and "XPhos" refers to 2-(dicyclohexylphosphino)-2',4',6'-tri-isopropyl-1,1'-biphenyl.

[0155] The compounds or salts thereof of this disclosure may be prepared by a variety of procedures, some of which are described in the following schemes, preparations, and examples. The specific synthetic steps of each described pathway may be combined in different ways, or combined with steps from different pathways, to prepare the compounds or salts thereof of this disclosure. [Examples]

[0156] Synthesis method The following schemes and examples illustrate various synthesis methods of the compounds disclosed herein.

[0157] [ka]

[0158] Scheme 1 shows the preparation of compound (26) used in the preparation of the free base compound (36) (e.g., formula II). Carbonylation of commercially available compound (13) yields compound (14). Palladium-catalyzed carbonylation can be achieved using a bidentate ligand such as XantPhos and a palladium source such as Pd(OAc)2 with a suitable base, such as triethylamine and a polar aprotic solvent such as acetonitrile. Subsequent nitration and bromination of compound (14) may yield compound (15). Nitration can be achieved with KNO3 or HNO3 in combination with a strong acid such as H2SO4. Bromination can be achieved with a variety of suitable reagents, including but not limited to NBS, POBr3, Br2, and DBDMH. Selective reduction of compound (15) may yield compound (16). Selective reduction can be achieved using a hydride reagent such as DIBAL-H to obtain the intermediate lactol compound, which is then further reduced with triethylsilane and TFA. Reduction of compound (16) can yield compound (17). Reduction can be achieved with iron or zinc powder and NH4Cl in a polar solvent such as THF, EtOH, or MeOH. Alternatively, reduction can also be achieved under hydrogenation conditions utilizing a sulfur-doped Pt / C catalyst. Thioacylation of compound (17) can yield compound (18). A suitable isothiocyanate, such as ethoxycarbonyl isothiocyanate, can facilitate the conversion. S-alkylation of compound (18) can yield compound (19). Alkylation can be achieved by slowly adding ethyl iodide in a polar solvent such as acetone using a weak base such as K2CO3. Ring closure of compound (19) can yield compound (20). Ring closure can be achieved at 175°C in a solvent such as anhydrous NMP or diphenyl ether. The hydroxyl moiety of compound (20) can be converted to a leaving group moiety such as a chloride, yielding compound (21). Chlorination can be achieved, for example, using oxalyl chloride, thionyl chloride, (chloromethylene)dimethyliminium chloride, or POCl3, and adjusting the solvent as necessary. Nucleophilic substitution of the chloride compound (21) can yield compound (23).This nucleophilic substitution can be performed using a suitable hindered base such as DIPEA in an aprotic solvent such as DMSO, and a substituted piperazine, such as the commercially available compound (22). Oxidation of compound (23) may yield compound (24). Oxidation can be achieved with mCPBA or other suitable oxidizing agents, along with a suitable solvent. Nucleophilic substitution of compound (24) may yield compound (25). Nucleophilic substitution can be achieved with a suitable nucleophile, such as a primary or secondary alcohol, such as the commercially available compound (25), using a strong base such as LiHMDS in THF.

[0159] [ka]

[0160] Scheme 2 shows the preparation of compound (32) used in the preparation of the free base compound (36) (i.e., the compound of formula II). Cyclization of compound (27) with an isothiocyanate yields compound (28). Suitable isothiocyanates, such as ethoxycarbonyl isothiocyanate, can facilitate the conversion. Deprotection of compound (28) may yield compound (29). Eoc deprotection can be achieved through basic hydrolysis using a hydroxide base such as NaOH. Protection of compound (29) may provide compound (30). Protection can be achieved in acetonitrile under reflux conditions using a catalytic amount of DMAP with a di-tert-butyl dicarbonate or Boc group. Other suitable carbamate protecting groups such as Alloc, Fmoc, and Cbz can also be used instead of the Boc protecting group. Oxidation of compound (30) may provide compound (31). Oxidation can be achieved with mCPBA or other suitable oxidizing agents, along with a suitable solvent. Chlorination of compound (31) may yield compound (32). Chlorination can be achieved, for example, using oxalyl chloride, thionyl chloride, (chloromethylene)dimethyliminium chloride, or POCl3, and by adjusting the solvent as necessary.

[0161] [ka]

[0162] Scheme 3 shows the preparation of the free base (36) (i.e., formula II) from compounds (26) and (32). The Suzuki coupling of (26) and compound (32) yields compound (33). The Suzuki coupling can be achieved by reacting a bromide such as compound (26) with bis(neopentyl glycolate)diborone in the presence of a weak base such as KOAc and a palladium complex such as dichloro[bis(2-(diphenylphosphinofino)phenyl) ether]palladium(II) to form a boronic acid ester, which is then coupled to a chlorothienopyridine such as compound (32). Deprotection of compound (33) may yield compound (34). Removal of the protecting group can be achieved by a method appropriate to the protecting group used, e.g., by Boc removal with TFA in DCM. N-alkylation of compound (34) may yield the free base compound (36). N-alkylation can be achieved with an alkylating agent such as an alkyl halide or an epoxide such as compound (35). Alternatively, reductive amination with a suitable reducing agent and aldehyde may provide the conversion.

[0163] [ka]

[0164] Scheme 4 shows the preparation of the dihydrofloxazoline disaccharate salt compound (38) (i.e., formula I) from the free base (36) (i.e., formula II). The combination of the free base (36) with compound (37) can yield the salt compound (38). Adding saccharin, i.e., compound (37), to a solution of the dihydrofloxazoline free base compound (36), and subsequent isolation yielded the dihydrofloxazoline disaccharate salt compound (38) (i.e., the compound of formula I). ​​An excess of saccharin is preferred relative to the amount of compound (36) used.

[0165] In one embodiment, the free base (36) (i.e., formula II) is suspended in a solvent, saccharin is added, and then a disaccharin salt is added. Furthermore, saccharin may be dissolved in the solvent before being added to the free base (36). In the embodiment, the preferred solvent is an alcohol. More preferably, the alcohol is a C1-C4 alcohol. In the embodiment, the alcohol solvent includes at least one of ethanol or methanol. In the embodiment, if saccharin is dissolved in the solvent, the resulting solution is gradually added to the free base (36), which is also in the solvent. In one embodiment, the free base (36) is dissolved in ethanol, and the resulting solution is heated to about 40-78°C, or about 47±5°C, after which saccharin is added. Alternatively, the solution is heated to about 70°C or below. As a general rule, saccharin is used in molar excess, such as 2 times, 2.1 times, 2.2 times, 2.3 times, 2.4 times, or 2.5 times or more relative to the free base (36).

[0166] [ka]

[0167] Scheme 5 shows an alternative method for preparing compound (23), which is used in the preparation of the free base compound (36) (i.e., formula II). In Scheme 8, cyclization of compound (39) may form compound (40). Cyclization can be achieved through benzyl bromination using a brominating agent such as NBS and a radiation source such as a blue LED, followed by lactone cyclization. Bromination of compound (40) may yield compound (15). Bromination can be achieved with a variety of suitable reagents, including but not limited to NBS, POBr3, Br2, and DBDMH. Reduction of compound (15) may yield compound (41). Selective reduction can be achieved using a hydride reagent such as DIBAL-H. Subsequent reduction of compound (41) may yield compound (16). Treatment with triethylsilane and TFA may accelerate the reduction. Hydrogenation of compound (16) may yield compound (17). Aromatic nitro reduction can be achieved using catalysts such as Rainy nickel, palladium on carbon, or platinum oxide. Thioacylation of compound (17) can yield compound (18). Suitable isothiocyanates, such as ethoxycarbonyl isothiocyanate, can facilitate the conversion. S-alkylation of compound (18) can yield compound (19). Alkylation can be achieved by slowly adding ethyl iodide in a polar solvent such as acetone using a weak base such as K2CO3. Ring closure of compound (19) can yield compound (20). Ring closure can be achieved at 175°C in a solvent such as anhydrous NMP or diphenyl ether. Alternatively, Eaton's reagent can be used to facilitate the conversion. The hydroxyl moiety of compound (20) can be converted to a leaving group such as a chloride, yielding compound (21). Chlorination can be achieved, for example, using oxalyl chloride, thionyl chloride, (chloromethylene)dimethyliminium chloride, or POCl3, and adjusting the solvent as necessary. Nucleophilic substitution of chloride compound (21) may result in compound (23). This nucleophilic substitution can be performed using a suitable hindered base such as DIPEA in an aprotic solvent such as DMSO, utilizing substituted piperazines such as commercially available compound (22).

[0168] [ka]

[0169] Scheme 6 shows the preparation of the free base compound (36) (i.e., formula II) from compounds (23) and (32). Boration of compound (23) may yield compound (42). Boration can be achieved by reacting a bromide such as compound (23) with a diboron complex such as B2pin2 in the presence of a weak base such as NaHCO3 and a palladium complex such as Pd-117, and subsequent transesterification of boronic acid with a suitable diol such as DEA may yield a boronic acid ester such as compound (42). Suzuki coupling of compound (42) and compound (32) may yield compound (43). Suzuki coupling can be achieved by reacting a boronic acid ester such as compound (42) with a chlorothienopyridine such as compound (32) in the presence of a weak base such as KOAc and a palladium complex such as Pd-170. Oxidation of compound (43) may yield compound (44). Oxidation can be achieved with mCPBA or other suitable oxidizing agents, along with a suitable solvent. Nucleophilic substitution of compound (44) may yield compound (33). Nucleophilic substitution is performed with LiO in a polar solvent such as DMA. t This can be achieved with a strong base such as Bu and a suitable nucleophile, such as a primary or secondary alcohol, like the commercially available compound (25). Deprotection of compound (33) may yield compound (34). Removal of the protecting group can be achieved by a method appropriate to the protecting group used, e.g., by Boc removal with TFA in DCM. N-alkylation of compound (34) may yield the free base compound (36). N-alkylation can be achieved through reductive amination of an aldehyde or an acetal such as (45) derived in situ with a suitable reducing agent such as STAB. Alternatively, N-alkylation can be achieved with an alkyl halide or epoxide.

[0170] Preparation 1 5-Fluorisobenzofuran-1(3H)-one

[0171] [ka]

[0172] A stirred mixture of (2-bromo-5-fluorophenyl)methanol (500 g, 2.44 mol) and TEA (474.6 mL, 3.41 mol, 1.4 equivalents) in ACN (2500 mL) was mixed with Pd(OAc)2 (10.95 g, 48.77 mmol, 0.02 equivalents) and XantPhos (42.33 g, 73.16 mmol, 0.03 equivalents) at room temperature, and the mixture was stirred at 120 °C under 10 atm carbon monoxide for 3 days. The reaction mixture was cooled to room temperature and concentrated. The residue was diluted with H2O (1,000 mL) and then extracted with HCl (2 × 2000 mL). The combined organic layers were washed with brine (2 × 1,000 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was ground with 10:1 hexane / HCl (1,100 mL) and then filtered. The filtered cake was dried at 50°C for approximately 18 hours to obtain the title compound (300g, 81%) as a yellow solid. MS(ES)m / z=153(M+1).

[0173] Preparation 2 4-Bromo-5-fluoro-6-nitroisobenzofuran-1(3H)-one

[0174] [ka]

[0175] 5-Fluoro-3H-isobenzofuran-1-one (300 g, 1.97 mol) was stirred in H2SO4 (1,500 mL) and HNO3 (273.38 g, 4.348 mol, 2.2 equivalents) was added dropwise at 65°C. The reaction mixture was stirred for 1 hour and then cooled to room temperature. 1,3-Dibromo-5,5-dimethylimidazolidin-2,4-dione (2,255.43 g, 7.88 mol, 4 equivalents) was added in small amounts over 20 minutes and stirred at room temperature for approximately 18 hours. The mixture was poured into ice / water (pre-treated with 3 kg of Na2SO3) and filtered. The filtered cake was dissolved in ELISA (3,000 mL), washed with saturated Na2CO3 aqueous solution (2 × 1,000 mL) and brine (2 × 1,000 mL), dried over anhydrous Na2SO4, and concentrated. The residue was ground with 10:1 hexane / siRNA (660 mL), filtered, and dried at 50°C for approximately 18 hours to obtain the title compound (270 g, 49%) as a yellow solid, which was used in the next step without further purification. 1 H NMR (400MHz, DMSO-d6) δ8.58 (s, 1H), 5.51 (s, 2H).

[0176] Preparation 3 4-Bromo-5-fluoro-6-nitro-1,3-dihydroisobenzofuran

[0177] [ka]

[0178] To a stirred mixture of 4-bromo-5-fluoro-6-nitro-3H-isobenzofuran-1-one (270 g, 978 mmol) in DCM (2,500 mL), DIBAL-H (1 M THF solution, 1,467 mL, 1.467 mol, 1.5 equivalents) was added dropwise under N2 at -78°C. The reaction mixture was stirred at -78°C for 5 hours, and then quenched with 5N NaOH (300 mL) at -78°C. The resulting mixture was warmed to room temperature and then concentrated. The residue was diluted with HCl (2,500 mL), washed with brine (2 × 1,000 mL), dried over anhydrous Na₂SO₄, and concentrated. The residue was ground with 10:1 hexane / HCl (550 mL) and filtered. The solid was dried (190 g, 683.4 mmol), then dissolved in DCM (1,500 mL), and treated with droplets of Et3SiH (662 mL, 4.10 mol, 6 equivalents) at 0°C. The reaction mixture was stirred at 0°C for 20 minutes. TFA (152 mL, 2.05 mol, 3 equivalents) was added dropwise at 0°C. The ice bath was removed, and the reaction mixture was stirred at room temperature for approximately 18 hours. The reaction mixture was concentrated to an oily substance, diluted with ELISA (2,000 mL), washed with saturated Na2CO3 aqueous solution (2 × 500 mL) and brine (2 × 500 mL), dried over anhydrous Na2SO4, filtered, and concentrated to obtain the title compound (110 g, 42%), which was used in the next step without further purification. 1 H NMR (400MHz, DMSO-d6) δ8.16 (d, J = 6.2 Hz, 1H), 5.18-5.15 (m, 2H), 5.11-5.06 (m, 2H).

[0179] Preparation 4 7-Bromo-6-fluoro-1,3-dihydroisobenzofuran-5-amine

[0180] [ka]

[0181] 4-Bromo-5-fluoro-6-nitro-1,3-dihydroisobenzofuran (110 g, 420 mmol) and NH4Cl (112.3 g, 2.10 mol, 5 equivalents) were stirred in EtOH (1,000 mL) and H2O (200 mL). Fe (117.22 g, 2.09 mol, 5 equivalents) was added in small amounts at room temperature, and the mixture was stirred at 80 °C for approximately 18 hours. The mixture was filtered and concentrated. The mixture was diluted with H2O (500 mL) and extracted with siRNA (2 × 1,000 mL). The combined organic layers were washed with brine (2 × 500 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified with silica (25%~50% siRNA / hexane) to obtain the title compound (70 g, 72%) as a yellow solid. MS(ES)m / z=231(M+1).

[0182] Preparation 5 N-[(7-bromo-6-fluoro-1,3-dihydroisobenzofuran-5-yl)carbamate ethyl]carbamate

[0183] [ka]

[0184] To a solution of 7-bromo-6-fluoro-1,3-dihydroisobenzofuran-5-amine (20.4 g, 87.9 mmol) in DCM (550 mL), ethoxycarbonyl isothiocyanate (9.7 mL, 82 mmol, 0.93 equivalents) was slowly packed using an additive funnel, and the mixture was stirred at room temperature for approximately 4 hours. The solid was filtered. The filtrate was concentrated and suspended in DCM (100 mL) and hexane (350 mL), and stirred at room temperature. The resulting filtered solid and the previously filtered solid were dried under vacuum at 50°C for 2 hours. The batches were combined to obtain the title compound (32.6 g, quantitatively) as a white solid. MS(ES)m / z=363(M+1).

[0185] Preparation 6 (NZ)-N-[[(7-bromo-6-fluoro-1,3-dihydroisobenzofuran-5-yl)amino]-ethylsulfanyl-methylene]carbamate ethyl, or (Z)-(((7-bromo-6-fluoro-1,3-dihydroisobenzofuran-5-yl)amino)(ethylthio)methylene)carbamate ethyl

[0186] [ka]

[0187] A 2 L three-necked RBF equipped with an overhead stirrer, dropping funnel, and thermocouple was filled with a suspension of N-[(7-bromo-6-fluoro-1,3-dihydroisobenzofuran-5-yl)carbamate ethyl (32.6 g, 89.8 mmol) and acetone (450 mL). Solid K2CO3 (37.2 g, 269 mmol, 3.00 equivalent) was added in several portions, followed by the dropwise addition of EtI (7.2 mL, 90 mmol, 1.0 equivalent) over 20 minutes. The mixture was stirred at room temperature for approximately 18 hours. The solid was filtered, the filtrate was concentrated, and partitioned between DCM (500 mL) and H2O (500 mL). The organic matter was further washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified with silica (0-30% toluene / hexane) to obtain the title compound (30.9 g, 85.6%) as a white solid. MS(ES)m / z=391(M+1).

[0188] Preparation 7 6-Bromo-3-(ethylthio)-5-fluoro-7,9-dihydrofl[3,4-f]quinazolin-1-ol

[0189] [ka]

[0190] A 2 L four-necked RBF was equipped with an overhead stirrer, dropping funnel, N2 inlet, and thermocouple, and purged with N2. NMP (anhydrous, 300 mL) was added. The mixture was heated to 175°C. In a second flask, ethyl (NZ)-N-[[(7-bromo-6-fluoro-1,3-dihydroisobenzofuran-5-yl)amino]-ethylsulfanyl-methylene]carbamate (22.63 g, 57.83 mmol) and NMP (anhydrous, 100 mL) were combined and stirred under N2 until a homogeneous solution was obtained. When the first flask reached 175°C, the contents of the second flask were poured into the dropping funnel and rapidly added dropwise to the hot NMP. After 30 minutes, the heat was turned off and the reaction mixture was cooled to 45°C. H2O (500 mL) was slowly added, and the mixture was stirred at room temperature for 1 hour. The solid was filtered, rinsed with H2O (300 mL), and dried under vacuum at 50°C for approximately 18 hours to obtain the title compound (15.2 g, 73%) as an off-white solid. MS(ES)m / z=363(M+1).

[0191] Preparation 8 6-Bromo-1-chloro-3-(ethylthio)-5-fluoro-7,9-dihydrofl[3,4-f]quinazoline

[0192] [ka]

[0193] A 5 L three-necked RBF equipped with a dropping funnel, thermocouple, and overhead stirrer was filled with a 1,000 mL solution of DCM (50 mL, 646 mmol, 4 equivalents) of DMF (100 mL) and placed in an ice / water bath to cool to approximately 4°C. Oxalyl chloride (50.0 mL, 576 mmol, 4 equivalents) was added dropwise using an addition funnel over approximately 40 minutes. After the addition was complete, the reaction mixture was stirred at approximately 4°C for 15 minutes. Solid 6-bromo-3-ethylsulfanyl-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-ol (50.4 g, 140 mmol) was added to the reaction mixture in several portions, and the resulting suspension was stirred at approximately 4°C for 30 minutes. The ice bath was removed, and the reaction mixture was allowed to warm to room temperature and stirred for 1 hour. Then, H2O (1 L) was added, and the mixture was stirred for 15 minutes. The mixture was partitioned, the organic layer was washed with brine (1 L), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified with silica eluted with DCM / hexane (60%-90%) to obtain the title compound (45.1 g, 89%) as a white solid. MS(ES)m / z=363(M+1).

[0194] Preparation 9 8-(6-bromo-3-(ethylthio)-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl

[0195] [ka]

[0196] To a suspension of 6-bromo-1-chloro-3-ethylsulfanyl-5-fluoro-7,9-dihydrofuro[3,4-f]quinazoline (21.0 g, 57.8 mmol) in ACN (580 mL) was charged tert-butyl 3,8-diazabicyclo[3.2.1]octane-3-carboxylate (15.2 g, 69.5 mmol, 1.20 eq.) and DIPEA (40 mL, 229 mmol, 4 eq.), and the mixture was stirred at room temperature for 90 minutes. H2O (1 L) was slowly added via an addition funnel, and the mixture was stirred at room temperature for 1 hour. The solid was filtered, rinsed with H2O (500 mL), and dried under vacuum at 50°C to give the title compound (31 g, quantitative yield) as a white solid, MS(ES) m / z=539 (M+1).

[0197] Preparation 10 tert-butyl 8-(6-bromo-3-(ethylsulfonyl)-5-fluoro-7,9-dihydrofuro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate

[0198]

Chemical Formula

[0199] To a solution of tert-butyl 8-(6-bromo-3-ethylsulfanyl-5-fluoro-7,9-dihydrofuro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate (4.3 g, 8.0 mmol) in DCM (40 mL) was charged mCPBA (6.21 g, 25.2 mmol, 3.2 eq.), and the mixture was stirred at room temperature for 90 minutes. The mixture was diluted with DCM, washed with saturated aqueous NaHCO3 and brine. The organic phase was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified on silica eluting with EtOAc / hexane (20-80%) to give the title compound (3.0 g, 66%) as a white solid. MS(ES) m / z=571 (M+1).

[0200] Preparation 11 8-(6-bromo-5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl

[0201] [ka]

[0202] To a mixture of [(2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolidine-8-yl]methanol (2.26 g, 13.8 mmol) in THF (110 mL), a THF solution of LiHMDS (13.7 mL, 13.7 mmol, 1.0 M) was added dropwise under N2 conditions and the mixture was stirred for 20 minutes. To this solution, a THF solution of 8-(6-bromo-3-ethylsulfonyl-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl (6.54 g, 11.4 mmol) in THF (40 mL) was added dropwise by syringe. After 35 minutes, the reaction mixture was diluted with ELISA (250 mL) and washed with brine. The aqueous layer was extracted with siRNA (2 × 200 mL), and the combined organic layer was dried over Na₂SO₄ and concentrated to obtain a brown solid. The crude material was purified with silica eluted with 0-10% MeOH / DCM to obtain the title compound (7.0 g, 96%) as a light brown solid. MS(ES)m / z=636(M+1).

[0203] Preparation 12 (3-Cyano-7-Fluorothieno[3,2-c]pyridine-2-yl)Ethyl Carbamate

[0204] [ka]

[0205] A solution of 2-(4-chloro-5-fluoropyridine-3-yl)acetonitrile (11.8 g, 56.1 mmol) in DMF (112 mL) was cooled to 0°C. Potassium tert-butoxide (7.00 g, 61.1 mmol) was added. After 15 minutes, ethoxycarbonyl isothiocyanate (7.45 mL, 61.8 mmol) was added dropwise. The reaction mixture was slowly warmed to room temperature overnight. The reaction mixture was poured into an ice / water mixture (1.5 L), stirred until all the ice had melted, and filtered through diatomaceous earth. The solid was dried overnight in a vacuum oven (60°C) and separated from the diatomaceous earth to obtain ethyl N-(3-cyano-7-fluorothieno[3,2-c]pyridine-2-yl)carbamate (11.9 g, 79%) as a solid. MS(ES)m / z=266(M+1).

[0206] Preparation 13 2-amino-7-fluorothieno[3,2-c]pyridine-3-carbonitrile

[0207] [ka]

[0208] A suspension of ethyl (3-cyano-7-fluorothieno[3,2-c]pyridine-2-yl)carbamate (11.9 g, 44.4 mmol) in DMSO (90 mL) was cooled to 0°C. NaOH (5 M aqueous solution, 90 mL) was added dropwise over 15 minutes. The reaction mixture was heated to 105°C over 1 hour and then cooled to room temperature. The reaction mixture was poured into an ice / water mixture (1.8 L), stirred until all the ice had melted, and filtered through diatomaceous earth. The solid was dried overnight in a vacuum oven (50°C) and separated from the diatomaceous earth to obtain crude 2-amino-7-fluorothieno[3,2-c]pyridine-3-carbonitride.

[0209] Preparation 14 (3-cyano-7-fluorothieno[3,2-c]pyridine-2-yl)carbamate tert-butyl

[0210] [ka]

[0211] A mixture of crude 2-amino-7-fluorothieno[3,2-c]pyridine-3-carbonitrile (8.6 g, 44.4 mmol), DCM (90 mL), DMF (90 mL), and N,N-diisopropylethylamine (15.5 mL, 88.9 mmol) was cooled to 0°C. 4-Dimethylaminopyridine (0.54 g, 4.42 mmol) and di-tert-butyl dicarbonate (14.6 g, 66.7 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. The solvent was removed under reduced pressure, and the remaining material was diluted with DCM (400 mL) and 5% aqueous citric acid solution (250 mL). The aqueous phase was washed twice with DCM. The combined organic phases were washed with saturated aqueous NaHCO₃ solution, dried over MgSO₄, filtered, and concentrated to give tert-butyl N-(3-cyano-7-fluoro-thieno[3,2-c]pyridin-2-yl)carbamate (7.5 g, 58%) as a brown solid. MS (ES) m / z = 294 (M+1).

[0212] Preparation 15 2-((tert-Butoxycarbonyl)amino)-3-cyano-7-fluorothieno[3,2-c]pyridine 5-oxide

[0213]

Chemical Formula

[0214] mCPBA (9.00 g, 40.2 mmol) was added to a solution of (3-cyano-7-fluorothieno[3,2-c]pyridine-2-yl)carbamate tert-butyl (7.85 g, 26.8 mmol) in DCM (180 mL). The reaction mixture was stirred overnight at room temperature and then cooled to 0°C over approximately 15 minutes. The solid was collected by filtration and dried in a vacuum oven (60°C). The filtrate was diluted with MeOH and silica gel, concentrated, and the residue was purified with silica eluted with 0-6% MeOH / DCM. The fraction containing the desired substance was combined with the solid obtained by filtration and concentrated to obtain N-(3-cyano-7-fluoro-5-oxidethieno[3,2-c]pyridine-5-ium-2-yl)carbamate tert-butyl (7.26 g, 88%) as an off-white solid. MS(ES)m / z = 310(M+1).

[0215] Preparation 16 (4-chloro-3-cyano-7-fluorothieno[3,2-c]pyridine-2-yl)carbamate tert-butyl

[0216] [ka]

[0217] A suspension of 2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorothieno[3,2-c]pyridine 5-oxide (5.27 g, 17.0 mmol) in 1,2-dichloroethane (34 mL) was cooled to 0°C. A solution of phosphoryl chloride (32 mL, 344 mmol) in 1,2-dichloroethane (34 mL) was added dropwise. The reaction mixture was stirred at room temperature for 30 minutes, then stirred at 45°C for 90 minutes, and cooled to room temperature. The reaction mixture was diluted with 1,2-dichloroethane (100 mL) and added to a mixture of saturated NaHCO3 aqueous solution (500 mL), NaOH (5M aqueous solution, 40 mL), and ice. Solid NaHCO3 was added to the stirred mixture to maintain the pH at approximately 6-7. The phases were separated when foaming ceased. The aqueous phase was extracted three times by DCM. The combined organic phase was dried over MgSO4 and filtered. The filtrate was diluted with MeOH and silica gel, concentrated, and the residue was purified with silica eluted with 50-100% DCM / hexane. The fraction containing the desired substance was concentrated to obtain the title compound (3.87 g, 69%) as a white solid. MS(ES)m / z=328(M+1).

[0218] Preparation 17 8-(6-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorothieno[3,2-c]pyridine-4-yl)-5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl

[0219] [ka]

[0220] K3PO4 (0.98 mL, 2.0 mmol, 2M) and 1,4-dioxane (12.5 mL) were sparged with N2 in a mixed H2O solution for 30 minutes. 8-(6-bromo-5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl (0.500 g, 0.786 mmol), B2pin2 (0.242 g, 0.943 mmol), XPhos (0.038 g, 0.078 mmol), and water (0.016 g, 0.040 mmol) were added, and the resulting mixture was heated at 100°C for about 18 hours. The reaction mixture was cooled to room temperature and partitioned between DCM and H2O. The layers were separated, and the aqueous layer was extracted once with DCM and twice with a 4:1 solution of CHCl3:IPA. The organic layers were combined, dried over Na2SO4, and concentrated under vacuum. The residue was purified with silica (0-60% in gradient DCM, 10% 1N NH3 / MeOH) to obtain the title compound (0.306 g, 61%) as an orange-brown solid. MS(ES)m / z=849(M+1).

[0221] Preparation 18 4-(1-(3,8-diazabicyclo[3.2.1]octan-8-yl)-5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)-2-amino-7-fluorothieno[3,2-c]pyridine-3-carbonitrile

[0222] [ka]

[0223] A solution of 8-(6-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorothieno[3,2-c]pyridine-4-yl)-5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl (0.301 g, 0.355 mmol) in DCM (4 mL) was filled with TFA (1.5 mL) and stirred at room temperature for 5 hours. The mixture was concentrated, DCM was added, and the mixture was concentrated again, and this process was repeated once. The mixture was filtered through an SCX column and eluted with MeOH (4 CV), followed by 1:1 2M ammonia / MeOH:DCM (4 CV) to obtain the crude product. This was purified on silica gel and eluted under a gradient of 20-100% [20% (1N NH3 MeOH) / DCM in DCM] to obtain the title compound (0.059 g, 12%) as a pale yellow solid. MS(ES)m / z=649(M+1).

[0224] Preparation 19 5-Fluoro-6-nitroisobenzofuran-1(3H)-one

[0225] [ka]

[0226] 4-Fluoro-2-methyl-5-nitrobenzoic acid was combined with (100 g, 502 mmol)ACN (3000 mL), NBS (69 g, 552 mmol), and Charge citric acid monohydrate (5.27 g, 25.1 mmol). The reaction mixture was continuously transferred through a flow system. The temperature of the flow system was maintained at 28-40°C while the reaction mixture was exposed to a blue LED (440-460 nm). The reaction mixture exited the flow system and entered aqueous NaHCO3 and aqueous Na2SO3 solutions. The solution was concentrated to 2000-2600 mL at below 35°C. Additional aqueous NaHCO3 solution was added. The aqueous layer was extracted with ELISA (2 × 700 vol). The combined organic layer was concentrated to 100-200 mL under vacuum at 40°C. Water (300 mL) was added, and the resulting solid was isolated by filtration. The solid was washed with water (150 mL). The solid was dried at 45-55°C to obtain the title compound (40.15 kg, 74%). MS(ES)m / z=196(M-1).

[0227] Preparation 20 4-Bromo-5-fluoro-6-nitroisobenzofuran-1(3H)-one

[0228] [ka]

[0229] A solution of 5-fluoro-6-nitroisobenzofuran-1(3H)-one (100 g, 507 mmol) in H2SO4 (796 mL) was continuously supplied at 25-30°C, and a solution of 1,3-dibromo-5,5-dimethylhydantoin (DMDBH, 243.68 g, 852 mmol) in H2SO4 (2506 mL) was continuously supplied at 25-30°C through a flow reaction system. The reaction mixture was supplied through a first flow reactor heated to 80-110°C. The reactants were supplied through a second flow reactor cooled to 2-8°C. The reaction mixture was supplied to a holding chamber at 2-8°C. The reaction mixture was then supplied dropwise to water (3000 mL) at 5-15°C to control the exothermic reaction. The mixture was cooled to -5-5°C over 1-4 hours. The resulting solid was collected by filtration and washed with water (500 mL). Next, the solid was stirred with water (500 mL) and stirred at 0–10°C for 2–4 hours. The solid was collected by filtration and washed with water (300 mL). Next, the solid was stirred with MTBE (300 mL) and stirred at 40–50°C for 2–4 hours. The temperature was cooled to 10–20°C over 2–4 hours and stirred for 2–4 hours. The obtained solid was collected by filtration and washed with MTBE (148 mL). The solid was dried at 45–55°C for 12 hours to obtain the title compound (99.36 g, 71%). MS(ES)m / z=276(M+1).

[0230] Preparation 21 4-bromo-5-fluoro-6-nitro-1,3-dihydroisobenzofuran-1-ol

[0231] [ka]

[0232] 4-Bromo-5-fluoro-6-nitroisobenzofuran-1(3H)-one (100g, 362 mmol) was combined with toluene (865 mL). The reaction mixture was sparged three times with N2. The reaction temperature was adjusted to -65°C. DIBAL-H (1M in toluene, 510 mL, 510 mol) was added. The reaction mixture was stirred at -65°C for 1 hour. While maintaining the reaction temperature below -60°C, pharmaceutically acceptable The layers were separated, the upper organic layer was removed, and the mixture was concentrated to 100-200 mL under vacuum at less than 45°C. HCl (500 mL) was added, and the resulting solution was concentrated to 100-200 mL under vacuum at less than 45°C. HCl (100 mL) and the resulting solution were stirred at 20°C for 2 hours. n-heptane (500 mL) was added, and the resulting solution was stirred at 20°C for 2 hours. The mixture was filtered, and the isolated solid was dried at 45°C for 20 hours to obtain the title compound (66.4 g, 66%). MS(ES)m / z=278(M+1).

[0233] Preparation 22 4-Bromo-5-fluoro-6-nitro-1,3-dihydroisobenzofuran

[0234] [ka]

[0235] 4-Bromo-5-fluoro-6-nitro-1,3-dihydroisobenzofuran-1-ol (100 g, 360 mmol) was combined with DCM (1000 mL) and triethylsilane (60 g, 516 mmol). The reaction mixture was cooled to 5°C. TFA (120 g, 1052 mmol) was added, and the reaction mixture was stirred at 5°C for 30 minutes. The temperature was adjusted to 25°C, and the reaction mixture was stirred at 25°C for 5 hours. While maintaining the temperature below 30°C, 10% NaOH aqueous solution (400 mL) was added. The mixture was stirred at 20°C for 1 hour. The mixture was left to stand for 1 hour and separated. The organic layer was removed, and the mixture was concentrated to 100-200 mL at below 30°C. Depositphotos (400 mL) was added, and the mixture was concentrated to 100-200 mL at below 45°C. Depositphotos (800 mL) was added, followed by water (500 mL). The mixture was stirred at 20°C for 1 hour. The mixture was allowed to stand for 1 hour and then separated. The organic layer was removed and the mixture was concentrated to 100-200 mL at below 45°C. 300 mL of n-heptane was added, and the mixture was concentrated twice more to 100-200 mL at below 45°C. 100 mL of n-heptane was added, followed by 50 mL of dimethyl phosphate. The temperature was adjusted to 0°C and the mixture was stirred for 3 hours. The resulting solid was isolated by filtration, washed with 100 mL of n-heptane, and dried at 45°C for 20 hours to obtain the title compound (56.9 g, 60.3%). MS(ES)m / z=(M+1).

[0236] Preparation 23 7-Bromo-6-fluoro-1,3-dihydroisobenzofuran-5-amine

[0237] [ka]

[0238] MS(ES)m / z = 232(M+1).

[0239] Preparation 24 N-[(7-bromo-6-fluoro-1,3-dihydroisobenzofuran-5-yl)carbamate ethyl]carbamate

[0240] [ka]

[0241] 7-Bromo-6-fluoro-1,3-dihydroisobenzofuran-5-amine (100 g, 431 mmol) was combined with toluene (1500 mL) and ethoxycarbonyl isothiocyanate (59.3 g, 452 mmol). The reaction mixture was stirred at 10-20°C for 18 hours. The reaction mixture was then concentrated, and toluene (500 mL), followed by heptane (1200 mL), was added. The mixture was stirred at 20-25°C for 3 hours. The mixture was filtered. The isolated solid was dried at 40-50°C for 16 hours to obtain the title compound (145.3 kg, 92.8%). MS(ES)m / z=363(M+1).

[0242] Preparation 25 (Z)-(((7-bromo-6-fluoro-1,3-dihydroisobenzofuran-5-yl)amino)(ethylthio)methylene)carbamate ethyl

[0243] [ka]

[0244] Ethyl N-[(7-bromo-6-fluoro-1,3-dihydroisobenzofuran-5-yl)carbamate (100 g, 275 mmol) was combined with acetone (761 mL) and K2CO3 (114 g, 825 mmol). The mixture was stirred at 25°C for 30 minutes. Iodoethane (45 g, 288.5 mmol), followed by acetone (241 mL), was added. The reaction mixture was stirred at 25°C for 7 hours. Water (700 mL) was added and the mixture was stirred at 25°C for 30 minutes. The mixture was allowed to stand for 1 hour and separated. The layers were separated. RINKAN (600 mL) was added to the bottom aqueous layer and the mixture was stirred at 25°C for 1 hour. The mixture was allowed to stand for 1 hour and separated. The layers were separated. The upper organic layers were combined. 25% NaCl solution (1330 g) was added to the combined organic layers and the mixture was stirred at 25°C for 30 minutes. The mixture was allowed to stand for 1 hour and separated. The layers were separated. The organic layer was concentrated to 100-200 mL under vacuum at a temperature below 50°C. HCl (600 mL) and water (600 mL) were added, and the mixture was stirred at 25°C for 30 minutes. The mixture was allowed to stand for 1 hour and separated. The layers were separated. The upper organic layer was concentrated to 100-300 mL under vacuum at a temperature below 50°C. n-heptane (300 mL) was added, and the mixture was concentrated three times under vacuum at a temperature below 50°C to 100-300 mL. HCl (100 mL) was added, and the mixture was stirred under N2 flow at 45°C for 10 minutes. n-heptane (700 mL) was added dropwise over 3 hours at 45°C. The temperature was adjusted to 25°C over 3 hours. The mixture was stirred at 25°C for 2 hours. The mixture was concentrated to 700-800 mL under vacuum at a temperature below 30°C. n-heptane (200 mL) was added, and the mixture was concentrated to 700-800 mL under vacuum at less than 30°C. Another 200 mL of n-heptane was added, and the mixture was stirred at 25°C for 3 hours. The mixture was filtered. The solid was washed with n-heptane (100 mL) and dried at 50°C for 12 hours to obtain the title compound (75 g, 70%). MS(ES)m / z=391(M+1).

[0245] Preparation 26 6-Bromo-3-(ethylthio)-5-fluoro-7,9-dihydrofl[3,4-f]quinazolin-1-ol

[0246] [ka]

[0247] (Z)-(((7-bromo-6-fluoro-1,3-dihydroisobenzofuran-5-yl)amino)(ethylthio)methylene)carbamate ethyl (100 g, 255.6 mmol) was combined with sulfolane (1000 mL) and Eaton's reagent (7.5 wt% in MSA, 677.47 g, 357.8 mmol). The solution was stirred at 30-40°C for 30-60 minutes to obtain a clear solution. The solution was filtered under vacuum. The solution and NMP (200 mL) were continuously supplied through the flow reaction system. Two streams were supplied through a first flow reactor heated to 140-170°C. The reaction products were supplied through a second flow reactor cooled to 40-60°C. The reaction mixture was supplied to a holding chamber. The reaction mixture was then supplied dropwise to water (2000 mL) at 0-30°C. The mixture was stirred at 0–30°C for 5–10 hours. The mixture was filtered, and the solid was washed with water (1000 mL). The solid was stirred with water (2000 mL) at 0–30°C for 5–10 hours. The mixture was filtered, and the solid was washed with water (1000 mL). The solid was stirred with water (1000 mL) at 0–30°C for 5–10 hours. The mixture was filtered, and the solid was washed with water (1000 mL). The mixture was filtered, and the solid was dried at 55°C for 22 hours. (64.4 g, 73%). MS(ES)m / z=345(M+1).

[0248] Preparation 27 6-Bromo-1-chloro-3-(ethylthio)-5-fluoro-7,9-dihydrofl[3,4-f]quinazoline

[0249] [ka]

[0250] Preparation 28 8-(6-bromo-3-(ethylthio)-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl

[0251] [ka]

[0252] Preparation 29 8-(6-(1,3,6,2-dioxazabolocan-2-yl)-3-(ethylthio)-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl

[0253] [ka]

[0254] 8-(6-bromo-3-(ethylthio)-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl (500 g, 927 mmol) was combined with B2pin2 (353 g, 1390 mmol), Pd-117 (79.8 g, 69.5 mmol), Me4NOAc (247 g, 1.854 mol), and CPME (5 L). The mixture was degassed with nitrogen. The mixture was stirred at 70 ± 5 °C for 15 hours. CPME (5 L) was added, and the mixture was stirred at 70 ± 5 °C for 10 minutes. The mixture was filtered through diatomaceous earth. The filtrate was distilled to 5 L, and the temperature was adjusted to 50 ± 5 °C. Diethanolamine (146 g, 1390 mmol) was added to IPA (500 mL) over 30 minutes at 50±5°C. 8-(6-(1,3,6,2-dioxazabolocan-2-yl)-3-(ethylthio)-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl seed crystal was added, followed by the addition of diethanolamine (146 g, 1390 mmol) in IPA (500 mL). The mixture was stirred at 50±5°C for 1 hour. The temperature was adjusted to 5±5°C, and the mixture was stirred at 5±5°C for 12 hours. The product was collected by filtration, dried on the filter for 1 hour, and then dried in a tray dryer to obtain the title compound (428 g, 80.5%). MS(ES)m / z=505 (M+1, boronic acid ester was decomposed into boronic acid by MS).

[0255] Preparation 30 8-(6-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorothieno[3,2-c]pyridine-4-yl)-3-(ethylthio)-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl

[0256] [ka]

[0257] 8-(6-(1,3,6,2-dioxazabolocan-2-yl)-3-(ethylthio)-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl (400g, 697.54 mmol) to 2-((tert-butoxycarbonyl)amino)-3 -Cyano-7-fluorothieno[3,2-c]pyridine 5-oxide (228.6 g, 697.5 mmol), Pd-170 (23.44 g, 34.9 mmol), Xphos (16.64 g, 34.9 mmol), NaHCO3 (1.758 g, 2092.5 mmol), and a solution of CPME / water (6 L / 4 L) pre-spared with nitrogen were combined. The mixture was stirred at 65 ± 5 °C for 4 hours, then adjusted to 20 ± 5 °C. The layers were separated, and the aqueous layer was transferred. The organic layer was washed with brine and added to the aqueous layer. The aqueous layer was back-extracted with CPME and added to the organic layer. The organic layer was concentrated to 100 mL and MeOH (3.5 L) was added. The organic layer was concentrated to 1.75 L and MeOH (3.15 L) was added. The organic layer was concentrated to 245 L and MeOH (2.25 L) was added. The organic layer was concentrated to 2.45 L and MeOH (1.75 L) was added. The mixture was stirred at 50±5°C for 1 hour. The temperature was adjusted to 20±5°C and stirred at 20±5°C for 2 hours. The product was collected by filtration, dried on the filter, and then dried in a tray dryer to obtain the title compound (462 g, 88.1%). MS(ES)m / z=752(M+1).

[0258] Preparation 31 8-(6-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorothieno[3,2-c]pyridine-4-yl)-3-(ethylsulfonyl)-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl

[0259] [ka]

[0260] 8-(6-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorothieno[3,2-c]pyridine-4-yl)-3-(ethylthio)-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl (300 g, 1.0 equivalent) was combined with DCM:THF (2700:150 mL) under nitrogen and stirred at 20°C for 15 minutes. The temperature was adjusted to 0°C. A solution of mCPBA (182.3 g, 1.056 mol) in DCM::THF (1425:75 mL) was added, and the temperature was maintained below 10°C. The temperature was adjusted to 10°C and stirred at 10°C for 4 hours. A solution of Na2S2O5 (54.1 g, 0.285 mol) in 600 mL of water was slowly added over 15 minutes, maintaining the temperature below 20°C. The temperature was adjusted to 10°C and the mixture was stirred for 16 hours. The mixture was distilled under vacuum at 40°C to 900 mL. The temperature was adjusted to 20°C. MeOH (1.5 L) was added over 15 minutes. The temperature was adjusted to 20°C and the mixture was stirred for 1 hour. While maintaining the temperature below 30°C, a solution of 5 wt% NaHCO3 (4.5 L) was slowly added over 45 minutes. The temperature was adjusted to 20°C, the mixture was stirred for 2 hours, and the mixture was filtered. The solid was washed with water (1.5 L x 2) and dried under nitrogen to obtain the title compound (292 g, 98%) as a pale yellow solid. MS(ES)m / z=784(M+1).

[0261] Preparation 32 8-(6-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorothieno[3,2-c]pyridine-4-yl)-5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl

[0262] [ka]

[0263] 8-(6-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorothieno[3,2-c]pyridine-4-yl)-3-(ethylsulfonyl)-5-fluoro-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate tert-butyl (50 g, 63.8 mmol) was combined with ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methanol (15.23 g, 0.957 mmol) and DMAc (250 mL), and stirred at 20±5°C for 15 minutes. The temperature was then adjusted to 10±5°C. While maintaining the temperature at ≤20°C, LiOtBu solution (2.2 M in THF, 58 mL, 127.6 mmol) was added. The reaction mixture was stirred at 10±5°C for 2 hours. The temperature was adjusted to 0±5°C. 3% citric acid aqueous solution (500 mL) was slowly added while maintaining the temperature at ≤10°C. The temperature was adjusted to 20±5°C. Depositphotos (500 mL) was added and stirred for 30 minutes. Stirring was stopped and the contents were allowed to settle for 15 minutes (layer separation was observed after 5 minutes). The lower aqueous layer and the upper organic layer were separated. Depositphotos (250 mL) was added to the aqueous layer and stirred at 20±5°C for 15 minutes. Stirring was stopped and the contents were allowed to settle for 15 minutes (layer separation was observed after 5 minutes). The upper organic layer was collected and added to the previously separated organic layer. 500 mL of 5% NaHCO3 solution was added to the combined organic layers and stirred at 20±5°C for 15 minutes. Stirring was stopped and the contents were allowed to settle for 15 minutes (layer separation was observed after 10 minutes). The lower aqueous layer was removed. 250 mL of 5% LiCl solution was added and stirred at 20±5°C for 15 minutes. Stirring was stopped and the contents were allowed to settle for 15 minutes (layer separation was observed after 5 minutes). The lower aqueous layer was removed. 250 mL of 5% LiCl solution was added and stirred at 20±5°C for 15 minutes. Stirring was stopped and the contents were allowed to settle for 15 minutes (layer separation was observed after 5 minutes). The lower aqueous layer was removed. The organic layer was concentrated to 150 mL, replaced with MTBE (250 mL), and concentrated to a final volume of 150 mL. MTBE (250 mL) was slowly added and the mixture was heated to 50°C. 500 mL of n-heptane was slowly added at 50°C, and a yellow precipitate was observed.The mixture was concentrated to a thick slurry of approximately 250 mL and replaced with n-heptane (500 mL) until the final volume reached 250 mL. n-heptane (350 mL) was added, and the slurry was heated to 50°C. The slurry was stirred at 50°C for 1 hour, cooled to ambient temperature, and stirred for 3 hours. The resulting mixture was filtered, the solid was washed with n-heptane (150 mL), and dried in a vacuum oven at 50°C for 20 hours with nitrogen bleed to obtain the title compound (50.4 g, 93%). MS(ES)m / z=849(M+1).

[0264] Preparation 33 4-(1-(3,8-diazabicyclo[3.2.1]octan-8-yl)-5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)-2-amino-7-fluorothieno[3,2-c]pyridine-3-carbonitrile

[0265] [ka]

[0266] 8-(6-(2-((tert-butoxycarbonyl)amino)-3-cyano-7-fluorothieno[3,2-c]pyridine-4-yl)-5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-7,9-dihydrofluoro[3,4-f]quinazolin-1-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl (5.0 g, 5.89 mmol) was combined with DCM (15 mL) and cooled to 0-5°C. TFA (13.4 g, 118 mmol) was added over 30 minutes. The mixture was stirred at 0-5°C for 4 hours. The temperature was adjusted to 20±5°C. The mixture was stirred at 20±5°C for 12 hours. The mixture was concentrated to a concentration of ≤15 g. The mixture was cooled to 0-5°C. DMAc (15 mL) was slowly added while maintaining the temperature below 20°C. The mixture was stirred at 20±5°C for 2 minutes to obtain a clear solution of the title compound TFA salt in DMAc. The crude solution of the title compound TFA salt was used directly in the next step. MS(ES)m / z=649(M+1)

[0267] Preparation 34 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (i.e., formula II)

[0268] [ka]

[0269] A solution of 4-(1-(3,8-diazabicyclo[3.2.1]octan-8-yl)-5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)-2-amino-7-fluorothieno[3,2-c]pyridine-3-carbonitrile (0.060 g, 0.093 mmol) in DCM (2 mL) and MeOH (2 mL) was treated with (R)-(+)-propylene oxide (0.110 g, 1.850 mmol). The mixture was stirred in a sealed tube at room temperature for approximately 60 hours, and then concentrated. The residue was purified by preparative HPLC on a Luna(registered trademark) C18, 50 mm × 250 mm × 10 μm, and the title compound (0.021 g, 24%) was obtained as a pale beige solid by elution with 25% to 40% ACN in a 10 mM NH4HCO3 aqueous solution containing 5% MeOH. MS(ES) m / z = 707 (M+1).

[0270] Alternative synthesis of Equation II 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (i.e., formula II)

[0271] [ka]

[0272] Step 1: Amberchrom (trademark) 50WX8 [H-resin] (4.5g) was added to a filter, washed with water (2 x 9mL), and combined with (R)-1,1-dimethoxypropan-2-ol (3.0g, 25.0 mmol) and water (4.5mL). The temperature was adjusted to 50±5℃ and the mixture was stirred for 2 hours. The mixture was cooled to 20±5℃. The resin was filtered and washed with DMAc (8mL). The concentration of the (R)-2-hydroxypropanal solution (aldehyde: 1.85g, 25 mmol) (total weight: 15g, approximately 12 wt% aldehyde solution) was calculated.

[0273] Step 2: NaOAc (9.66 g, 117.8 mmol) was added to DMAc (10 mL). The mixture was cooled to 0 ± 5 °C. While maintaining the internal temperature below 10 °C, a DMAc solution of 4-(1-(3,8-diazabicyclo[3.2.1]octan-8-yl)-5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)-2-amino-7-fluorothieno[3,2-c]pyridine-3-carbonitrile TFA salt was added at 0-5 °C. The temperature was adjusted to 0-5 °C. STAB (5g, 23.6 mmol) was added, followed by (R)-2-hydroxypropanal solution (aldehyde: 1.85g, 25 mmol) (total weight: 15g, approximately 12 wt% aldehyde solution) over 15 minutes. The mixture was stirred at 0-5°C for 30 minutes. The temperature was adjusted to 20±5°C and stirred for 2 hours. While maintaining the internal temperature below 30°C, water (30 mL) was slowly added over 30 minutes. While maintaining the internal temperature below 20°C, 10% NH4OH aqueous solution (30 mL) was slowly added over 30 minutes. The temperature was set to 20±5°C and stirred for 2 hours. The solid was collected by filtration and washed with water (2 × 20 mL). The solid was dried in a vacuum oven at ≤50°C to obtain the title compound (4.1g, 97.8%). MS(ES)m / z=707(M+1).

[0274] Example 1 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharate (i.e., formula I)

[0275] [ka]

[0276] 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (0.177 g, 0.250 mmol) was suspended in EtOH (2 mL). The slurry was heated to 47°C while stirring at 500 rpm. 2 mL of saccharin (0.103 g, 0.562 mmol) in EtOH (4 mL) at 47°C was added to the slurry. All solids dissolved, and a clear solution was obtained. After stirring for several minutes, a white slurry was formed. The remaining 2 mL of saccharin solution was added. The solution was stirred at 47°C for 15 minutes. The solution was cooled to room temperature. A thick slurry of white solid was formed. The solid was isolated using filter paper and dried at 50°C to obtain the title compound (0.209 g, 78%) as a bright white solid.

[0277] Alternative synthesis of formula I 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharate (i.e., formula I)

[0278] [ka]

[0279] Amberlite IRN78 hydroxide resin (425 g) was pre-washed with THF (2125 mL). 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (425 g, 420.92 mmol) was combined with the pre-washed Amberlite IRN78 hydroxide resin (425 g) and THF (4250 mL). The slurry was heated to 50°C and stirred for 2 hours. The slurry was filtered to remove the resin. The filtrate was heated to 50°C and stirred. Saccharin (220.30 g, 1050 mmol) was added to the solution. The reaction mixture was stirred for 10 minutes. 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharinate seed crystal (425 mg, 0.0004 mmol) was added. The slurry was aged at 50°C for 16 hours. The slurry was cooled to 22°C over 2 hours, and then aged at 22°C for 4 hours. The slurry was filtered. The solid was washed with 2×THF (850 mL), followed by washing with heptane (850 mL). The solid was dried in a vacuum oven at 22°C to a certain weight to obtain the title compound (384.4 g, 85.1%). 1H NMR(400MHz,DMSO-d6)δ 8.47(s,2H),8.44(s,1H),7.73-7.67(m,2H),7.66-7.57(m,6H),5.66-5.46(m,3H),5.08(br d,J=12.35°Hz,1H),4.74(br d,J=12.35Hz,1H),4.67-4.53(m,3H),4.44(br s,1H),4.05(br s,1H),4.00-3.70(m,4H),3.55-3.05(m,5H),2.92(br d,J=19.07Hz,1H),2.70-2.45(m,2H),2.40-2.30(m,1H),2.25-2.00(m,6H),1.98-1.80(m,2H),1.13(d,J=6.24Hz,3H).

[0280] Alternative synthesis of formula I 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharate (i.e., formula I)

[0281] [ka]

[0282] A mixture of saccharin (34.2 kg) and acetone (854.1 kg) was placed in a reactor. The temperature was adjusted to 20°C, and the mixture was stirred until a clear solution was formed. The contents were transferred to a second reactor through a 0.45 micron filter. The first reactor was rinsed with acetone (47.5 kg), and the rinse solution was transferred to the second reactor through a 0.45 micron filter. 603.8 g of 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitride disaccharinate seed crystal was added to the second reactor. The temperature of the second reactor was adjusted to 35°C, and the reaction mixture was stirred for 30 minutes. 60.0 kg of 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (249.6 kg) and benzyl alcohol (249.6 kg) were added to the first reactor. The temperature of the first reactor was adjusted to 40°C, and the mixture was stirred until a clear solution was formed, at which point the temperature was adjusted to 20°C. The contents were transferred to the second reactor through a 0.45 micron filter over at least 4 hours while maintaining the temperature of the second reactor at 35°C. The first reactor was rinsed with acetone (47.5 kg), and while maintaining the temperature of the second reactor at 35°C, the rinse solution was transferred to the second reactor through a 0.45 micron filter. The mixture in the second reactor was stirred for at least 2 hours, the temperature was adjusted to 5°C over at least 3 hours, and the mixture was stirred for at least 12 hours. The solid was recovered by filtration, washed with acetone (2 × 190 kg), and dried under a nitrogen stream for at least 1 hour. The solid was combined with acetone (474 ​​kg, passed through a 0.45 micron filter). The temperature was adjusted to 50°C.The mixture was stirred for at least 5 hours, the temperature was adjusted to 20°C over at least 3 hours, and the mixture was stirred for at least 2 hours. The solid was recovered by filtration, washed with acetone (2 × 190 kg), dried by a nitrogen stream for at least 1 hour, and dried in a tray dryer at a maximum temperature of 65°C for 16 hours to obtain the title compound.

[0283] Alternative synthesis of formula I 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharate (i.e., formula I)

[0284] [ka]

[0285] 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (2.7 g, 3.9 mmol) was suspended in tetrahydrofuran (60 mL) in a reactor. The mixture was heated to a jacket temperature of 50°C and the complete dissolution of the solid was observed. Saccharin (1.6 g, 8.7 mmol) was suspended in tetrahydrofuran (30 mL) in a second reactor. The mixture was heated to a jacket temperature of 50°C and the complete dissolution of the solid was observed. The third reactor was heated to a jacket temperature of 50°C, and a portion of the mixture from the first reactor (20 mL) was added over 10 minutes. Crystalline 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharinate (seed crystal material; 0.2 g, 1.0 mmol) was added to the third reactor. After 4 hours, the remaining mixture from the first reactor (40 mL) and the mixture from the second reactor (30 mL) were simultaneously added to the third reactor over 14 hours. After 48 hours, the solid was collected by filtration and then dried under vacuum at 60°C to obtain the title compound (3.61 g, 87%).

[0286] Alternative synthesis of formula I 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharate (i.e., formula I)

[0287] [ka]

[0288] 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (2.7 g, 3.9 mmol) was suspended in tetrahydrofuran (60 mL) in a reactor. The mixture was heated to a jacket temperature of 50°C and the complete dissolution of the solid was observed. Saccharin (3.2 g, 17.6 mmol) was suspended in tetrahydrofuran (30 mL) in a second reactor. The mixture was heated to a jacket temperature of 50°C and the complete dissolution of the solid was observed. The third reactor was heated to a jacket temperature of 50°C, and a portion of the mixture from the first reactor (20 mL) was added over 10 minutes. Crystalline 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharinate (seed crystal material; 0.2 g, 1.0 mmol) was added to the third reactor. After 4 hours, the remaining mixture from the first reactor (40 mL) and the mixture from the second reactor (30 mL) were simultaneously added to the third reactor over 14 hours. After 48 hours, the solid was collected by filtration and then dried under vacuum at 60°C to obtain the title compound (3.76 g, 91%).

[0289] Example 2 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile 1.25 acetate 0.75 hydrate

[0290] [ka]

[0291] The THF (5 mL) stock solution of seed crystal material: 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (151.2 mg) was prepared by stirring the suspension at 50°C until a clear solution was formed. To 0.50 mL of this solution, acetic acid (0.020 mL) and n-heptane (0.50 mL) were added. After 48 hours, the supernatant solvent was drained from the solid. The solid was washed with n-heptane (1 mL) and dried under vacuum at 40°C for 24 hours to obtain seed crystal material.

[0292] A mixture of 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (1.01 g) in THF (20 mL) was stirred at 300 rpm at 50°C until a solution was formed. Acetic acid (1.44 mL) was added, and the temperature was adjusted to 20°C over 10 minutes. Seed crystal material (10.9 mg) was added. n-heptane (20 mL) was added over 6 hours. The temperature was adjusted to 5°C over 6 hours, and the mixture was stirred for 4.5 hours. The solid was recovered by filtration, washed with n-heptane (10 mL), and dried in a vacuum oven at 40°C for 24 hours to obtain the title compound (0.92 g, 81%) as an off-white solid.

[0293] Example 3 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile monocinnamate

[0294] [ka]

[0295] 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (1.07 g, 1.51 mmol) was combined with trans-cinnamic acid (saturated 2-propanol solution, 6 mL) in 2-propanol (4 mL). The suspension was slurryed at 55°C and stirred overnight at 500 rpm. The slurry was filtered. The solid was washed with 2-propanol (2 × 5 mL). The solid was dried under nitrogen and vacuum until it reached a certain weight, yielding the title compound (1.18 g, 91%) as a white solid.

[0296] Example 4 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile 1.5 saccharic acid 1.5 THF 0.25 hydrate

[0297] [ka]

[0298] 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile (100 g, 141 mmol) was combined with THF (1750 mL) at 60°C. The substance was filtered at this temperature, and the solid was rinsed with THF (50 mL). The combined filtrate was adjusted to 50°C. A solution of saccharin (65 g, 355 mmol) in THF (400 mL + 50 mL rinse solution) was added over 10 minutes through a 0.45 micron filter. 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharinate seed crystal (0.100 g) was added. The mixture was stirred at 50°C for 12 hours, then cooled to 20°C and stirred for 2 hours. The solid was recovered by filtration and dried overnight in a vacuum oven at 40°C to obtain the crude product.

[0299] Saccharin (0.60 mL, 0.057 mmol; from a methanol (5 mL) stock solution of 87.0 mg of saccharin) was added to a portion of the crude product (0.0675 g, 0.069 mmol). Acetone (1 mL) was added, and the solution was cooled to 5°C. 2-Amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrilate 1.5 saccharinate 1.5 THF Mixed crystals of 0.25 hydrate and 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile disaccharinate (formula I) were obtained in about two weeks.

[0300] Powder X-ray diffraction (XRPD) XRPD patterns were obtained using a Bruker D8 Endeavor X-ray powder diffractometer operating at 40 kV and 40 mA, equipped with a CuKα (1.5418 Å) source and a Linxeye detector. Samples were scanned at 4–42 2θ° with a step size of 0.009 2θ° and a scanning speed of 0.5 sec / step, using a 0.3° primary slit aperture and a 3.9° PSD aperture. Dry powder was packed into a quartz or silicon sample holder, and a smooth surface was obtained using a glass slide. Crystalline morphological diffraction patterns were recovered at ambient temperature and relative humidity. Crystalline peak positions were determined by MDI-Jade after overall pattern shift based on an internal NIST 675 standard with peaks at 8.853 and 26.774 2θ°.

[0301] In the field of crystallography, it is well known that for any crystalline form, the relative intensity of diffraction peaks may vary due to preferred orientations resulting from factors such as crystalline form and crystal habit. While peak intensity changes when a preferred orientation effect is present, the characteristic peak positions of the polymorph remain constant. See, for example, The United States Pharmacopeia #23, National Formulary #18, pages 1843-1844, 1995. Furthermore, in the field of crystallography, it is also well known that for any crystalline form, the angular peak positions may vary slightly. For example, peak positions may shift due to variations in the temperature at which the sample is analyzed, displacement of the sample, or the presence or absence of an internal standard. In this case, a peak position variation of ±0.2 2θ° is estimated to account for these potential variations without hindering the clear identification of the indicated crystalline form. Confirmation of the crystalline form can be performed based on any unique combination of characteristic peaks.

[0302] XRPD of Example 1 (Formula I) Prepared samples in the crystalline disaccharic acid form (i.e., formula I) are characterized by having diffraction peaks (2-theta values) as described in Table 2 below, as determined by XRPD patterns using CuKα radiation, particularly having peaks at 4.4±0.2 and 11.3±0.2 in combination with one or more peaks selected from the group consisting of 12.9 and 16.2. The tolerance for diffraction angles is 0.2 degrees.

[0303] In some embodiments, the crystalline form of the compound of formula I is characterized by having an X-ray powder diffraction (XRPD) pattern containing peaks at 2θ values ​​of 4.4±0.2 and 11.3±0.2. The crystalline form may be further characterized by containing peaks at 2θ values ​​of 12.9±0.2 and 16.2±0.2. The crystalline form may be further characterized by containing peaks at 2θ values ​​of 17.5±0.2, 20.1±0.2 and 20.8±0.2. The crystalline morphology can be further characterized by the presence of peaks at 2θ values ​​of 4.4±0.2, 11.3±0.2, 17.5±0.2, 20.1±0.2, and 20.8±0.2 or 4.4±0.2, 11.3±0.2, 12.9±0.2, 16.2±0.2, 17.5±0.2, 20.1±0.2, and 20.8±0.2.

[0304] [Table 2]

[0305] XRPD (crystalline 1.25 acetate 0.75 hydrate) of Example 2 Prepared samples in the form of crystalline 1.25 acetate 0.75 hydrate are characterized by an XRPD pattern using CuKα radiation, having diffraction peaks (2-theta values) as described in Table 3 below, and in particular having a peak at 6.0 in combination with one or more peaks selected from the group consisting of 14.9, 17.0, and 19.5, with a tolerance of 0.2 degrees for the diffraction angle.

[0306] In some embodiments, the crystalline morphology of the compound of Example 2 is characterized by having an X-ray powder diffraction (XRPD) pattern containing a peak at a 2θ value of 6.0 ± 0.2. The crystalline morphology may be further characterized by containing peaks at 2θ values ​​of 14.9 ± 0.2, 17.0 ± 0.2, and 19.5 ± 0.2. The crystalline morphology may also be further characterized by containing peaks at 2θ values ​​of 6.0 ± 0.2, 9.2 ± 0.2, 11.8 ± 0.2, and 18.4 ± 0.2 or 6.0 ± 0.2, 14.9 ± 0.2, 17.0 ± 0.2, 19.5 ± 0.2, 9.2 ± 0.2, 11.8 ± 0.2, and 18.4 ± 0.2.

[0307] [Table 3]

[0308] XRPD (crystalline monocinnamate) of Example 3 Prepared samples of crystalline monocinnamate are characterized by an XRPD pattern using CuKα radiation, having diffraction peaks (2-theta values) as described in Table 4 below, and in particular, having a peak at 7.4 in combination with one or more peaks selected from the group consisting of 4.3, 8.9, and 13.8, with a tolerance of 0.2 degrees for the diffraction angle.

[0309] In some embodiments, the crystalline morphology of the compound of Example 3 is characterized by having an X-ray powder diffraction (XRPD) pattern containing a peak at a 2θ value of 7.4 ± 0.2. The crystalline morphology may be further characterized by containing peaks at 2θ values ​​of 4.3 ± 0.2, 8.9 ± 0.2, and 13.8 ± 0.2. The crystalline morphology may also be further characterized by containing peaks at 2θ values ​​of 7.4 ± 0.2, 13.0 ± 0.2, 19.5 ± 0.2, and 20.6 ± 0.2 or 7.4 ± 0.2, 4.3 ± 0.2, 8.9 ± 0.2, 13.8 ± 0.2, 13.0 ± 0.2, 19.5 ± 0.2, and 20.6 ± 0.2.

[0310] [Table 4]

[0311] Example 5 In vitro dissolution test of API / polymer film casts 1. Set the centrifuge to 37°C. 2. Approximately 10 minutes before starting the dissolution experiment, place the sample in a 37°C Vortemp vortex mixer. 3. Pipette 1.5 mL of SGF at approximately 37°C into each vial. The theoretical concentration of the API is 1.5 mg / mL. 4. Mix the samples for 15 minutes. 5. Filter 0.170 mL of each sample using a 0.45 μm centrifuge filter. 6. Dilute 0.1 mL of the filtrate of each sample directly in an HPLC vial with 0.4 mL of 10% water / 90% MeOH. This is the dilution factor for 5. 7. After 20 minutes, dilute the sample in Vortemp with 3 mL of FaSIF at approximately 37°C. This is time zero. Theoretically, the API concentration is 0.5 mg / mL. 8. At 10, 30, 60, 120, and 240 minutes, 0.3 mL of sample is collected and filtered using a 0.45 centrifuge filter. 0.1 mL of the filtrate is directly diluted in an HPLC vial with 0.4 mL of 10% water / 90% MeOH. This is a dilution factor of 5. 9. All samples are subjected to HPLC analysis. 10. Plot the concentration versus time for each evaluated sample. Report the result as supersaturation.

[0312] In this example, 0.01N HCl or water was used as the stomach. 4 mg / mL of the active substance, i.e., formula I or formula II, was the target concentration in 2.25 mL of 0.01N HCl or DIW. 0.01N HCl = pH 2, which is the pH of a normal, healthy human stomach. DIW = approximately pH 5.5, which represents the pH of a patient taking acid reducing agents, who is ill, elderly, or has gastric acid deficiency.

[0313] [Table 5]

[0314] Table 5 above contains comparative solubility data for Formula II and Formula I. Compared to Formula II, the improved solubility of Formula I is expected to facilitate the administration of higher doses of the compound of Formula I, if necessary. The compound of Formula I dissolves immediately and completely in the stomach.

[0315] Example 6 Beagle dogs and untreated cynomolgus monkeys were orally administered 30 mg / kg or 60 mg / kg of Formula I, filled in capsules (enteric-coated and non-enteric-coated), in a crossover animal pharmacokinetic study. Multiple capsules were administered (if necessary) to achieve the target dose based on each animal's body weight. For fasting studies, animals were fasted overnight and fed 2 hours after administration. For non-fasting studies, animals were given sterile water for injection (WFI) immediately after capsule administration and then fed.

[0316] Blood samples were collected at 0.25, 0.5, 1, 2, 4, 8, 24, 48, and 72 hours after oral administration. The sides of the cages were observed twice daily, and irregularly as needed. These observations included monitoring for mortality, morbidity, general health status, vomiting, and signs of toxicity. Clinical observations and body weight were measured before administration.

[0317] The free plasma concentration in Equation I was analyzed using LC-MS / MS. Pharmacokinetic calculations were performed using WinNonlin (Phoenix™, version 8.3). The following pharmacokinetic parameters were analyzed using plasma concentration versus time data (AUC). last and AUC inf ) was calculated from.

[0318] The results are shown in Tables 6-8 and Figures 4-6.

[0319] [Table 6] Male beagle dogs were administered formula I (30 mg / kg, orally) in capsule form. Non-enteric coated capsules = HPMC capsules Enteric-coated capsules = Eudracaps® capsules The amount of vomit is approximately

[0320] Table 6 above shows that vomiting was significantly reduced in fasted dogs administered enteric-coated capsules containing formula I compared to non-enteric-coated capsules containing formula I. Specifically, none of the eight animals administered enteric-coated capsules containing formula I exhibited vomiting.

[0321] [Table 7] Untreated female cynomolgus monkeys were administered formula I (60 mg / kg, orally) in capsule form. Non-enteric coated capsules = HPMC capsules Enteric-coated capsules = Eudracaps® capsules

[0322] Table 7 above shows that no vomiting events were observed in fasted monkeys administered enteric-coated capsules containing formula I. In contrast, vomiting was observed in 2 of 3 fasted monkeys administered non-enteric-coated capsules containing formula I.

[0323] [Table 8] Untreated female cynomolgus monkeys in a fasting state were administered formula I (60 mg / kg, orally) in capsule form. Non-enteric coated capsules = HPMC capsules Enteric-coated capsules = Eudracaps® capsules

[0324] Table 8 above shows that in fasted monkeys administered enteric-coated capsules containing formula I, the results were higher AUC exposure and lower variability compared to non-enteric-coated capsules containing formula I.

[0325] Figures 4, 5, and 6 show that fasted monkeys administered enteric-coated capsules containing formula I exhibit higher AUC exposure and lower variability compared to non-enteric-coated capsules containing formula I.

Claims

1. The following saccharin salt compounds: 【Chemistry 1】 A saccharinate compound in which a solvent may be optionally present.

2. formula: 【Chemistry 2】 A compound in which a solvent may be optionally present.

3. The crystalline form of the compound according to claim 1 or 2, characterized by having an X-ray powder diffraction (XRPD) pattern that includes peaks at 2θ values ​​of 4.4 ± 0.2 and 11.3 ± 0.

2.

4. The crystalline form according to claim 3, characterized by having an X-ray powder diffraction (XRPD) pattern further including peaks at 2θ values ​​of 12.9 ± 0.2 and 16.2 ± 0.

2.

5. The crystalline form according to claim 3 or 4, characterized by having an X-ray powder diffraction (XRPD) pattern further including peaks at 2θ values ​​of 17.5±0.2, 20.1±0.2, and 20.8±0.

2.

6. A pharmaceutical composition comprising a compound or crystalline form according to any one of claims 1 to 5, and at least one excipient.

7. A method for treating a patient with cancer, comprising administering an effective amount of the pharmaceutical composition described in claim 6 to a patient in need of the treatment.

8. The method according to claim 7, wherein the cancer is a KRAS G12D-related cancer.

9. The method according to claim 7 or 8, wherein the cancer includes lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, or colorectal cancer.

10. A compound or crystalline form according to any one of claims 1 to 5, for use in therapeutic purposes.

11. A compound or crystalline form according to any one of claims 1 to 5, for use in the treatment of the aforementioned cancer.

12. The compound or crystalline form for use according to claim 11, wherein the cancer is a KRAS G12D-related cancer.

13. The compound or crystalline form for use according to claim 11 or 12, wherein the cancer is selected from lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer.

14. The compound or crystalline form for use according to claim 12, wherein the cancer is selected from colorectal cancer, pancreatic cancer, and gastric cancer.

15. A compound or crystalline form according to any one of claims 1 to 5, for use in combination with a PD-1 inhibitor simultaneously, separately, or in succession in the treatment of the aforementioned cancer.

16. The compound or crystalline form for use according to claim 15, wherein the cancer is a KRAS G12D-related cancer.

17. The compound or crystalline form for use according to claim 15 or 16, wherein the cancer is selected from lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer.

18. The compound or crystalline form for use according to claim 16 or 17, wherein the cancer is selected from colorectal cancer, pancreatic cancer, and non-small cell lung cancer.

19. A compound or crystalline form according to any one of claims 1 to 5, for use in combination with a PD-1 inhibitor simultaneously, separately, or sequentially in the treatment of the aforementioned cancer, wherein the cancer is selected from colorectal cancer, pancreatic cancer, and gastric cancer.

20. The PD-1 inhibitor comprises pembrolizumab, wherein the compound or crystalline form for use according to any one of claims 15 to 19.

21. A compound or crystalline form according to any one of claims 1 to 5, for use in combination with an EGFR inhibitor simultaneously, separately, or in succession in the treatment of the aforementioned cancer.

22. The compound or crystal form according to claim 21, wherein the cancer is a KRAS G12D-related cancer.

23. The compound or crystalline form according to claim 21 or 22, wherein the cancer is selected from lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer.

24. The compound or crystalline form according to claim 23, wherein the cancer is selected from colorectal cancer, pancreatic cancer, and non-small cell lung cancer.

25. The compound or crystalline form according to claim 21, wherein the cancer is selected from colorectal cancer, pancreatic cancer, and gastric cancer.

26. The EGFR inhibitor comprises cetuximab, wherein the compound or crystalline form for use according to any one of claims 21 to 25.

27. A method for preparing a compound or crystalline form according to any one of claims 1 to 5, comprising suspending 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile in a solvent, adding saccharin, and isolating the disaccharin salt.

28. The method according to claim 27, wherein the saccharin is dissolved in the solvent and then added to 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile in the solvent.

29. The method according to claim 27 or 28, wherein the solvent is an alcohol.

30. The aforementioned alcohol is C １ ~C ４ The method according to claim 29, wherein the alcohol is used.

31. The method according to claim 29 or 30, wherein the solvent comprises at least one of ethanol or methanol.

32. The method according to any one of claims 27 to 31, wherein the saccharin is dissolved in the solvent, and the resulting solution is gradually added to 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile in the solvent.

33. The method according to any one of claims 27 to 32, wherein the solvent is ethanol, and 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile in ethanol is heated to about 47±5°C, after which the saccharin in ethanol is added.

34. The method according to any one of claims 27 to 33, wherein the saccharin is used in a molar excess with respect to 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile.

35. Formula I: 【Transformation 3】 A compound wherein the solvent may be optionally present, and which can be obtained by treating 2-amino-7-fluoro-4-(5-fluoro-3-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidine-7a(5H)-yl)methoxy)-1-((1R,5S)-3-((R)-2-hydroxypropyl)-3,8-diazabicyclo[3.2.1]octan-8-yl)-7,9-dihydrofluoro[3,4-f]quinazolin-6-yl)thieno[3,2-c]pyridine-3-carbonitrile with saccharin.

36. a) a core composition comprising a compound or crystalline form described in any one of claims 1 to 5; b) a capsule containing the core composition and having a body and a cap; c) a polymer seal covering the transition between the capsule cap and the body; and d) an enteric coating coating the polymer seal and the capsule.

37. The aforementioned compound, 【Chemistry 4】 The dosage form according to claim 36, which is a saccharin salt.

38. The aforementioned compound, 【Transformation 5】 The dosage form according to claim 36.

39. A process for preparing a dosage form according to any one of claims 36 to 38, comprising: a) blending all components contained in the core composition in a suitable blender; b) weighing the amount of the blend formed in step (a) that will become the core composition; c) compressing the blend weighed in step (b) using a capsule slag mold; d) placing the slag formed in step (c) inside the capsule; e) covering the transition between the capsule cap and the body with the polymer seal; and f) coating the capsule with the enteric coating.

40. A solid pharmaceutical composition comprising: (1) a compound or crystalline form according to any one of claims 1 to 5 constituting about 30 to 67% of the composition; (2) microcrystalline cellulose constituting about 20 to 55% of the composition; (3) mannitol constituting about 0 to 10% of the composition; (4) crospovidone constituting about 2% to about 5% of the composition; (5) colloidal silicon dioxide constituting a maximum of about 0.5 to 2% of the composition; (6) magnesium stearate constituting about 1 to 3% of the composition; and (7) an enteric coating, all percentages being weight percentages and the total weight being 100%.

41. The aforementioned compound, 【Transformation 6】 The pharmaceutical composition according to claim 40, wherein the saccharinate compound is...

42. The aforementioned compound, 【Transformation 7】 The pharmaceutical composition according to claim 40.

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