Crystal form of mutant BRAF degradation agent and preparation method thereof

Abstract

The present invention provides advantageous (3R)-3-[6-[2-cyano-3-[[ethyl (meth) sulfamoyl] amino]-6-fluorophenoxy]-4-oxoquinazolin-3-yl]-8-[2-[1-[3-(2, 4-dioxo-1, 3, 4-triazole-3-yl)-4-oxoquinazolin-3-yl]-5-(4-fluorophenyl)-4-oxo-7-oxo-7-oxo-7-oxo-7-oxo-7-oxo-7-oxo-7-oxo-7-oxo-7- The present invention relates to isolated crystalline forms of 1, 3-diazinane-1-yl)-5-fluoro-1-methylindazol-6-yl]-4-hydroxypiperidin-4-yl] acetyl]-1-oxa-8-azaspiro [4.5] decane (Compound 1), as well as methods of preparing crystalline forms of Compound 1 for therapeutic applications. The invention also provides an improved process for the synthesis of Compound 1, a novel pharmaceutical composition comprising Compound 1 and a novel use of Compound 1.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese Patent Application No. 202211627337.6, filed on December 2, 2022, U.S. Provisional Application No. 63 / 438,422, filed on January 11, 2023, and U.S. Provisional Application No. 63 / 599,450, filed on November 15, 2023. The entire contents of each of these applications are incorporated herein by reference for all purposes. Technical Field

[0003] The present invention provides an isolated crystalline form of (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluorophenoxy]-4-oxoquinazolin-3-yl]-8-[2-[1-[3-(2,4-dioxo-1,3-diazinane-1-yl)-5-fluoro-1-methylindazol-6-yl]-4-hydroxypiperidin-4-yl]acetyl]-1-oxa-8-azaspiro[4.5]decane (Compound 1) that is advantageous as a mutant BRAF degrader, and methods for preparing the crystalline form of Compound 1 for therapeutic applications as further described herein. The present invention also provides improved methods for synthesizing Compound 1 and its isotopic (e.g., deuterated) derivatives, new pharmaceutical compositions containing Compound 1, and new uses of Compound 1. Background Art

[0004] BRAF is a serine / threonine protein kinase, a member of the signal transduction protein kinases. BRAF plays a key role in the mitogen-activated protein kinase (MAPK) signaling pathway and is mutated in approximately 8% of all human cancers, including melanoma (approximately 60%), colorectal cancer (approximately 10%), and lung adenocarcinoma (approximately 5%). BRAF mutations have also been identified in thyroid cancer and other cancers. The most common mutation in BRAF is V600E (class I), which occurs in half of malignant melanomas. This mutation causes ERK to overactivate and function as a RAF inhibitor-sensitive monomer. Other common activating mutations include class II mutations such as G469A and class III mutations such as G466V. Class II and class III mutations activate ERK by promoting RAF homodimerization or heterodimerization.

[0005] The BRAF protein provides a mechanism for signal propagation that requires protein homodimerization (BRAF-BRAF) or heterodimerization with other RAF proteins (BRAF-RAF1 or BRAF-ARAF). When BRAF is mutated, as observed in oncology indications with BRAFV600E / K substitutions, BRAF signaling becomes independent of homodimers and / or heterodimers. The kinase activity becomes overactivated as a monomeric protein and drives cell proliferation signals.

[0006] Several BRAF inhibitors have been described that inhibit monomeric BRAF but not dimeric BRAF, including vemurafenib, dabrafenib, and conafenib. However, resistance typically develops within a year and includes RAS mutations, BRAFV600E amplification, and BRAFV600E intragenic deletions or splice variants. These inhibitors are also ineffective against non-V600 BRAF mutants (class II and class III) that activate ERK by promoting RAF homodimerization or heterodimerization.

[0007] Despite the therapeutic benefit of available BRAF inhibitors, the duration of antitumor responses to these drugs can be limited by the emergence of resistance.

[0008] Examples of BRAF inhibitors are described in patent applications filed by Hoffman La Roche AG, including WO2021 / 116055, WO2021 / 116050, WO2022 / 129259, WO2022 / 129260, WO2022 / 258612, and WO2022 / 258600.

[0009] Patent application WO2022 / 261250 filed by C4 Therapeutics Inc. describes BRAF degradation compounds. Additionally, non-limiting examples of BRAF degradation compounds include those described in WO2018 / 119448, WO2019 / 199816, WO2020 / 051564, WO2021 / 255212, and WO2022 / 047145.

[0010] Despite these efforts, there remains a need for new therapeutic agents and approaches to treat BRAF-mediated cancers, particularly mutant BRAF-mediated cancers. Summary of the Invention

[0011] Stable and crystalline forms of Compound 1 were discovered, including Compound 1 Form B and Compound 1 sodium salt Form F.

[0012]

[0013] Due to its high stability, scalability and reproducibility, Form B is superior to other crystalline forms of Compound 1. For example, when Compound 1 Form B was tested for stability in an open container at 25 ° C / 92% relative humidity, in an open container at 40 ° C / 75% RH and in a closed container at 60 ° C for one week, Compound 1 Form B exhibited excellent chemical and physical stability without any change in purity, crystal form and crystallinity (see Example 15, volume stability). When Compound 1 Form B was tested in a water adsorption and desorption experiment at 25 ° C (see Example 16), Compound 1 Form B exhibited excellent stability and no change in crystallinity, with only slight hygroscopicity (water absorption 1.6% at 95% RH). Compound 1 Form B also showed excellent crystalline stability under compression (2 MPa and 10 MPa, Example 17), dry milling and wet granulation conditions (Examples 18-19). Wet granulation experiments in the presence of water or ethanol also showed that the crystallinity did not change (Example 19). These favorable and beneficial stability characteristics of Compound 1 Form B are useful properties that extend the shelf life of Compound 1 Form B and its pharmaceutical composition, and allow for improved purity and repeatability during production-scale preparation. The high stability of Compound 1 Form B under high humidity conditions is useful and beneficial for the development and preparation of pharmaceutical compositions. These properties can also be used to adjust the delivery rate of Compound 1 for improving therapeutic effects.

[0014] Sodium salt Form F also shows good physicochemical properties, including good crystallinity, stoichiometry, high dehydration temperature, and good counterion safety. When Compound 1 sodium salt Form F was tested for stability in an open container at 25°C / 92% relative humidity, in an open container at 40°C / 75% RH, and in a closed container at 60°C for one week, Compound 1 sodium salt Form F exhibited excellent crystal stability (no crystal change) (see Example 25). After one week of intensive testing at 25°C / 92% RH, Form F showed only a slight decrease in chemical purity (1.2%), and after one week of intensive testing at 60°C in a closed container, the crystallinity decreased slightly. In water adsorption and desorption experiments at 25°C (Example 27), Compound 1 sodium salt Form F exhibited excellent stability and no change in crystallinity. These favorable and beneficial stability characteristics of Compound 1 sodium salt Form F are useful properties for increasing the shelf life of Compound 1 sodium salt Form F and its pharmaceutical compositions, and allow for improved purity and reproducibility during manufacturing scale production. These properties can also be used to prepare pharmaceutical compositions containing Compound 1 sodium salt Form F and to adjust the delivery rate of Compound 1 for improved therapeutic effects.

[0015] Compound 1 is a small molecule mutant BRAF degrader that degrades mutant BRAF, such as class I, class II and / or class III mutant BRAF, through the ubiquitin proteasome pathway. Compound 1 binds to the ubiquitously expressed E3 ligase protein cereblon (CRBN) and changes the substrate specificity of the CRBN E3 ubiquitin ligase complex, resulting in the recruitment and ubiquitination of mutant BRAF (e.g., such as BRAF V600E). Compound 1 effectively degrades class I mutant BRAF such as V600E, class II mutant BRAF such as G469A, class III mutant BRAF such as G466V mutation, and splice variants such as p61-BRAF. V600E For example, in A375 cells, compound 1 potently degraded BRAF V600E (E max = 26% (ie, 74% BRAF protein degradation); 24 h DC 50 =14 nM) and inhibited ERK phosphorylation (24 h IC 50 =11 nM) and cell growth (96 h GI 50 =94 nM).

[0016] In other aspects, a novel advantageous pharmaceutical composition suitable for administration to humans is provided, comprising compound 1 or a crystalline form of compound 1 according to the present invention. The advantageous pharmaceutical composition comprises compound 1 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition is a tablet comprising intragranular and extragranular particles, and wherein the intragranular particles comprise compound 1. In certain embodiments, compound 1 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients are spray-dried to form a spray-dried dispersion, which is then processed into intragranular particles. In certain embodiments, the intragranular excipients added prior to the spray-drying step include solubility enhancers, such as hypromellose acetate succinate; and permeation enhancers, such as d-alpha tocopheryl polyethylene glycol succinate. In certain embodiments, one or more additional intragranular excipients are added to the spray-dried dispersion. In certain embodiments, the intragranular excipients added to the spray-dried dispersion include, but are not limited to, fillers such as mannitol; mucoadhesives / disintegrants such as croscarmellose sodium; flow aids such as colloidal silicon dioxide; and binders / glidants such as microcrystalline cellulose. In certain embodiments, the spray-dried dispersion and the intragranular excipients added after the spray-drying step are blended, ground, and optionally rolled together into ribbons, and a lubricant such as magnesium stearate is optionally added as a further intragranular excipient. In certain embodiments, the material or ribbon is subsequently ground and blended with an extragranular excipient.

[0017] Non-limiting examples of extragranular excipients include, but are not limited to, binders / glidants, such as microcrystalline cellulose; mucoadhesives / disintegrants, such as croscarmellose sodium; and lubricants, such as magnesium stearate. In certain embodiments, the addition of a disintegrant as an extragranular excipient will help the tablet disintegrate into small fragments more efficiently. In certain embodiments, the presence of a disintegrant as both an extragranular and intragranular excipient will ensure more efficient disintegration of the tablet.

[0018] Such pharmaceutical formulations comprising intragranular and extragranular excipients have improved pharmaceutical properties, such as improved tablet disintegration, drug release, dissolution profile and / or mechanical strength.

[0019] In certain embodiments, a pharmaceutical composition comprising compound 1 is prepared from a crystalline form as described herein, such as compound 1 crystalline form B. In certain embodiments, compound 1 crystalline form B can be dissolved and then spray-dried to form a solid spray-dried dispersion with one or more pharmaceutically acceptable excipients. In some aspects, the pharmaceutical composition comprises compound 1, a dissolution enhancer, a penetration enhancer, a filler, one or more adhesives and / or glidants, and one or more flow aids. Non-limiting examples of pharmaceutically acceptable excipients include hypromellose (e.g., hypromellose acetate succinate), vitamin E (e.g., d-α-tocopheryl polyethylene glycol succinate), mannitol, cellulose (e.g., microcrystalline cellulose), cross-linked sodium carboxymethyl cellulose, silicon dioxide (e.g., untreated pyrogenic colloidal silicon dioxide), and magnesium stearate. In certain embodiments, the pharmaceutical composition according to the present invention is formulated into a unit dosage form, such as an oral unit dosage form. In certain embodiments, the pharmaceutical composition according to the present invention is formulated into a tablet dosage form.

[0020] In certain non-limiting embodiments, pharmaceutical compositions comprising Compound 1 comprise the following excipients.

[0021]

[0022]

[0023] Also provided is a method for preparing a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of compound 1 according to the present invention. In certain non-limiting aspects, a method for preparing a pharmaceutical composition comprising compound 1 comprises: (i) a spray drying step to provide a spray-dried intermediate (SDI) containing compound 1 and a pharmaceutically acceptable excipient; (ii) a granulation step to provide particles containing compound 1 and one or more pharmaceutically acceptable excipients having a desired bulk density of about 0.4 to 0.6 g / mL (e.g., about 0.48 to 0.54 g / mL); and (iii) a tableting step to provide a pharmaceutical composition in the form of an oral unit dosage comprising compound 1 and a pharmaceutically acceptable excipient.

[0024] Pharmaceutical compositions comprising compound 1 or a crystalline form of compound 1 of the present invention can be used to treat mutant BRAF-mediated cancers, such as, but not limited to, melanoma, lung cancer (including, for example, non-small cell lung cancer), colorectal cancer (including, for example, microsatellite-stable colorectal cancer), thyroid cancer (including, for example, anaplastic thyroid cancer), or ovarian cancer. In certain non-limiting embodiments, pharmaceutical compositions comprising compound 1 or a crystalline form of compound 1 of the present invention are used to treat solid tumors mediated by V600X mutant BRAF. Other non-limiting examples of treatable conditions include solid tumor malignancies with mutant BRAF drivers.

[0025] In certain embodiments, a pharmaceutical composition comprising Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of Compound 1 of the present invention can be administered to treat cancer that has become resistant to BRAF inhibitors.

[0026] The present invention also provides an improved method for producing Compound 1 or a pharmaceutically acceptable salt thereof or an isotopic derivative thereof, such as a deuterated derivative, on a production scale.

[0027] In certain aspects, production-scale preparation of Compound 1 or a pharmaceutically acceptable salt thereof comprises reacting Compound A with Compound B according to the following Reaction Scheme 1, resulting in the formation of an amide bond. In non-limiting embodiments, deuterium is placed on an atom at the alpha, beta, or gamma position of the metabolic site.

[0028] Option 1:

[0029]

[0030] In certain embodiments, in order to promote the formation of an amide bond between compound A and compound B, the carboxylic acid group in compound B is activated by converting the carboxylic acid hydroxyl group into a more reactive group. In certain embodiments, the activation of the carboxylic acid group in compound B is carried out by reacting compound B with a coupling agent that activates carboxylic acid. In certain embodiments, the coupling agent is TSTU (2-succinimidyl-1,1,3,3-tetramethyluronium tetrafluoroborate). TSTU is used as an effective coupling agent by rapidly activating carboxylic acid to form an N-succinimidyl active ester, which reacts with primary amines to form carboxamides. This reaction is suitable for amplification and is effective in the presence of water and is selective to amines in the presence of alcohol groups.

[0031] In certain embodiments, the reaction between Compound A and Compound B to produce Compound 1 can be carried out using a separate activating reagent, resulting in the formation of an amide bond. In certain embodiments, the reaction between Compound A and Compound B is carried out using a combination of N-hydroxysuccinimide (NHS) and EDCl (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) or a combination of NHS and DCC (dicyclohexylcarbodiimide).

[0032] In certain embodiments, compound A and / or compound B has at least one desired atomic isotope substitution in an amount higher than the natural abundance of the isotope, i.e., enriched. In certain embodiments, isotopic derivatives of compound A and / or compound B are used for the synthesis described in Scheme 1. In certain non-limiting embodiments, compound A and / or compound B include a deuterium atom or multiple deuterium atoms. In certain embodiments, compound A has formula A-d3, and compound B has formula B-d4:

[0033]

[0034] In certain embodiments, Compound A or an isotopic derivative thereof (eg, a deuterated derivative) is prepared from Compound C of the following formula:

[0035]

[0036] where R X is hydrogen or an amine protecting group. In certain embodiments, the amine protecting group R X Non-limiting examples of include, but are not limited to, benzyloxycarbonyl (Cbz), p-methoxybenzylcarbonyl (Moz or MeOZ), tert-butyloxycarbonyl (BOC), 9-fluorenylmethoxycarbonyl (Fmoc), benzoyl (Bz), benzyl (Bn), carbamate, p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP), and tosyl (Ts). In certain embodiments, the amine protecting group R XIt is tert-butoxycarbonyl (BOC).

[0037] The improved synthetic methods described herein include methods for preparing chirally pure Compound C. Compound C is used to synthesize Compound A as a mixture of (R)- and (S)-stereoisomers or as a single (R)-stereoisomer:

[0038]

[0039] In certain embodiments, the (R)-stereoisomer of Compound C is prepared using a transaminase-catalyzed reductive amination of the corresponding ketone precursor:

[0040]

[0041] In this asymmetric synthesis reaction, for example, isopropylamine can be used as a sacrificial amine donor source.

[0042] In other embodiments, the (R)-stereoisomer of Compound C is prepared from racemic Compound C by diastereomeric recrystallization of the salt formed between racemic Compound C and a chiral resolving acid, followed by liberation of the desired enantiomer with a base. In certain embodiments, the chiral resolving acid is pyroglutamic acid. In certain embodiments, the chiral resolving acid is D-mandelic acid, and the base used to recover the separated individual (R)- and (S)-enantiomers of Compound C from the diastereomeric salt of Compound C with D-mandelic acid is a suitable inorganic base, such as sodium hydroxide, as shown in Scheme 2. In other aspects, the base used to recover the separated enantiomers is an organic base.

[0043] Option 2:

[0044]

[0045] In certain aspects, Compound (R)-C is used to prepare Compound A or an isotopic derivative thereof, as shown in Scheme 3.

[0046] These two methods allow the preparation of chiral pure substances without the use of expensive chiral chromatography. The improved method also avoids the loss of a large amount of high-grade synthetic materials (which have undesirable chirality). Therefore, the throughput and scalability of the synthetic sequence are significantly improved.

[0047] Option 3:

[0048]

[0049] In certain embodiments, step 1 in Scheme 3 comprises reacting 5-hydroxyanthranilic acid (2-amino-5-hydroxybenzoic acid) or an isotopic derivative thereof (e.g., a deuterated derivative) with compound (R)-C and trialkyl orthoformate CH(OAlk)3 (e.g., triethyl orthoformate CH(OEt)3) to generate compound D containing a quinazolin-4-one ring system or an isotopic derivative thereof. In certain embodiments, the isotopic derivative of 2-amino-5-hydroxybenzoic acid is 2-amino-3,4-6-trideuterated-5-hydroxybenzoic acid. In certain embodiments, step 2 in Scheme 3 comprises a nucleophilic aromatic substitution reaction (S) between 2,3,6-trifluorobenzonitrile and the hydroxyl group of the quinazolinone fragment in compound D in the presence of a base (e.g., an organic or inorganic base, such as potassium carbonate). N Ar) to obtain compound E or an isotopic derivative thereof as a single enantiomer. In certain embodiments, in step 3 of Scheme 3, compound E or an isotopic derivative thereof is reacted with [ethyl(methyl)sulfamoyl]amine in the presence of a base (e.g., an organic or inorganic base, such as cesium carbonate) in the presence of a nucleophilic aromatic substitution (S N Ar) under conditions to obtain compound A or its isotopic derivative without losing its enantiomeric purity (e.g., >99% ee). In certain embodiments, in an optional step, when the nitrogen atom in compound A or its isotopic derivative (e.g., deuterated derivative) is protected by a nitrogen protecting group R X When protected, the protecting group, such as benzyloxycarbonyl (Cbz), p-methoxybenzylcarbonyl (Moz or MeOZ), tert-butyloxycarbonyl (BOC), 9-fluorenylmethoxycarbonyl (Fmoc), benzoyl (bz), benzyl (Bn), carbamate, p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP) or tosyl (Ts), can be removed to release the free amino group in compound A. In certain embodiments, when the amino protecting group R X In the case of BOC, the deprotection reaction is carried out under acidic conditions, such as but not limited to hydrochloric acid in an aqueous medium and / or an organic solvent, such as hydrochloric acid in acetone or ethyl acetate.

[0050] This synthetic sequence can be performed on a production scale to produce chirally pure isomers (eg, greater than 90%, 95%, or 99% enantiomeric purity).

[0051] In certain embodiments, Compound B or an isotopically enriched derivative thereof, such as a deuterated derivative, is prepared according to the reaction sequence depicted in Scheme 4.

[0052] Option 4:

[0053]

[0054] In certain embodiments, the synthetic route of compound B or its isotopic derivative or salt according to the present invention shown in Scheme 4 comprises four steps. In certain embodiments, the first step comprises a nucleophilic aromatic substitution reaction (S N Ar) under the conditions of optionally protected group R Y The protected carboxyl group of Compound F or its isotopic derivative is reacted with 2,4,5-trifluorobenzonitrile to obtain Compound G. In certain embodiments, the isotopic derivative of Compound F is tert-butyl 2-(3,3,5,5-tetradeuterated-4-hydroxy-4-piperidinyl)acetate. Y Examples of tert-butyl ( t In certain embodiments, the carboxylic acid protecting group R Y is tert-butyl ( t Bu). In certain embodiments, the nucleophilic aromatic substitution reaction of step 1 is carried out in the presence of a base. In certain embodiments, the base is an organic or inorganic base and includes but is not limited to sodium carbonate, potassium carbonate, cesium carbonate or N,N-diisopropylethylamine (DIEA, Hünig's base).

[0055] The resulting intermediate compound G is converted to a compound H containing an aminoindazole ring system by heating with methylhydrazine in a suitable solvent (e.g., an organic or inorganic solvent, including water and NMP (N-methyl-2-pyrrolidone)). In certain embodiments, in step 3 of Scheme 4, compound K is prepared from compound H in a reaction of compound H with acrylic acid (or a derivative thereof, such as an ester, such as a methyl ester or an ethyl ester) under Michael addition conditions. In certain embodiments, the Michael addition conditions include basic or acidic conditions. In certain embodiments, the Michael addition conditions include the use of dilute aqueous hydrochloric acid as a medium for the Michael addition reaction. In certain embodiments, compound K is converted to compound B via intermediate formation of a urea compound L in a condensation reaction with a cyanate, optionally followed by removal (deprotection) of the protecting group R Y To obtain compound B according to the present invention (steps 4a-b in Scheme 4). In certain embodiments, the cyanate used in step 4a of Scheme 4 is sodium cyanate or potassium cyanate used in the presence of acetic acid. In certain embodiments, the cyclization and deprotection step 4b in Scheme 4 is carried out under acidic conditions, including but not limited to the use of dilute hydrochloric acid.

[0056] The synthetic methods according to the present invention illustrated and described with reference to Schemes 1-4 have several advantageous and beneficial aspects. Advantageous features of the improved methods according to the present invention for producing compound 1 and / or its isotopes, such as deuterated derivatives, include high yields and / or high purity of compound 1. Also provided are isotopes, such as deuterated derivatives, and synthetic intermediates thereof, including chemical and chiral purity. Advantageous features of the improved methods according to the present invention for producing compound 1 also include high reliability, reproducibility, scalability, and / or atom efficiency.

[0057] Another advantageous and useful aspect of the improved synthetic method according to the present invention is that it is the palladium-free synthesis of synthetic intermediate and compound 1 and / or its isotope (such as deuterium) derivative.When synthesizing intermediate compound and precursor for the preparation of compound 1 provided by the invention or its isotope (such as deuterium) derivative, use palladium-free condition to allow to avoid final drug substance from being polluted by trace toxic and undesirable palladium (this palladium may be difficult to remove due to the complexing of palladium with intermediate compound and precursor, and it is brought to synthetic downstream and / or final step), and therefore other purification step can be eliminated. Palladium is the heavy metal of known toxicity, and it may damage bone marrow, kidney or liver, and when possible, avoids in pharmaceutical composition and drug substance.By avoiding the use of palladium catalyst, avoid the demand of expensive and time-consuming palladium removal.

[0058] Therefore, the palladium-free synthesis of Compound 1 and its isotopic (eg, deuterium) derivatives according to the present invention facilitates the subsequent use of Compound 1, its crystalline forms and derivatives as pharmaceutical substances and in pharmaceutical compositions.

[0059] In other aspects, a method for treating a mutant BRAF-mediated cancer that has metastasized to the brain or central nervous system (CNS) is provided, comprising administering an effective amount of Compound 1, or a pharmaceutically acceptable salt or crystalline form thereof, to a patient in need thereof. In certain embodiments, the cancer that has metastasized to the brain or CNS is colorectal cancer, melanoma, or non-small cell lung cancer. In certain embodiments of this aspect, Compound 1, or a pharmaceutically acceptable salt thereof, is administered to a patient in need thereof. In other embodiments of this aspect, a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition prepared from a crystalline form of Compound 1 is administered to a patient in need thereof.

[0060] Compound 1 has a high level of blood-brain barrier penetration (see Figure 52A When tested in an intracranial melanoma model, Compound 1 significantly reduced the bioluminescent signal relative to vehicle control or enfamil-treated mice, indicating a reduction in tumor burden (see Figure 40 A significant reduction in bioluminescent signal, an indicator of tumor burden growth, resulted in a significant increase in the survival time of mice bearing intracranial melanoma compared with enfamil (see Figure 41).

[0061] In certain aspects, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat a mutant BRAF-mediated cancer that has metastasized to the brain or CNS, wherein BRAF has mutated from wild-type. There are many possibilities for BRAF mutations. In certain non-limiting embodiments, the mutation is a Class I mutation, a Class II mutation, or a Class III mutation, or any combination thereof. Non-limiting examples of Class I mutations include V600 mutations, such as V600E, V600K, V600R, V600D, V600M, and V600N. Non-limiting examples of Class II mutations include G469A, G469V, G469L, G469R, L597Q, and K601E. Non-limiting examples of Class III mutations include G466A, G466E, G466R, G466V, S467L, G469E, N581I, D594E, D594G, and D594N. In certain embodiments, the BRAF mutation is a V600 mutation, e.g., V600E BRAF, which mediates cancers that have metastasized to the brain or CNS.

[0062] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, can treat mutant BRAF-mediated cancers that have metastasized to the brain or CNS, wherein the mutation is not a Class I, Class II, or Class III mutation. Non-limiting examples of mutations include G464I, G464R, N581T, L584F, E586K, G593D, G596C, L597R, L597S, S605I, S607F, N684T, E26A, V130M, L745L, and D284E.

[0063] In certain embodiments, Compound 1 or a pharmaceutically acceptable salt thereof can treat mutant BRAF-mediated cancers that have metastasized to the brain or CNS, wherein the mutation is a splice variant, such as p61-BRAF V600E .

[0064] In certain embodiments, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat mutant BRAF-mediated cancers that have metastasized to the brain or CNS, wherein the cancer is mediated by two or more mutant proteins, e.g., BRAF V600E / NRAS Q61K BRAF V600E / NRAS Q61R Double mutant mediated cancers. Non-limiting examples of double mutant cancers include colorectal cancer, which is caused by BRAF mutations (e.g., BRAF V600E ) and mutations in NRAS, MEK1, or phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) (e.g., BRAF V600E / MAP2K1P124S BRAF V600E / PIK3CA H1047R BRAF V600E / PIK3CA P449T ) mediated.

[0065] In certain embodiments, compound 1 or its pharmaceutically acceptable salt is used to treat mutant BRAF-mediated cancer that has been transferred to the brain or CNS, wherein cancer is resistant to at least one BRAF inhibitor, for example, cancer is resistant to or has acquired resistance to a BRAF inhibitor selected from dabrafenib, vemurafenib and connefenib. In certain aspects, there is a RAF protein homodimerization or heterodimerization that promotes mutations to be treated with BRAF inhibitors. For example, in certain embodiments, compound 1 or its pharmaceutically acceptable salt is used to treat mutant BRAF-mediated cancer that has been transferred to the brain or CNS, wherein cancer has one or more mutations that promote RAF protein dimerization. In certain embodiments, there is a cancer that promotes one or more RAF protein dimerization that is mutated that is resistant to treatment with BRAF inhibitors (such as dabrafenib, vemurafenib or connefenib). In certain embodiments, RAF protein dimers are BRAF-BRAF homodimers. In other embodiments, the RAF protein dimer is a heterodimer with other RAF proteins (BRAF-RAF1 or BRAF-ARAF).

[0066] In other embodiments, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat mutant BRAF-mediated cancers that have metastasized to the brain or CNS, where the cancer may have developed escape mutations such as BRAF V600E / NRAS Q61K or BRAF V600E / NRAS Q61R Double mutation cancer.

[0067] In other embodiments, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat melanoma that has metastasized to the brain or CNS. In other embodiments, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat colorectal cancer or lung cancer that has metastasized to the brain or CNS.

[0068] In alternative aspects, the S-enantiomer or a mixture of enantiomers, or a pharmaceutically acceptable salt thereof, is used in place of Compound 1 in the therapeutic or pharmaceutical compositions described herein.

[0069]

[0070] Other features and benefits of the present application will be apparent from the following detailed description.

[0071] Therefore, the present invention includes at least the following features:

[0072] (a) a crystalline form of Compound 1 as described herein;

[0073] (b) Compound 1, Form B;

[0074] (c) Compound 1 sodium salt Form F;

[0075] (d) a pharmaceutical composition comprising the crystalline form of Compound 1 according to any one of embodiments (a) to (c) and one or more pharmaceutically acceptable excipients;

[0076] (e) a pharmaceutical composition prepared from a crystalline form of Compound 1 (e.g., Form B) according to any one of embodiments (a) to (c);

[0077] (f) The pharmaceutical composition of (e), wherein the pharmaceutical composition is prepared from a spray-dried dispersion of Compound 1 and one or more pharmaceutically acceptable excipients;

[0078] (g) A pharmaceutical composition comprising Compound 1 or a pharmaceutically acceptable salt thereof, a dissolution enhancer, a permeation enhancer, a filler, a binder / glidant and / or a mucoadhesive / disintegrant.

[0079] (h) a method for treating a disorder mediated by BRAF, comprising administering an effective amount of a crystalline form of Compound 1 or a pharmaceutical composition according to any one of embodiments (a) to (g);

[0080] (i) Use of the Compound 1 crystalline form or pharmaceutical composition according to any one of embodiments (a) to (g) for treating a disorder mediated by BRAF;

[0081] (j) Use of a compound or pharmaceutical composition according to any one of embodiments (a) to (g) for the preparation of a medicament for treating a disorder mediated by BRAF;

[0082] (k) a method for treating cancer, comprising administering an effective amount of the crystalline form or pharmaceutical composition of Compound 1 according to any one of Embodiments (a) to (g);

[0083] (1) the crystalline form or pharmaceutical composition of Compound 1 according to any one of embodiments (a) to (g) for use in treating cancer;

[0084] (m) Use of the Compound 1 crystalline form or pharmaceutical composition according to any one of embodiments (a) to (g) for treating cancer;

[0085] (n) Use of the crystalline form or pharmaceutical composition of Compound 1 according to any one of Embodiments (a) to (g) in the preparation of a medicament for treating cancer;

[0086] (o) a method for preparing Compound 1 Form B as described herein;

[0087] (p) A method for preparing Compound 1 sodium salt Form F as described herein.

[0088] (q) a method for treating mutant BRAF-mediated cancer that has metastasized to the brain or CNS, comprising administering an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof;

[0089] (r) The method of (q), wherein the mutant BRAF-mediated cancer is melanoma;

[0090] (s) Use of Compound 1 or a pharmaceutically acceptable salt thereof for treating mutant BRAF-mediated cancers that have metastasized to the brain or CNS;

[0091] (t) use of Compound 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating mutant BRAF-mediated cancer that has metastasized to the brain or CNS; and

[0092] (u) (s) or (t) wherein the mutant BRAF-mediated cancer is melanoma. BRIEF DESCRIPTION OF THE DRAWINGS

[0093] Figure 1 Depicted is the XRPD pattern of Compound 1 Form B. The experiments were performed as described in Examples 8 and 28.

[0094] Figure 2 Depicted is the DSC thermogram of Compound 1 Form B. The experiment was performed as described in Example 28.

[0095] Figure 3 Depicted is the TGA thermogram of Compound 1 Form B. The experiment was performed as described in Example 28.

[0096] Figure 4 Depicted is the DVS isotherm of Compound 1 Form B at 25° C. The experiment was performed as described in Example 28.

[0097] Figure 5 Depicted is the DVS mass change graph of Compound 1 Form B at 25° C. The experiment was performed as described in Example 28.

[0098] Figure 6 Depicted are XRPD overlays of Compound 1 Form B before and after DVS testing. The experiment was performed as described in Example 28.

[0099] Figure 7 Depicted is the XRPD pattern of Compound 1 Form A. The experiments were performed as described in Examples 8 and 28.

[0100] Figure 8Depicted is the DSC thermogram of Compound 1 Form A. The experiment was performed as described in Example 28.

[0101] Figure 9 Depicted is the TGA thermogram of Compound 1 Form A. The experiment was performed as described in Example 28.

[0102] Figure 10 Depicted is the XRPD pattern of Compound 1 Form C. The experiments were performed as described in Examples 8 and 28.

[0103] Figure 11 Depicted is the DSC thermogram of Compound 1 Form C. The experiment was performed as described in Example 28.

[0104] Figure 12 Depicted is the TGA thermogram of Compound 1 Form C. The experiment was performed as described in Example 28.

[0105] Figure 13 Depicted are overlays of XRPD patterns of Compound 1 Form B samples before compression, after compression at 2 MPa, and after compression at 10 MPa. The experiment was performed as described in Example 28.

[0106] Figure 14 Depicted are overlays of XRPD patterns of Compound 1 Form B samples before dry milling and after 1, 2, and 5 minutes of dry milling. The experiment was performed as described in Example 28.

[0107] Figure 15 Depicted are overlays of XRPD patterns of Compound 1 Form B samples before and after wet granulation in the presence of water or ethanol. The experiments were performed as described in Example 28.

[0108] Figure 16 Described is an XRPD pattern of Compound 1 Form C, a comparison of the XRPD pattern obtained after about 8 equilibration cycles of Compound 1 Form C in 2-methyltetrahydrofuran, and the XRPD pattern of Compound 1 Form B. As a result of equilibration, Form C converts to the more stable Form B. The experiment was performed as described in Example 28.

[0109] Figure 17 Depicted is the XRPD pattern of Compound 1 sodium salt Form F. The experiment was performed as described in Example 28.

[0110] Figure 18 Depicted is the DSC thermogram of Compound 1 sodium salt Form F. The experiment was performed as described in Example 28.

[0111] Figure 19 Depicted is the TGA thermogram of Compound 1 sodium salt Form F. The experiment was performed as described in Example 28.

[0112] Figure 20 Depicted is the DVS isotherm of Compound 1 sodium salt Form F at 25° C. The experiment was performed as described in Example 28.

[0113] Figure 21 Depicted is a graph of the DVS mass change of Compound 1 sodium salt Form F at 25° C. The experiment was performed as described in Example 28.

[0114] Figure 22 Depicted are XRPD overlays of Compound 1 sodium salt Form F before and after DVS testing. The experiment was performed as described in Example 28.

[0115] Figure 23 Depicted is the XRPD pattern of Compound 1 sodium salt Form D. The experiment was performed as described in Example 28.

[0116] Figure 24 Depicted is the DSC thermogram of Compound 1 sodium salt Form D. The experiment was performed as described in Example 28.

[0117] Figure 25 Depicted is the TGA thermogram of Compound 1 sodium salt Form D. The experiment was performed as described in Example 28.

[0118] Figure 26 Depicted is the XRPD pattern of Compound 1 sodium salt Form E. The experiment was performed as described in Example 28.

[0119] Figure 27 Depicted is the DSC thermogram of Compound 1 sodium salt Form E. The experiment was performed as described in Example 28.

[0120] Figure 28 Depicted is the TGA thermogram of Compound 1 sodium salt Form E. The experiment was performed as described in Example 28.

[0121] Figure 29 Depicted is the XRPD pattern of Compound 1 potassium salt Form G. The experiment was performed as described in Example 28.

[0122] Figure 30 Depicted is the DSC thermogram of Compound 1 potassium salt Form G. The experiment was performed as described in Example 28.

[0123] Figure 31 Depicted is the TGA thermogram of Compound 1 potassium salt Form G. The experiment was performed as described in Example 28.

[0124] Figure 32 Depicted is the XRPD pattern of Compound 1 potassium salt Form H. The experiment was performed as described in Example 28.

[0125] Figure 33 Depicted is the DSC thermogram of Compound 1 potassium salt Form H. The experiment was performed as described in Example 28.

[0126] Figure 34 Depicted is the TGA thermogram of Compound 1 potassium salt Form H. The experiment was performed as described in Example 28.

[0127] Figure 35 Comparisons are described of the XRPD patterns of Compound 1 sodium salt Form D obtained in a slurry equilibration of Compound 1 Form A with 1 equivalent of sodium hydroxide in methanol, the amorphous XRPD patterns obtained in a slurry equilibration of Compound 1 Form A with 1 equivalent of sodium hydroxide in acetone or acetonitrile, and the XRPD patterns of Compound 1 Form A. The experiments were performed as described in Example 21.

[0128] Figure 36 Described are the XRPD patterns of Compound 1 sodium salt Form E (prepared in methanol) and Form F (prepared in acetone or acetonitrile) obtained in a slurry equilibrium of Compound 1 Form A with 1 equivalent of sodium bicarbonate, NaHCO, compared to the XRPD patterns of Form A and Form B. The experiments were performed as described in Example 21.

[0129] Figure 37 Described is an XRPD pattern of Compound 1 Form I obtained by slurry equilibration of Form A in the presence of 1 equivalent of ammonia in methanol and acetonitrile compared to the XRPD pattern of Form A. The experiment was performed as described in Example 21.

[0130] Figure 38 The single crystal structure of (R)-8-(tert-butoxycarbonyl)-1-oxa-8-azaspiro[4.5]decan-3-amine (R)-2-hydroxy-2-phenylacetate is described. The experiments were performed as described in Example 32.

[0131] Figure 39 is a line graph of the viability of BRAF V600E mutant human colorectal adenocarcinoma HT29 cells after 72 hours of treatment with Compound 1. The X-axis is the concentration of Compound 1 in nM and the Y-axis is the cell viability %. The experiment was performed as described in Example 33.

[0132] Figure 40It is a line graph showing the change over time of the luciferase bioluminescence signal representing tumor load in the BRAF V600E mutant A375-luciferase intracranial tumor model after administering the test compound (carrier, connefenib administered at 35 mg / kg, compound 1 administered at 10 mg / kg and 30 mg / kg). The X-axis is the number of days after treatment begins, and the Y-axis is the bioluminescence (photons / second) corresponding to the luciferase signal. The error bars represent the standard error of the mean (SEM). Experiments were performed as described in Example 34.

[0133] Figure 41 Is a Kaplan-Meier survival curve, describing the survival rate in the BRAF V600E A375-luciferase intracranial tumor model after treatment with test compound (vehicle, connefenib at 35 mg / kg, compound 1 at 10 mg / kg and 30 mg / kg). The X-axis is the number of days after treatment started, and the Y-axis is the probability of survival. Experiments were performed as described in Example 34.

[0134] Figure 42 It is a line graph describing the change in plasma concentration over time of the test compound (administered with 35 mg / kg of conafenib and compound 1 at 10 mg / kg and 30 mg / kg) in the BRAF V600E mutant A375-luciferase intracranial model. The X-axis is the time after treatment, and the Y-axis is the compound concentration in the plasma in ng / mL. The error bars represent the standard error of the mean (SEM). Experiments were performed as described in Example 34.

[0135] Figure 43 It is a line graph describing the brain concentration change over time of the test compound (conafenib administered at 35 mg / kg and compound 1 administered at 10 mg / kg and 30 mg / kg) in the BRAF V600E mutant A375-luciferase intracranial model. The X-axis is the time after treatment, and the Y-axis is the compound concentration in the plasma in ng / g. The error bars represent the standard error of the mean (SEM). The experiment was performed as described in Example 34.

[0136] Figure 44 It is a line graph describing the change in BRAF V600E protein degradation over time in A375 CNS tumors after administration of the test compound (conafenib at 35 mg / kg and compound 1 at 10 mg / kg and 30 mg / kg). The X-axis is the time after treatment and the Y-axis is the mutant BRAF level (normalized). The error bars represent the standard error of the mean (SEM). The experiment was performed as described in Example 34.

[0137] Figure 45It is a line graph depicting the change in tumor size over time in human colorectal adenocarcinoma (HT-29CDX model of CRC) after administration of test compounds (10 μL / kg vehicle, 11 mg / kg cetuximab, 10 mg / kg compound 1, 10 mg / kg compound 1 + 11 mg / kg cetuximab, 35 mg / kg enemafenib, and 35 mg / kg enemafenib + 11 mg / kg cetuximab). The X-axis is the number of days after treatment and the Y-axis is in mm. 3 The tumor size was calculated using the PCR amplification assay. Error bars represent the standard error of the mean (SEM). The experiment was performed as described in Example 35.

[0138] Figure 46 Graph depicting the change in tumor size over time in a BRAF V600E mutant NSCLC xenograft model (PDX model of NSCLC) following administration of test compounds (vehicle control, 0.1 mg / kg trametinib, 100 mg / kg dabrafenib, 100 mg / kg dabrafenib + 0.1 mg / kg trametinib, 10 mg / kg compound 1, and 10 mg / kg compound 1 + 0.1 mg / kg trametinib). On day 28, mice treated with compound 1 with or without trametinib were discontinued, and tumor growth was monitored until day 45. The X-axis is days after treatment initiation, and the Y-axis is in mm. 3 The tumor size was calculated using the RT-PCR method. Error bars represent the standard error of the mean (SEM). The experiment was performed as described in Example 36.

[0139] Figure 47 Graph depicting the change in tumor size over time in a melanoma PDX model of BRAF inhibitor-resistant melanoma with BRAF kinase domain duplications obtained during combination treatment with the BRAF inhibitor dabrafenib and the MEK inhibitor trametinib following administration of test compounds (vehicle control, 0.1 mg / kg trametinib, 100 mg / kg dabrafenib, 100 mg / kg trametinib + 0.1 mg / kg trametinib, 10 mg / kg compound 1, and 10 mg / kg compound 1 + 0.1 mg / kg trametinib). On day 21, mice treated with compound 1 + trametinib stopped treatment and tumor growth was monitored until day 38. The X-axis is the number of days after treatment initiation and the Y-axis is in mm. 3 The experiment was performed as described in Example 37.

[0140] Figure 48is a line graph depicting the change in tumor size over time after administration of test compounds (10 μL / g vehicle control, 100 mg / kg dabrafenib, 35 mg / kg conafenib, and 10 mg / kg compound 1) in a xenograft CDX model derived from the A2058 human melanoma cancer cell line, a BRAF inhibitor-resistant melanoma with a MEK1 P124S mutation and BRAF V600E. The X-axis is days after treatment and the Y-axis is mm 3 The tumor volume / size was calculated using the PCR amplification assay. Error bars represent the standard error of the mean (SEM). The experiment was performed as described in Example 38.

[0141] Figure 49 Figure 3 is a line graph depicting changes in body weight following administration of a test compound to female BALB / c nude mice bearing A2058 melanoma xenografts harboring the oncogenic mutations BRAF V600E and MEK1 P124S. The X-axis represents the number of days post-treatment, and the Y-axis represents the mean body weight in grams per group. Data points represent the mean body weight per group. Error bars represent the standard error of the mean (SEM). The experiment was performed as described in Example 39.

[0142] Figure 50 Graph depicting the percentage change in body weight (BW) over time in mice implanted with A2058 melanoma CDX. Change in BW was calculated based on the weight of the animals on the first day of grouping. The X-axis is the number of days post-treatment, and the Y-axis is the percentage change in body weight. Data points represent the average percentage change in BW per group. Error bars represent the standard error of the mean (SEM). Experiments were performed as described in Example 39.

[0143] Figure 51 Figure 3 is a line graph depicting tumor volume in cubic millimeters following administration of a test compound in female BALB / c nude mice bearing A2058 xenografts. The X-axis represents the number of days post-treatment, and the Y-axis represents tumor size in millimeters. Data points represent the mean tumor volume per group. Error bars represent the standard error of the mean (SEM). Experiments were performed as described in Example 39.

[0144] Figure 52A 1 h, 6 h, 12 h, 24 h, and 48 h after administration, the concentration of Compound 1 in plasma and tumors collected from mice implanted with A2058 melanoma CDX over time. The Y axis is the concentration of Compound 1 in ng / g. The error bars represent the standard error of the mean (SEM). The experiment was performed as described in Example 39.

[0145] Figure 52B1 hour, 6 hours, 12 hours, 24 hours and 48 hours after compound administration, the concentration of connefenib in plasma and tumors collected from mice implanted with A2058 melanoma CDX changes over time. The Y axis is the concentration of connefenib in ng / g. The error bars represent the standard error of the mean (SEM). The experiment was performed as described in Example 39.

[0146] Figure 52C 6 hours, 12 hours, 24 hours and 48 hours after administration, in the plasma and tumors collected from mice implanted with A2058 melanoma CDX over time. The Y axis is the concentration of dabrafenib in ng / g. The error bars represent the standard error of the mean (SEM). The experiment was performed as described in Example 39.

[0147] Figure 53A : is a bar graph depicting Western blot analysis of p-ERK and ERK expression levels in A2058 tumors at 6, 12, 24, and 48 hours following a single dose of 10 mg / kg Compound 1. The expression level of the vehicle was used as a control at 6 hours post-administration. The experiment was performed as described in Example 39.

[0148] Figure 53B Figure 2 is a bar graph depicting Western blot analysis of p-ERK and ERK expression levels in A2058 tumors at 6, 12, 24, and 48 hours after administration of 35 mg / kg conafenib. The expression level of the vector was used as a control at 6 hours post-administration. The experiment was performed as described in Example 39.

[0149] Figure 53C Figure 2 is a bar graph depicting Western blot analysis of p-ERK and ERK expression levels in A2058 tumors at 6, 12, 24, and 48 hours after administration of 100 mg / kg dabrafenib. The expression level of the vector was used as a control at 6 hours post-administration. The experiment was performed as described in Example 39.

[0150] Figure 54A 1 is a bar graph depicting Western blot analysis of BRAF (V600E) expression levels in A2058 tumors at 6, 12, 24, and 48 hours after administration of 10 mg / kg Compound 1. The expression level of the vector was used as a control at 6 hours after administration. The experiment was performed as described in Example 39.

[0151] Figure 54B: This is a bar graph depicting Western blot analysis of BRAF (V600E) expression levels in A2058 tumors at 6, 12, 24, and 48 hours after administration of 35 mg / kg conafenib. The expression levels of the vector were used as a control at 6 hours after administration. The experiment was performed as described in Example 39.

[0152] Figure 54C Graph depicting Western blot analysis of BRAF (V600E) expression levels in A2058 tumors at 6, 12, 24, and 48 hours after administration of 100 mg / kg dabrafenib. The expression level of the vector was used as a control at 6 hours post-administration. The experiment was performed as described in Example 39. Specific implementation plan

[0153] definition

[0154] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which this application belongs. In the specification, the singular also includes the plural, unless the context clearly provides otherwise. Although methods and materials similar or equivalent to the methods and materials described in this application can be used in the practice and testing of this application, suitable methods and materials are described below. All publications, patent applications, patents and other references mentioned herein are incorporated herein by reference. The references cited here are not considered to be prior art of the application for protection. In the event of conflict, this specification (including definitions) will be the standard. In addition, materials, methods and examples are only illustrative, not restrictive.

[0155] The compounds are described using standard nomenclature. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0156] The terms "a" and "an" do not represent a quantitative limitation, but rather represent the presence of at least one of the items mentioned. The term "or" refers to "and / or". Unless otherwise stated herein, the enumeration of numerical ranges is intended only to serve as a shorthand method for individually referring to each individual numerical value falling within the range, and each individual numerical value is incorporated into this specification as if it were individually listed herein. The endpoints of all ranges are included within the range and are independently combinable. Unless otherwise stated herein or the context is otherwise clearly contradictory, all methods described herein can be performed in the appropriate order. Unless otherwise stated, the use of examples or exemplary language (e.g., "such as") is only intended to better illustrate the present invention and does not limit the scope of the present invention.

[0157] The term "alkyl" alone or in combination means a straight chain, branched or cyclic alkyl group having 1 to 8 carbon atoms, particularly a straight chain, branched or cyclic alkyl group having 1 to 6 carbon atoms, more particularly a straight chain or branched alkyl group having 1 to 4 carbon atoms. Examples of straight chain, branched or cyclic C1-C6 alkyl groups are methyl, ethyl, propyl, cyclopropyl, isopropyl, butyl, cyclobutyl, isobutyl, tert-butyl, pentyl, cyclopentyl, isopentyl, hexyl, isohexyl and cyclohexyl. In certain embodiments, "alkyl" is methyl. In certain embodiments, "alkyl" is ethyl.

[0158] The present invention includes compounds described herein having at least one desired atomic isotope substituted in an amount above the natural abundance of the isotope, i.e., enriched. Isotopes are atoms having the same atomic number but different mass numbers, i.e., the same number of protons but different numbers of neutrons. If isotopic substitution is used, typically the substitution is at least one deuterium for a hydrogen.

[0159] More generally, examples of isotopes that may be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen and fluorine, for example, 2 H. 3 H. 11 C. 13 C. 14 C. 15 N. 17 O. 18 O. 18 F and 35 In one non-limiting embodiment, isotope-labeled compounds can be used in metabolic studies (e.g., 14 C), reaction kinetics studies (e.g., 2 H or 3 H), detection or imaging techniques (e.g., positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution analysis) or radiotherapy of patients. In addition, any hydrogen atoms present in the compounds of the invention may be 18 F atom substitutions may be particularly desirable for PET or SPECT studies. Isotopically labeled compounds of this invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the following Schemes or Examples and Preparations by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.

[0160] By way of general example and not limitation, isotopes of hydrogen, such as deuterium ( 2 H) and tritium ( 3 H) can be used anywhere in the structure to achieve the desired result. Alternatively or additionally, isotopes of carbon can be used, for example, 13 C and 14 C.

[0161] Isotope substitution, such as deuterium substitution, can be partial or complete. Partial deuterium substitution refers to that at least one hydrogen is replaced by deuterium. In certain embodiments, at any target position, the isotope is 90%, 95% or 99% or more isotopically enriched. In a non-limiting embodiment, deuterium is 90%, 95% or 99% enriched at the desired position.

[0162] In a non-limiting embodiment, hydrogen atom is replaced with deuterium atom and can be provided in any compound described herein.For example, when any group is methyl, ethyl or methoxy, or when comprising methyl, ethyl or methoxy by substitution, alkyl residue can be by deuterated (in non-limiting embodiments, CDH2, CD2H, CD3, CH2CD3, CD2CD3, CHDCH2D, CH2CD3, CHDCHD2, OCDH2, OCD2H or OCD3 etc.).In certain other embodiments, when two substituents are combined to form a ring, unsubstituted carbon can be by deuterated.This substitution can improve the property of compound 1, for example, deuterium substitution at metabolic site can reduce metabolic rate (for example, kinetic isotope effect).Similarly, deuterium substitution near metabolic site can also reduce metabolic rate.In certain embodiments, deuterium is substituted at metabolic site or at α, β or γ position.

[0163] The compounds of the present invention can form solvates with solvents (including water). Therefore, in a non-limiting embodiment, the present invention includes solvated forms of the compounds described herein. The term "solvate" refers to a molecular complex of a compound of the present invention (including its salt) and one or more solvent molecules. Non-limiting examples of solvents are water, ethanol, isopropanol, dimethyl sulfoxide, acetone and other common organic solvents. The term "hydrate" refers to a molecular complex comprising a compound of the present invention and water. Pharmaceutically acceptable solvates according to the present invention include those in which the solvent can be isotopically substituted, for example, D2O, acetone-d6, DMSO-d6. Solvate can be in liquid or solid form.

[0164] "Dosage form" refers to the unit of administration of an active agent. Examples of dosage forms include tablets, capsules, injections, suspensions, liquids, emulsions, granules, pellets, creams, ointments, suppositories, inhalable forms, transdermal forms, oral, sublingual, topical, gel, mucosal, and the like. "Dosage form" may also include implants, such as those inserted into a tumor or abnormal cell growth.

[0165] "Parenteral" administration of a compound includes, for example, subcutaneous (sc), intravenous (iv), intramuscular (im) or intrasternal injection or infusion techniques.

[0166] The term "pharmaceutically acceptable salt" refers to a salt suitable for contact with human and animal tissues. The salts of the compounds of the present invention can be synthesized by conventional chemical methods from a parent compound containing an alkaline or acidic portion. Typically, such salts can be prepared by reacting the free acid form of these compounds with a stoichiometric amount of a suitable base (e.g., Na, Ca, Mg or K hydroxide, carbonate, bicarbonate, etc.), or by reacting the free base form of the compound with a stoichiometric amount of a suitable acid. Such reactions are typically carried out in water, an organic solvent, or a mixture thereof. Typically, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are typical where feasible. The salts of the compounds of the present invention can also optionally include a solvate of the compound or its salt.

[0167] Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues. For example, conventional non-toxic alkaline salts include those derived from inorganic bases such as sodium hydroxide, sodium bicarbonate, sodium carbonate, potassium hydroxide, potassium bicarbonate, and potassium carbonate. A list of other suitable salts can be found, for example, in Remington's Pharmaceutical Sciences, 17th edition, Mack Publishing Company, Easton, Pa., p. 1418 (1985).

[0168] The term "carrier" refers to a diluent, excipient, or vehicle in which the active agent is used or delivered.

[0169] The term "pharmaceutically acceptable excipient" refers to ingredients used to prepare dosage forms and are suitably non-toxic, such as disintegrants, binders, fillers, solvents, buffers, tonicity agents, stabilizers, antioxidants, surfactants or lubricants used to formulate drug products.

[0170] "Patient" or "host" or "subject" refers to a human or non-human animal in need of treatment for any of the conditions described in detail herein (e.g., regulated by BRAF, particularly mutant BRAF). Typically, the host is a human. "Host" may alternatively refer to, for example, a mammal, a primate (e.g., a human), a horse, a dog, a cat, a cow, a sheep, a goat, a bird, etc.

[0171] A "therapeutically effective amount" refers to an amount of a compound that, when administered to a host in need thereof, is sufficient to treat a disease state. A "therapeutically effective amount" will vary depending on the disease state being treated, the severity of the disease being treated, the age and relative health of the host, the route and form of administration, the judgment of the attending physician or veterinarian, and other factors.

[0172] Throughout this disclosure, various aspects of the present invention can be presented in range format. It should be understood that the description of range format is for convenience only and should not be construed as limiting the scope of the invention. The description of a range should be considered to have clearly disclosed all possible subranges and single numerical values ​​within the range. For example, the description of a range (e.g., from 1 to 6) should be considered to be specific disclosure of each subrange (e.g., from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc.) and a single numeral (e.g., 1, 2, 2.7, 3, 4, 5, 5.3, and 6) within the range. Regardless of the width of the range, this applies.

[0173] The term "inhibitor" refers to a compound that competes with a specific ligand, reduces or prevents binding to a specific receptor, or reduces or prevents the function of a specific protein.

[0174] If one of the starting materials or compounds of the present invention contains one or more functional groups that are unstable or reactive under the reaction conditions of one or more reaction steps, suitable protecting groups (e.g., as described in "Protective Groups in Organic Chemistry" PGM Wuts, 5th edition, 2007, Wiley, New York) can be introduced before the key step using methods well known in the literature. Such protecting groups can be removed later in the synthesis using standard methods described in the literature. Examples of protecting groups are tert-butyl ( t The following are some of the following: 1-butyloxycarbonyl (Bu), tert-butyloxycarbonyl (Boc), 9-fluorenylmethylcarbamate (Fmoc), 2-trimethylsilylethylcarbamate (Teoc), benzyloxycarbonyl (Cbz), and p-methoxybenzyloxycarbonyl (Moz).

[0175] If not otherwise indicated in the context, the compounds of the invention may contain asymmetric centers and may exist as optically pure enantiomers, mixtures of enantiomers (eg racemates), mixtures of diastereomers, diastereomeric racemates or mixtures of diastereomeric racemates.

[0176] The term "asymmetric carbon atom" refers to a carbon atom having four different substituents. According to the Cahn-Ingold-Prelog rules, an asymmetric carbon atom can be in the "R" or "S" configuration.

[0177] In any case where a chiral carbon is present in a chemical structure, unless the context or drawings indicate otherwise, the structure is intended to encompass all stereoisomers associated with that chiral carbon as pure stereoisomers and as mixtures thereof.

[0178] In embodiments where chirally pure isomers (or optically pure enantiomers) are provided, chirally pure means that the compound contains greater than 90% by mole of the desired isomer, particularly greater than 95% by mole of the desired isomer, greater than 98% by mole of the desired isomer, greater than 99% by mole of the desired isomer, the molar percentages being based on the total moles of isomers in the compound. Chirally pure or chirally enriched compounds can be prepared by chiroselective synthesis or by separation of enantiomers.

[0179] Compound 1 crystal form

[0180] Compound 1 itself (free form) and salts of Compound 1 (salt forms) were studied for the crystalline forms of Compound 1. A total of three crystalline free forms of Compound 1 were identified as polymorphs, including Form A (which is a hydrate) and two anhydrates (identified as Form B and Form C). Five salts and their polymorphs were identified from the salt studies, including Compound 1 sodium salt Form D, Compound 1 sodium salt Form E, Compound 1 sodium salt Form F, Compound 1 potassium salt Form G, and Compound 1 potassium salt Form H.

[0181] Free compound 1 crystal form

[0182] The formation of crystalline forms of Compound 1 was investigated under various crystallization conditions. The conditions tested included equilibrium, slow cooling, slow evaporation, precipitation by addition of antisolvent, and vapor diffusion.

[0183] Three crystalline forms of compound 1 were identified: Form A, the starting material, and two anhydrates, Form B and Form C.

[0184] Compound 1 Form A

[0185] Form A is a low crystalline form. It was obtained from IPA, MTBE, EtOH, IPAc and toluene by equilibration at 25°C, from IPA, IPAc, toluene, DMSO / water (v:v=23:77) and DMSO / water (v:v=57:43) by temperature cycling, and from acetone by anti-solvent addition experiments. No improvement in crystallinity was observed after 2 weeks of equilibration. According to the KF results, it contains about 1.9 wt% of water. DCS shows a broad endothermic peak starting from about 13°C. Its T onset It melts at an enthalpy of 18 J / g at about 160°C. TGA shows a weight loss of about 2.7% at about 160°C. 1 H NMR showed no detectable residual solvent.

[0186] In certain embodiments, Form A is characterized by an XRPD pattern having one or more peaks within + / - 0.4, 0.3, or 0.2 degrees 2-theta of the peaks listed in Peak List #1 (see Example 28 for a description of the XRPD method).

[0187] Peak List #1

[0188]

[0189] 1. In certain embodiments, Compound 1 Form A is characterized by an XRPD pattern having at least three peaks selected from 15.3, 18.4, 24.9, and 25.3 + / - 0.4° 2θ. In other embodiments, Compound 1 Form A is characterized by an XRPD pattern having at least three peaks selected from 15.3, 18.4, 24.9, and 25.3 + / - 0.3° 2θ. Or in other embodiments, Compound 1 Form A is characterized by an XRPD pattern having at least three peaks selected from 15.3, 18.4, 24.9, and 25.3 + / - 0.2° 2θ.

[0190] In certain embodiments, Form A is characterized by a melting onset temperature of about 167.9°C ± 10°C and / or a melting enthalpy of about 18 J / g ± 10 J / g on a DSC thermogram. In certain embodiments, Form A is characterized by a weight loss of about 2.7% at about 160.0°C ± 10°C as measured by TGA.

[0191] In certain embodiments, as described in Example 7, Form A is characterized by a solubility in DMSO of at least about 250 mg / mL when dissolved at about 25°C ± 5°C.

[0192] Figure 7 The XRPD pattern of Compound 1 Form A is described. Figure 8 Depicted is the DSC thermogram of Compound 1 Form A. Figure 9 The TGA thermogram of Compound 1 Form A is depicted.

[0193] Compound 1 Form B

[0194] Form B is an anhydrate. It was obtained by equilibrium from the solvent system tested. Form B has moderate crystallinity. DSC showed a T of 194.4°C. onset Form B decomposes upon melting. TGA shows a weight loss of approximately 1.1% at approximately 170°C. 1 H NMR showed no detectable residual solvent. Since Form B can also be isolated from organic solvents, such as methanol or acetonitrile, and water mixtures with a water activity above about 0.9, this anhydrate should also be stable in aqueous media.

[0195] Due to its high stability, scalability and repeatability, Form B is superior to other crystalline forms of Compound 1. For example, when Compound 1 Form B was subjected to stability testing for one week at 25°C / 92% relative humidity in an open container, at 40°C / 75% RH in an open container, and at 60°C in a closed container, Compound 1 Form B exhibited excellent chemical and physical stability without any change in purity, crystal form and / or crystallinity. When Compound 1 Form B was tested in a water adsorption and desorption experiment at 25°C with a 40-0-95-0-40% RH cycle, Compound 1 Form B exhibited excellent stability without any change in crystallinity and only slight hygroscopicity (1.6% water absorption at 95% RH). Compound 1 Form B also showed excellent crystalline stability under compression (2MPa and 10MPa), dry milling and wet granulation conditions. Wet granulation experiments in the presence of water or ethanol also showed no change in crystallinity. Compound 1 Form B also exhibits crystalline stability in aqueous media over a wide pH range of 1.2-7.0.

[0196] In certain embodiments, Form B is characterized by an XRPD pattern having peaks within + / - 0.4, 0.3, or 0.2 degrees 2-theta of one or more peaks listed in Peak List #2 (see Example 28 for a description of the XRPD method).

[0197] Peak List #2

[0198]

[0199]

[0200] 1. In certain embodiments, Compound 1 Form B is characterized by an XRPD pattern having at least three peaks selected from 7.5, 8.8, 10.0, 10.5, 12.6, 14.7, 15.4, 16.4, 16.7, 18.6, 22.7, and 25.3 + / - 0.4° 2θ.

[0201] Other embodiments include but are not limited to the following:

[0202] 2. The crystalline form of embodiment 1, wherein Compound 1 Form B is characterized by an XRPD pattern having at least three peaks selected from 7.5, 8.8, 10.0, 10.5, 12.6, 14.7, 15.4, 16.4, 16.7, 18.6, 22.7, and 25.3 + / - 0.3° 2θ.

[0203] 3. The crystalline form of embodiment 1, wherein Compound 1 Form B is characterized by an XRPD pattern having at least three peaks selected from 7.5, 8.8, 10.0, 10.5, 12.6, 14.7, 15.4, 16.4, 16.7, 18.6, 22.7, and 25.3 + / - 0.2° 2θ.

[0204] 4. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least four of the listed peaks.

[0205] 5. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least five of the listed peaks.

[0206] 6. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least six of the listed peaks.

[0207] 7. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least seven of the listed peaks.

[0208] 8. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least eight of the listed peaks.

[0209] 9. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least nine of the listed peaks.

[0210] 10. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least ten of the listed peaks.

[0211] 11. The crystalline form of any one of embodiments 1-10, wherein the XRPD comprises a peak at 16.4 + / - 0.2° 2θ.

[0212] 12. The crystalline form of any one of embodiments 1-11, wherein the XRPD comprises a peak at 10.0 + / - 0.2° 2θ.

[0213] 13. The crystalline form of any one of embodiments 1-12, wherein the XRPD comprises a peak at 15.4 + / - 0.2° 2θ.

[0214] 14. The crystalline form of any one of embodiments 1-13, wherein the XRPD comprises a peak at 16.7 + / - 0.2° 2θ.

[0215] 15. The crystalline form of any one of embodiments 1-14, wherein the XRPD comprises a peak at 8.8 + / - 0.2° 2θ.

[0216] 16. The crystalline form of any one of embodiments 1-15, wherein the XRPD comprises a peak at 12.6 + / - 0.2° 2θ.

[0217] 17. The crystalline form of any one of embodiments 1-16, wherein the XRPD comprises a peak at 14.7 + / - 0.2° 2θ.

[0218] 18. The crystalline form of any one of embodiments 1-17, wherein the XRPD comprises a peak at 10.5 + / - 0.2° 2θ.

[0219] 19. The crystalline form of any one of embodiments 1-18, wherein the XRPD comprises a peak at 25.3 + / - 0.2° 2θ.

[0220] 20. The crystalline form of any one of embodiments 1-19, wherein the XRPD comprises a peak at 22.7 + / - 0.2° 2θ.

[0221] In certain embodiments, Compound 1 Form B is characterized by a melting onset temperature of about 194.4°C ± 10°C and / or a melting enthalpy of about 71.3 J / g ± 10 J / g on a DSC thermogram. In certain embodiments, Form B is characterized by a weight loss of about 1.1% at about 170.0°C ± 10°C as measured by TGA.

[0222] In certain embodiments, as described in Example 7, Compound 1 Form B is characterized by a solubility in DMSO of at least about 250 mg / mL when dissolved at about 25°C ± 5°C.

[0223] Figure 1 The XRPD pattern of Compound 1 Form B is described. Figure 2 Depicted is the DSC thermogram of Compound 1 Form B. Figure 3 The TGA thermogram of Compound 1 Form B is depicted.

[0224] Compound 1 Form C

[0225] Form C is an anhydrate. It is obtained from ethyl acetate and 2-methyltetrahydrofuran (2-MeTHF) by equilibration at 25°C, and from 2-MeTHF by temperature cycling. Form C has moderate crystallinity. DSC shows a broad endothermic peak starting at 25.9°C and a T of 185.3°C. onset It decomposes upon melting. TGA shows a weight loss of about 2.2% at about 180°C. 1 H NMR showed about 2.7 wt% ethyl acetate remaining (0.2 equivalent molar ratio). Form C is a metastable anhydrate. For example, after about 8 temperature cycles, it converts to anhydrous Form B.

[0226] In certain embodiments, Form C is characterized by an XRPD pattern having one or more peaks within + / - 0.4, 0.3, or 0.2 degrees 2-theta of the peaks listed in Peak List #3 (see Example 28 for a description of the XRPD method).

[0227] Peak List #3

[0228]

[0229] 1. In certain embodiments, Compound 1 Form C is characterized by an XRPD pattern having at least three peaks selected from 7.2, 13.4, 14.5, 14.9, 15.8, 16.5, 17.9, 18.4, 19.7, 20.6, 22.2, and 29.4 + / - 0.4° 2θ.

[0230] 2. The crystalline form of embodiment 1, wherein Compound 1 Form C is characterized by an XRPD pattern having at least three peaks selected from 7.2, 13.4, 14.5, 14.9, 15.8, 16.5, 17.9, 18.4, 19.7, 20.6, 22.2, and 29.4 + / - 0.3° 2θ.

[0231] 3. The crystalline form of embodiment 1, wherein Compound 1 Form C is characterized by an XRPD pattern having at least three peaks selected from 7.2, 13.4, 14.5, 14.9, 15.8, 16.5, 17.9, 18.4, 19.7, 20.6, 22.2, and 29.4 + / - 0.2° 2θ.

[0232] Other embodiments include but are not limited to the following:

[0233] 4. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least four of the listed peaks.

[0234] 5. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least five of the listed peaks.

[0235] 6. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least six of the listed peaks.

[0236] 7. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least seven of the listed peaks.

[0237] 8. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least eight of the listed peaks.

[0238] 9. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least nine of the listed peaks.

[0239] 10. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least ten of the listed peaks.

[0240] 11. The crystalline form of any one of embodiments 1-10, wherein the XRPD comprises a peak at 20.6 + / - 0.2° 2θ.

[0241] 12. The crystalline form of any one of embodiments 1-11, wherein the XRPD comprises a peak at 18.4 + / - 0.2° 2θ.

[0242] 13. The crystalline form of any one of embodiments 1-12, wherein the XRPD comprises a peak at 15.8 + / - 0.2° 2θ.

[0243] 14. The crystalline form of any one of embodiments 1-13, wherein the XRPD comprises a peak at 19.7 + / - 0.2° 2θ.

[0244] 15. The crystalline form of any one of embodiments 1-14, wherein the XRPD comprises a peak at 17.9 + / - 0.2° 2θ.

[0245] 16. The crystalline form of any one of embodiments 1-15, wherein the XRPD comprises a peak at 14.9 + / - 0.2° 2θ.

[0246] 17. The crystalline form of any one of embodiments 1-16, wherein the XRPD comprises a peak at 14.5 + / - 0.2° 2θ.

[0247] 18. The crystalline form of any one of embodiments 1-17, wherein the XRPD comprises a peak at 16.5 + / - 0.2° 2θ.

[0248] 19. The crystalline form of any one of embodiments 1-18, wherein the XRPD comprises a peak at 22.2 + / - 0.2° 2θ.

[0249] 20. The crystalline form of any one of embodiments 1-19, wherein the XRPD comprises a peak at 13.4 + / - 0.2° 2θ.

[0250] In certain embodiments, Form C is characterized by a melting onset temperature of about 185.3°C ± 10°C and / or a melting enthalpy of about 33.9 J / g ± 10 J / g on a DSC thermogram. In certain embodiments, Form C is characterized by a weight loss of about 2.2% at about 180.0°C ± 10°C as measured by TGA.

[0251] Figure 10 The XRPD pattern of Compound 1 Form C is described. Figure 11 Depicted is the DSC thermogram of Compound 1 Form C. Figure 12 The TGA thermogram of Compound 1 Form C is depicted.

[0252] Compound 1 salt crystal form

[0253] The formation of crystalline salt forms of Compound 1 was studied under various crystallization conditions. The conditions tested included slurry equilibration, precipitation by addition of antisolvent, and reslurry.

[0254] Five salt forms of compound 1 were identified: sodium salt form D, sodium salt form E, sodium salt form F, potassium salt form G, and potassium salt form H. Among these salt forms DH, sodium salt form F exhibited good physicochemical properties, including good crystallinity, reasonable stoichiometry, high dehydration initiation, and good counterion safety.

[0255] Compound 1 sodium salt form F

[0256] Compound 1 sodium salt form F is a hydrate. Sodium salt form F has medium crystallinity. DSC shows dehydration peaks at 13.7°C, 64.7°C, 116.5°C and T at 194.7°C. onset The melting point of the product was 1.547 nm. TGA showed a weight loss of approximately 4.8% at approximately 180°C. HPLC indicated a chemical purity of 98.9%. IC and HPLC indicated a stoichiometric ratio of 1:1.2 between the form and the sodium salt. 1 HNMR showed no detectable residual solvent. KF (Karl Fischer analysis) showed that it contained about 4.6% by weight of water, equivalent to 2.5 water molecules. The hydrated form was stable under vacuum drying at about 30°C.

[0257] Sodium salt Form F showed a decrease in chemical purity of approximately 1% after photostability evaluation. Form F showed a slight decrease in chemical purity (approximately 1.2%) after stress at 25°C / 92% RH. Form F also showed a slight decrease in crystallinity after stress at 60°C in a closed container for one week, likely due to partial dehydration.

[0258] The hygroscopicity of sodium salt Form F was evaluated by dynamic vapor sorption (DVS) testing at 25°C. Sodium salt Form F was slightly hygroscopic below 80% RH. It then became hygroscopic and showed a water absorption of 12.6% from 80% RH to 95% RH at 25°C. After the DVS test, sodium salt Form F showed no change in crystalline form and no decrease in crystallinity.

[0259] In certain embodiments, Form F is characterized by an XRPD pattern having one or more peaks within + / - 0.4, 0.3, or 0.2° 2Θ of the peaks listed in Peak List #4 (see Example 28 for a description of the XRPD method).

[0260] Peak List #4

[0261]

[0262]

[0263] 1. In certain embodiments, Compound 1 sodium salt Form F is characterized by an XRPD pattern having at least three peaks selected from 13.6, 15.0, 15.5, 16.0, 17.5, 18.3, 19.7, 20.3, 21.7, 24.0, 24.5, 30.3, 30.4, and 34.5 + / - 0.4° 2θ.

[0264] 2. The crystalline form of embodiment 1, wherein Compound 1 sodium salt Form F is characterized by an XRPD pattern having at least three peaks selected from 13.6, 15.0, 15.5, 16.0, 17.5, 18.3, 19.7, 20.3, 21.7, 24.0, 24.5, 30.3, 30.4, and 34.5 + / - 0.3° 2θ.

[0265] 3. The crystalline form of embodiment 1, wherein Compound 1 sodium salt Form F is characterized by an XRPD pattern having at least three peaks selected from 13.6, 15.0, 15.5, 16.0, 17.5, 18.3, 19.7, 20.3, 21.7, 24.0, 24.5, 30.3, 30.4 and 34.5 + / - 0.2° 2θ.

[0266] Other embodiments include but are not limited to the following:

[0267] 4. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least four of the listed peaks.

[0268] 5. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least five of the listed peaks.

[0269] 6. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least six of the listed peaks.

[0270] 7. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least seven of the listed peaks.

[0271] 8. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least eight of the listed peaks.

[0272] 9. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least nine of the listed peaks.

[0273] 10. The crystalline form according to any one of embodiments 1 to 3, which exhibits at least ten of the listed peaks.

[0274] 11. The crystalline form of any one of embodiments 1-10, wherein the XRPD comprises a peak at 19.7 + / - 0.2° 2θ.

[0275] 12. The crystalline form of any one of embodiments 1-11, wherein the XRPD comprises a peak at 20.3 + / - 0.2° 2θ.

[0276] 13. The crystalline form of any one of embodiments 1-12, wherein the XRPD comprises a peak at 15.0 + / - 0.2° 2θ.

[0277] 14. The crystalline form of any one of embodiments 1-13, wherein the XRPD comprises a peak at 21.7 + / - 0.2° 2θ.

[0278] 15. The crystalline form of any one of embodiments 1-14, wherein the XRPD comprises a peak at 30.4 + / - 0.2° 2θ.

[0279] 16. The crystalline form of any one of embodiments 1-15, wherein the XRPD comprises a peak at 13.6 + / - 0.2° 2θ.

[0280] 17. The crystalline form of any one of embodiments 1-16, wherein the XRPD comprises a peak at 24.0 + / - 0.2° 2θ.

[0281] 18. The crystalline form of any one of embodiments 1-17, wherein the XRPD comprises a peak at 18.3 + / - 0.2° 2θ.

[0282] 19. The crystalline form of any one of embodiments 1-18, wherein the XRPD comprises a peak at 24.5 + / - 0.2° 2θ.

[0283] 20. The crystalline form of any one of embodiments 1-19, wherein the XRPD comprises a peak at 34.5 + / - 0.2° 2θ.

[0284] In certain embodiments, Form F is characterized by any one of the following peaks: 59.8°C ± 10°C, 132.5°C ± 10°C, and 197.7°C ± 10°C, and / or a melting enthalpy of about 26.4 J / g ± 10 J / g and 41.8 J / g ± 10 J / g (as measured by DSC). In certain embodiments, Form F is characterized by a weight loss of about 2.7% at about 100.0°C ± 10°C, and a weight loss of about 3.2% over a temperature range of about 100.0°C to 170.0°C ± 10°C, as measured by TGA.

[0285] Figure 17 The XRPD pattern of Compound 1 sodium salt Form F is described. Figure 18 The DSC thermogram of Compound 1 sodium salt Form F is described. Figure 19 The TGA thermogram of Compound 1 sodium salt Form F is described.

[0286] Compound 1 sodium salt form D

[0287] Compound 1 sodium salt form D is a hydrate with medium crystallinity. DSC shows a dehydration peak at 4.5°C, a peak at 67.5°C, an endothermic peak at 117.9°C, and a T peak at 229.5°C. onset The melting point of the compound was 1.5477 nm. TGA showed a weight loss of about 6.9% at about 150°C. HPLC showed a chemical purity of 97.4%. IC and HPLC showed that the compound was + The stoichiometric ratio of the salts was 1:1. 1 HNMR showed no detectable residual solvent.

[0288] In certain embodiments, Form D is characterized by an XRPD pattern having one or more peaks within + / - 0.4, 0.3, or 0.2 degrees 2-theta of the peaks listed in Peak List #5 (see Example 28 for a description of the XRPD method).

[0289] Peak List #5

[0290]

[0291] 1. In certain embodiments, Compound 1 sodium salt Form D is characterized by an XRPD pattern having at least three peaks selected from 6.2, 7.7, 11.7, 12.5, 13.6, 15.3, 15.7, 17.5, 20.8, and 27.5 + / - 0.4° 2θ.

[0292] 2. The crystalline form of embodiment 1, wherein Compound 1 sodium salt Form D is characterized by an XRPD pattern having at least three peaks selected from 6.2, 7.7, 11.7, 12.5, 13.6, 15.3, 15.7, 17.5, 20.8, and 27.5 + / - 0.3° 2θ.

[0293] 3. The crystalline form of embodiment 1, wherein Compound 1 sodium salt Form D is characterized by an XRPD pattern having at least three peaks selected from 6.2, 7.7, 11.7, 12.5, 13.6, 15.3, 15.7, 17.5, 20.8, and 27.5 + / - 0.2° 2θ.

[0294] Figure 23 The XRPD pattern of Compound 1 sodium salt Form D is described. Figure 24 The DSC thermogram of Compound 1 sodium salt Form D is described. Figure 25 The TGA thermogram of Compound 1 sodium salt Form D is described.

[0295] Compound 1 sodium salt form E

[0296] Compound 1 crystalline sodium salt E is a hydrate. The sodium salt crystalline E has medium crystallinity. DSC shows a dehydration peak at 5.6 ° C, a peak at 61.1 ° C and a T of 228.7 ° C.onset The melting point of the compound was 10.8%. TGA showed a weight loss of about 10.8% at about 180°C. HPLC showed a chemical purity of about 98%. IC and HPLC showed that the compound was + The stoichiometric ratio of the salts was 1:1. 1 H NMR showed no detectable residual solvent. According to KF analysis, the water content was 10.2 wt%.

[0297] In certain embodiments, Form E is characterized by an XRPD pattern having one or more peaks within + / - 0.4, 0.3, or 0.2 degrees 2-theta of the peaks listed in Peak List #6 (see Example 28 for a description of the XRPD method).

[0298] Peak List #6

[0299]

[0300] 1. In certain embodiments, Compound 1 sodium salt Form E is characterized by an XRPD pattern having at least three peaks selected from 5.8, 6.3, 7.8, 12.5, 13.9, 15.7, 17.8, 20.2, and 25.3 + / - 0.4° 2θ.

[0301] 2. The crystalline form of embodiment 1, wherein Compound 1 sodium salt Form E is characterized by an XRPD pattern having at least three peaks selected from 5.8, 6.3, 7.8, 12.5, 13.9, 15.7, 17.8, 20.2, and 25.3 + / - 0.3° 2θ.

[0302] 3. The crystalline form of embodiment 1, wherein Compound 1 sodium salt Form E is characterized by an XRPD pattern having at least three peaks selected from 5.8, 6.3, 7.8, 12.5, 13.9, 15.7, 17.8, 20.2, and 25.3 + / - 0.2° 2θ.

[0303] Figure 26 The XRPD pattern of Compound 1 sodium salt Form E is described. Figure 27 The DSC thermogram of Compound 1 sodium salt Form E is described. Figure 28 The TGA thermogram of Compound 1 sodium salt Form E is described.

[0304] Compound 1 potassium salt form G

[0305] Compound 1 potassium salt form G is a solvate. Potassium salt form G has low crystallinity. DSC shows multiple thermal events ( Figure 30). TGA showed a weight loss of about 4.5% at about 170°C and a weight loss of about 4.8% in the temperature range of about 170°C to 230°C. HPLC showed a chemical purity of about 99%. IC and HPLC showed that the compound was + The stoichiometric ratio of the salts was 1:1. 1 H NMR showed 2.5 wt% acetone residual solvent (0.42 equivalents molar ratio).

[0306] Figure 29 The XRPD pattern of Compound 1 potassium salt Form G is described. Figure 30 The DSC thermogram of Compound 1 potassium salt Form G is described. Figure 31 The TGA thermogram of Compound 1 potassium salt Form G is described.

[0307] Compound 1 potassium salt form H

[0308] Compound 1 potassium salt crystal form H is a hydrate. Potassium salt crystal form H has medium crystallinity. DSC shows a dehydration peak at 4.2°C, a peak at 87.2°C, an endothermic peak at 123.0°C, and a T of 221.7°C. onset The melting point of the compound was 1.5477 W / m. TGA showed a weight loss of about 6.6% at about 170°C. HPLC showed a chemical purity of about 99%. IC and HPLC showed that the compound was + The stoichiometric ratio of the salts was 1:1.2. 1 H NMR showed no detectable residual solvent.

[0309] Figure 32 The XRPD pattern of Compound 1 potassium salt Form H is described. Figure 33 The DSC thermogram of Compound 1 potassium salt Form H is depicted. Figure 34 The TGA thermogram of Compound 1 potassium salt Form H is depicted.

[0310] Other implementation plans

[0311] 1. The isolated crystalline form B of the compound having the following structure:

[0312]

[0313] Characterized in that, it comprises an X-ray powder diffraction (XRPD) pattern comprising at least five 2θ values ​​selected from 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2° and 25.3±0.2°.

[0314] 2. The isolated crystalline Form B according to embodiment 1, wherein the XRPD pattern comprises at least six 2θ values ​​selected from 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

[0315] 3. The isolated crystalline Form B according to embodiment 1, wherein the XRPD pattern comprises at least seven 2θ values ​​of 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

[0316] 4. The isolated crystalline Form B according to embodiment 1, wherein the XRPD pattern comprises at least eight 2θ values ​​selected from the group consisting of 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

[0317] 5. The isolated crystalline Form B according to embodiment 1, wherein the XRPD pattern comprises at least nine 2θ values ​​selected from the group consisting of 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

[0318] 6. The isolated crystalline Form B according to embodiment 1, wherein the XRPD pattern comprises at least ten 2θ values ​​selected from the group consisting of 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

[0319] 7. The isolated crystalline Form B according to embodiment 1, wherein the XRPD pattern comprises at least eleven 2θ values ​​selected from the group consisting of 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

[0320] 8. The isolated crystalline Form B according to any one of embodiments 1-7, wherein the XRPD pattern comprises at least three 2θ values ​​selected from 7.5±0.1°, 8.8±0.1°, 10.0±0.1°, 10.5±0.1°, 12.6±0.1°, 14.7±0.1°, 15.4±0.1°, 16.4±0.1°, 16.7±0.1°, 18.6±0.1°, 22.7±0.1°, and 25.3±0.1°.

[0321] 9. The isolated crystalline Form B according to any one of embodiments 1-8, wherein the XRPD pattern comprises a 2θ value of at least 16.4±0.2°.

[0322] 10. The isolated crystalline Form B according to any one of embodiments 1-9, wherein the XRPD pattern comprises a 2θ value of at least 10.0±0.2°.

[0323] 11. The isolated crystalline Form B according to any one of embodiments 1-10, wherein the XRPD pattern comprises a 2θ value of at least 15.4±0.2°.

[0324] 12. The isolated crystalline Form B according to any one of embodiments 1-11, wherein the XRPD pattern comprises a 2θ value of at least 16.7±0.2°.

[0325] 13. The isolated crystalline Form B according to any one of embodiments 1-12, wherein the XRPD pattern comprises a 2θ value of at least 8.8±0.2°.

[0326] 14. The isolated crystalline Form B according to any one of embodiments 1-13, wherein the XRPD pattern comprises a 2θ value of at least 12.6±0.2°.

[0327] 15. The isolated crystalline Form B according to any one of embodiments 1-14, wherein the XRPD pattern comprises a 2θ value of at least 14.7±0.2°.

[0328] 16. The isolated crystalline Form B according to any one of embodiments 1-15, wherein the XRPD pattern comprises a 2θ value of at least 10.5±0.2°.

[0329] 17. The isolated crystalline Form B according to any one of embodiments 1-16, wherein the XRPD pattern comprises a 2θ value of at least 25.3±0.2°.

[0330] 18. The isolated crystalline Form B according to any one of embodiments 1-17, wherein the XRPD pattern comprises a 2θ value of at least 22.7±0.2°.

[0331] 19. The isolated crystalline form B according to any one of embodiments 1-18, characterized in that Figure 1 XRPD pattern of characteristic 2θ values.

[0332] 20. The isolated crystalline Form B according to any one of embodiments 1-19, having a differential scanning calorimetry (DSC) onset endotherm of about 194±20°C.

[0333] 21. The isolated crystalline Form B according to any one of embodiments 1-20, having a differential scanning calorimetry (DSC) onset endotherm of about 194±10°C.

[0334] 22. A pharmaceutical composition comprising the isolated crystalline Form B of any one of embodiments 1-21 in a pharmaceutically acceptable carrier.

[0335] 23. A pharmaceutical composition comprising Compound 1, a solubility enhancer, a permeation enhancer, a filler, a binder / glidant, and a mucoadhesive / disintegrant, wherein Compound 1 has the following structure:

[0336]

[0337] or a pharmaceutically acceptable salt thereof.

[0338] 24. The pharmaceutical composition according to embodiment 23, wherein:

[0339] a. The solubility enhancer is hypromellose acetate succinate;

[0340] b. The penetration enhancer is vitamin E;

[0341] c. The filler is mannitol;

[0342] d. The binder / glidant is cellulose; and

[0343] e. The mucoadhesive is cross-linked sodium carboxymethyl cellulose.

[0344] 25. A method of treating a mutant BRAF-mediated disorder, comprising administering to a patient in need thereof an effective amount of the isolated crystalline Form B of any one of embodiments 1-21 or the pharmaceutical composition of any one of embodiments 22-24.

[0345] 26. The method according to embodiment 25, wherein the patient is a human.

[0346] 27. The method according to embodiment 25 or 26, wherein the mutant BRAF-mediated disorder is cancer.

[0347] 28. The method according to embodiment 27, wherein the mutant BRAF-mediated cancer is melanoma.

[0348] 29. The method according to embodiment 27, wherein the mutant BRAF-mediated cancer is lung cancer.

[0349] 30. The method according to embodiment 27, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

[0350] 31. The method according to embodiment 27, wherein the mutant BRAF-mediated cancer is colorectal cancer.

[0351] 32. The method according to embodiment 27, wherein the mutant BRAF-mediated cancer is microsatellite stable colorectal cancer.

[0352] 33. The method of embodiment 27, wherein the mutant BRAF-mediated cancer is thyroid cancer.

[0353] 34. The method of embodiment 27, wherein the mutant BRAF-mediated cancer is ovarian cancer.

[0354] 35. The method of embodiment 25, wherein the mutant BRAF-mediated disorder is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans' histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoastrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid carcinoma, anaplastic thyroid carcinoma, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

[0355] 36. The method according to any one of embodiments 25-35, wherein the patient also receives an additional active agent.

[0356] 37. The method according to embodiment 36, wherein the additional active agent is a MEK inhibitor.

[0357] 38. The method according to embodiment 37, wherein the MEK inhibitor is trametinib.

[0358] 39. The method of embodiment 36, wherein the additional active agent is an immune checkpoint inhibitor.

[0359] 40. The method according to embodiment 39, wherein the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, cemiplimab, ipilimumab, relalizumab, atezolizumab, avelumab, and durvalumab.

[0360] 41. The method according to embodiment 36, wherein the additional active agent is cetuximab or panitumumab.

[0361] 42. The isolated crystalline form B according to any one of embodiments 1-21 or the pharmaceutical composition according to any one of embodiments 22-24 for use in the therapeutic treatment of a mutant BRAF-mediated disorder.

[0362] 43. The isolated crystalline Form B according to embodiment 42, wherein the mutant BRAF-mediated disorder is cancer.

[0363] 44. The isolated crystalline Form B according to embodiment 43, wherein the mutant BRAF-mediated cancer is melanoma.

[0364] 45. The isolated crystalline Form B according to embodiment 43, wherein the mutant BRAF-mediated cancer is lung cancer.

[0365] 46. ​​The isolated crystalline Form B according to embodiment 43, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

[0366] 47. The isolated crystalline Form B according to embodiment 43, wherein the mutant BRAF-mediated cancer is colorectal cancer.

[0367] 48. The isolated crystalline Form B according to embodiment 43, wherein the mutant BRAF-mediated cancer is microsatellite stable colorectal cancer.

[0368] 49. The isolated crystalline Form B according to embodiment 43, wherein the mutant BRAF-mediated cancer is thyroid cancer.

[0369] 50. The isolated crystalline Form B according to embodiment 43, wherein the mutant BRAF-mediated cancer is ovarian cancer.

[0370] 51. The isolated crystalline Form B according to embodiment 42, wherein the mutant BRAF-mediated disorder is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoastrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid carcinoma, anaplastic thyroid carcinoma, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

[0371] 52. The isolated crystalline form B according to any one of embodiments 1-21 or the pharmaceutical composition according to any one of embodiments 22-24, for use in the treatment of a mutant BRAF-mediated disorder.

[0372] 53. The isolated crystalline Form B according to embodiment 52, wherein the mutant BRAF-mediated disorder is cancer.

[0373] 54. The isolated crystalline Form B according to embodiment 53, wherein the mutant BRAF-mediated cancer is melanoma.

[0374] 55. The isolated crystalline Form B according to embodiment 53, wherein the mutant BRAF-mediated cancer is lung cancer.

[0375] 56. The isolated crystalline Form B according to embodiment 53, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

[0376] 57. The isolated crystalline Form B according to embodiment 53, wherein the mutant BRAF-mediated cancer is colorectal cancer.

[0377] 58. The isolated crystalline Form B according to embodiment 53, wherein the mutant BRAF-mediated cancer is microsatellite stable colorectal cancer.

[0378] 59. The isolated crystalline Form B according to embodiment 53, wherein the mutant BRAF-mediated cancer is thyroid cancer.

[0379] 60. The isolated crystalline Form B according to embodiment 53, wherein the mutant BRAF-mediated cancer is ovarian cancer.

[0380] 61. The isolated crystalline Form B according to embodiment 52, wherein the mutant BRAF-mediated disorder is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoastrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid carcinoma, anaplastic thyroid carcinoma, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

[0381] 62. Use of the isolated crystalline Form B according to any one of embodiments 1-21 or the pharmaceutical composition according to any one of embodiments 22-24 for the preparation of a medicament for treating a mutant BRAF-mediated disorder.

[0382] 63. The use according to embodiment 62, wherein the mutant BRAF-mediated disorder is cancer.

[0383] 64. The use according to embodiment 63, wherein the mutant BRAF-mediated cancer is melanoma.

[0384] 65. The use according to embodiment 63, wherein the variant BRAF-mediated cancer is lung cancer.

[0385] 66. The use according to embodiment 63, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

[0386] 67. The use according to embodiment 63, wherein the mutant BRAF-mediated cancer is colorectal cancer.

[0387] 68. The use according to embodiment 63, wherein the mutant BRAF-mediated cancer is microsatellite stable colorectal cancer.

[0388] 69. The use according to embodiment 63, wherein the mutant BRAF-mediated cancer is thyroid cancer.

[0389] 70. The use according to embodiment 63, wherein the mutant BRAF-mediated cancer is ovarian cancer.

[0390] 71. The use according to embodiment 62, wherein the mutant BRAF-mediated disorder is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans' histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoastrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid cancer, anaplastic thyroid cancer, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

[0391] 72. The isolated crystalline form B according to any one of embodiments 1-21 for use as a therapeutically active substance.

[0392] 73. A method for preparing a compound of the formula:

[0393]

[0394] It involves the transaminase-catalyzed reductive amination of a compound of the formula:

[0395]

[0396] To obtain a compound of formula A-1;

[0397] where R X is selected from hydrogen and an amino protecting group.

[0398] 74. The method according to embodiment 73, wherein:

[0399] Amino protecting group R X is tert-butoxycarbonyl (BOC);

[0400] The first base is sodium hydroxide or potassium hydroxide; and

[0401] The isopropylamine salt is isopropylamine hydrochloride.

[0402] 75. A method for preparing a compound of the formula:

[0403]

[0404] The method comprises the following steps:

[0405] (i) making the following compound:

[0406]

[0407] reacting with a chiral acid to obtain a mixture of diastereomeric salts formed between the compound of formula A-3 and the chiral acid;

[0408] (ii) crystallizing the mixture of diastereomeric salts prepared in step (i) from a first solvent to provide the individually separated diastereomeric salts formed between the anion of the chiral acid and the cation of the N-protected (R)-1-oxa-8-azaspiro[4.5]decan-3-ammonium of the formula:

[0409] and

[0410] (iii) treating the individually isolated diastereomeric salt containing a cation of formula A-4 isolated in step (ii) with a second base to liberate the compound of formula A-1;

[0411] where R X is selected from hydrogen and an amino protecting group.

[0412] 76. The method according to embodiment 75, wherein:

[0413] Amino protecting group R X is tert-butoxycarbonyl (BOC);

[0414] The chiral acid is (R)-mandelic acid;

[0415] The first solvent is acetonitrile; and

[0416] The second base is sodium hydroxide or potassium hydroxide.

[0417] 77. A method for preparing the compound of the formula:

[0418]

[0419] The method comprises reacting 2-amino-5-hydroxybenzoic acid with a compound of the formula:

[0420]

[0421] wherein the compound of formula A-1 is prepared in a method according to any one of embodiments 73-76; and

[0422] R X is selected from hydrogen and an amino protecting group.

[0423] 78. The method according to embodiment 77, wherein:

[0424] Amino protecting group R X is tert-butoxycarbonyl (BOC);

[0425] the trialkyl orthoformate is triethyl orthoformate; and

[0426] The second solvent is n-butanol.

[0427] 79. A method for preparing a compound of the formula:

[0428]

[0429] It comprises reacting 2,3,6-trifluorobenzonitrile with a compound of the formula:

[0430]

[0431] in

[0432] R X is selected from hydrogen and an amino protecting group; and

[0433] The compound of formula A-5 is prepared in a process according to any one of embodiments 77-78.

[0434] 80. The method according to embodiment 79, wherein:

[0435] The third base is potassium carbonate, and the third solvent is acetonitrile; and

[0436] Amino protecting group R X It is tert-butoxycarbonyl (BOC).

[0437] 81. A method for preparing a compound of the formula:

[0438]

[0439] It includes the following steps:

[0440] (i) reacting [methyl(sulfamoyl)amino]ethane NH2SO2N(Me)Et with a compound of the formula:

[0441]

[0442] To obtain a compound of formula A-8:

[0443]

[0444] in:

[0445] The compound of formula A-6 is prepared in the method of any one of embodiments 79-80;

[0446] R X is selected from hydrogen and an amino protecting group; and

[0447] (ii) When R X When it is an amino protecting group, the amino protecting group connected to the piperidine nitrogen atom of the compound of formula A-8 is removed in the presence of the first acid.

[0448] 82. The method according to embodiment 81, wherein:

[0449] The fourth base is cesium carbonate, and the fourth solvent is N,N-dimethylacetamide;

[0450] Amino protecting group R X is tert-butoxycarbonyl (BOC); and

[0451] The first acid is hydrochloric acid.

[0452] 83. A method for preparing a compound of the formula:

[0453]

[0454] It comprises reacting 2,4,5-trifluorobenzonitrile with a compound of the formula:

[0455]

[0456] where R Y is hydrogen or a carboxylic acid protecting group.

[0457] 84. The method according to embodiment 83, wherein:

[0458] The fifth base is N,N-diisopropylethylamine, and the fifth solvent is N,N-dimethylformamide; and

[0459] R Y It is tert-butyl (tBu).

[0460] 85. A method for preparing a compound of the formula:

[0461]

[0462] It comprises reacting methylhydrazine with a compound of the formula:

[0463]

[0464] wherein the compound of formula B-1 is prepared in a method according to any one of embodiments 83-84;

[0465] where R Y is hydrogen or a carboxylic acid protecting group.

[0466] 86. The method according to embodiment 85, wherein:

[0467] The sixth solvent is N-methyl-2-pyrrolidone; and

[0468] R Y It is tert-butyl.

[0469] 87. A method for preparing a compound of the formula:

[0470]

[0471] It comprises reacting acrylic acid with a compound of the formula:

[0472]

[0473] in:

[0474] The compound of formula B-3 is prepared in a method according to any one of embodiments 85-86; and

[0475] R Y is hydrogen or a carboxylic acid protecting group.

[0476] 88. The method according to embodiment 87, wherein:

[0477] The seventh solvent is aqueous hydrochloric acid; and

[0478] R Y It is tert-butyl.

[0479] 89. A method for preparing a compound of the formula:

[0480]

[0481] It includes the following steps:

[0482] (i) First, a metal cyanate is reacted with a compound of the formula:

[0483]

[0484] To obtain a reaction mixture containing a first intermediate compound of the following formula:

[0485]

[0486] in:

[0487] The compound of formula B-4 is prepared in a method according to any one of embodiments 87-88; and

[0488] R Y is hydrogen or a carboxylic acid protecting group;

[0489] (ii) the first intermediate compound of formula B-6 from step (i) is then cyclized, optionally in the presence of a third acid, to provide a second intermediate compound of formula:

[0490] and

[0491] (iii) Then, when R Y When it is a carboxylic acid protecting group, the carboxylic acid protecting group R is removed from the second intermediate compound of formula B-7 from step (ii) in the presence of a fourth acid. Y , to obtain the compound of formula B-5.

[0492] 90. The method according to embodiment 89, wherein

[0493] R Y is tert-butyl;

[0494] The metal cyanate is sodium cyanate or potassium cyanate;

[0495] The second acid is acetic acid;

[0496] The third acid is hydrochloric acid or acetic acid; and

[0497] The fourth acid is hydrochloric acid.

[0498] 91. A method for preparing a compound of the formula:

[0499]

[0500] The method comprises the following steps:

[0501] (i) First, in the presence of a sixth base and an eighth solvent, a compound of the formula:

[0502]

[0503] React with a carboxylic acid hydroxyl activating reagent to obtain a third intermediate compound of the following formula:

[0504]

[0505] The carboxylic acid hydroxyl group is converted into a more reactive group -OR Z ;

[0506] wherein the compound of formula B-5 is prepared in a method according to any one of embodiments 89-90; and

[0507] (ii) then reacting (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluoro-phenoxy]-4-oxo-quinazolin-3-yl]-1-oxa-8-azaspiro[4.5]decane of the formula:

[0508]

[0509] reacting with the third intermediate compound of formula B-8 prepared in step (ii) to obtain a compound of formula 1,

[0510] wherein the compound of formula A-7 is prepared in a process according to any one of embodiments 81-82.

[0511] 92. The method according to embodiment 91, wherein

[0512] (i) the sixth base is N,N-diisopropylethylamine;

[0513] The eighth solvent is acetonitrile;

[0514] The carboxylic acid hydroxyl activation reagent is 2-succinimidyl-1,1,3,3-tetramethyluronium tetrafluoroborate, and R Z The group is 2,5-dioxopyrrolidin-1-yl; and

[0515] (ii) The seventh base is triethylamine, and the ninth solvent is N,N-dimethylformamide.

[0516] 93. A method for treating brain or central nervous system (CNS) metastasis of mutant BRAF-mediated cancer comprising administering to a human patient in need thereof an effective amount of a compound of the structure

[0517]

[0518] or a pharmaceutically acceptable salt thereof.

[0519] 94. The method according to embodiment 93, wherein the cancer has metastasized to the brain.

[0520] 95. The method according to embodiment 93 or 94, wherein the cancer has metastasized to the CNS.

[0521] 96. The method according to any one of embodiments 93-95, wherein the mutant BRAF-mediated cancer is melanoma.

[0522] 97. The method according to any one of embodiments 93-95, wherein the mutant BRAF-mediated cancer is lung cancer.

[0523] 98. The method according to any one of embodiments 93-95, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

[0524] 99. The method according to any one of embodiments 93-95, wherein the mutant BRAF-mediated cancer is colorectal cancer.

[0525] 100. The method according to any one of embodiments 93-95, wherein the mutant BRAF-mediated cancer is microsatellite stable colorectal cancer.

[0526] 101. The method according to any one of embodiments 93-95, wherein the mutant BRAF-mediated cancer is thyroid cancer.

[0527] 102. The method according to any one of embodiments 93-95, wherein the mutant BRAF-mediated cancer is ovarian cancer.

[0528] 103. The method of any one of embodiments 93-95, wherein the mutant BRAF-mediated cancer is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoastrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid cancer, anaplastic thyroid cancer, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

[0529] 104. The method according to any one of embodiments 93-103, wherein the patient also receives an additional active agent.

[0530] 105. The method according to embodiment 104, wherein the additional active agent is a MEK inhibitor.

[0531] 106. The method according to embodiment 105, wherein the MEK inhibitor is trametinib.

[0532] 107. The method of embodiment 104, wherein the additional active agent is an immune checkpoint inhibitor.

[0533] 108. The method according to embodiment 107, wherein the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, cemiplimab, ipilimumab, relalizumab, atezolizumab, avelumab, and durvalumab.

[0534] 109. The method according to embodiment 104, wherein the additional active agent is cetuximab or panitumumab.

[0535] 110. The method according to any one of embodiments 93-109, wherein the mutant BRAF has a V600 mutation.

[0536] 111. The method according to embodiment 110, wherein the V600 mutation is V600E, V600K, V600R, V600D, V600M or V600N.

[0537] 112. The method according to embodiment 110, wherein the V600 mutation is V600E.

[0538] 113. The method according to any one of embodiments 93-112, wherein the mutant BRAF has a mutation selected from the group consisting of: G469A, G469V, G469L, G469R, L597Q and K601E.

[0539] 114. The method according to any one of embodiments 93-113, wherein the mutant BRAF has a mutation selected from the group consisting of: G466A, G466E, G466R, G466V, S467L, G469E, N581I, D594E, D594G and D594N.

[0540] 115. The method according to any one of embodiments 93-114, wherein the mutant BRAF has a mutation selected from the group consisting of: G464I, G464R, N581T, L584F, E586K, G593D, G596C, L597R, L597S, S605I, S607F, N684T, E26A, V130M, L745L and D284E.

[0541] 116. The method according to any one of embodiments 93-115, wherein the mutant BRAF has a splicing mutation.

[0542] 117. The method according to embodiment 116, wherein the splicing mutation is a p61-BRAF V600 splicing mutation.

[0543] 118. The method according to any one of embodiments 93-117, wherein the mutant BRAF-mediated cancer has a NRAS, MEK1 or PI3K mutation.

[0544] 119. The method of embodiment 118, wherein the cancer has a NRAS mutation.

[0545] 120. The method of embodiment 119, wherein the NRAS mutation is Q61K.

[0546] 121. The method of embodiment 119, wherein the NRAS mutation is Q61R.

[0547] 122. The method of embodiment 118, wherein the cancer has a MEK1 mutation.

[0548] 123. The method of embodiment 118, wherein the cancer has a PI3K mutation.

[0549] 124. The method according to embodiment 123, wherein the PI3K mutation is PIK3CA H1047R or PIK3CA P449T .

[0550] 125. The method according to any one of embodiments 93-124, wherein the mutant BRAF-mediated cancer has one or more mutations that promote homodimerization or heterodimerization of the RAF protein.

[0551] 126. The method of embodiment 125, wherein the one or more mutations promote RAF protein homodimerization.

[0552] 127. The method of embodiment 125, wherein the one or more mutations promote RAF protein heterodimerization.

[0553] Treatment of BRAF-mediated disorders

[0554] Compound 1 is a small molecule that degrades mutant BRAF (e.g., class I, class II, and / or class III mutant BRAF) through the ubiquitin proteasome pathway. Compound 1 binds to the ubiquitously expressed E3 ligase protein cereblon (CRBN) and alters the substrate specificity of the CRBN E3 ubiquitin ligase complex, leading to the recruitment and ubiquitination of mutant BRAF (e.g., BRAF V600E). Compound 1 effectively degrades class I mutant BRAF such as V600E, class II mutant BRAF such as G469A, and class III mutant BRAF such as G466V mutation.

[0555] Therefore, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of Compound 1 of the present invention can be used to treat mutant BRAF-mediated cancers, such as melanoma, lung cancer including, for example, non-small cell lung cancer, colorectal cancer including, for example, microsatellite stable colorectal cancer, thyroid cancer including, for example, anaplastic thyroid cancer, or ovarian cancer. In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form thereof is used to treat solid tumors mediated by V600X mutant BRAF. Non-limiting examples of conditions that can be treated with a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form thereof include melanoma, non-small cell lung cancer, thyroid cancer, colorectal cancer, and other solid tumor malignancies with mutant BRAF drivers.

[0556] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form thereof, can treat cancers that have become resistant to BRAF inhibitors. For example, Compound 1 is effective in treating G466V mutant BRAF lung tumor cell lines. In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a crystalline form thereof, is orally bioavailable.

[0557] In certain aspects, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form thereof is used to treat BRAF-mediated cancer, wherein BRAF is mutated from wild type. In certain non-limiting embodiments, the mutation is a Class I mutation, a Class II mutation, or a Class III mutation, or any combination thereof. Non-limiting examples of Class I mutations include V600 mutations, such as V600E, V600K, V600R, V600D, V600M, and V600N. Non-limiting examples of Class II mutations include G469A, G469V, G469L, G469R, L597Q, and K601E. Non-limiting examples of Class III mutations include G466A, G466E, G466R, G466V, S467L, G469E, N581I, D594E, D594G, and D594N.

[0558] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof treats a condition mediated by a BRAF mutant, wherein the mutation is not a Class I, Class II, or Class III mutation. Non-limiting examples of mutations include G464I, G464R, N581T, L584F, E586K, G593D, G596C, L597R, L597S, S605I, S607F, N684T, E26A, V130M, L745L, and D284E.

[0559] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof treats a condition mediated by a BRAF mutant, wherein the mutation is a splice variant, such as p61-BRAF V600E .

[0560] Compound 1 or a pharmaceutical composition thereof, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof, or a pharmaceutically acceptable salt of Compound 1 or a pharmaceutical composition thereof can be used to treat patients suffering from any disease mediated by mutant BRAF.

[0561] BRAF is a serine / threonine protein kinase, a member of the signal transduction protein kinase family. BRAF V600X mutations, particularly BRAF V600E / K mutations, are commonly observed in a variety of human tumors, including melanoma, thyroid cancer, colorectal cancer, and lung cancer. Non-limiting examples of V600X mutations include V600E, V600K, V600R, V600D, and V600N. Although clinically available BRAF inhibitors have demonstrated therapeutic efficacy in many of these indications, the duration of antitumor responses to these drugs is limited by the development of drug resistance.

[0562] The BRAF protein presents a signal propagation mechanism that requires protein homodimerization (BRAF-BRAF) or heterodimerization with other RAF proteins (BRAF-RAF1 or BRAF-ARAF). When BRAF is mutated, as observed in oncology indications with BRAF V600X substitutions, BRAF signaling becomes independent of the production of homodimers and / or heterodimers. In this case, the kinase is overactivated as a monomeric protein and drives cell proliferation signals.

[0563] Because currently available inhibitors block the activity of only monomeric BRAF forms and are ineffective against BRAF homodimers or heterodimers, it is not surprising that many BRAF resistance-inducing mechanisms operate by restoring signaling mediated by RAF homodimerization and heterodimerization.

[0564] Targeted protein degradation induces target ubiquitination by recruiting E3 ligases, thereby promoting proteasome-mediated destruction of bound targets. BRAF degradation through targeted degradation offers advantages over conventional inhibition because it eliminates the scaffolding activity of BRAF V600E / K and specifically induces BRAF protein elimination. This activity prevents dimerization-mediated resistance mechanisms.

[0565] Ablation of the BRAF protein could represent a strategy to delay the onset of drug resistance and target tumors that have acquired resistance to available inhibitors. This offers new therapeutic opportunities for treating tumors harboring the BRAF V600X mutation, such as melanoma, colorectal cancer, and lung cancer.

[0566] Another aspect of the present invention provides a crystalline form of Compound 1 or an isotopic derivative of Compound 1 as described herein, a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition thereof, for use in the preparation of a medicament for treating or preventing cancer in a patient in need thereof; wherein BRAF inhibition is required to treat or prevent cancer.

[0567] In certain aspects, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used to treat BRAF-mediated cancer, wherein BRAF is mutated from wild type. There are multiple possibilities for BRAF mutations. In certain non-limiting embodiments, the mutation is a Class I mutation, a Class II mutation, or a Class III mutation, or any combination thereof. Non-limiting examples of Class I mutations include V600 mutations, such as V600E, V600K, V600R, V600D, V600M, and V600N. Non-limiting examples of Class II mutations include G469A, G469V, G469L, G469R, L597Q, and K601E. Non-limiting examples of Class III mutations include G466A, G466E, G466R, G466V, S467L, G469E, N581I, D594E, D594G, and D594N.

[0568] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof treats a condition mediated by a BRAF mutant, wherein the mutation is not a Class I, Class II, or Class III mutation. Non-limiting examples of mutations include G464I, G464R, N581T, L584F, E586K, G593D, G596C, L597R, L597S, S605I, S607F, N684T, E26A, V130M, L745L, and D284E.

[0569] In certain embodiments, the BRAF mutation is an exon 11 mutation.

[0570] In certain embodiments, the BRAF mutation is an exon 15 mutation.

[0571] In certain embodiments, the BRAF mutation is a G464 mutation.

[0572] In certain embodiments, the BRAF mutation is a G466 mutation.

[0573] In certain embodiments, the BRAF mutation is a G466R mutation.

[0574] In certain embodiments, the BRAF mutation is a G466E mutation.

[0575] In certain embodiments, the BRAF mutation is a G469 mutation.

[0576] In certain embodiments, the BRAF mutation is a G469E mutation.

[0577] In certain embodiments, the BRAF mutation is a D594 mutation.

[0578] In certain embodiments, the BRAF mutation is a D594A mutation.

[0579] In certain embodiments, the BRAF mutation is an L597 mutation.

[0580] In certain embodiments, the BRAF mutation is an L597R mutation.

[0581] In certain embodiments, the BRAF mutation is an L597S mutation.

[0582] In certain embodiments, the BRAF mutation is an L597Q mutation.

[0583] In certain embodiments, the BRAF mutation is a V600 mutation.

[0584] In certain embodiments, the BRAF mutation is a V600E mutation.

[0585] In certain embodiments, the BRAF mutation is a V600K mutation.

[0586] In certain embodiments, the BRAF mutation is a V600R mutation.

[0587] In certain embodiments, the BRAF mutation is a V600D mutation.

[0588] In certain embodiments, the BRAF mutation is a V600M mutation.

[0589] In certain embodiments, the BRAF mutation is a V600N mutation.

[0590] In certain embodiments, the BRAF mutation is a K601 mutation.

[0591] In certain embodiments, the BRAF mutation is a K601E mutation.

[0592] In certain embodiments, the BRAF mutation is a K601N mutation.

[0593] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof treats a condition mediated by a BRAF mutant, wherein the mutation is a splice variant, such as p61-BRAF V600E or BRAF kinase domain duplication or BRAF amplification.

[0594] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used to treat a disorder mediated by two or more mutant proteins, such as BRAF. V600E / NRAS Q61K BRAF V600E / NRAS Q61R Double mutant-mediated cancer.

[0595] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used to treat a cancer that is resistant to at least one BRAF inhibitor, such as a cancer that is resistant or has acquired resistance to a BRAF inhibitor selected from dabrafenib, trametinib, vemurafenib, and contrafenib.

[0596] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used to treat cancers that have escape mutations, such as BRAF V600E NRAS Q61K Double mutation cancer.

[0597] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used to treat melanoma.

[0598] Non-limiting examples of melanoma include nonacral cutaneous melanoma, acral melanoma, mucosal melanoma, uveal melanoma, and leptomeningeal melanoma, each of which can be primary or metastatic.

[0599] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used to treat triple-negative breast cancer, such as triple-negative breast cancer having a G464V BRAF mutant.

[0600] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used to treat lung cancer, such as lung adenocarcinoma having a G466V BRAF mutant.

[0601] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used to treat melanoma having a V600 BRAF mutant.

[0602] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form thereof treats a condition mediated by a BRAF mutant, wherein the mutation is a splice variant, such as p61-BRAF V600E .

[0603] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form thereof is used to treat a disorder mediated by two or more mutant proteins, such as BRAF.V600E / NRAS Q61K BRAF V600E / NRAS Q61R Double mutant-mediated cancer.

[0604] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form thereof is used to treat cancers that have escape mutations, such as BRAF V600E NRAS Q61K Double mutant cancers.

[0605] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form thereof is used to treat a cancer that is resistant to at least one BRAF inhibitor, such as a cancer that is resistant or has acquired resistance to a BRAF inhibitor selected from dabrafenib, vemurafenib, and contrafenib.

[0606] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat melanoma.

[0607] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat triple-negative breast cancer, such as triple-negative breast cancer with a G464V BRAF mutant.

[0608] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat lung cancer, such as lung adenocarcinoma having a G466V BRAF mutant.

[0609] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat melanoma with a V600 BRAF mutant.

[0610] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat bile duct cancer.

[0611] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat Erdeheim-Chester disease.

[0612] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat Langerhans' histiocytosis.

[0613] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat ganglioglioma.

[0614] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or a crystalline form thereof is used to treat glioma.

[0615] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat GIST.

[0616] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat glioblastoma.

[0617] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat hairy cell leukemia.

[0618] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat multiple myeloma.

[0619] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or a crystalline form thereof is used to treat non-small cell lung cancer.

[0620] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat ovarian cancer.

[0621] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat pilomyxoid astrocytoma.

[0622] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat anaplastic pleomorphic xanthoastrocytoma.

[0623] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat astrocytoma.

[0624] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or a crystalline form thereof is used to treat thyroid cancer.

[0625] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or a crystalline form thereof is used to treat papillary thyroid cancer.

[0626] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat anaplastic thyroid cancer.

[0627] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat pancreatic cancer.

[0628] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat clear cell sarcoma of the breast.

[0629] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat salivary gland cancer.

[0630] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or crystalline form thereof is used to treat colorectal cancer.

[0631] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt or a crystalline form thereof is used to treat microsatellite-stable colorectal cancer.

[0632] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form thereof is used to treat a condition selected from the group consisting of bile duct cancer, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, GIST, glioblastoma, hairy cell leukemia, multiple myeloma, lung cancer, non-small cell lung cancer, ovarian cancer, pilomyxoid astrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, thyroid cancer, papillary thyroid cancer, anaplastic thyroid cancer, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, colorectal cancer, and microsatellite stable colorectal cancer.

[0633] In certain aspects, a method for treating a mutant BRAF-mediated cancer that has metastasized to the brain or central nervous system (CNS) is provided, comprising administering an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form thereof to a patient in need thereof. In certain embodiments, the cancer that has metastasized to the brain or CNS is colorectal cancer, melanoma, non-small cell lung cancer, or other solid tumors carrying BRAF V600E. In certain embodiments of this aspect, Compound 1, or a pharmaceutically acceptable salt thereof, is administered to a patient in need thereof. In other embodiments of this aspect, a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition prepared from a crystalline form of Compound 1 is administered to a patient in need thereof.

[0634] In certain aspects, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat a mutant BRAF-mediated cancer that has metastasized to the brain or CNS, wherein BRAF has mutated from wild-type. There are many possibilities for BRAF mutations. In certain non-limiting embodiments, the mutation is a Class I mutation, a Class II mutation, or a Class III mutation, or any combination thereof. Non-limiting examples of Class I mutations include V600 mutations, such as V600E, V600K, V600R, V600D, V600M, and V600N. Non-limiting examples of Class II mutations include G469A, G469V, G469L, G469R, L597Q, and K601E. Non-limiting examples of Class III mutations include G466A, G466E, G466R, G466V, S467L, G469E, N581I, D594E, D594G, and D594N. In certain embodiments, the BRAF mutation is a V600 mutation, such as V600E BRAF, which mediates cancers that have metastasized to the brain or CNS.

[0635] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, can treat mutant BRAF-mediated cancers that have metastasized to the brain or CNS, wherein the mutation is not a Class I, Class II, or Class III mutation. Non-limiting examples of mutations include G464I, G464R, N581T, L584F, E586K, G593D, G596C, L597R, L597S, S605I, S607F, N684T, E26A, V130M, L745L, and D284E.

[0636] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is an exon 11 mutation.

[0637] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is an exon 15 mutation.

[0638] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a G464 mutation.

[0639] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a G466 mutation.

[0640] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a G466R mutation.

[0641] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a G466E mutation.

[0642] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a G469 mutation.

[0643] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a G469E mutation.

[0644] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a D594 mutation.

[0645] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a D594A mutation.

[0646] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is an L597 mutation.

[0647] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is an L597R mutation.

[0648] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is an L597S mutation.

[0649] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a L597Q mutation.

[0650] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a V600 mutation.

[0651] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a V600E mutation.

[0652] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a V600K mutation.

[0653] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a V600R mutation.

[0654] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a V600D mutation.

[0655] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a V600M mutation.

[0656] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a V600N mutation.

[0657] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a K601 mutation.

[0658] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a K601E mutation.

[0659] In certain embodiments, the disorder is cancer that has metastasized to the brain or CNS and the BRAF mutation is a K601N mutation.

[0660] In certain embodiments, Compound 1 or a pharmaceutically acceptable salt thereof can treat mutant BRAF-mediated cancers that have metastasized to the brain or CNS, wherein the mutation is a splicing mutation, such as p61-BRAF V600E .

[0661] In certain embodiments, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat mutant BRAF-mediated cancers that have metastasized to the brain or CNS, where the cancer is mediated by two or more mutant proteins, such as BRAF V600E / NRAS Q61K BRAF V600E / NRAS Q61R Double mutant mediated cancers. Non-limiting examples of double mutant cancers include colorectal cancer, which is caused by BRAF mutations (e.g., BRAF V600E ), NRAS, MEK1, or PI3K mutations (e.g., BRAF V600E / PIK3CA H1047R BRAF V600E / PIK3CA P449T ) mediated.

[0662] In certain embodiments, compound 1 or a pharmaceutically acceptable salt thereof is used to treat a mutant BRAF-mediated cancer that has been transferred to the brain or CNS, wherein the cancer is resistant to at least one BRAF inhibitor, such as cancer is resistant to or has acquired resistance to a BRAF inhibitor selected from dabrafenib, vemurafenib and connefenib. In certain aspects, cancers resistant to treatment with BRAF inhibitors have RAF protein homodimerization or heterodimerization that promotes mutations. For example, in certain embodiments, compound 1 or a pharmaceutically acceptable salt thereof is used to treat a mutant BRAF-mediated cancer that has been transferred to the brain or CNS, wherein the cancer has one or more mutations that promote RAF protein dimerization. In certain embodiments, cancers with one or more RAF protein dimerizations that promote mutations are resistant to treatment with BRAF inhibitors (such as dabrafenib, vemurafenib or connefenib). In certain embodiments, the RAF protein dimer is a homodimer (BRAF-BRAF) or a heterodimer with another RAF protein (BRAF-RAF1 or BRAF-ARAF).

[0663] In certain embodiments, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat cancer that has metastasized to the brain or CNS, where the cancer has developed an escape mutation, such as BRAF V600E / NRAS Q61K or BRAF V600E / NRAS Q61R Double mutant cancers.

[0664] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat cancer that has metastasized to the brain or CNS, where the cancer has developed amplification of a driver mutation.

[0665] In other embodiments, compound 1 reduces phosphorylated ERK signaling, indicating inhibition of the EGFR-mediated MAPK pathway. In another embodiment, compound 1 is used to treat BRAF V600X colorectal cancer (CRC). In certain embodiments, compound 1 is used to treat BRAF-V600X non-small cell lung cancer (NSCLC). In certain embodiments, compound 1 shows activity against drug-resistant tumor mutations, such as point mutations in MEK, PI3K, splice variants (such as p61-BRAF-V600E), BRAF kinase domain duplications, and BRAF amplification.

[0666] In certain embodiments, Compound 1 or a pharmaceutically acceptable salt thereof can be used to treat colorectal cancer with a V600 mutation. For example, in certain embodiments, Compound 1 or a pharmaceutically acceptable salt thereof, rather than a combination of enacetinib and cetuximab, can be used to reduce tumor growth in a colorectal cancer model with a BRAF V600 mutation.

[0667] In certain embodiments, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat melanoma that has metastasized to the brain or CNS. In other embodiments, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat colorectal cancer that has metastasized to the brain or CNS.

[0668] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat melanoma that has metastasized to the brain or CNS.

[0669] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat triple-negative breast cancer that has metastasized to the brain or CNS, such as triple-negative breast cancer with the G464V BRAF mutant that has metastasized to the brain or CNS.

[0670] In certain embodiments, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat lung cancer that has metastasized to the brain or CNS, such as lung adenocarcinoma with a G466V BRAF mutant that has metastasized to the brain or CNS.

[0671] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat melanoma with a V600 BRAF mutant that has metastasized to the brain or CNS.

[0672] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat bile duct cancer that has metastasized to the brain or CNS.

[0673] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat Erdeheim-Chester disease that has metastasized to the brain or CNS.

[0674] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat Langerhans' histiocytosis that has metastasized to the brain or CNS.

[0675] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat ganglioglioma that has metastasized to the brain or CNS.

[0676] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat gliomas that have metastasized to the brain or CNS.

[0677] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat GIST that has metastasized to the brain or CNS.

[0678] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat glioblastoma that has metastasized to the brain or CNS.

[0679] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat hairy cell leukemia that has metastasized to the brain or CNS.

[0680] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat multiple myeloma that has metastasized to the brain or CNS.

[0681] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat non-small cell lung cancer that has metastasized to the brain or CNS.

[0682] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat ovarian cancer that has metastasized to the brain or CNS.

[0683] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat pilocytic astrocytoma that has metastasized to the brain or CNS.

[0684] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat anaplastic pleomorphic xanthoastrocytoma that has metastasized to the brain or CNS.

[0685] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat astrocytomas that have metastasized to the brain or CNS.

[0686] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat thyroid cancer that has metastasized to the brain or CNS.

[0687] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat papillary thyroid cancer that has metastasized to the brain or CNS.

[0688] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat anaplastic thyroid cancer that has metastasized to the brain or CNS.

[0689] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat pancreatic cancer that has metastasized to the brain or CNS.

[0690] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat clear cell sarcoma of the chest that has metastasized to the brain or CNS.

[0691] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat salivary gland cancer that has metastasized to the brain or CNS.

[0692] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat colorectal cancer that has metastasized to the brain or CNS.

[0693] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is used to treat microsatellite stable colorectal cancer that has metastasized to the brain or CNS.

[0694] In certain aspects, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat mutant BRAF-mediated cancers that have metastasized to the brain. In other aspects, Compound 1 or a pharmaceutically acceptable salt thereof is used to treat mutant BRAF-mediated cancers that have metastasized to the CNS.

[0695] Another aspect of the present invention provides a method for treating or preventing a proliferative disease. The method comprises administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form of Compound 1, or an enantiomer, diastereomer, stereoisomer, or isotopic derivative of Compound 1, or a pharmaceutically acceptable salt, hydrate, or solvate thereof, and optionally a pharmaceutically acceptable carrier.

[0696] In certain embodiments, the disease or disorder is cancer or a proliferative disease.

[0697] In certain embodiments, the BRAF-mediated disorder is abnormal cell proliferation, including but not limited to solid or hematological cancers.

[0698] In certain embodiments, the hematological cancer is acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), lymphoblastic T-cell leukemia, chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia, chronic neutrophilic leukemia (CNL), acute lymphoblastic T-cell leukemia, acute monocytic leukemia, plasmacytoma, immunoblastic large cell leukemia, mantle cell leukemia, multiple myeloma, megakaryoblastic leukemia, acute megakaryocytic leukemia, leukemia, promyelocytic leukemia, mixed lineage leukemia (MLL), erythroleukemia, malignant lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, lymphoblastic T-cell lymphoma, Burkitt's lymphoma, follicular lymphoma, acute lymphoblastic leukemia, diffuse large B-cell lymphoma, Myc and B-cell leukemia (BCL) 2 and / or BCL6 rearrangement / overexpression [double and triple hit lymphoma], myelodysplastic / myeloproliferative neoplasms, mantle cell lymphoma, including bortezomib-resistant mantle cell lymphoma.

[0699] Solid tumors that are sensitive to BRAF or mutant BRAF inhibition or degradation and thus can be treated with the compounds described herein include, but are not limited to, lung cancer (including small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC)), breast cancer (including inflammatory breast cancer, ER-positive breast cancer (including tamoxifen-resistant ER-positive breast cancer and triple-negative breast cancer), colon cancer, midline cancer, liver cancer, kidney cancer, prostate cancer (including castration-resistant prostate cancer (CRPC)), brain cancer (including gliomas, glioblastomas, neuroblastomas and medulloblastomas, including MYC-amplified medulloblastomas), colorectal cancer, Wilms' cell carcinoma, Ewing's sarcoma, rhabdomyosarcoma, ependymoma, head and neck cancer, melanoma, squamous cell carcinoma, ovarian cancer, pancreatic cancer (including pancreatic ductal adenocarcinoma (PDAC) and pancreatic neuroendocrine tumors (PanNET)), osteosarcoma, giant cell tumor of bone, thyroid cancer, bladder cancer, urothelial carcinoma, vulvar cancer, cervical cancer, endometrial cancer, mesothelioma, esophageal cancer, salivary gland cancer, gastric cancer, nasopharyngeal cancer, oral cancer, cancers of the mouth, GIST (gastrointestinal stromal tumor), NUT-midline cancer, testicular cancer, squamous cell carcinoma, hepatocellular carcinoma (HCC), MYCN-driven solid tumors, and NUT-midline carcinoma (NMC).

[0700] In further embodiments, the disease or disorder is a sarcoma of bone, muscle, tendon, cartilage, nerve, fat, or blood vessels.

[0701] In further embodiments, the disease or condition is soft tissue sarcoma, skeletal sarcoma, or osteosarcoma.

[0702] In further embodiments, the disease or condition is angiosarcoma, fibrosarcoma, liposarcoma, leiomyosarcoma, Kaposi's sarcoma, osteosarcoma, gastrointestinal stromal tumor, synovial sarcoma, pleomorphic sarcoma, chondrosarcoma, Ewing's sarcoma, reticulum cell sarcoma, angiosarcoma, botryoid sarcoma, rhabdomyosarcoma, or embryonal rhabdomyosarcoma.

[0703] In certain embodiments, the disorder is bone, muscle, tendon, cartilage, nerve, fat, or angiosarcoma.

[0704] In other embodiments, the disease or condition is a cancer that is sensitive to BRAF or mutant BRAF inhibition or degradation. In further embodiments, the cancer is lung cancer, colon cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, brain cancer, kidney cancer, ovarian cancer, stomach cancer, skin cancer, bone cancer, stomach cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, hepatocellular carcinoma, papillary renal carcinoma, head and neck squamous cell carcinoma, leukemia, lymphoma, myeloma, solid tumor, blood cancer or solid cancer.

[0705] The term "cancer" as used herein refers to cancers that can be treated with Compound 1, and refers to any cancer caused by the proliferation of malignant cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, lymphomas, etc., wherein the disease is sensitive to BRAF or mutant BRAF inhibition or degradation. For example, cancers include, but are not limited to, mesothelioma, leukemias, and lymphomas, such as cutaneous T-cell lymphoma (CTCL), non-cutaneous peripheral T-cell lymphoma, lymphomas associated with human T-cell lymphotropic virus (HTLV), such as adult T-cell leukemia / lymphoma (ATLL), B-cell lymphomas, acute non-lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, lymphomas and multiple myeloma, non-Hodgkin's lymphoma, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), Hodgkin's lymphoma, Burkitt's lymphoma, adult T-cell leukemia lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), or hepatocellular carcinoma. Further examples include myelodysplastic syndrome, solid tumors of childhood (such as brain tumors, neuroblastoma, retinoblastoma, Wilms' tumor, bone tumors and soft tissue sarcomas), common solid tumors in adults such as head and neck cancers (such as oral, laryngeal, nasopharyngeal and esophageal cancers), genitourinary cancers (such as prostate, bladder, kidney, uterus, ovary, testis), lung cancer (such as small cell and non-small cell), breast cancer, pancreatic cancer, melanoma and other skin cancers, stomach cancer, brain tumors, tumors associated with Gorlin's syndrome (such as medulloblastoma or meningioma) and liver cancer.

[0706] One aspect of the present application provides a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form of Compound 1, which can be used to treat diseases, disorders, and conditions characterized by excessive or abnormal cell proliferation that is sensitive to BRAF or mutant BRAF inhibition or degradation. Such diseases include, but are not limited to, proliferative or hyperproliferative diseases. Examples of proliferative and hyperproliferative diseases include, but are not limited to, cancer. The term "cancer" includes, but is not limited to, the following cancers: breast; ovary; cervix; prostate; testis, genitourinary tract; esophagus; larynx, glioblastoma; neuroblastoma; stomach; skin, keratoacanthoma; lung, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung adenocarcinoma; bone; colon; colorectal; adenoma; pancreas, adenocarcinoma; thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma; seminoma; melanoma; sarcoma; bladder cancer; liver cancer and biliary tract cancer; kidney cancer; bone marrow disorders; lymphoid disorders, Hodgkin's, hairy cell; oral cavity and pharynx (mouth), lip, tongue, mouth, pharynx; small intestine; colon, large intestine, rectum, brain and central nervous system; chronic myeloid leukemia (CML) and leukemia. The term "cancer" includes but is not limited to the following cancers: myeloma, lymphoma, or a cancer selected from stomach, kidney, or the following: head and neck, oropharyngeal, non-small cell lung cancer (NSCLC), endometrial cancer, liver cancer, non-Hodgkin's lymphoma, and lung.

[0707] Other exemplary forms of cancer include, but are not limited to, bone or smooth muscle cancer, stomach cancer, small intestine cancer, rectum cancer, salivary gland cancer, endometrial cancer, adrenal cancer, anal cancer, rectal cancer, parathyroid cancer, and pituitary cancer.

[0708] Other cancers that can be treated by the present invention include colon cancer, familial adenomatous polyposis cancer and hereditary non-polyposis colorectal cancer or melanoma. In addition, cancers include, but are not limited to, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, stomach cancer, adenocarcinoma, thyroid cancer (medullary and papillary thyroid cancer), kidney cancer, renal parenchymal cancer, cervical cancer, uterine corpus cancer, endometrial cancer, choriocarcinoma, testicular cancer, urethral cancer, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermal tumor, gallbladder cancer, bronchogenic carcinoma, multiple myeloma, basal cell tumor, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma and plasmacytoma. In one aspect of the present application, the present application provides use of one or more compounds described herein in the preparation of a medicament for treating cancer (including but not limited to various types of cancer disclosed herein).

[0709] In some embodiments, the pharmaceutical compositions or crystal forms described herein can be used to treat cancers that are sensitive to BRAF or mutant BRAF inhibition or degradation, such as colorectal, thyroid, breast, and lung cancer; and myeloproliferative disorders, such as polycythemia vera, thrombocythemia, myeloid metaplasia with myelofibrosis, chronic myeloid leukemia, chronic myelomonocytic leukemia, hypereosinophilic syndrome, juvenile myelomonocytic leukemia, and systemic mast cell disease. In some embodiments, the compound 1 crystal form described herein can be used to treat hematopoietic disorders that are sensitive to BRAF or mutant BRAF inhibition or degradation, particularly acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute promyelocytic leukemia, and acute lymphoblastic leukemia (ALL).

[0710] In certain embodiments, the pharmaceutical compositions or crystalline forms described herein can be used in an effective amount to treat a host, such as a human, suffering from a lymphoma or a lymphocytic or myeloproliferative disorder or abnormality. For example, the crystalline form of Compound 1 described herein can be administered to a host suffering from Hodgkin's lymphoma or non-Hodgkin's lymphoma. For example, the host may have a non-Hodgkin's lymphoma, such as, but not limited to, ADIS-associated lymphoma; anaplastic large cell lymphoma; angioimmunoblastic lymphoma; blastic NK-cell lymphoma; Burgit's lymphoma; Burkitt-like lymphoma (small non-cleaved cell lymphoma); diffuse small cleaved cell lymphoma (DSCCL); chronic lymphocytic leukemia / small lymphocytic lymphoma; cutaneous T-cell lymphoma; diffuse large B-cell lymphoma; enteropathy-type T-cell lymphoma; follicular lymphoma; hepatosplenic gamma-delta T-cell lymphoma; lymphoblastic lymphoma; mantle cell lymphoma; marginal zone lymphoma; nasal T-cell lymphoma; childhood lymphoma; peripheral T-cell lymphoma; primary central nervous system lymphoma; T-cell leukemia; transformed lymphoma; therapy-related T-cell lymphoma; Langerhans cell histiocytosis; or Waldenstrom's macroglobulinemia.

[0711] In other embodiments, the pharmaceutical compositions or crystalline forms described herein can be used in an effective amount to treat a patient, e.g., a human, suffering from Hodgkin's lymphoma, such as, but not limited to: nodular sclerosis classical Hodgkin's lymphoma (CHL); mixed cellularity CHL; lymphocyte-depleted CHL; lymphocyte-rich CHL; lymphocyte-predominant Hodgkin's lymphoma; or nodular lymphocyte-predominant HL.

[0712] The present application further includes treating or preventing cell proliferative disorders, such as hyperplasia, dysplasia and precancerous lesions that are sensitive to BRAF or mutant BRAF inhibition or degradation. Dysplasia is the earliest form of precancerous lesions identified by pathologists during biopsy. The pharmaceutical compositions or crystalline forms described herein can be used to prevent the hyperplasia, dysplasia or precancerous lesions from continuing to expand or becoming cancer. Examples of precancerous lesions may occur in the skin, esophageal tissue, breast and cervical epithelial tissue.

[0713] In certain aspects, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used to treat BRAF-mediated cancer, wherein BRAF is mutated from wild type. There are multiple possibilities for BRAF mutation. In certain non-limiting embodiments, the mutation is a Class I mutation, a Class II mutation, or a Class III mutation, or any combination thereof. Non-limiting examples of Class I mutations include V600 mutations, such as V600E, V600K, V600R, V600D, V600M, and V600N. Non-limiting examples of Class II mutations include G469A, G469V, G469L, G469R, L597Q, and K601E. Non-limiting examples of Class III mutations include G466A, G466E, G466R, G466V, S467L, G469E, N581I, D594E, D594G, and D594N.

[0714] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof treats a BRAF mutant-mediated condition, wherein the mutation is not a Class I, Class II, or Class III mutation. Non-limiting examples of mutations include G464I, G464R, N581T, L584F, E586K, G593D, G596C, L597R, L597S, S605I, S607F, N684T, E26A, V130M, L745L, and D284E.

[0715] Another aspect of the present invention provides a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof, for preparing a medicament for treating or preventing a disease mediated by BRAF.

[0716] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof can be used to treat a disorder comprising abnormal cell proliferation, such as a tumor or cancer, wherein BRAF is an oncogenic protein or signaling mediator of an abnormal cell proliferation pathway and its degradation reduces abnormal cell growth.

[0717] Combination therapy

[0718] Pharmaceutical compositions comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 described herein, can be used alone or in combination, or administered simultaneously or sequentially with another biologically active agent or a second therapeutic agent to treat patients, such as humans, suffering from mutant BRAF-mediated disorders, including but not limited to those described herein.

[0719] The term "biologically active agent" or "additional active agent" is used to describe an agent other than Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of Compound 1 according to the present invention, which can be used in combination or alternation with a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of Compound 1 to achieve the desired therapeutic result. In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of Compound 1 and a biologically active agent is administered in such a manner that the active agent is administered over an overlapping period of time (e.g., with overlapping C max 、T max In another embodiment, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 and a biologically active agent is administered to a patient in need thereof, wherein the pharmaceutical composition does not have overlapping pharmacokinetic parameters, but one has a therapeutic effect on the therapeutic effect of the other.

[0720] In certain aspects, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used in combination with a second active agent as described herein to treat mutant BRAF-mediated cancers. Non-limiting examples of molecular classes that can be used in combination with a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof include MEK inhibitors, immune checkpoint inhibitors, and EGFR antibodies. In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used in combination with trametinib to treat mutant BRAF-mediated cancers, such as melanoma or non-small cell lung cancer. In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used in combination with an immune checkpoint inhibitor to treat mutant BRAF-mediated cancers. In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used in combination with cetuximab or panitumumab to treat mutant BRAF-mediated cancers, such as colorectal cancer. In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used in combination with nivolumab, pembrolizumab, cemiplimab, ipilimumab, relimab, atezolizumab, avelumab, or durvalumab to treat mutant BRAF-mediated cancers, such as colorectal cancer, melanoma, or non-small cell lung cancer.

[0721] In other aspects, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used in combination with two or more additional active agents as described herein to treat mutant BRAF-mediated cancers. In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a crystalline form thereof is used in combination with a MEK inhibitor and an immune checkpoint inhibitor to treat melanoma or non-small cell lung cancer.

[0722] In some embodiments, Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of Compound 1 is administered with another BRAF inhibitor such as sorafenib, vemurafenib, or Dabrafenib or Conafenib

[0723] In certain embodiments, the bioactive agent is a MEK inhibitor. MEK inhibitors are well known and include, for example, trametinib / GSK1120212 (N-(3-{3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H-yl)phenyl)acetamide), selumetinib (6-(4-bromo-2-chloroanilino)-7-fluoro-N-(2-hydroxyethoxy)-3-methylbenzimidazole-5-carboxamide), pimatitinib / AS703026 / MSC 1935369 ((S)-N-(2,3-dihydroxypropyl)-3-(2-fluoro-4-iodophenyl)amino)isonicotinamide), XL-518 / GDC-0973 (l-({3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl}carbonyl)-3-[(2S)-piperidin-2-yl]azetidin-3-ol), rifatinib / BAY869766 / RDEAl 19 (N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide), PD-0325901 (N-[(2R)-2,3-dihydroxypropyloxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide), TAK733 (I-3-(2, 3-dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione), MEK162 / ARRY438162 (5-[(4-bromo-2-fluorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H-benzimidazole-6-carboxamide), R051267 66(3-[[3-fluoro-2-(methylsulfonamido)-4-pyridinyl]methyl]-4-methyl-7-pyrimidin-2-yloxychromen-2-one), WX-554, R04987655 / CH4987655(3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-5-((3-oxo-1,2-oxazin-2-yl) methyl)benzamide) or AZD8330 (2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carboxamide), U0126-EtOH, PD184352 (CI-1040), GDC-0623, BI-847325, cobimetinib, PD98059, BIX 02189, BIX 02188, bimetinib, SL-327, TAK-733, PD318088.

[0724] In certain embodiments, the MEK inhibitor is trametinib.

[0725] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 is used in combination with cetuximab or trametinib to treat colorectal cancer. In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 is used in combination with cetuximab and BYL719 to treat colorectal cancer. In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 is used in combination with cetuximab and irinotecan to treat colorectal cancer.

[0726] In certain embodiments, the bioactive agent is a SHP2 inhibitor. In certain embodiments, the SHP2 inhibitor is SHP099.

[0727] In certain embodiments, the bioactive agent is a RAF inhibitor. Non-limiting examples of Raf inhibitors include, for example, vemurafenib (N-[3-[[5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl]-1-propanesulfonamide), sorafenib tosylate (4-[4-[[4-chloro-3-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-N-methylpyridine-2-carboxamide; 4-methylbenzenesulfonate), AZ628 (3-(2-cyanophenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl]-1-propanesulfonamide), benzamide), NVP-BHG712 (4-methyl-3-(1-methyl-6-(pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino)-N-(3-(trifluoromethyl)phenyl)benzamide), RAF-265 (1-methyl-5-[2-[5-(trifluoromethyl)-1H -imidazol-2-yl]pyridin-4-yl]oxy-N-[4-(trifluoromethyl)phenyl]benzimidazol-2-amine), 2-Bromoaldisine (2-bromo-6,7-dihydro-1H,5H-pyrrolo[2,3-c]azepine-4,8-dione), Raf kinase inhibitor IV (2-chloro-5-(2-phenyl-5-(pyridin-4-yl)-1H-imidazol-4-yl)phenol), sorafenib N-oxide (4-[4-[ [[[4-chloro-3-(trifluoromethyl)phenyl]amino]carbonyl]amino]phenoxy]-N-methyl-2-pyridinecarboxamide 1-oxide), PLX-4720, dabrafenib (GSK2118436), GDC-0879, RAF265, AZ628, SB590885, ZM336372, GW5074, TAK-632, CEP-32496, LY3009120, and GX818 (conafenib ( ).

[0728] In certain embodiments, the RAF inhibitor is conafenib.

[0729] In certain embodiments, the RAF inhibitor is vemurafenib.

[0730] In certain embodiments, the RAF inhibitor is dabrafenib.

[0731] In certain embodiments, the bioactive agent is an EGFR inhibitor, including, for example, gefitinib. Erlotinib Lapatinib Osimertinib Lenaticin Vandetanib Dacomitinib Cirotinib (XEGAFRI TM ), afatinib ( GIOTRIFF TM AFANIX TM ), lazertinib, or neratinib.

[0732] Other examples of EGFR inhibitors include sirolimus (CO-1686), omodinib (OLITA TM ), naquintinib (ASP8273), nazartinib (EGF816), PF-06747775, icotinib (BPI-2009), neratinib (HKI-272; PB272); avitinib (AC0010), EAI045, talotinib (TH-4000; PR-610), PF-06459988 (Pfizer), tevatinib (XL647; EXEL-7647; KD-019), transtinib, WZ-3146, WZ8040, CNX-2006, dacomitinib (PF-00299804; Pfizer), brigatinib Lorlatinib and PF-06747775 (PF7775).

[0733] In certain embodiments, the bioactive agent is a first generation EGFR inhibitor, such as erlotinib, gefitinib, or lapatinib. In certain embodiments, the bioactive agent is a second generation EGFR inhibitor, such as afatinib and / or dacomitinib. In certain embodiments, the bioactive agent is a third generation EGFR inhibitor, such as osimertinib.

[0734] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 is administered in combination with osimertinib to a patient in need thereof.

[0735] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 is administered in combination with cilottinib to a patient in need thereof.

[0736] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 is administered in combination with Avitinib to a patient in need thereof.

[0737] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 is administered in combination with lazertinib to a patient in need thereof.

[0738] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 is administered in combination with neratinib to a patient in need thereof.

[0739] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1, is administered to a patient in need thereof in combination with an EGFR antibody (e.g., cetuximab, panitumumab, or necituzumab).

[0740] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1, is administered to a patient in need thereof in combination with cetuximab.

[0741] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1, is administered to a patient in need thereof in combination with panitumumab.

[0742] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1, is administered to a patient in need thereof in combination with necituzumab.

[0743] In certain embodiments, bioactive agent is an immunomodulator, including but not limited to checkpoint inhibitors, including PD-1 inhibitors, PD-L1 inhibitors, PD-L2 inhibitors, CTLA-4 inhibitors, LAG-3 inhibitors, TIM-3 inhibitors, V domain Ig inhibitors of T cell activation (VISTA) inhibitors, small molecules, peptides, nucleotides or another inhibitor as non-limiting examples. In some aspects, immunomodulator is an antibody, such as a monoclonal antibody.

[0744] PD-1 inhibitors, such as nivolumab, inhibit immunosuppression by blocking the interaction between PD-1 and PD-L1 by binding to the PD-1 receptor. Pembrolizumab Pidilizumab, AMP-224 (AstraZeneca and MedImmune), PF-06801591 (Pfizer), MEDI0680 (AstraZeneca), PDR001 (Novartis), REGN2810 (Regenron), SHR-12-1 (Jiangsu Hengrui Medicine Company and Incyte Corporation), TSR-042 (GlaxoSmithKline plc) and PD-L1 / VISTA inhibitor CA-170 (Curis Inc.). PD-L1 inhibitors that inhibit the interaction between PD-1 and PD-L1 by binding to the PD-L1 receptor and thereby inhibiting immunosuppression include, for example, atezolizumab. Durvalumab (AstraZeneca and MedImmune), KN035 (Alphamab Co. Ltd.) and BMS-936559 (Bristol-Myers Squibb). CTLA-4 checkpoint inhibitors that bind to CTLA-4 and inhibit immunosuppression include, but are not limited to, ipilimumab, tremelimumab (AstraZeneca and MedImmune), AGEN1884 and AGEN2041 (Agenus). LAG-3 checkpoint inhibitors include, but are not limited to, BMS-986016 (Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline plc), IMP321 (Prima BioMed), LAG525 (Novartis) and dual PD-1 and LAG-3 inhibitor MGD013 (MacroGenics). An example of a TIM-3 inhibitor is TSR-022 (GlaxoSmithKline plc).

[0745] In certain embodiments, the checkpoint inhibitor is selected from nivolumab. Pembrolizumab and pidilizumab / CT-011, MPDL3280A / RG7446; MEDI4736; MSB0010718C; BMS 936559, PDL2 / lg fusion proteins (such as AMP 224) or inhibitors of B7-H3 (e.g., MGA271), B7-H4, BTLA, HVEM, TIM3, GAL9, LAG 3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK 1, CHK2, A2aR, B-7 family ligands, or a combination thereof.

[0746] In yet other embodiments, an effective amount of Compound 1 as described herein, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1, can be used in combination or alternating administration with an effective amount of an estrogen inhibitor (including but not limited to SERMs (selective estrogen receptor modulators), SERDs (selective estrogen receptor degraders), complete estrogen receptor degraders, or another form of partial or complete estrogen antagonists or agonists) to treat abnormal tissues of the female reproductive system (e.g., breast cancer, ovarian cancer, endometrial cancer, or uterine cancer). Some anti-estrogens such as raloxifene and tamoxifen retain some estrogen-like effects, including estrogen-like stimulation of uterine growth, and in some cases, estrogen-like effects during the progression of breast cancer that actually stimulates tumor growth. In contrast, fulvestrant (a complete anti-estrogen) has no estrogen-like effect on the uterus and is effective for tamoxifen-resistant tumors.

[0747] Non-limiting examples of anti-estrogen compounds are provided in WO 2014 / 19176 to Astra Zeneca, WO 2013 / 090921, WO 2014 / 203129, WO 2014 / 203132, and US 2013 / 0178445 to Olema Pharmaceuticals, and U.S. Patent Nos. 9,078,871, 8,853,423, and 8,703,810, and US 2015 / 0005286, WO 2014 / 205136, and WO 2014 / 205138.

[0748] Other non-limiting examples of anti-estrogen compounds include: SERMS, such as anordrin, bazedoxifene, bromostriene, chlorfenestrol, clomiphene citrate, cyclofenil, lasofoxifene, ormeloxifene, raloxifene, tamoxifen, toremifene, and fulvestrant; aromatase inhibitors, such as aminoglutethimide, testolactone, anastrozole, exemestane, fadrozole, formestane, and letrozole; and antigonadotropins, such as leuprolide, cetrorelix, allylestradiol, chlormadinone acetate, cyproterone acetate, delmadinone acetate, dehydrogenated progesterone acetate, medroxyprogesterone acetate, megestrol acetate, nomegestrol acetate, norethisterone acetate, progesterone, and spironolactone.

[0749] U.S. Patent Nos. 4,418,068; 5,478,847; 5,393,763; and 5,457,117, WO 2011 / 156518, U.S. Patent Nos. 8,455,534 and 8,299,112, U.S. Patent Nos. 9,078,871; 8,853,423; 8,703,810; US 2015 / 0005286; and WO 2014 / 205138, US 2016 / 0175289, US 2015 / 0258080, WO 2014 / 191726, WO 2012 / 084711; WO 2002 / 013802; WO 2002 / 004418; WO 2002 / 003992; WO 2002 / 003991; WO 2002 / 003990; WO 2002 / 003989; WO2002 / 003988; WO 2002 / 003986; WO 2002 / 003977; WO 2002 / 003976; WO 2002 / 003975; WO 2006 / 078834; US 6821989; US 2002 / 0128276; US 6777424; US2002 / 0016340; US 6326392; US6756401; US2002 / 0013327; US 6512002; US Other estrogen ligands that may be used according to the present invention are described in US 6,632,834; US 2001 / 0056099; US 6,583,170; US 6,479,535; WO 1999 / 024027; US 6,005,102; EP 0,802,184; US 5,998,402; US 5,780,497, US 5,880,137, WO 2012 / 048058 and WO 2007 / 087684.

[0750] In another embodiment, an effective amount of a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of Compound 1 can be administered in combination or alternating with an effective amount of an androgen (e.g., testosterone) inhibitor (including but not limited to a selective androgen receptor modulator, a selective androgen receptor degrader, a complete androgen receptor degrader, or another form of a partial or complete androgen antagonist) for the treatment of abnormal tissues of the male reproductive system, such as prostate cancer or testicular cancer. In certain embodiments, prostate cancer or testicular cancer is androgen-resistant.

[0751] Non-limiting examples of anti-androgen compounds are provided in WO 2011 / 156518 and U.S. Patent Nos. 8,455,534 and 8,299, 112. Other non-limiting examples of anti-androgen compounds include enzalutamide, apalutamide, cyproterone acetate, chlormadinone acetate, spironolactone, canrenone, drospirenone, ketoconazole, topiramide, abiraterone acetate, and cimetidine.

[0752] In certain embodiments, the bioactive agent is an ALK inhibitor. Examples of ALK inhibitors include, but are not limited to, crizotinib Alectinib Ceritinib, TAE684 (NVP-TAE684), GSK1838705A, AZD3463, ASP3026, PF-06463922, entrectinib (RXDX-101), and AP26113.

[0753] In certain embodiments, the bioactive agent is a HER-2 inhibitor. Examples of HER-2 inhibitors include trastuzumab, lapatinib, trastuzumab emtansine, and pertuzumab.

[0754] In certain embodiments, the bioactive agent is a CD20 inhibitor. Examples of CD20 inhibitors include obinutuzumab Rituximab Ofatumumab, ibritumomab tiuxetan, tositumomab, and ocrelizumab.

[0755] In certain embodiments, the bioactive agent is a JAK3 inhibitor. Examples of JAK3 inhibitors include tofacitinib.

[0756] In certain embodiments, the bioactive agent is a BCL-2 inhibitor. Examples of BCL-2 inhibitors include venetoclax, ABT-199 (4-[4-[[2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-en-1-yl]methyl]piperazin-1-yl]-N-[[3-nitro-4-[(tetrahydro-2H-pyran-4-yl)methyl]amino]phenyl]sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridin-5-yl)oxy]benzamide), ABT-737 (4-[4-[[2-(4-chlorophenyl)phenyl]methyl] piperazin-1-yl]-N-[4-[[(2R)-4-(dimethylamino)-1-phenylsulfanylbutan-2-yl]amino]-3-nitrophenyl]sulfonylbenzamide)(navitoclax), ABT-263 (I-4-(4-(4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-morpholinyl-1-(phenylsulfanyl)butan-2-yl)amino)-3-( (trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide), GX15-070 (obaklav mesylate, (2Z)-2-[(5Z)-5-[(3,5-dimethyl-1H-pyrrol-2-yl)methylene]-4-methoxypyrrol-2-ylidene)indole; methanesulfonic acid), 2-methoxy-antimycin A3, YC137 (4-(4,9-dioxo-4,9-dihydronaphtho[2,3-d]thiazol-2-ylamino)-phenyl ester), gossypol, 2-amino-6-bromo- 4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylic acid ethyl ester, nilotinib-d3, TW-37 (N-[4-[[2-(1,1-dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2-(1-methylethyl)phenyl]methyl]benzamide), apogossypolone (ApoG2), HA14-1, AT101, sabutoclax, gambogic acid, or G3139 (oblimersen).

[0757] In certain embodiments, the bioactive agent is a kinase inhibitor. In certain embodiments, the kinase inhibitor is selected from a phosphoinositide 3-kinase (PI3K) inhibitor, a Bruton's tyrosine kinase (BTK) inhibitor, or a spleen tyrosine kinase (Syk) inhibitor, or a combination thereof.

[0758] Examples of PI3 kinase inhibitors include, but are not limited to, wortmannin, demethylchloroisothiazolin, perifosine, idelalisib, pitilisib, palomid 529, ZSTK474, PWT33597, CUDC-907 and AEZS-136, dovilice, GS-9820, BKM120, GDC-0032 (Taselisib) (2-[4-(2-isopropyl-5-methyl-1,2,4-triazol-3-yl-5,6-dihydroimidazo[1,2-d][1,4]benzoxazepin-9-yl]pyrazol-1-yl]-2-methylpropanamide), MLN-1117 ((2R)-1-phenoxy-2-butyl hydrogen (S)-methylphosphonate; or methyl(oxo){[(2R)-1- [phenoxy-2-butyl]oxy}phosphine)), BYL-719 ((2S)-N1-[4-methyl-5-[2-(2,2,2-trifluoro-1,1-dimethylethyl)-4-pyridinyl]-2-thiazolyl]-1,2-pyrrolidinedicarboxamide), GSK2126458 (2,4-difluoro-N-{2-(methyloxy)-5-[4-(4-pyridazinyl)-6-quinolinyl]-3-pyridinylbenzenesulfonamide) (omipalisib), TGX-221 ((±)-7-methyl-2-(morpholin-4-yl)-9-(l-phenylaminoethyl)-pyrido[1, 2-a]-pyrimidin-4-one), GSK2636771 (2-methyl-1-(2-methyl-3-(trifluoromethyl)benzyl)-6-morpholino-1H-benzo[d]imidazole-4-carboxylic acid dihydrochloride), KIN-193 (1-2-((1-(7-methyl-2-morpholino-4-oxo-4H-pyrido[1,2-a]pyrimidin-9-yl)ethyl)amino)benzoic acid), TGR-1202 / RP5264, GS-9820 ((S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-monohydroxypropan-1-one), GS- 1101 (5-fluoro-3-phenyl-2-([S]]-1-[9H-purin-6-ylamino]-propyl)-3H-quinazolin-4-one), AMG-319, GSK-2269557, SAR245409 (N-(4-(N-(3-(3,5-dimethoxyphenyl)amino)quinoxalin-2-yl)sulfonyl)phenyl)-3-methoxy-4-methylbenzamide), BAY80-6946 (2-amino-N-(7-methoxy-8-(3-morpholinopropoxy)-2,3-dihydroimidazo[1,2-c]quinazole), AS 252424 (5-[1-[5-(4-fluoro-2-hydroxyphenyl)-furan-2-yl]-m-(Z)-ylidene]-thiazolidine-2,4-dione), CZ 24832 (5-(2-amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-tert-butylpyridine-3-sulfonamide), Bupanisib (5-[2,6-bis(4-morpholinyl)-4-pyrimidinyl]-4-(trifluoromethyl)-2-pyridinamine), GDC-0941 (2-(1H-indazol-4-yl)-6-[[4-(methylsulfonyl)-l-piperazinyl]methyl]-4-(4-morpholinyl)thieno[3,2-d]pyrimidine), GDC-0980 ((S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidine) ]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (also known as RG7422)), SF1126 ((8S,14S,17S)-14-(carboxymethyl)-8-(3-guanidinopropyl)-17-(hydroxymethyl)-3,6,9,12,15-pentaoxo-1-(4-(4-(4-oxo-8-phenyl-4H-chromen-2-yl)morpholin-4-ium)-2-oxa-7,10,13,16-tetraazaoctadecane-18-oic acid), PF-05212384 (N-[4-[[4-(dimethyl [amino]-1-piperidinyl]carbonyl]phenyl]-N'-[4-(4,6-di-4-morpholino-1,3,5-triazin-2-yl)phenyl]urea)(gedatolisib), LY3023414, BEZ235 (2-methyl-2-{4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl]phenyl}propionitrile)(dactolisib), XL-765 (N-(3-(3,5-dimethoxyphenylamino)-1-piperidinyl)carbonyl]phenyl)urea )quinoxalin-2-yl)sulfonyl)phenyl)-3-methoxy-4-methylbenzamide) and GSK1059615 (5-[[4-(4-pyridyl)-6-quinoxalin-2-yl)methylene]-2,4-thiazolidinedione), PX886 ([(3aR,6E,9S,9aR,10R,11aS)-6-[[bis(prop-2-enyl)amino]methylene]-5-hydroxy-9-(methoxymethyl)-9a,11a-dimethyl-1,4,7-trioxo-2,3,3a,9,10,11-hexahydroindeno[4,5h]isochromen-10-yl] acetate (also known as sonolisib), LY294002, AZD8186, PF-4989216, pilaralisib, GNE-317, PI-3065, PI-103, NU7441 (KU-57788), HS173, VS-5584 (SB2343), CZC24832, TG100-115, A66, YM201636, CAY10505, PIK-75, PIK-93, AS-605240, BGT226 (NVP-BGT226), AZD6482, vodesime, apellisib, IC-87114, TGI100713, CH5132799, PKI-402, kupanisib (BAY 80-6946), XL 147, PIK-90, PIK-293, PIK-294, 3-MA (3-methyladenine), AS-252424, AS-604850, apitolisib (GDC-0980; RG7422).

[0759] Examples of BTK inhibitors include ibrutinib (also known as PCI-32765) (1-[(3R)-3-[4-amino-3-(4-phenoxy-phenyl)pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl]prop-2-en-1-one), diphenylaminopyrimidine-based inhibitors such as AVL-101 and AVL-291 / 292 (N-(3-((5-fluoro-2-((4-(2-methoxyethoxy)phenyl)amino)pyrimidin-4-yl)amino)phenyl)acrylamide) (Avila Therapeutics) (see U.S. Patent Publication No. 2011 / 0117073, incorporated herein in its entirety), dasatinib ([N-(2-chloro-6-methylphenyl)-2-(6-(4-(2-hydroxyethyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide), LMF-A13 (α-cyano-β-hydroxy-β-methyl-N-(2,5-dibromophenyl)acrylamide), GDC-0834 ([RN-(3-(6-(4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenylamino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide), CGI-560 4-(tert-butyl)-N-(3-(8-(phenylamino)imidazo[1,2-a]pyrazin-6-yl)phenyl)benzamide, CGI-1746(4-(tert-butyl)-N-(2-methyl-3-(4-methyl-6-(4-(morpholine-4-carbonyl)phenyl)amino)-5-oxo-4,5-dihydropyrazin-2-yl)phenyl)benzamide), CNX-774(4-(4 -((4-((3-acrylamidophenyl)amino)-5-fluoropyrimidin-2-yl)amino)phenoxy)-N-methylpicolinamide), CTA056 (7-benzyl-1-(3-(piperidin-1-yl)propyl)-2-(4-(pyridin-4-yl)phenyl)-1H-imidazo[4,5-g]quinoxalin-6(5H)-one), GDC-0834 (IN-(3-(6-(( 4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenyl)amino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide), GDC-0837 (IN-(3-(6-((4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenyl)-4-methyl-5-oxo- 4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide), HM-71224, ACP-196, ONO-4059 (Ono Pharmaceuticals), PRT062607 (4-(3-(2H-1,2,3-triazol-2-yl)phenyl)amino)-2-(((1R,2S)-2-aminocyclohexyl)amino)pyrimidine-5-carboxamide hydrochloride), QL-47 (1-(1-acryloylindolin-6-yl)-9-(1-methyl-1H-pyrazol-4-yl)benzo[h][1,6]naphthalen-2(1H)-one), and RN486 (6-cyclopropyl-8-fluoro-2-(2-hydroxymethyl-3-{1-methyl-5-[5-(4-methyl-piperazin-1-yl)-pyridin-2-ylamino]-6-oxo-1,6-dihydropyridin-3-yl}-phenyl)-2H-isoquinolin-1-one), as well as other molecules capable of inhibiting BTK activity, such as those disclosed in Akinleye et al., Journal of Hematology & Oncology, 2013, 6:59, which is incorporated herein by reference in its entirety.

[0760] Syk inhibitors include, but are not limited to, selutinib (4-(cyclopropylamino)-2-((4-(4-(ethylsulfonyl)piperazin-1-yl)phenyl)amino)pyrimidine-5-carboxamide), entrectinib (6-(1H-indazol-6-yl)-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazin-8-amine), fotantinib ([6-({5-fluoro-2-[(3,4,5-trimethoxyphenyl)amino]-4-pyrimidinyl}amino 6-({5-fluoro-2-[(3,4,5-trimethoxyphenyl)amino)-4-pyrimidinyl)amino)-2,2-dimethyl-3-oxo-2H-pyrido[3,2-b][1,4]oxazin-4(3H)-yl)methyl phosphate sodium salt), BAY 61-3606 (2-(7-(3,4-(3,4-dimethoxyphenyl)imidazo[1,2-c]pyrimidin-5-ylamino)nicotinamide HCl), RO9021 (6-[(1R,2S)-2-amino-cyclohexylamino]-4-(5,6-dimethyl-pyridin-2-ylamino)-pyridazine-3-carboxamide), imatinib ( 4-[(4-methylpiperazin-1-yl)methyl]-N-(4-methyl-3-{[4-(pyridin-3-yl)pyrimidin-2-yl]amino}phenyl)benzamide), staurosporine, GSK143 (2-((3R,4R)-3-aminotetrahydro-2H-pyran-4-yl)amino)-4-(p-tolylamino)pyrimidine-5-carboxamide), PP2 (1-(tert-butyl)-3-(4-chlorophenyl)-1H-pyrazolo[3-d]pyrimidin-4-amine), PRT-060318 (2-(((1R,2S)-2-aminocyclohexyl)amino)-4-(m-tolylamino)pyrimidine-5-carboxamide), PRT-062607 (4-(3-

[0015] R112 (3,3'-(5-fluoropyrimidine-2,4-diyl)bis(azanediyl)diphenol), R348 (3-ethyl-4-methylpyridine), R406 (6-((5-fluoro-2-((3,4,5-trimethoxyphenyl)amino)pyrimidin-4-yl)amino)-2,2-dimethyl-2H-pyrido[3,2-b][1,4]oxazin-3(4H)-one), piceatannol (3-hydroxyresveratrol), YM193306 (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643), 7-azaindole, piceatannol, ER-27319 (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), Compound D (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), PRT060318 (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem.2012, 55, 3614-3643, incorporated herein in its entirety), luteolin (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), apigenin (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), quercetin (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), fisetin (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), myricetin (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), morin (see Singh et al., Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety).

[0761] In certain embodiments, the bioactive agent is a c-MET inhibitor, for example, crizotinib ( CRIZONIX™), tepotinib (XL880, EXEL-2880, GSK1363089, GSK089), or tivatinib (ARQ197).

[0762] In certain embodiments, the bioactive agent is an AKT inhibitor, including but not limited to MK-2206, GSK690693, perifosine, (KRX-0401), GDC-0068, triciribine, AZD5363, honokiol, PF-04691502 and miltefosine, a FLT-3 inhibitor, including but not limited to P406, dovitinib, quizartinib (AC220), amvatinib (MP-440), tandutinib (MLN518), ENMD-2076 and KW-2449, or a combination thereof.

[0763] In certain embodiments, the active agent is an mTOR inhibitor. Examples of mTOR inhibitors include, but are not limited to, rapamycin and its analogs, everolimus, Temsirolimus, defolimus, sirolimus, and defolimus.

[0764] In certain embodiments, the bioactive agent is a RAS inhibitor. Examples of RAS inhibitors include, but are not limited to, Reolysin and siG12D LODER.

[0765] In certain embodiments, the bioactive agent is a HSP inhibitor.HSP inhibitors include, but are not limited to, geldanamycin or 17-N-allylamino-17-demethoxygeldanamycin (17AAG) and radicicol.

[0766] Other biologically active compounds include, for example, everolimus, trabectedin, nab-paclitaxel, TLK286, AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, FLT-3 inhibitors, VEGFR inhibitors, Aurora kinase inhibitors, PIK-1 modulators, HDAC inhibitors, c-MET inhibitors, PARP inhibitors, Cdk inhibitors, IGFR-TK inhibitors, anti-HGF antibodies, focal adhesion kinase inhibitors, Map kinase (MEK) inhibitors, VEGF TRAP antibody, pemetrexed, panitumumab, amrubicin, ogavuzumab, Lep-etu, nolatrexate, azd2171, batabulin, ofatumumab, zalimumab, edotecarin, tetrandrine, rubitecan, patozuzuril, oblimersen, tremelimumab, ipilimumab, gossypol, Bio 111, 131-I-TM-601, ALT-110, BIO 140, CC 8490, cilengitide, gimatidecan, IL13-PE38QQR, INO 1001, IpdR1 KRX-0402, methionine, LY317615, neuradiab, vitespan, Rta 744, Sdx 102, talampanel, atrasentan, Xr 311, romidepsin, ADS-100380, sunitinib, 5-fluorouracil, vorinostat, etoposide, gemcitabine, doxorubicin, liposomal doxorubicin, 5'-deoxy-5-fluorouracil, vincristine, temozolomide, ZK-304709, seliciclib, PD0325901, AZD-6244, capecitabine, L-glutamic acid, N-[4-[2-(2-amino)- [4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-, disodium salt, heptahydrate, camptothecin, PEG-labeled irinotecan, tamoxifen, toremifene citrate, anastrozole, exemestane, letrozole, DES (diethylstilbestrol), estradiol, estrogen, conjugated estrogens, bevacizumab, IMC-1C11, CHIR-258);3-[5-(Methylsulfonylpiperidinylmethyl)-indolyl]-quinolone, vatalanib, AG-013736, AVE-0005, goserelin acetate, leuprorelin acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, megestrol acetate, raloxifene, bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714; TAK-165, HKI-272, lapatinib, calatoninib, ABX-EGF antibody, Erbitux, EKB-569, PKI-166, GW-572016, lonafarnib, BMS-214662, tebuconazole Pifarnib, amifostine, NVP-LAQ824, suberoylanilide hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951, aminoglutethimide, arnsacrine, anagrelide, L-asparaginase, Bacillus Calmette-Guérin (BCG) vaccine, adriamycin, bleomycin, buserelin, busulfan, carboplatin, carmustine, levofloxacin, cisplatin, cladribine, clodronate, cyproterone acetate, cytarabine, dacarbazine, dactinomycin D, daunorubicin, diethylstilbestrol, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, Gemcitabine, hydroxyurea, idarubicin, ifosfamide, imatinib, leuprorelin, levamisole, lomustine, nitrogen mustard, melphalan, 6-mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxaliplatin, pamidronate, pentostatin, plicamycin, porfimer, procarbazine, raltitrexed, rituximab , streptozotocin, teniposide, testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine, 13-cis retinoic acid, phenylalanine mustard, uramustine, estramustine, hexamethylmelamine, floxuridine, 5-deoxyuridine, cytarabine, 6-mercaptopurine, deoxycodone, calcitriol, valrubicin, mithramycin, vinblastine, vinorelbine, topotecan, razoxane, and marimastatin He, COL-3, nevasastatin, BMS-275291, squalamine, endostatin, SU5416, SU6668, EMD121974, interleukin-12, IM862, angiostatin, vitamin, droloxifene, idoxifene, spironolactone, finasteride, cimetidine, trastuzumab, denileukin, gefitinib, bortezomib, paclitaxel, without Polyethylene castor oil-based paclitaxel, docetaxel, epothilone B, BMS-247550, BMS-310705, droloxifene, 4-hydroxytamoxifen, pinzazoxifene, ERA-923, arzopfen, fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE-424, HMR-3339, ZK186619, topotecan, PTK787 / ZK 222584, VX-745, PD 184352, rapamycin, 40-O-(2-hydroxyethyl)-rapamycin, temsirolimus, AP-23573, RAD001, ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646, wortmannin, ZM336372, L-779,450, PEG-filgrastim, darbepoietin, erythropoietin, granulocyte colony-stimulating factor, zoledronic acid, prednisone, cetuximab, granulocyte macrophage colony-stimulating factor, histrelin, pegylated interferon alfa-2a, interferon alfa-2a, pegylated interferon alfa-2b, interferon alfa-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane, alemtuzumab, all-trans retinoic acid, ketoconazole, interleukin-2, megestrol acetate, immune globulin, nitrogen mustard, methylprednisolone, ibritumomab tiuxetan, and androgens , decitabine, altretamine, bexarotene, tositumomab, arsenic trioxide, cortisone, edidronic acid, mitotane, cyclosporine, liposomal daunorubicin, Edwina-asparaginase, strontium 89, casopitant, netupitant, NK-1 receptor antagonist, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, methylprednisolone, prochlorperazine, granisetron, ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin, epoetin alfa, darbepoetin alfa, and mixtures thereof.

[0767] In certain embodiments, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 is administered in combination with ifosfamide.

[0768] In certain embodiments, the bioactive agent is selected from, but not limited to, imatinib mesylate. Dasatinib Nilotinib Bosutinib Trastuzumab Trastuzumab-DM1, pertuzumab Lapatinib Gefitinib Erlotinib Cetuximab Panitumumab Vandetanib Vemurafenib Vorinostat Romidepsin Besarodine alitretinoin retinoic acid Carfilzomib Pralatrexate Bevacizumab ziv-aflibercept Sorafenib Sunitinib Pazopanib Regorafenib and cabozantinib

[0769] In certain aspects, the bioactive agent is an anti-inflammatory agent, a chemotherapeutic agent, a radiotherapeutic agent, an adjunctive therapeutic agent, or an immunosuppressant.

[0770] Suitable chemotherapeutic bioactive agents include, but are not limited to, radioactive molecules, toxins (also known as cytotoxins or cytotoxic agents, which include any agent that is detrimental to cell viability), and liposomes or other vesicles containing chemotherapeutic compounds. Common anticancer agents include: vincristine or liposomal vincristine Daunorubicin (daunorubicin or ) or doxorubicin Cytosine arabinoside (ara-C or ), L-asparaginase or PEG-L-asparaginase (PEG-L-asparaginase or ), etoposide (VP-16), teniposide 6-Mercaptopurine (6-MP or ), methotrexate, cyclophosphamide Prednisone, dexamethasone Imatinib Dasatinib Nilotinib Bosutinib and ponatinib

[0771] Examples of additional suitable chemotherapeutic agents include, but are not limited to, 1-dehydrotestosterone, 5-fluorouracil ponatinib, 6-mercaptopurine, 6-thioguanine, actinomycin D, doxorubicin, aldesleukin, alkylating agents, allopurinol sodium, hexamethylmelamine, amifostine, anastrozole, anthracyclin (AMC), antimitotic agents, cisplatin (II) (DDP) (cisplatin), diaminodichloroplatinum, anthracyclines, antibiotics, antimetabolites, asparaginase, live BCG (intravesical), betamethasone sodium phosphate and betamethasone acetate, bicalutamide, bleomycin sulfate, busulfan, calcium leucovorin, calicinomycin, capecitabine, avena cava, carboplatin, lomustine (CCNU), carmustine (BSNU), chlorambucil, cisplatin, cladribine, colchicine, conjugated estrogens, cyclophosphamide, cyclothosphamide, cytarabine, cytochalasin B, carboplatin, dacarbazine, dactinomycin, dactinomycin D (formerly actinomycin), daunorubicin hydrochloride, daunorubicin citrate, denileukin, dexrazoxane, mannitol, dihydroxyanthraquinone, docetaxel, dolasetron mesylate, doxorubicin hydrochloride, dronabinol, Escherichia coli L-asparaginase, emetine, epoetin-α, Erwinia lactamase L-asparaginase, esterified estrogens, estradiol, estrogen Sodium phosphate of ciprofloxacin, ethidium bromide, ethinyl estradiol, etidronic acid, etoposide citrororum factor, etoposide phosphate, filgrastim, floxuridine, fluconazole, fludarabine phosphate, fluorouracil, flutamide, folinic acid, gemcitabine hydrochloride, glucocorticoids, goserelin acetate, gramicidin D, granisetron hydrochloride, hydroxyurea, idarubicin hydrochloride, ifosfamide, interferon α-2b, irinotecan hydrochloride, letrozole, leucovorin calcium, leuprorelin hydrochloride, levamisole hydrochloride, lidocaine, lomustine, maytansine, nitrogen mustard hydrochloride, medroxyprogesterone acetate, megestrol acetate, melphalan hydrochloride, mercaptopurine, mesna, methotrexate, methyltestosterone, Mithramycin, mitomycin C, mitotane, mitoxantrone, nilutamide, octreotide acetate, ondansetron hydrochloride, paclitaxel, pamidronate disodium, pentostatin, pilocarpine hydrochloride, pirlimycin, polyphenylpropan 20 implant containing carmustine, porfimer sodium, procaine, procarbazine hydrochloride, propranolol, rituximab, sargramostim, streptozotocin, tamoxifen, paclitaxel, teniposide, testolactone, tetracaine, thiotepa chlorambucil, thioguanine, thiotepa, topotecan hydrochloride, toremifene citrate, trastuzumab, tretinoin, valrubicin, vinblastine sulfate, vincristine sulfate, and vinorelbine tartrate.

[0772] In some embodiments, a pharmaceutical composition comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of Compound 1 is administered in combination with a chemotherapeutic agent (e.g., a cytotoxic agent or other compounds useful for treating cancer). Examples of chemotherapeutic agents include alkylating agents, antimetabolites, folic acid analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodophyllotoxins, antibiotics, L-asparaginase, topoisomerase inhibitors, interferon, platinum coordination complexes, anthracenedione-substituted urea, methylhydrazine derivatives, adrenocortical inhibitors, adrenocortical steroids, progesterone, estrogens, antiestrogens, androgens, antiandrogens and gonadotropin-releasing hormone analogs. Also included are 5-fluorouracil (5-FU), folinic acid (LV), binutuzum, oxaliplatin, capecitabine, paclitaxel and docetaxel. Non-limiting examples of chemotherapeutic agents include alkylating agents, such as thiotepa and cyclophosphamide; alkyl sulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodepa, carboquinone, metodepa, and uredepa; ethyleneimines and methylmelamines, including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trishydroxymethylmelamine; annonaceous lactones (particularly bratacin and bratacinone); camptothecins (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its synthetic analogues adolesine, carzelesin, and biszelesin); candidiasis (particularly candidiasis 1 and candidiasis 8); dolastatin; Duocarmycins (including synthetic analogs, KW-2189 and CB1-TM1); acanthopanax; hyoscyamine; stoloniferol; spongestatin; nitrogen mustards, such as chlorambucil, naphthiazolin, clofosamide, estramustine, ifosfamide, dichloromethyldiethylamine, nitrogen oxide mustard hydrochloride, melphalan, nembicevicin, phenylephrine, prednimustine, trolofosamide, uramustine; nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics, such as estradiol antibiotics (e.g., calicheamicins, especially calicheamicin gammall and calicheamicin omegall (see, e.g., Agnew, Chem. Int. Ed., 1996). Engl. 33:183-186 (1994); danonomycins, including danonomycin A; bisphosphonates, such as clodronate; espamycin; and the neocarcin chromophore and related chromoprotein estradiyne antibiotic chromophores), aclarubicin, actinomycin, anthramycin, azaserine, bleomycin, actinomycin, carabicin, caminomycin, carbophilin, chromomycin, actinomycin D, daunorubicin, detoxibacin, 6-diazo-5-oxo-L-norleucine, (doxorubicin, including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolidino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, mexilomycin, mitomycins such as mitomycin C, mycophenolic acid, nogamycin, olivomycin, peplomycin, potfimycin, puromycin, triferric doxorubicin, rhodorubicin, streptomycin, streptozotocin, tuberculin antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs, such as leucovorin, methotrexate, pteropterin, trimetrexate; purine analogs, such as fludarabine, 6-mercaptopurine, thiopurine, thioguanine; pyrimidine analogs, such as ancitabine, obinutuzumab, 6-azauridine, carmofur, cytarabine, dideoxyuridine, glycosides, doxifluridine, enocitabine, floxuridine; androgens, such as calutosterone, drostanolone propionate, cyclothiocarbamate, melastane, and testolactone; antiadreners, such as aminoglutethimide, mitotane, and trilostane; folic acid supplements, such as folic acid; aceglucuronolide; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; amustine; bisantrene; binutuzumab; defofamine; colcemid; diazocine; elfomithine; elixirsium acetate; epothilones; ethoxydimidine; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids, such as maytansine and ansamitocin; mitoguanidine; mitoxantrone; mopidarol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; Polysaccharide complex (JHS Natural Products, Eugene, OR); razoxane; rhizophyllin; schizophyllin; spirogermanamine; tenuisporic acid; triazoline; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A, and anguidine); urethane; vindesine; dacarbazine; mannomustine; dibromomannitol; dibromodulcitol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; obinutuzumab, for example (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ), Polyethylene-free castor oil albumin-engineered paclitaxel nanoparticle formulation (American Pharmaceutical Partners, Schaumberg, IL) and docetaxel (Rhone-Poulenc Rorer, Antony, France), binutizumab; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum coordination compounds, such as cisplatin, oxaliplatin, and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; Vinorelbine; can kill tumors; teniposide; edatrexate; daunorubicin; aminopterin; xeloda; ibandronate; irinotecan (e.g., CPT-11); topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; and any of the above-mentioned pharmaceutically acceptable salts, acids, or derivatives. Two or more chemotherapeutic agents can be used in a mixture for combined administration with a pharmaceutical composition comprising an effective amount of compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of compound 1. Suitable dosing regimens for combined chemotherapy are known in the art. For example, Saltz et al., Proc.Am.Soc.Clin.Oncol.18:233a (1999) and Douillard et al., Lancet 355 (9209): 1041-1047 (2000) describe combined dosing regimens.

[0773] Additional therapeutic agents that can be administered in combination with a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 may include bevacizumab, sunitinib, sorafenib, 2-methoxyestradiol or 2ME2, finasunate, vatalanib, vandetanib, aflibercept, volociximab, etaracizumab (MEDI-522), cilengitide, erlotinib, cetuximab, panitumumab, gefitinib, trastuzumab, dovitinib, fentolimumab, atacicept, rituximab, alemtuzumab, aldesleukin, tadalafil, tocilizumab, temsirolimus, edema, tadalafil ... Viromax, rukalimumab, dacillinam, HLL1, huN901-DM1, atilimod, natalizumab, bortezomib, carfilzomib, marizomib, tanemycin, saquinavir mesylate, ritonavir, nelfinavir mesylate, indinavir sulfate, belinostat, panobinostat, lexalimumab, dulanamine, ABT-737, oblimersen, pratazone, tapimod, P276-00, enzastaurin, tipifarnib, perifosine, imatinib, dasatinib, lenalidomide, thalidomide, simvastatin, celecoxib, bazedoxifene, AZD4547, rilotumumab, oxaliplatin PD0332991, ribociclib (LEE011), amebaciclib (LY2835219), HDM201, fulvestrant Exemestane PIM447, ruxolitinib (INC424), BGJ398, nexitozumab, pemetrexed and ramucirumab (IMC-1121B).

[0774] In certain embodiments, additional therapy is a monoclonal antibody (Mab). Some monoclonal antibodies stimulate an immune response that destroys cancer cells. Similar to antibodies naturally produced by B cells, these monoclonal antibodies may "coat" the surface of cancer cells, thereby triggering their destruction by the immune system. For example, bevacizumab targets vascular endothelial growth factor (VEGF), a protein secreted by tumor cells and other cells in the tumor microenvironment that promotes tumor blood vessel development. When combined with bevacizumab, VEGF cannot interact with its cell receptors, thereby preventing the signal transduction that causes new blood vessel growth. Monoclonal antibodies that bind to cell surface growth factor receptors prevent the targeted receptors from sending normal growth-promoting signals. They can also trigger apoptosis and activate the immune system to destroy tumor cells.

[0775] In one aspect of the invention, the bioactive agent is an immunosuppressant. The immunosuppressant may be a calcineurin inhibitor, such as cyclosporine or an ascomycin, such as cyclosporine A. FK506 (tacrolimus), pimecrolimus, mTOR inhibitors such as rapamycin or its derivatives such as sirolimus Everolimus Temsirolimus, zotarolimus, biolipid-7, biolipid-9, rapamycin and its analogs, such as defotilimus, azathioprine, campath1H, S1P receptor modulators, such as fingolimod or its analogs, anti-IL-8 antibodies, mycophenolic acid or its salts, such as sodium salt, or its prodrugs, such as mycophenolate mofetil OKT3 (ORTHOCLONE ),prednisone, Buquina sodium, OKT4, T10B9.A-3A, 33B3.1, 15-deoxyspergualin, treperolimus, leflunomide CTLAI-Ig, anti-CD25, anti-IL2R, basiliximab Daclizumab Mizoribine, methotrexate, dexamethasone, ISAtx-247, SDZ ASM 981 (pimecorimus, ), CTLA4lg (abatacept), belatacept, LFA3lg, etanercept (by Immunex as Sold), Adalimumab Infliximab Anti-LFA-1 antibody, natalizumab Envoy, gavilimumab, antithymocyte immunoglobulin, cilizumab, afacept, efalizumab, podesan, mesalamine, ansacral, codeine phosphate, benolate, fenbufen, naproxen, diclofenac, etodolac, and indomethacin, aspirin, and ibuprofen.

[0776] In some embodiments, the bioactive agent is a therapeutic agent that is a biologic such as a cytokine (e.g., an interferon or an interleukin (e.g., IL-2)) used for cancer treatment. In some embodiments, the biologic is an anti-angiogenic agent, such as an anti-VEGF agent, such as bevacizumab. In some embodiments, biologics are immunoglobulin-based biologics, such as monoclonal antibodies (e.g., humanized antibodies, fully human antibodies, Fc fusion proteins, or functional fragments thereof), that agonize targets to stimulate anti-cancer responses or antagonize antigens important for cancer. Such agents include (rituximab); (daclizumab); (basiliximab); (palivizumab); (infliximab); (trastuzumab); (gemtuzumab); (alemtuzumab); (ibritumomab tiuxetan); (adalimumab); (omalizumab); (tositumomab-I-131); (efalizumab); (cetuximab); (bevacizumab); (natalizumab); (tocilizumab); (panitumumab); (ranibizumab); (eculizumab); (certolizumab); (golimumab); (canakinumab); (Ustekinumab); (ofatumumab); (denosumab); (motavizumab); (Raxicumab); (Belimumab); (ipilimumab); (Bentuximab); (Pertuzumab); (enmetazuzumab); and (Obinutuzumab). Also included are antibody-drug conjugates.

[0777] Combination therapy may include non-drug therapeutic agents. For example, a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1, may be administered in addition to radiation therapy, cryotherapy, hyperthermia, and / or surgical resection of tumor tissue.

[0778] Pharmaceutical composition

[0779] Provided are novel and advantageous pharmaceutical compositions suitable for administration to humans, comprising an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of Compound 1 of the present invention. In certain embodiments, pharmaceutical compositions comprising Compound 1 or a pharmaceutically acceptable salt thereof are prepared from a crystalline form described herein, such as Compound 1 Form B. For example, Compound 1 Form B can be dissolved and then spray-dried to form a solid spray-dried dispersion with one or more pharmaceutically acceptable excipients.

[0780] Pharmaceutically acceptable excipients suitable for use as carriers or diluents are known to those skilled in the art and can be used in a variety of formulations. For example, see Remington's Pharmaceutical Sciences, 23rd edition, A. Adejare, ed., Academic Press (2020); Handbook of Pharmaceutical Excipients, 6th edition, RC Rowe, PJ Sheskey, ME Quinn, eds., American Pharmaceutical Association, and Pharmaceutical Press (2009); and Handbook of Pharmaceutical Additives, 3rd edition, compiled by Michael and Irene Ash, Synapse Information Resources (2007).

[0781] The pharmaceutical composition can be formulated into any pharmaceutically useful form, such as a solid dosage form, liquid, aerosol, cream, gel, pill, injection or infusion solution, capsule, tablet, syrup, transdermal patch, subcutaneous patch, dry powder, inhalation preparation, in a medical device, suppository, oral or sublingual preparation, parenteral preparation or ophthalmic solution. Some dosage forms, such as tablets and capsules, are subdivided into appropriately sized unit doses containing an appropriate amount of the active ingredient, such as an effective amount to achieve the desired purpose.

[0782] The carrier includes an excipient and a diluent and should be of sufficiently high purity and sufficiently low toxicity to make it suitable for administration to the patient being treated in an effective amount. The carrier may be inert, or it may have its own pharmaceutical benefits. The amount of carrier used in conjunction with Compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of Compound 1 is sufficient to provide the actual amount of material administered per unit dose of the compound.

[0783] Types of carriers include, but are not limited to, binders, buffers, colorants, diluents, disintegrants, emulsifiers, flavorings, glidants, lubricants, preservatives, stabilizers, surfactants, tableting agents, and wetting agents. Some carriers may be listed in more than one category; for example, a vegetable oil may be used as a lubricant in some formulations and as a diluent in others. Exemplary pharmaceutically acceptable carriers include sugars, starches, cellulose, tragacanth powder, malt, gelatin; talc, and vegetable oils. Optional active agents may be included in the pharmaceutical composition that do not substantially interfere with the activity of Compound 1.

[0784] An effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or a crystalline form of Compound 1 disclosed herein, can be administered orally, topically, systemically, parenterally, by inhalation or spray, sublingually, by implant (e.g., an implant inserted into a tumor or abnormal cell proliferation), transdermally, orally, rectally, as an ophthalmic solution, by injection (including intravenous, intraaortic, intracranial, subdermal, intraperitoneal, subcutaneous), nasally, sublingually, or rectally, or by other means, in the form of a unit dosage formulation containing a conventional pharmaceutically acceptable carrier.

[0785] In certain embodiments, pharmaceutical compositions prepared from or comprising Compound 1 or a pharmaceutically acceptable salt or crystalline form of Compound 1 of the present invention are provided in solid, gel, or liquid dosage forms for oral delivery.

[0786] In certain embodiments, the pharmaceutical composition comprises compound 1 or a pharmaceutically acceptable salt thereof, a dissolution enhancer, a penetration enhancer, a filler, one or more adhesives and / or glidants and one or more flow aids. Non-limiting examples of pharmaceutically acceptable excipients include hypromellose (e.g., hypromellose acetate succinate), vitamin E (e.g., d-α-tocopheryl polyethylene glycol succinate), mannitol, cellulose (e.g., microcrystalline cellulose), cross-linked sodium carboxymethyl cellulose, silicon dioxide (e.g., untreated pyrogenic colloidal silicon dioxide) and magnesium stearate. In certain embodiments, the pharmaceutical composition according to the present invention is formulated into a unit dosage form, such as an oral unit dosage form. In certain embodiments, the pharmaceutical composition according to the present invention is formulated into a tablet dosage form.

[0787] In certain embodiments, pharmaceutical compositions comprising Compound 1 or a pharmaceutically acceptable salt thereof comprise one or more of the following excipients.

[0788]

[0789]

[0790] In certain embodiments, methods for preparing pharmaceutical compositions comprising compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of compound 1 formed according to the present invention are also provided. In certain embodiments, a method for preparing a pharmaceutical composition comprising compound 1 comprises: (i) a spray drying step to provide a spray-dried intermediate (SRI) comprising compound 1 and a pharmaceutically acceptable excipient; (ii) a granulation step to provide particles containing compound 1 and a pharmaceutically acceptable excipient, the particles having a desired bulk density of about 0.4 to 0.6 g / mL (e.g., about 0.48 to 0.54 g / mL); and (iii) a tableting step to provide a pharmaceutical composition in the form of an oral unit dosage comprising compound 1 and a pharmaceutically acceptable excipient.

[0791] In certain embodiments, the dosage strength of the pharmaceutical composition is at least about 10 mg, 20 mg, 40 mg, 80 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg, 280 mg, 300 mg, 320 mg, 340 mg, 360 mg, 380 mg, 400 mg, 440 mg, 480 mg, 520 mg, 560 mg, 600 mg, 640 mg, 680 mg, 720 mg, 760 mg, or 800 mg, which can be administered (in a non-limiting manner) once a week, twice a week, three times a week, four times a week, five times a week, six times a week, once a day (QD), or twice a day (BID), optionally on days 1-7, 1-14, 1-21, or 1-28 of a 28-day treatment cycle. In some embodiments, compound 1 or a pharmaceutically acceptable salt thereof or a crystalline form of compound 1 of the present invention is provided as a soft shell capsule or tablet for oral administration. In certain embodiments, the dosage strength of the solid or gel dosage form is at least about 10 mg, 20 mg, 40 mg, 80 mg, 140 mg, 240 mg, 360 mg or 480 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 800 mg, which can be administered once a week, twice a week, three times a week, four times a week, five times a week, six times a week, once a day (QD) or twice a day (BID) in a non-limiting manner, optionally on days 1-7, 1-14, 1-21 or 1-28 of a 28-day treatment cycle.

[0792] In certain aspects, the pharmaceutical composition is administered orally once or twice daily (BID) as needed.

[0793] Embodiments of the pharmaceutical composition include:

[0794] 1. In certain embodiments, Compound 1 is present in an effective amount in a pharmaceutical composition comprising at least three, four, or all of a dissolution enhancer, a permeation enhancer, a filler, a binder / glidant, and / or a mucoadhesive / disintegrant.

[0795] 2. The pharmaceutical composition according to embodiment 1, wherein the pharmaceutical composition is provided in tablet dosage form.

[0796] 3. The pharmaceutical composition according to any one of embodiments 1-2, wherein the pharmaceutical composition is prepared from a crystalline form of Compound 1, for example, by dissolving and then spray drying Compound 1 Form B.

[0797] 4. The pharmaceutical composition according to any one of embodiments 1 to 3, wherein the pharmaceutical composition further comprises a flow aid and a lubricant.

[0798] 5. The pharmaceutical composition according to any one of embodiments 1-4, wherein the pharmaceutical composition comprises about 5% to about 20% by weight of Compound 1.

[0799] 6. The pharmaceutical composition according to any one of embodiments 1 to 5, wherein the dissolution enhancer is hypromellose or a hypromellose derivative.

[0800] 7. The pharmaceutical composition according to embodiment 6, wherein the hypromellose derivative is hypromellose acetate succinate.

[0801] 8. The pharmaceutical composition according to any one of embodiments 1 to 7, wherein the penetration enhancer is vitamin E or a vitamin E derivative.

[0802] 9. The pharmaceutical composition according to embodiment 8, wherein the vitamin E derivative is d-α-tocopheryl polyethylene glycol succinate.

[0803] 10. The pharmaceutical composition according to any one of embodiments 1 to 9, wherein the filler is mannitol.

[0804] 11. The pharmaceutical composition according to any one of embodiments 1 to 10, wherein the binder / glidant is cellulose.

[0805] 12. The pharmaceutical composition according to embodiment 11, wherein the binder / glidant is microcrystalline cellulose.

[0806] 13. The pharmaceutical composition according to any one of embodiments 1 to 12, wherein the mucoadhesive / disintegrant is croscarmellose or a croscarmellose derivative.

[0807] 14. The pharmaceutical composition according to embodiment 13, wherein the croscarmellose derivative is croscarmellose sodium.

[0808] 15. The pharmaceutical composition according to any one of embodiments 4-14, wherein the flow aid is colloidal silicon dioxide.

[0809] 16. The pharmaceutical composition according to embodiment 15, wherein the colloidal silicon dioxide is untreated fumed colloidal silicon dioxide.

[0810] 17. The pharmaceutical composition according to any one of embodiments 4-16, wherein the lubricant is magnesium stearate.

[0811] 18. The pharmaceutical composition according to any one of embodiments 1-17, wherein the pharmaceutical composition comprises from about 15% to about 55% by weight of the solubility enhancer.

[0812] 19. The pharmaceutical composition according to any one of embodiments 1-18, wherein the pharmaceutical composition comprises from about 1 wt% to about 10 wt% of the penetration enhancer.

[0813] 20. The pharmaceutical composition according to any one of embodiments 1-19, wherein the pharmaceutical composition comprises from about 20% to about 40% by weight of filler.

[0814] 21. The pharmaceutical composition according to any one of embodiments 1-20, wherein the pharmaceutical composition comprises from about 10% to about 30% by weight of the binder / glidant.

[0815] 22. The pharmaceutical composition according to any one of embodiments 1-21, wherein the pharmaceutical composition comprises from about 0.5% to about 5% by weight of the mucoadhesive / disintegrant.

[0816] 23. The pharmaceutical composition according to any one of embodiments 1-22, wherein the pharmaceutical composition comprises about 10% by weight of Compound 1.

[0817] 24. The pharmaceutical composition of any one of embodiments 1-23, wherein the pharmaceutical composition comprises about 35% by weight of the solubility enhancer.

[0818] 25. The pharmaceutical composition according to any one of embodiments 1-24, wherein the pharmaceutical composition comprises about 5% by weight of the permeation enhancer.

[0819] 26. The pharmaceutical composition according to any one of embodiments 1-25, wherein the pharmaceutical composition comprises about 28% by weight of filler.

[0820] 27. The pharmaceutical composition according to any one of embodiments 1-26, wherein the pharmaceutical composition comprises about 20% by weight of the binder / glidant.

[0821] 28. The pharmaceutical composition according to any one of embodiments 1-27, wherein the pharmaceutical composition comprises about 2% by weight of the mucoadhesive / disintegrant.

[0822] 29. The pharmaceutical composition according to any one of embodiments 4-28, wherein the pharmaceutical composition comprises about 0.5% by weight of a flow aid.

[0823] 30. The pharmaceutical composition according to any one of embodiments 4-29, wherein the pharmaceutical composition comprises about 0.6% by weight of a lubricant.

[0824] Pharmaceutical composition comprising a crystalline form of compound 1

[0825] Compound 1 crystalline form can be administered as a neat chemical, but is typically administered as a pharmaceutical composition comprising an effective amount of Compound 1 crystalline form to a host, typically a human, in need of such treatment for any of the conditions described herein. Accordingly, the present disclosure provides pharmaceutical compositions comprising an effective amount of Compound 1 crystalline form and at least one pharmaceutically acceptable carrier or excipient as described herein. The pharmaceutical composition may comprise Compound 1 crystalline form as the sole active agent, or in alternative embodiments, comprise Compound 1 crystalline form and at least one additional active agent.

[0826] In certain embodiments, the pharmaceutical composition is a dosage form comprising about 0.001 mg to about 1000 mg, about 0.01 mg to about 800 mg, about 1 mg to about 800 mg, or about 200 mg to about 600 mg of a crystalline form of Compound 1 in a unit dosage form, and optionally comprising about 0.1 mg to about 2000 mg, about 10 mg to about 1000 mg, about 100 mg to about 800 mg, or about 200 mg to about 600 mg of an additional active agent. Examples are dosage forms having at least about or no more than 0.001, 0.005, 0.010, 0.10, 1, 5, 10, 25, 50, 100, 200, 250, 300, 400, 500, 600, 700, or 750 mg of a crystalline form of Compound 1.

[0827] In certain embodiments, the dosage form of the pharmaceutical composition comprises about 50 mg of a crystalline form of Compound 1. In certain embodiments, the dosage form of the pharmaceutical composition comprises about 100 mg of a crystalline form of Compound 1. In certain embodiments, the dosage form of the pharmaceutical composition comprises about 150 mg of a crystalline form of Compound 1. In certain embodiments, the dosage form of the pharmaceutical composition comprises about 200 mg of a crystalline form of Compound 1. In certain embodiments, the dosage form of the pharmaceutical composition comprises about 250 mg of a crystalline form of Compound 1. In certain embodiments, the dosage form of the pharmaceutical composition comprises about 300 mg of a crystalline form of Compound 1. In certain embodiments, the dosage form of the pharmaceutical composition comprises about 400 mg of a crystalline form of Compound 1. In certain embodiments, the dosage form of the pharmaceutical composition comprises about 500 mg of a crystalline form of Compound 1. In certain embodiments, the dosage form of the pharmaceutical composition comprises about 600 mg of a crystalline form of Compound 1. In certain embodiments, the dosage form of the pharmaceutical composition comprises about 700 mg of a crystalline form of Compound 1. In certain embodiments, the dosage form of the pharmaceutical composition comprises about 800 mg of a crystalline form of Compound 1.

[0828] In certain embodiments, the crystalline form of Compound 1 is administered to a patient in need thereof once or twice daily.

[0829] In certain embodiments, pharmaceutical compositions according to the present invention include a crystalline form of Compound 1 and a pharmaceutically acceptable excipient. In certain embodiments, pharmaceutically acceptable excipients include, but are not limited to, hypromellose acetate succinate, vitamin E TPGS (d-α-tocopheryl polyethylene glycol succinate), mannitol, microcrystalline cellulose, croscarmellose sodium, untreated pyrogenic colloidal silicon dioxide, and magnesium stearate.

[0830] Pharmaceutical compositions can be formulated for oral administration. These compositions can contain any amount of Compound 1 crystalline form to achieve the desired result, for example, 0.1 to 99 weight percent (wt.%), typically at least about 5 wt% of Compound 1 crystalline form. Some embodiments contain about 5 wt% to about 30 wt%, about 25 wt% to about 50 wt%, or about 5 wt% to about 75 wt% Compound 1 crystalline form.

[0831] Example

[0832] General synthesis

[0833] The compounds described herein can be prepared by methods known to those skilled in the art. In one non-limiting example, the disclosed compounds can be prepared using the following schemes.

[0834] For convenience, compounds of the present invention having stereocenters may be drawn without stereochemistry. Those skilled in the art will recognize that pure enantiomers and diastereomers can be prepared by methods known in the art. Examples of methods for obtaining optically active materials include at least the following:

[0835] i) Physical separation of crystals - a technique that artificially separates macroscopic crystals of individual enantiomers. This technique can be used if crystals of individual enantiomers exist, i.e., if the material is an aggregate and the crystals are visually distinct;

[0836] ii) Simultaneous crystallization - a technique in which the individual enantiomers are crystallized separately from a solution of the racemate, which is possible only when the enantiomers are aggregates in the solid state;

[0837] iii) Enzymatic resolution - a technique that partially or completely separates racemates by virtue of the different reaction rates of the enantiomers with enzymes;

[0838] iv) Enzymatic asymmetric synthesis - a synthetic technique in which at least one step in the synthesis uses an enzymatic reaction to obtain an enantiomerically pure or enriched synthetic precursor of the desired enantiomer;

[0839] v) Chemical asymmetric synthesis - a synthetic technique in which the desired enantiomer is synthesized from achiral precursors under conditions that produce asymmetry (i.e., chirality) in the product, which can be achieved by chiral catalysts or chiral auxiliaries;

[0840] vi) Diastereomeric separation - a technique in which a racemic compound is reacted with an enantiomerically pure reagent (chiral auxiliary) to convert the individual enantiomers into diastereomers. The resulting diastereomers are then separated by chromatography or crystallization due to their now more distinct structural differences, and the chiral auxiliary is subsequently removed to yield the desired enantiomer;

[0841] vii) First- and second-order asymmetric transformations - techniques whereby diastereomers from the racemate rapidly equilibrate to produce a preponderance of the desired enantiomer in a diastereomeric solution, wherein preferential crystallization of the diastereomer from the desired enantiomer disturbs the equilibrium so that, in principle, all of the material is eventually converted to the crystalline diastereomer from the desired enantiomer. The desired enantiomer is then liberated from the diastereomer;

[0842] viii) Kinetic resolution - This technique involves the partial or complete resolution of a racemate (or further resolution of a partially resolved compound) by the different reaction rates of the enantiomers with a chiral, non-racemic reagent or catalyst under kinetic conditions;

[0843] ix) Enantiomer-specific synthesis from non-racemic precursors - synthetic techniques that obtain the desired enantiomer from achiral starting materials and in which the stereochemical integrity is not compromised or is only minimally compromised during the synthesis;

[0844] x) Chiral liquid chromatography - a technique whereby the enantiomers of a racemate are separated in a liquid mobile phase by their differential interactions with the stationary phase (including vial chiral HPLC). The stationary phase may be made of a chiral material, or the mobile phase may contain additional chiral materials to induce this differential interaction;

[0845] xi) Chiral gas chromatography - a technique in which the racemate is volatilized and the enantiomers are separated by differential interactions of a gaseous mobile phase with a column containing an immobilized non-racemic chiral adsorbent phase;

[0846] xii) Extraction with chiral solvents - a technique for separating enantiomers by preferentially dissolving one enantiomer in a specific chiral solvent;

[0847] xiii) Transport across chiral membranes - a technique where the racemate is in contact with a membrane barrier. The barrier separates two miscible fluids, one containing the racemate, and a driving force (e.g., concentration or pressure differential) results in preferential transport across the barrier. The separation occurs due to the non-racemic chiral nature of the membrane, which allows only one enantiomer of the racemate to pass through.

[0848] xiv) In one embodiment simulated moving bed chromatography is used.A wide variety of chiral stationary phases are commercially available.

[0849] The term "nucleophilic aromatic substitution" (S N Ar) refers to the substitution reaction of a nucleophile to replace a good leaving group on an aromatic ring, such as halogen (F, Cl, Br, I), tosylate, mesylate, triflate, etc. N Examples of nucleophiles in the Ar reaction include, but are not limited to, amines (including aromatic amines), alcohols, phenols, and aromatic compounds comprising hydroxyl groups attached to aromatic rings. Nucleophilic aromatic substitution can be carried out at reduced temperatures (below 0°C), room temperature (about 25°C), or elevated temperatures (about 30-200°C). The reaction is carried out in any suitable solvent, including but not limited to DMSO, N,N-dimethylformamide, THF, N,N-dimethylacetamide, 1,4-dioxane, and acetonitrile. The reaction is carried out in the presence of a base (e.g., an inorganic or organic base), including but not limited to NaOH, KOH, Na2CO3, Na3PO4, Cs2CO3, Li2CO3, DIEA (N,N-diisopropylethylamine or Hünig's base).

[0850] The synthesis of the compounds according to the present invention may include steps of protecting and deprotecting some reactive groups (e.g., amino and carboxylic acid hydroxyl groups). Methods for protecting and deprotecting amino and hydroxyl groups and corresponding protecting groups are known to those skilled in the art, for example, Greene's Protective Groups in Organic Synthesis, 5th edition, PGM Wuts, Wiley (2014).

[0851] Example 1: Synthesis of (R)-3-(6-(2-cyano-3,6-difluorophenoxy)-4-oxoquinazolin-3(4H)-yl)-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester - Route 1 (via transamination)

[0852]

[0853] Step 1 can be carried out on a production scale to prepare large quantities of 1-2 for use in the preparation of compound 1. A non-limiting example of such a scaled-up reaction and raw material inputs is provided below.

[0854]

[0855]

[0856]

[0857] 1 H NMR(400MHz,CDCl3),δ,ppm:3.94(m,1H)3.46-3.67(m,4H)3.26-3.39(m,2H)2.06(m,1 H)1.69-1.76(m,1H)1.65(m,1H)1.54-1.62(m,1H)1.48-1.52(m,2H)1.41-1.48(m,11H)

[0858] Step 2 can be carried out on a production scale to prepare large quantities of 1-4 for use in the preparation of compound 1. A non-limiting example of such scale-up reactions and raw material inputs is provided below.

[0859]

[0860]

[0861]

[0862] Step 3 can be performed on a production scale to prepare large quantities of 1-6 for use in the preparation of compound 1. A non-limiting example of this scale-up reaction and raw material input is provided below.

[0863]

[0864]

[0865]

[0866] Step 4 can be carried out on a production scale to prepare large quantities of 1-7 for use in the preparation of compound 1. A non-limiting example of this scale-up reaction and raw material input is provided below.

[0867]

[0868]

[0869]

[0870] Example 2: Synthesis of (R)-3-(6-(2-cyano-3,6-difluorophenoxy)-4-oxoquinazolin-3(4H)-yl)-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester - Route 2 (by diastereomeric crystallization)

[0871]

[0872] Step 1: Synthesis of tert-butyl 3-(hydroxyimino)-1-oxa-8-azaspiro[4.5]decane-8-carboxylate (2-1). To a solution of tert-butyl 3-oxo-1-oxa-8-azaspiro[4.5]decane-8-carboxylate (1-1, 30 g, 117.50 mmol) in ethanol (450 mL) was added sodium acetate (14.46 g, 176.26 mmol) and hydroxylamine hydrochloride (12.25 g, 176.26 mmol). The mixture was stirred at 70 ° C for 1 hour. The reaction mixture was concentrated to remove EtOH and diluted with ethyl acetate (300 mL) and water (300 mL). The aqueous layer was extracted with ethyl acetate (2 x 150 mL) and the combined organic phases were washed with saturated sodium bicarbonate (aqueous, 300 mL), water (300 mL), brine (300 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give tert-butyl 3-(hydroxyimino)-1-oxa-8-azaspiro[4.5]decane-8-carboxylate (2-1, 32 g, crude) as a light yellow oil.

[0873] 1H NMR (400MHz, CDCl3) δ8.71-7.60(m,1H),4.52(s,1H),4.38(s,1H),3.74-3.57(m,2H),3.41 -3.26(m,2H),2.58(s,1H),2.50(s,1H),1.77-1.66(m,2H),1.65-1.54(m,2H),1.46(s,9H).

[0874] Step 2: Synthesis of tert-butyl 3-amino-1-oxa-8-azaspiro[4.5]decane-8-carboxylate (2-2). To a solution of tert-butyl (3E)-3-hydroxyimino-1-oxa-8-azaspiro[4.5]decane-8-carboxylate (2-1, 32 g, 118.38 mmol) in ethanol (600 mL) was added Raney-Ni (6.00 g, 70.03 mmol) and ammonium acetate (19.1 g, 247.79 mmol). The mixture was stirred at 70 ° C for 2 h under a hydrogen atmosphere (15 psi). The reaction mixture was filtered through celite and the filtrate was concentrated. The residue was dissolved in aqueous hydrochloric acid (1 M, 150 mL), and the resulting mixture was washed with methyl tert-butyl ether (2×150 mL), followed by addition of aqueous NaOH (4 M) and adjusted to pH=12. The aqueous layer was extracted with dichloromethane (3 x 150 mL), and the combined organic phases were washed with brine (100 mL), dried over sodium sulfate, filtered and concentrated to give tert-butyl 3-amino-1-oxa-8-azaspiro[4.5]decane-8-carboxylate (2-2, 22.2 g, 86.60 mmol, 73.16% yield) as a light yellow solid.

[0875] 1 H NMR (400MHz, CDCl3) δ4.02-3.85(m,1H),3.70-3.43(m,4H),3.39-3.21(m,2H),2.05(br dd,J=7.5,12.7Hz,1H),1.78-1.53(m,3H),1.51-1.33(m,13H)

[0876] Step 3: Synthesis of (R)-8-(tert-Butoxycarbonyl)-1-oxa-8-azaspiro[4.5]decane-3-amine (R)-2-hydroxy-2-phenylacetate (2-4) (D-mandelate). To a mixture of tert-butyl 3-amino-1-oxa-8-azaspiro[4.5]decane-8-carboxylate (2-2, 25 g, 97.53 mmol) in acetonitrile (350 mL) was added portionwise (2R)-2-hydroxy-2-phenyl-acetic acid (2-3, 7.42 g, 48.76 mmol) at 15° C. (ambient temperature). A precipitate formed and the mixture was stirred at 70° C. for 2 h. After slowly cooling to 15° C., the suspension was stirred at 15° C. for an additional 12 h. The precipitate was filtered, washed with acetonitrile (100 mL), and dried in vacuo. (R)-8-(tert-Butoxycarbonyl)-1-oxa-8-azaspiro[4.5]decan-3-amine (R)-2-hydroxy-2-phenyl acetate (2-4, 19.5 g) was obtained.

[0877] 1 H NMR (400MHz, methanol-d4)δ=7.52-7.45(m,2H),7.35-7.28(m,2H),7.28-7.21(m,1H),4.89(br s,1H),4.01(dd,J=5.4,10.1Hz,1H),3.91-3.79(m,2H),3.65(tdd,J=4.5,8.7,13.1Hz,2H),3.3 2-3.21(m,2H),2.24(dd,J=8.0,13.9Hz,1H),1.80-1.62(m,4H),1.55-1.49(m,1H),1.48(s,9H).

[0878] 1 H NMR (400MHz, DMSO-d6) δ = 7.37 (d, J = 7.3Hz, 2H), 7.28-7.21 (m, 2H), 7.20-7.13 (m, 1H), 4.57 (s, 1H), 3.86 (dd, J = 6.0 ,9.5Hz,2H),3.71(ddd,J=5.6,8.0,10.9Hz,2H),3.62(dd,J=4.9,9.5Hz,2H),3.42(qd,J=4.2,13.1Hz,3H),3.19(br d,J=10.3Hz,3H),2.05(dd,J=8.1,13.4Hz,1H),1.66-1.54(m,3H),1.54-1.45(m,1H),1.45-1.40(m,1H),1.39(s,9H).

[0879] Step 4: Synthesis of (3R)-3-amino-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (1-2). (R)-8-(tert-Butoxycarbonyl)-1-oxa-8-azaspiro[4.5]decane-3-amine (R)-2-hydroxy-2-phenyl acetate salt 2-4 was suspended in water (200 mL) and neutralized with aqueous sodium hydroxide solution (2M, 25 mL). The mixture was extracted with ethyl acetate (3×250 mL), and the combined organic layers were washed with brine (3×100 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to give (3R)-3-amino-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (1-2, 10.3 g, 95.3% ee) as a white solid.

[0880] Steps 3 and 4 can be repeated to obtain 1-2 in higher enantiomeric purity.

[0881] To a mixture of (3R)-3-amino-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (1-2, 10.3 g, 39 mmol) in acetonitrile (350 mL) was added portionwise (2R)-2-hydroxy-2-phenyl-acetic acid (2-3, 5.76 g, 37.8 mmol) at 15°C (ambient temperature) to form a precipitate, and the mixture was then stirred at 70°C for 2 h. After slowly cooling to 15°C, the suspension was stirred at 15°C for an additional 12 h. The precipitate was filtered, washed with acetonitrile (40 mL), and dried under vacuum. (3 g was retained). The remaining D-mandelate salt was suspended in water (80 mL), neutralized with aqueous sodium hydroxide solution (2 M, 20 mL), and extracted with ethyl acetate (3×150 mL). The combined organic layers were washed with brine (3 x 50 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give tert-butyl (3R)-3-amino-1-oxa-8-azaspiro[4.5]decane-8-carboxylate (1-2, 6.5 g, 23.80 mmol, 99.5% ee) as a white solid.

[0882] 1 H NMR (400MHz, CDCl3) δ3.93 (dd, J=5.8, 8.8Hz, 1H), 3.72-3.43 (m, 4H), 3.41-3.2 3(m,2H),2.05(dd,J=7.3,12.6Hz,1H),1.79-1.53(m,4H),1.52-1.34(m,13H).

[0883] Stereochemistry determination of compounds 2-4.

[0884] The mandelate salt 2-4 in Example 2 was determined by X-ray crystallography to be (R)-8-(tert-butoxycarbonyl)-1-oxa-8-azaspiro[4.5]decan-3-amine (R)-2-hydroxy-2-phenylacetate. (For single crystal X-ray crystallography, see Example 32.) This intermediate 2-4 was used in the synthesis of 1-7, which, based on the mechanism of subsequent reactions, is also the R-isomer.

[0885] The stereochemistry of compound 1-7 obtained in Example 2 (Scheme 2) was then compared with that obtained in Example 1 (Scheme 1) using chiral SFC and optical rotation determination.

[0886] Chiral SFC method:

[0887] column: AD-3, 50 × 4.6 mm ID, 3 μm;

[0888] Mobile phase: CO2 phase A, and MeOH (0.05% DEA) phase B;

[0889] Gradient elution: 40% MeOH (0.05% DEA) in CO2;

[0890] Flow rate: 3 mL / min;

[0891] Detector: PDA;

[0892] Column temperature: 35°C;

[0893] Back pressure: 100 bar.

[0894] Retention time of 1-7 (route 2) = 1.70 min (late eluting peak).

[0895] Retention time of 1-7 (route 1) = 1.69 min (late eluting peak).

[0896] Racemic 1-7 (control) had retention times = 0.96 min (early eluting peak) and 1.77 min (late eluting peak).

[0897] Optical rotation of 1-7 (route 2)

[0898] Optical rotation of 1-7 (route 1)

[0899] Since intermediates 1-7 from routes 1 and 2 were late eluting peaks on chiral SFC (same method) and had the same negative specific rotation, compound 1-7 in route 1 was determined to be the R-isomer. This also confirmed that the absolute stereochemistry of compound 1 was R as described below.

[0900]

[0901] Example 3: Synthesis of 2-(1-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-5-fluoro-1-methyl-1H-indazol-6-yl)-4-hydroxypiperidin-4-yl)acetic acid

[0902]

[0903] Step 1 can be carried out on a production scale to prepare large quantities of 3-3 for use in the preparation of compound 1. A non-limiting example of this scale-up reaction and raw material input is provided below.

[0904]

[0905]

[0906]

[0907]

[0908] Step 2 can be carried out on a production scale to prepare large quantities of 3-4 for use in the preparation of compound 1. A non-limiting example of this scale-up reaction and raw material input is provided below.

[0909]

[0910]

[0911]

[0912]

[0913] Step 3 can be carried out on a production scale to prepare large quantities of 3-6 for use in the preparation of compound 1. A non-limiting example of this scale-up reaction and raw material input is provided below.

[0914]

[0915]

[0916]

[0917] Step 4 can be carried out on a production scale to prepare large quantities of 3-7 for use in the preparation of compound 1. A non-limiting example of such scale-up reactions and raw material inputs is provided below.

[0918]

[0919]

[0920]

[0921] Example 4: Synthesis of (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluorophenoxy]-4-oxoquinazolin-3-yl]-8-[2-[1-[3-(2,4-dioxo-1,3-diazinane-1-yl)-5-fluoro-1-methylindazol-6-yl]-4-hydroxypiperidin-4-yl]acetyl]-1-oxa-8-azaspiro[4.5]decane (Compound 1)

[0922]

[0923]

[0924] Compound 1 was prepared from three starting materials 4-1, 1-7, and 3-7 in seven steps, as shown in the above scheme. Starting material 4-1 was treated with sulfuryl chloride (SO2Cl2) in the presence of triethylamine (TEA) to give sulfamoyl chloride 4-2, which was reacted with ammonia in the presence of methanol (NH3 / MeOH) to produce sulfonamide 4-3. 1-7 (>99% ee) was substituted with the nucleophilic aromatic substituent (S N Ar) to give intermediate 4-4 without loss of its enantiomeric purity (>99% ee). Deprotection of the Boc group in 4-4 using HCl gave intermediate 4-5 as the free base. Treatment of 3-7 with TSTU produced its activated form 4-6. In the presence of DIEA, an amide bond was formed between 4-5 and 4-6 to give compound 1 as an amorphous solid. Recrystallization of amorphous compound 1 from a mixture of acetone and water gave compound 1 drug substance with greater than>99% ee as a crystalline solid.

[0925] Compound 1 drug substance is produced in seven steps, the general process is described below.

[0926] Step 1: Starting material 4-1 is treated with sulfuryl chloride and triethylamine in dichloromethane at -10 to 5°C to generate sulfamoyl chloride 4-2. After the reaction is complete, water is slowly added to the reaction mixture. The resulting organic layer is collected, washed with water, and concentrated in vacuo to afford 4-2 as a dichloromethane solution.

[0927] Step 2: Sulfamoyl chloride 4-2 in dichloromethane is added to a solution of ammonia in methanol at -10°C to 0°C, followed by warming the reaction to 15°C to 25°C to produce sulfonamide 4-3. After completion of the reaction, the reaction mixture is concentrated under vacuum and subjected to an aqueous workup with ethyl acetate and water. A solvent switch is performed to isolate 4-3 as a dimethylacetamide solution.

[0928] Step 3: A mixture of starting material 1-7, sulfonamide 4-3 and cesium carbonate in dimethylacetamide is stirred at 55-65°C to generate intermediate 4-4. After the reaction reaches the target conversion, water is added to the reaction mixture at 20-30°C and filtered to remove residual solids. The resulting aqueous solution is washed with methyl tert-butyl ether, neutralized with 1N hydrochloric acid, and extracted with ethyl acetate. The ethyl acetate layer containing the product is passed through a CUNO filter. The filtered solution is concentrated under vacuum to obtain 4-4 as an ethyl acetate solution with chiral purity maintained (>99%).

[0929] Step 4: Intermediate 4-4 in ethyl acetate was treated with aqueous hydrochloric acid at 45-55°C to give the deprotected form 4-5. After completion of the reaction, water was added to the reaction mixture followed by a 20% aqueous sodium carbonate solution to adjust the pH of the reaction mixture to approximately 9. The free base 4-5 was isolated as a solid by filtration.

[0930] Step 5: The starting material 3-7 is treated with N,N,N',N'-tetramethyl-O-(N-succinimidyl)uronium tetrafluoroborate (TSTU) in acetonitrile in the presence of diisopropylethylamine (DIEA), first at -5 to 5°C and then warmed to 20 to 30°C, to produce the activated form of 4-6. After the reaction reaches the target conversion, the resulting solid is collected by filtration. The filtered solid is washed with water and then with acetonitrile. The washed solid is dried to obtain 4-6 as a white solid.

[0931] Step 6: A mixture of intermediates 4-5 and 4-6 in dimethylformamide is treated with triethylamine and stirred at 20-30°C to produce compound 1. After the reaction reaches the target conversion, the reaction mixture is added to a second reactor containing water. The resulting solid is first collected by filtration and redissolved in dichloromethane. The dichloromethane layer containing the product is passed through a CUNO filter. The filtered solution is concentrated under vacuum. The concentrated material is triturated from the dimethylformamide-water mixture to provide compound 1 as an amorphous solid.

[0932] Step 7: Amorphous Compound 1 is recrystallized from a mixture of acetone, water, and ethanol to obtain crystalline Compound 1 drug substance. Amorphous Compound 1 is first dissolved in a mixture of acetone and water at 48-58°C and then seeded with a small amount of crystalline Compound 1 at 35-45°C. After stirring for several hours, ethanol is slowly added to the mixture. The resulting mixture is cooled to -3 to 3°C and stirred at this temperature for several hours. Filtered, then washed with ethanol and dried to obtain Compound 1 drug substance as a crystalline solid.

[0933] Step 1 can be performed on a production scale to prepare large quantities of 4-2 for use in the preparation of compound 1. A non-limiting example of such a scaled-up reaction is provided below.

[0934]

[0935] Process Description Steps <![CDATA[1. Load SO2Cl2 (7.53 kg, 1.0 eq.) into R1.]]> 2. Load DCM (16.5L, 5V) into R1. 3. Adjust R1 to -10~0℃. 4. Load DCM (16.5L, 5V) into R2. 5. Load 4-1 (3.30 kg, 1.0 eq.) into R2. 6. Load TEA (5.65 kg, 1.0 eq.) into R2. 7. Add the mixture solution in R2 dropwise to R1 at -10 to 0°C over 8 hours. 8. Stir R1 at -5 to 5°C for 4 hours. <![CDATA[9. Add H2O (16.5 L, 5V) dropwise to R1 within 30 min.]]> 10. Stir R1 at 15-25°C for 1 hr. 11. Separation. <![CDATA[12. Wash the organic liquid with H2O (9.9 L, 3 V).]]> 13. Concentrate R1 to 2-3V in vacuo at 20-30°C. 14. 11.38 kg of 4-2 in DCM were obtained.

[0936] R1 and R2 are glass-lined reactors with mechanical stirrers.

[0937] Step 2 can be performed on a production scale to prepare large quantities of 4-3 for use in the preparation of compound 1. A non-limiting example of such a scaled-up reaction is provided below.

[0938]

[0939]

[0940]

[0941] R1 is a stainless steel reactor and R2 is a glass-lined reactor, both equipped with a mechanical stirrer.

[0942] Step 3 can be performed on a production scale to prepare large quantities of 4-4 for use in the preparation of compound 1. A non-limiting example of such a scaled-up reaction is provided below.

[0943]

[0944]

[0945]

[0946] R1 and R2 are glass-lined reactors with mechanical stirrers.

[0947] Step 4 can be performed on a production scale to prepare large quantities of 4-5 for use in the preparation of compound 1. A non-limiting example of such a scaled-up reaction is provided below.

[0948]

[0949]

[0950]

[0951] R1 is a glass-lined reactor with a mechanical stirrer.

[0952] Step 5 can be performed on a production scale to prepare large quantities of 4-6 for use in the preparation of compound 1. A non-limiting example of such a scaled-up reaction is provided below.

[0953]

[0954]

[0955]

[0956] R1 is a glass-lined reactor with a mechanical stirrer.

[0957] 1 H NMR (400MHz, DMSO-d6): δppm 1.70-2.01(m,4H)2.66-2.93(m,8H)2.99-3.22(m,4H)3.76-4.09(m,5H)4.96(br s,1H)7.13(br d,J=1.10Hz,1H)7.33(br d,J=11.49Hz,1H)10.54(br s,1H).

[0958] Step 6 can be performed on a production scale to prepare large quantities of Compound 1 (amorphous form) for use in the preparation of Compound 1. A non-limiting example of such a scaled-up reaction is provided below.

[0959]

[0960]

[0961]

[0962] R1 and R3 were glass-lined reactors with mechanical stirrers, and R2 was a stainless steel reactor with mechanical stirrer.

[0963] 1H NMR (400MHz, DMSO-d6): δppm 10.53 (s, 1H), 10.20 (br s,1H),8.36(s,1H),7.95-7.75(m,2H),7.70(dd,J=3.2,8.8Hz,1H),7.50(dd,J=4. 4,9.6Hz,1H),7.41-7.26(m,2H),7.11(d,J=7.2Hz,1H),5.40-5.19(m,1H),5.03(br s,1H),4.18-4.08(m,2H),3.94(s,3H),3.89(t,J=6.7Hz,2H),3.78(br dd,J=4.8,12.8Hz,1H),3.68-3.58(m,1H),3.56-3.39(m,2H),3.19-3.14(m,4H),3.10-3.00(m,2H),2.79(s,3H),2.73(t,J=6.7Hz,2H),2.57(br s,2H),2.42-2.32(m,1H),2.14-1.97(m,1H),1.86-1.49(m,8H),1.05(t,J=7.2Hz,3H)

[0964] Step 7 can be performed on a production scale to prepare large quantities of crystalline Compound 1. A non-limiting example of such a scale-up reaction is provided below.

[0965]

[0966]

[0967] R1 is a stainless steel reactor and R2 is a Hastelloy reactor, both equipped with a mechanical stirrer.

[0968] In certain aspects, step 7 is modified, for example, in certain embodiments, the crystallization is performed at a lower temperature, such as a temperature between about 0° C. and about 35° C., a temperature between about 20° C. and about 35° C., or a temperature between about 0° C. and about 20° C., or a temperature between about 10° C. and about 30° C. In certain embodiments, the temperature is about 0° C., about 5° C., about 10° C., about 15° C., about 20° C., about 25° C., or about 30° C.

[0969] In certain embodiments, crystallization is carried out with slow stir speed (S.S.), for example, between about 50rpm and about 300rpm, between about 150rpm and about 300rpm, or between about 200rpm and about 300rpm. In certain embodiments, stirring is about 50rpm, about 60rpm, about 70rpm, about 80rpm, about 90rpm, about 100rpm, about 110rpm, about 120rpm, about 130rpm, about 140rpm, about 150rpm, about 160rpm, about 170rpm, about 180rpm, about 190rpm, about 200rpm, about 210rpm, about 220rpm, about 230rpm, about 240rpm, about 250rpm, about 260rpm, about 270rpm, about 280rpm, about 290rpm or about 300rpm.

[0970] In certain embodiments, crystallization is carried out for a period of more than ten hours, e.g., a crystallization time of between about 10 hours to about 30 hours, a crystallization time of between about 15 hours to about 30 hours, a crystallization time of between about 20 hours to about 30 hours, a crystallization time of between about 10 hours to about 25 hours, or a crystallization time of between about 15 hours to about 25 hours. In certain embodiments, the crystallization time is about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 25 hours, about 26 hours, about 27 hours, about 28 hours, about 29 hours, about 30 hours, about 31 hours, about 32 hours, about 33 hours, about 34 hours, or about 35 hours.

[0971] Example 5: Synthesis of (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluorophenoxy]-4-oxo-quinazolin-3-yl]-8-[2-[3,3,5,5-tetradeutero-1-[3-(2,4-dioxohexahydropyrimidin-1-yl)-5-fluoro-1-methyl-indazol-6-yl]-4-hydroxy-4-piperidinyl]acetyl]-1-oxa-8-azaspiro[4.5]decane

[0972]

[0973] Step 1: To a solution of 1-benzylpiperidin-4-one (10 g, 52.84 mmol, 9.80 mL, 1 eq.) in CDCl (50 mL) and D2O (50 mL) was added 3,4,6,7,8,9-hexahydro-2H-pyrimido[1,2-a]pyrimidine (1.40 g, 10.06 mol, 0.19 eq.). The mixture was stirred in a sealed tube at 15°C for 16 hours. Aqueous HCl (1 M) was added to the reaction mixture to adjust the pH to approximately 7. The separated organic phase was washed with water (20 mL) and brine (20 mL), dried over Na2SO4, filtered, and concentrated to give compound 1-benzyl-3,3,5,5-tetradeutero-piperidin-4-one (9.8 g, crude) as a yellow oil. 1 H NMR (400 MHz, chloroform-d) δ = 7.53-7.14 (m, 5H), 3.64 (s, 2H), 2.75 (s, 4H).

[0974] Step 2: To a solution of LDA (2M, 30.11 mL, 1.2 eq.) was added a solution of tert-butyl acetate (7.00 g, 60.22 mmol, 8.08 mL, 1.2 eq.) in THF (75 mL) at -65°C, and the mixture was stirred at -60°C for 0.5 h, followed by the addition of a solution of 1-benzyl-3,3,5,5-tetradeutero-piperidin-4-one (9.7 g, 50.19 mmol, 1.76 mL, 1 eq.) in THF (10 mL) at -60°C. The mixture was stirred at -60°C for 1.5 h, and then at 10°C for 0.5 h. The reaction mixture was poured into an NH4Cl aqueous solution (100 mL), extracted with EtOAc (100 mL x 2), and the combined organic phases were washed with water (100 mL), brine (100 mL), dried over Na2SO4, filtered, and concentrated to give tert-butyl 2-(1-benzyl-3,3,5,5-tetradeutero-4-hydroxy-4-piperidinyl)acetate (17 g, crude) as a yellow solid. LCMS m / z (ESI): 310.2 [M+H] + . 1 H NMR (400 MHz, CHLOROFORM-d) δ = 7.28-7.14 (m, 5H), 3.53 (s, 1H), 3.45 (s, 2H), 2.49 (br d, J = 11.4 Hz, 2H), 2.34 (br d, J = 11.4 Hz, 2H), 2.30 (s, 2H), 1.39 (s, 9H).

[0975] Step 3: To a solution of tert-butyl 2-(1-benzyl-3,3,5,5-tetradeutero-4-hydroxy-4-piperidinyl)acetate (17 g, 54.94 mmol, 1 eq.) in EtOH (170 mL) was added Pd(OH)2 / C (3.40 g, 20% purity) and HCOOH (5.28 g, 109.88 mmol, 2 eq.). The mixture was stirred at 50°C for 2 hours. The reaction mixture was filtered and concentrated. The residue was triturated with petroleum / EtOAc (10 / 1, 5 V) at 25°C for 2 hours, the suspension was filtered, and the filter cake was collected. Tert-butyl 2-(3,3,5,5-tetradeutero-4-hydroxy-4-piperidinyl)acetate (10 g, crude) was obtained as a yellow solid. 1 H NMR (400 MHz, CHLOROFORM-d) δ = 3.02 (br d, J = 12.4 Hz, 2H), 2.82 (d, J = 12.4 Hz, 2H), 2.38 (s, 2H), 1.47 (s, 9H).

[0976] Step 4: To a solution of tert-butyl 2-(3,3,5,5-tetradeutero-4-hydroxy-4-piperidinyl)acetate (10 g, 45.60 mmol, 1 eq.) in DMF (40 mL) was added DIEA (11.79 g, 91.19 mmol, 15.88 mL, 2 eq.) and 2,4,5-trifluorobenzonitrile (6.80 g, 43.32 mmol, 0.95 eq.). The mixture was stirred at 70 ° C for 1 hr. The reaction mixture was cooled to 25 ° C and poured into water (250 mL). The resulting mixture was stirred at 25 ° C for 2 hours, filtered, and the filter cake was collected. The compound tert-butyl 2-[1-(4-cyano-2,5-difluoro-phenyl)-3,3,5,5-tetradeutero-4-hydroxy-4-piperidinyl]acetate (12 g, crude) was obtained as a yellow solid. LCMS m / z(ESI):357.1[M+H] + . 1 H NMR (400 MHz, chloroform-d) δ = 7.17 (dd, J = 6.0, 12.4 Hz, 1H), 6.66 (dd, J = 7.0, 11.0 Hz, 1H), 3.87 (s, 1H), 3.46-3.35 (m, 2H), 3.31-3.22 (m, 2H), 2.43 (s, 2H), 1.49 (s, 9H)

[0977] Step 5: To a solution of tert-butyl 2-[1-(4-cyano-2,5-difluoro-phenyl)-3,3,5,5-tetradeutero-4-hydroxy-4-piperidinyl]acetate (12 g, 33.67 mmol, 1 eq.) in NMP (100 mL) was slowly added methylhydrazine (19.43 g, 168.70 mmol, 22.21 mL, 40% purity, 5.0 eq.) at 55°C. The mixture was stirred at 90°C for 16 hours. The reaction mixture was quenched by adding aqueous citric acid at 25°C to adjust the pH to 6-7, and then water was added to the mixture to 600 mL. The resulting mixture was stirred at 20°C for 2 hours and filtered to collect the filter cake. The compound tert-butyl 2-[1-(3-amino-5-fluoro-1-methyl-indazol-6-yl)-3,3,5,5-tetradeutero-4-hydroxy-4-piperidinyl]acetate (11 g, crude product) was obtained as a yellow solid. LCMS m / z (ESI): 383.2 [M+H] + . 1 H NMR (400 MHz, CHLOROFORM-d) δ = 7.12 (d, J = 11.8 Hz, 1H), 6.73-6.65 (m, 1H), 3.81 (s, 1H), 3.79 (s, 3H), 3.27-3.22 (m, 2H), 3.18-3.09 (m, 2H), 2.46 (s, 2H), 1.50 (s, 9H).

[0978] Step 6: To a solution of tert-butyl 2-[1-(3-amino-5-fluoro-1-methyl-indazol-6-yl)-3,3,5,5-tetradeutero-4-hydroxy-4-piperidinyl]acetate (11 g, 28.76 mmol, 1 eq.) in dioxane (110 mL) was added acrylic acid (6.22 g, 86.28 mmol, 5.92 mL, 3 eq.). The mixture was stirred at 100°C for 30 hr. The reaction mixture was diluted with water (300 mL) and extracted with 2-MeTHF / MeOH (5 / 1, 200 x 3). The organic phase was washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The residue was triturated with petroleum ether / EtOAc (1 / 1, 110 mL) at 20°C for 2 hr, then filtered. The filter cake was collected and further ground with i-PrOH / i-PrOAc (5 / 1, 10V) at 50°C for 2h. The suspended mixture was filtered at 50°C. The compound 3-[[6-[4-(2-tert-butoxy-2-oxo-ethyl)-3,3,5,5-tetradeutero-4-hydroxy-1-piperidinyl]-5-fluoro-1-methyl-indazol-3-yl]amino]propionic acid (5.2g, 10.53mmol, 36.60% yield, 92% purity) was obtained as a white solid. LCMSm / z(ESI):455.3[M+H] + . 1H NMR (400MHz, DMSO-d6) δ = 7.36 (d, J = 12.6Hz, 1H), 6.86 (d, J = 7.0Hz, 1H), 3.70 (s, 3H), 3.40 (br t,J=6.8Hz,2H),3.13-3.05(m,2H),3.02-2.95(m,2H),2.56(t,J=6.8Hz,2H),2.37(s,2H),1.42(s,9H).

[0979] Step 7: To a solution of 3-[[6-[4-(2-tert-butoxy-2-oxo-ethyl)-3,3,5,5-tetradeutero-4-hydroxy-1-piperidinyl]-5-fluoro-1-methyl-indazol-3-yl]amino]propanoic acid (5.2 g, 11.44 mmol, 1 eq.) in AcOH (50 mL) was added sodium cyanate (1.49 g, 22.88 mmol, 2 eq.). The mixture was stirred at 60°C for 12 h, then HCl (2M, 52.00 mL, 9.09 eq.) was added and the mixture was stirred at 60°C for 16 h. Saturated Na2CO3 and NaHCO3 were added to the reaction mixture to adjust the pH to approximately 5. The mixture was stirred at 15°C for 2 h, whereupon a solid precipitated and was filtered. The filter cake was collected, triturated with water (10 V) at 100°C for 16 h, and then filtered. The compound 2-[3,3,5,5-tetradeutero-1-[3-(2,4-dioxohexahydropyrimidin-1-yl)-5-fluoro-1-methyl-indazol-6-yl]-4-hydroxy-4-piperidinyl]acetic acid (2.4 g, 5.64 mmol, 49.30% yield, 99.5% purity) was obtained as a gray solid. LCMS m / z (ESI): 424.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ = 10.53 (s, 1H), 7.33 (br d, J = 12.8Hz, 1H), 7.12 (br d, J = 6.8Hz, 1H), 4.03-3.81 (m, 5H), 3.15 (br d, J = 11.6Hz, 2H), 3.04 (br d,J=11.6Hz,2H),2.74(br t,J=6.4Hz,2H),2.43(s,2H).

[0980] Step 8: To a solution of 2-[3,3,5,5-tetradeutero-1-[3-(2,4-dioxohexahydropyrimidin-1-yl)-5-fluoro-1-methyl-indazol-6-yl]-4-hydroxy-4-piperidinyl]acetic acid (2.4 g, 5.67 mmol, 1 eq.) in ACN (24 mL) was added TSTU (3.41 g, 11.34 mmol, 2 eq.) and DIEA (1.47 g, 11.34 mmol, 1.97 mL, 2 eq.) at 15° C. The mixture was stirred at 15° C. for 2 hr. The reaction mixture was filtered and the filter cake was washed with MeCN (10 mL), water (10 mL), and MeCN (10 mL). The compound (2,5-dioxopyrrolidin-1-yl) 2-[3,3,5,5-tetradeutero-1-[3-(2,4-dioxohexahydropyrimidin-1-yl)-5-fluoro-1-methyl-indazol-6-yl]-4-hydroxy-4-piperidinyl] acetate (2.4 g, crude product) was obtained as a white solid. LCMS m / z (ESI): 521.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ=10.54(s,1H),7.33(br d,J=12.8Hz,1H),7.14(br d,J=7.0Hz,1H),4.94(s,1H),4.01-3.78(m,5H),3.18(br d, J=11.6Hz, 2H), 3.04 (br d, J=11.6Hz, 2H), 2.83 (br d, J=9.0Hz, 6H), 2.73 (br t, J=6.4Hz, 2H).

[0981] Step 9: To a solution of (2,5-dioxopyrrolidin-1-yl) 2-[3,3,5,5-tetradeutero-1-[3-(2,4-dioxohexahydropyrimidin-1-yl)-5-fluoro-1-methyl-indazol-6-yl]-4-hydroxy-4-piperidinyl] acetate (1 g, 1.92 mmol, 1 eq.) in DMF (10 mL) was added (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluoro-phenoxy]-4-oxo-quinazolin-3-yl]-1-oxa-8-azaspiro[4.5]decane (1.07 g, 1.92 mmol, 1 eq.) and TEA (97.20 mg, 960.61 μmol, 133.71 μL, 0.5 eq.). The mixture was stirred at 25°C for 18 hours. The reaction mixture was slowly added to water (80 mL), and then the pH was adjusted to 6 with aqueous HCl (1 M). The mixture was filtered. The filter cake was collected and triturated with i-PrOH / i-PrOAc (5 / 1, 20 mL) at 25°C for 2 hours, and then further triturated with MeOH (14 mL) at 25°C for 12 hours. Compound (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluoro-phenoxy]-4-oxo-quinazolin-3-yl]-8-[2-[3,3,5,5-tetradeutero-1-[3-(2,4-dioxohexahydropyrimidin-1-yl)-5-fluoro-1-methyl-indazol-6-yl]-4-hydroxy-4-piperidinyl]acetyl]-1-oxa-8-azaspiro[4.5]decane (1.4 g, 1.43 mmol, 74.69% yield, 98.6% purity) was obtained as a gray solid. LCMS m / z (ESI): 962.5 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ = 10.52 (s, 1H), 10.18 (br s, 1H), 8.35 (s, 1H), 7.92-7.75 (m, 2H), 7.69 (dd, J = 2.8, 9.0Hz, 1H), 7.49 (br dd,J=3.9,9.0Hz,1H),7.42-7.26(m,2H),7.11(br d,J=7.0Hz,1H),5.30(br d,J=6.4Hz,1H),5.00(br s,1H),4.23-4.06(m,2H),4.01-3.83(m,5H),3.82-3.71(m,1H),3.68-3.58(m,1H),3.51(br d,J=7.4Hz,1H),3.46-3.38(m,1H),3.20-3.10(m,4H),3.04(br d,J=11.6Hz,2H),2.84-2.69(m,5H),2.56(br s,2H),2.41-2.34(m,1H),2.14-2.00(m,1H),1.84-1.48(m,4H),1.05(t,J=7.0Hz,3H).

[0982] Example 6: Synthesis of (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluoro-phenoxy]-5,7,8-trideutero-4-oxo-quinazolin-3-yl]-8-[2-[1-[3-(2,4-dioxohexahydropyrimidin-1-yl)-5-fluoro-1-methyl-indazol-6-yl]-4-hydroxy-4-piperidinyl]acetyl]-1-oxa-8-azaspiro[4.5]decane

[0983]

[0984]

[0985] Step 1: To a solution of 1,2,3,4,5-pentadeuterated-6-nitro-benzene (25 g, 195.10 mmol, 4.17 mL, 1 eq.) in H2SO4 (140 mL, purity %) was added TCCA (15.42 g, 66.33 mmol, 0.34 eq.), and the mixture was stirred at 80 ° C for 3 h. The reaction mixture was cooled to room temperature, poured into ice / water (200 mL), and extracted with EtOAc (100 mL×3). The combined organic layer was washed with water (2×40 mL), brine (2×40 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( 220g Purification by silica flash column, 0% ethyl acetate / petroleum ether eluent, gradient: 50 mL / min) gave 1-chloro-2,3,4-6-tetradeutero-5-nitro-benzene (32 g, crude) as a light brown oil. HPLC Rt = 2.386 min. 2 H NMR (400MHz, DMSO) δ = 8.72-8.21 (m, 2H).

[0986] Step 2: To a solution of 1-chloro-2,3,4-6-tetradeutero-5-nitro-benzene (37 g, 228.99 mmol, 7.19 mL, 1 eq.) and NH4Cl in MeOH (480 mL) and H2O (80 mL) was added Fe (76.73 g, 1.37 mol, 6 eq.) (97.99 g, 1.83 mol, 8 eq.) in portions, and the mixture was stirred at 80 ° C for 1 h. The mixture was filtered through celite and the filter cake was washed with MeOH (4×50 mL) and EtOAc (4×50 mL). The filtrate was concentrated under reduced pressure to obtain a residue. The residue was diluted with H2O (50 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine (30 mL×2), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (0.1% FA conditions; H2O / ACN = 1 / 0 to 1 / 1) to give 3-chloro-2,4,5,6-tetradeutero-phenylamine (17.8 g, 132.56 mmol, 57.89% yield, 98% purity) as a light brown oil. LCMS m / z (ESI): 132.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ = 5.36 (s, 2H). 2 H NMR (400MHz, DMSO) δ = 7.53 (s, 1H), 7.12-7.04 (m, 3H).

[0987] Step 3: A solution of NaNO2 (9.23 g, 133.74 mmol, 1.1 eq.) in H2O (48 mL) was added to a solution of 3-chloro-2,4,5,6-tetradeuterated-aniline (16 g, 121.58 mmol, 1 eq.) in H2SO4 (258.51 g, 843.44 mmol, 140.50 mL, 32% purity) at 0°C, and the mixture was stirred at 0°C for 1 h. A solution of KI (30.27 g, 182.38 mmol, 1.5 eq.) in H2O (80 mL) was then added at 0°C, and the resulting mixture was extracted with EtOAc (80 mL×3). The combined organic layers were washed with brine (50 mL×2), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography ( The product was purified by silica flash column; eluent: petroleum ether) to give 1-chloro-2,3,4-6-tetradeutero-5-iodo-benzene (20 g, 78.77 mmol, 64.79% yield, 95.5% purity) as a light brown oil. 2 H NMR (400MHz, DMSO) δ = 7.71 (s, 1H), 7.58 (s, 1H), 7.31 (s, 1H), 7.702 (s, 1H).

[0988] Step 4: To a solution of 1-chloro-2,3,4-6-tetradeutero-5-iodo-benzene (19 g, 78.36 mmol, 518.13 μL, 1 eq.) in THF (190 mL) was added n-BuLi (2.5 M, 34.48 mL, 1.1 eq.) at -65°C, and the mixture was stirred at -65°C for 1 h. The reaction mixture was poured into a large amount of dry ice and stirred at -65°C for 1 h. The mixture was then quenched by basification with saturated NH4Cl (50 mL) to pH = 11 and extracted with MBTE (3 x 40 mL). The aqueous phase was adjusted to pH approximately 4 with aqueous HCl and extracted with EtOAc (3 x 150 mL). The combined organic layers were washed with brine (2×40 mL) and dried under vacuum to give 3-chloro-2,4,5,6-tetradeutero-benzoic acid (9.5 g, 57.26 mmol, yield 73.08%, purity 96.8%) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ = 13.46 (s, 1H). 2 H NMR (400MHz, DMSO) δ = 8.44 -8.08 (m, 4H).

[0989] Step 5: To a solution of 3-chloro-2,4,5,6-tetradeutero-benzoic acid (8.6 g, 53.55 mmol, 103.63 μL, 1 eq.) in H2SO4 (15 mL, 98% purity) was added a mixture of HNO3 (7.60 g, 82.06 mmol, 5.43 mL, 68%, 1.53 eq.) and H2SO4 (10.72 g, 107.10 mmol, 5.83 mL, 98%, 2 eq.) at 25°C. The resulting mixture was stirred at 25°C for 2 h and then at 40°C for 1 h. The mixture was poured into ice / H2O (180 mL) and extracted with DCM (3 x 200 mL). The organic layer was washed with 0.1% aqueous HCl (160 mL), dried over Na2SO4 and evaporated to give 3-chloro-2,4,5-trideuterio-6-nitro-benzoic acid (10.2 g, 43.5 mmol, 79.68% yield, 87.3% purity) as a light yellow solid. LCMS m / z (ESI): 203.0 [MH]- . 2 H NMR (400MHz, DMSO) δ = 8.84-8.70 (m, 3H).

[0990] Step 6: A mixture of 3-chloro-2,4,5-trideuterio-6-nitro-benzoic acid (8 g, 39.10 mmol, 103.63 μL, 1 eq.) and NaOH (12.51 g, 312.83 mmol, 8 eq.) in D2O (156.4 mL) was stirred at 100°C (internal) for 20 h. The mixture was cooled to room temperature, acidified with 6N HCl, and extracted with EtOAc (3 x 80 mL). The combined organic layers were dried over Na2SO4 and evaporated to dryness. The crude product was triturated with EtOAc / Tol (1 / 10, 10 V) at 25°C for 12 h to afford 2,4,5-trideuterio-3-hydroxy-6-nitro-benzoic acid (6.1 g, 30.48 mmol, 77.94% yield, 93% purity) as a pale yellow solid. LCMS m / z (ESI): 185.1 [M+H] - . 1 H NMR (400MHz, DMSO-d6) δ = 14.30-13.13 (m, 1H), 11.91-10.84 (m, 1H). 2 H NMR (400MHz, DMSO) δ = 8.80-8.29 (m, 1H), 7.72-7.31 (m, 2H).

[0991] Step 7: To a solution of 2,4,5-trideuterio-3-hydroxy-6-nitro-benzoic acid (2 g, 10.74 mmol, 1 eq.) in CD3OD (20 mL) was added Pd / C (0.4 g, 10% purity). The mixture was stirred at 25° C. under a D2 (15 psi) atmosphere for 16 hr. The reaction mixture was filtered and the filtrate was concentrated to give 2-amino-3,4-6-trideuterio-5-hydroxybenzoic acid (1.2 g, crude) as a black solid. 1 H NMR (400MHz, DMSO-d6) δ = 8.60 (s, 1H), 8.50-7.76 (m, 2H).

[0992] Step 8: To a solution of 2-amino-3,4-6-trideuterio-5-hydroxy-benzoic acid (1.2 g, 7.68 mmol, 1 eq.) in n-BuOH (12 mL) was added tert-butyl (3R)-3-amino-1-oxa-8-azaspiro[4.5]decane-8-carboxylate (1.97 g, 7.68 mmol, 1 eq.) in n-BuOH (12 mL) at 120°C. The mixture was stirred at 125°C for 15 min, followed by the addition of diethoxymethoxyethane (2.73 g, 18.44 mmol, 3.07 mL, 2.4 eq.). The mixture was stirred at 125°C for 16 hr. The reaction mixture was diluted with water (50 mL) and EtOAc (50 mL), then filtered. The filtrate was washed with aqueous HCl (1M, 20 mL), aqueous NaHCO3 (20 mL), water (20 mL) and brine (20 mL), dried over Na2SO4, filtered and concentrated. The residue was triturated with petroleum / EtOAc (3 / 1, 15 mL) at 20°C for 2 h, then filtered to collect the filter cake. Compound (3R)-3-(5,7,8-trideuterated-6-hydroxy-4-oxo-quinazolin-3-yl)-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (1.5 g, 3.40 mmol, yield 44.30%, purity 91.8%) was obtained as a brown solid. LCMS m / z (ESI): 405.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ=10.49-9.97(m,1H),8.19(s,1H),5.34(br s,1H),4.10(br d,J=5.0Hz,2H),3.42(br s,2H),2.37(br dd,J=9.0,13.0Hz,1H),1.98(br dd,J=5.2,13.4Hz,1H),1.73-1.61(m,3H),1.54(br d,J=7.8Hz,1H),1.39(br s,9H).

[0993] Step 9: To a solution of 2,3,6-trifluorobenzonitrile (704.22 mg, 4.08 mmol, 91% purity, 1.1 eq.) in ACN (15 mL) was added CsCO (3.02 g, 9.27 mmol, 2.5 eq.) and (3R)-3-(5,7,8-trideuterio-6-hydroxy-4-oxo-quinazolin-3-yl)-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (1.5 g, 3.71 mmol, 1 eq.). The mixture was stirred at 20° C. for 2 hr. The reaction mixture was concentrated and then diluted with EtOAc (20 mL) and water (20 mL). The organic phase was washed with water (20 mL), brine (20 mL), dried over Na2SO4, filtered and concentrated in vacuo to give compound (3R)-3-[6-(2-cyano-3,6-difluoro-phenoxy)-5,7,8-trideuterio-4-oxo-quinazolin-3-yl]-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (1.4 g, crude) as a brown foam solid. LCMS m / z (ESI): 542.3 [M+H] + . 1 H NMR (400MHz, chloroform-d) δ = 8.33 (s, 1H), 7.46 (dt, J = 5.0, 9.4Hz, 1H), 7.12 (ddd, J = 3.6,7.6,9.2Hz,1H),5.50(dt,J=4.0,9.0Hz,1H),4.27-4.10(m,2H),3.66(br s,2H),3.45-3.16(m,2H),2.48(dd,J=8.9,14.0Hz,1H),1.89(dd,J=4.2,14.0Hz,1H),1.83-1.69(m,3H),1.60-1.51(m,1H),1.46(s,9H).

[0994] Step 10: To a solution of [methyl(sulfamoyl)amino]ethane (535.85 mg, 3.88 mmol, 1.5 eq.) in DMAc (12 mL) was added CsCO (2.53 g, 7.76 mmol, 3 eq.) and (3R)-3-[6-(2-cyano-3,6-difluoro-phenoxy)-5,7,8-trideuterio-4-oxo-quinazolin-3-yl]-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (1.4 g, 2.59 mmol, 1 eq.). The mixture was stirred in a sealed tube at 65°C for 16 hours. The reaction mixture was poured into water (60 mL) and washed with MTBE (20 mL x 3). The aqueous phase was adjusted to pH 6-7 with HCl (aq., 1 M) and extracted with EtOAc (20 mL x 2). The organic phase was washed with brine (20 mL x 2), dried over Na2SO4, filtered and concentrated to give compound (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluoro-phenoxy]-5,7,8-trideuterio-4-oxo-quinazolin-3-yl]-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (1.1 g, 1.47 mmol, yield 56.76%, purity 88%) as a brown foam solid. LCMS m / z (ESI): 660.3 [M+H] + . 1 H NMR (400MHz, chloroform-d) δ = 8.32 (s, 1H), 7.58-7.50 (m, 1H), 7.45-7.34 (m, 1H), 5.48 (br dd, J = 4.6, 8.6Hz, 1H), 4.22-4.04 (m, 2H), 3.64 (br s,2H),3.39-3.23(m,4H),3.00(s,3H),2.46(dd,J=8.8,14.0Hz,1H),1.86(dd,J=4.2 ,14.0Hz,1H),1.81-1.66(m,3H),1.59-1.49(m,1H),1.44(s,9H),1.20-1.11(m,3H).

[0995] Step 11: To a solution of (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluoro-phenoxy]-5,7,8-trideuterated-4-oxo-quinazolin-3-yl]-1-oxa-8-azaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (1.1 g, 1.67 mmol, 1 eq.) in acetone (6 mL) was added HCl (12 M, 486.30 μL, 3.5 eq.). The mixture was stirred at 50°C for 3 hours. The reaction mixture was concentrated. The residue was diluted with water (3 mL) and EtOAc (3 mL), then adjusted to pH 7-8 with aqueous NaHCO₃ solution, resulting in the precipitation of a large amount of solid. The suspended mixture was stirred at 25°C for 1 hour and filtered to collect the filter cake. The compound (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluoro-phenoxy]-5,7,8-trideuterated-4-oxo-quinazolin-3-yl]-1-oxa-8-azaspiro[4.5]decane (0.63 g, 1.07 mmol, yield 64.14%, purity 95%) was obtained as a gray solid. LCMS m / z (ESI): 560.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ = 8.29 (s, 1H), 7.34-7.13 (m, 2H), 5.28 (br s, 1H), 4.27-4.01 (m, 2H), 3.09 (br d,J=1.2Hz,4H),2.96(q,J=7.0Hz,2H),2.53(s,3H),2.43-2.32(m,1H),2.13(br d,J=8.8Hz,1H),2.01-1.68(m,4H),1.01(br t,J=7.0Hz,3H).

[0996] Step 12: To a solution of (2,5-dioxopyrrolidin-1-yl) 2-[1-[3-(2,4-dioxohexahydropyrimidin-1-yl)-5-fluoro-1-methyl-indazol-6-yl]-4-hydroxy-4-piperidinyl]acetate (273.49 mg, 519.99 μmol, purity 98.2%, 1 eq.) in DMF (3 mL) was added (3R)-3-[6-[2 [0.3 g, 519.99 μmol, 97% purity, 1 eq.] and TEA (26.31 mg, 259.99 μmol, 36.19 μL, 0.5 eq.) were added to a mixture of 1-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluoro-phenoxy]-5,7,8-trideuterio-4-oxo-quinazolin-3-yl]-1-oxa-8-azaspiro[4.5]decane (0.3 g, 519.99 μmol, 97% purity, 1 eq.). The mixture was stirred at 25°C for 16 hours. The reaction mixture was slowly added to water (20 mL), then the pH was adjusted to 6 with aqueous HCl (1 M). The suspended mixture was stirred at 20°C for 0.5 hours and filtered. The filter cake was triturated with i-PrOH / i-PrOAc (5 / 1, 3 mL) at 25°C for 2 hours, filtered, and further triturated with MeOH (3 mL) at 25°C for 2 hours and filtered. Compound (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluoro-phenoxy]-5,7,8-trideuterated-4-oxoquinazolin-3-yl]-8-[2-[1-[3-(2,4-dioxohexahydropyrimidin-1-yl)-5-fluoro-1-methyl-indazol-6-yl]-4-hydroxy-4-piperidinyl]acetyl]-1-oxa-8-azaspiro[4.5]decane (250 mg, 242.71 μmol, 46.68% yield, 93.3% purity) was obtained as a gray solid. LCMS m / z (ESI): 961.5 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ = 10.52 (brs, 1H), 10.18 (br d, J = 0.9Hz, 1H), 8.35 (br s, 1H), 7.85 (br t, J = 9.1Hz, 1H), 7.49 (br d,J=5.6Hz,1H),7.40-7.24(m,1H),7.11(br d,J=6.0Hz,1H),5.29(br s,1H),5.03(br s,1H),4.14(br s,2H),4.02-3.83(m,5H),3.78(br d,J=4.8Hz,1H),3.62(br d,J=3.6Hz,1H),3.56-3.44(m,2H),3.16(br d,J=6.4Hz,4H),3.07(br d,J=10.0Hz,2H),2.86-2.67(m,5H),2.56(br s,2H),2.37(br s,1H),2.07(br dd,J=5.4,6.8Hz,1H),1.88-1.46(m,8H),1.05(br t,J=6.4Hz,3H).

[0997] Evaluation of the free crystalline form of compound 1

[0998] The following techniques were used to characterize the free crystalline form of Compound 1.

[0999] Example 7: Approximate solubility at 25°C and 50°C

[1000] Weigh approximately 5 mg of Compound 1 Form A into a 2 mL glass vial. Add a 20 μL aliquot of each solvent to dissolve the drug substance at 25°C. Weigh approximately 10 mg of Compound 1 Form A into a 2 mL glass vial. Add a 20 μL aliquot of each solvent to dissolve the drug substance at 50°C. Vortex and sonicate to aid dissolution. The maximum volume of each solvent added is 1 mL. Determine approximate solubility by visual inspection.

[1001] Weigh approximately 5 mg of Compound 1 Form B into a 2 mL glass vial. Add a 20 μL aliquot of each solvent to dissolve the drug substance at 25°C. Weigh approximately 10 mg of Compound 1 Form B into a 2 mL glass vial. Add a 20 μL aliquot of each solvent to dissolve the drug substance at 50°C. Vortex and sonicate to aid dissolution. The maximum volume of each solvent added is 1 mL. Determine approximate solubility by visual inspection.

[1002] The results are summarized in Table 1.

[1003] Table 1: Approximate solubility of Form A and Form B at 25°C and 50°C

[1004]

[1005] Example 8: Equilibration with solvent at 25°C for 4 weeks

[1006] Approximately 50 mg of Compound 1 free crystalline Form A was equilibrated in 0.1-1 mL of solvent at 25° C. on a magnetic stirring plate with a stirring bar at 300 rpm for 4 weeks. The resulting suspension was filtered through a 0.45 μm nylon membrane filter by centrifugation at 14,000 rpm. The solid portion (wet cake) was studied by XRPD. The results are summarized in Tables 2 and 3.

[1007] Table 2: Form A equilibrated with solvents for 4 weeks at 25°C

[1008] solvent XRPD Notes Methanol Form B Low crystallinity acetone Form B Medium crystallinity Methyl ethyl ketone Form B Medium crystallinity Acetonitrile Form B Medium crystallinity THF / heptane (v:v=1:1) Form B Medium crystallinity Ethyl acetate Form C* Medium crystallinity Isopropyl alcohol Form A Low crystallinity MTBE Form A Low crystallinity 1,4-Dioxane / heptane (v:v=1:1) Not conducted Sticky materials DMF Not conducted Clear solution (approximately 500 mg / mL) 2-MeTHF Form C Medium crystallinity ethanol Form A Low crystallinity Isopropyl acetate Form A Low crystallinity Toluene Form A Low crystallinity

[1009] * Additional analysis of Form C by DSC: endothermic peak starting at 25.9°C; T onset 185.3°C; TGA: 2.2% at 180°C; and 1 H NMR: 2.7 wt% ethyl acetate (0.2 equivalents molar ratio).

[1010] Table 3: Form A equilibrated with solvent for 4 weeks at 25°C

[1011]

[1012]

[1013] *The water activity of binary solvent systems is calculated according to the UNIFAC method (UNIQUAC functional group activity coefficients).

[1014] Figure 1 The XRPD pattern of Compound 1 Form B is described. Figure 7 The XRPD pattern of Compound 1 Form A is described, and Figure 10 The XRPD pattern of Compound 1 Form C is described.

[1015] Example 9: Equilibrium under temperature cycling

[1016] Approximately 50 mg of Compound 1 Form A was equilibrated in 0.1-1 mL of solvent for 16 cycles at a heating / cooling rate of 0.1°C / min under a temperature cycle from 5°C to 40°C. Equilibration was performed on a magnetic stirring plate with a stirring bar at a speed of 300 rpm. The obtained suspension was filtered through a 0.45 μm nylon membrane filter by centrifugation at a speed of 14,000 rpm. The solid portion (wet cake) was studied by XRPD. The results are summarized in Tables 4 and 5.

[1017] Table 4: Equilibrium of Compound 1 Form A under temperature cycling

[1018]

[1019] Table 5: Equilibrium of Compound 1 Form A under temperature cycling

[1020] solvent XRPD Notes DCM / methanol (v:v=1:1) Not conducted Clear solution (approximately 500 mg / mL) Methanol / water (v:v=22:78) aw=0.9* Almost amorphous form - Acetone / water (v:v=36:64) aw=0.9* Not conducted Sticky materials Acetonitrile / water (v:v=42:58) aw=0.9* Not conducted Sticky materials THF / water (v:v=82:18) aw=0.9* Not conducted Clear solution (approximately 500 mg / mL) DMSO / water(v:v=23:77)aw=0.9* Form A Low crystallinity DMF / water (v:v=30:70) aw=0.9* Form B Low crystallinity Methanol / water (v:v=69:31) aw=0.6* Form B Low crystallinity Acetone / water (v:v=86:14) aw=0.6* Not conducted Clear solution (approximately 500 mg / mL) Acetonitrile / water (v:v=96:4) aw=0.65* Form B** Medium crystallinity THF / water (v:v=92:8) aw=0.6* Not conducted Clear solution (approximately 500 mg / mL) DMSO / water(v:v=57:43)aw=0.6* Form A Low crystallinity DMF / water (v:v=73:27) aw=0.6* Form B Medium crystallinity

[1021] *The water activity of binary solvent systems at 5°C is calculated according to the UNIFAC method (UNIQUAC functional group activity coefficients).

[1022] **After about 8 cycles, XRPD: moderate crystallinity, Form B; DSC: melting T onset 195.4℃; TGA: 1.9% at 170℃; 1 H NMR: no residual solvent was detected.

[1023] Example 10: Crystallization at room temperature by slow evaporation

[1024] Approximately 30 mg of Compound 1 Form A was dissolved in 0.2-10 mL of solvent. The resulting solution was filtered through a 0.45 μm syringe filter. The clear solution was slowly evaporated under ambient conditions (approximately 20-25° C., 40-80% RH). The solid residue was studied by XRPD. The results are summarized in Table 6.

[1025] Table 6: Crystallization at room temperature by slow evaporation

[1026] solvent XRPD Notes Methanol Almost amorphous form - acetone Not conducted glassy materials Methyl ethyl ketone Not conducted glassy materials Acetonitrile Almost amorphous form - Ethyl acetate Almost amorphous form - THF Not conducted glassy materials 1,4-Dioxane Not conducted glassy materials DCM Almost amorphous form - DCM / methanol (v:v=1:1) Almost amorphous form -

[1027] Example 11: Crystallization from hot saturated solution by slow cooling

[1028] At 50°C, approximately 50 mg of Compound 1 Form A was dissolved in a minimum amount of the selected solvent. The resulting solution was filtered through a 0.45 μm syringe filter. The clear solution was cooled to 5°C at 0.1°C / min. The sample without precipitation at 5°C was further cooled to -20°C. The precipitate was collected by centrifugation through a 0.45 μm nylon membrane filter at 14,000 rpm. The solid portion (wet cake) was studied by XRPD.

[1029] In order to determine whether Compound 1 Form B can be obtained from a clear solution, cooling was also attempted to prepare Form B. At 50°C, approximately 50 mg of Compound 1 Form A was dissolved in a minimum amount of ACN or acetone. The obtained solution was filtered through a 0.45 μm syringe membrane filter. The clear solution was cooled to 5°C at 0.1°C / min. When cooled to approximately 24°C, approximately 2 mg of seed crystals were added to the clear solution. The obtained suspension was cooled to 5°C at 0.1°C / min and kept at 5°C for approximately 12 hours. The precipitate was collected by centrifugation at 14,000 rpm through a 0.45 μm nylon membrane filter. The solid portion (wet cake) was studied by XRPD. The results showed that Form B can be obtained directly from a clear acetone solution by cooling with the addition of Form B seed crystals. The results are summarized in Table 7.

[1030] Table 7: Crystallization from hot saturated solutions by slow cooling

[1031]

[1032] Example 12: Crystallization by adding antisolvent

[1033] At ambient temperature (about 20-25 ° C), about 50 mg of Compound 1 Form A was dissolved in a minimum amount of a good solvent. The obtained solution was filtered through a 0.45 μm syringe membrane filter. 1-4 times the anti-solvent was slowly added to the clear solution until a large amount of solid precipitated out. The precipitate was collected by centrifugation at a speed of 14,000 rpm through a 0.45 μm nylon membrane filter. The solid portion (wet cake) was studied by XRPD. The results are summarized in Table 8.

[1034] Table 8: Crystallization by Addition of Antisolvent

[1035] Solvent (mL) Antisolvent (mL) XRPD Results (after 2 hours) DMSO (0.2) Water (0.2) Almost amorphous form - Acetone (3) Water (4) Not conducted From suspensions to viscous materials Acetone (3) Heptane (2) Crystal A Low crystallinity Acetonitrile (3) Water (5) Not conducted From suspension to oil material THF(0.2) Heptane (0.2) Almost amorphous form Viscous materials + suspensions THF(0.2) Water (0.2) Not conducted Oil material DCM(0.2) Heptane (0.2) Not conducted Sticky materials 2-MeTHF(8) MTBE(8) Almost amorphous form - 1,4-Dioxane (0.1) Water (0.2) Not conducted Sticky materials

[1036] Example 13: Crystallization by Vapor Diffusion

[1037] At ambient temperature (about 20-25 ° C), about 50 mg of Compound 1 Form A was dissolved in a minimum amount of selected solvent. The obtained solution was filtered through a 0.45 μm syringe membrane filter. The clear solution was transferred to an 8 mL uncovered glass vial. These 8 mL uncovered vials were then placed in 40 mL glass vials. Antisolvent was added to the 40 mL vial. The 40 mL sample vials were then tightly capped and placed under ambient conditions for 21 days. The precipitate was collected by centrifugation through a 0.45 μm nylon membrane filter at 14,000 rpm. The solid portion (wet cake) was studied by XRPD. The results are summarized in Table 9.

[1038] Table 9: Crystallization by Vapor Diffusion

[1039] Solvent (mL) Antisolvent (mL) XRPD result DMSO (0.2) Ethanol (0.8) Not conducted Oil material Acetone (2.5) MTBE (10.0) Not conducted Oil material Acetonitrile (3.0) MTBE (12.0) Not conducted Oil material THF (0.2) MTBE (0.8) Almost amorphous form - 2-MeTHF (8.0) MTBE (32.0) Not conducted Very few solids 1,4-Dioxane (0.2) MTBE (0.8) Almost amorphous form -

[1040] Example 14: Crystallization by heating-cooling DSC

[1041] The polymorphic behavior of Compound 1, Form A, was investigated using two different heat-cool DSC cycles. A Tzero pan and a Tzero sealing lid with a pinhole were used. No new crystalline forms were observed by heat-cool DSC. The results are summarized in Table 10.

[1042] Table 10: Crystallization by hot-cooling DSC

[1043]

[1044] Example 15: Volume stability

[1045] Compound 1 Form B was placed in an open container at 25°C / 92% RH, an open container at 40°C / 75% RH, and a sealed container at 60°C for one week. The samples after fortification were characterized by XRPD and HPLC, and color changes were examined. The results are summarized in Table 11.

[1046] Table 11: Volume stability

[1047]

[1048] Example 16: Water adsorption and desorption experiments

[1049] The water adsorption and desorption behavior of Compound 1 Form B was investigated by DVS at 25°C using a cycle of 40% to 95% to 0% to 40% RH, a dm / dt of 0.002, a minimum equilibrium time of 60 min, and a maximum equilibrium time of 360 min. XRPD measurements were performed after the DVS test to confirm the crystalline form changes. The results are summarized in Table 12.

[1050] Table 12: Water adsorption and desorption experiments

[1051]

[1052] *N / A: Not applicable

[1053] Example 17: Compression simulation experiment

[1054] Approximately 20 mg of Compound 1 Form B was compressed using a hydraulic press at 2 MPa and 10 MPa for 5 minutes. XRPD was used to evaluate potential crystalline form changes and crystallinity. The results are summarized in Table 13.

[1055] Table 13: Compression simulation experiment

[1056] pressure XRPD Notes 2MPa Form B Slight decrease in crystallinity; slight discoloration. 10MPa Form B Slight decrease in crystallinity; slight discoloration.

[1057] Example 18: Dry grinding simulation experiment

[1058] Approximately 20 mg of Compound 1 Form B was manually ground using a mortar and pestle for 1, 2, and 5 minutes. XRPD was performed to assess potential polymorphic changes and crystallinity. The results are summarized in Table 14.

[1059] Table 14: Dry grinding simulation experiment

[1060]

[1061]

[1062] Example 19: Wet granulation simulation experiment

[1063] Water or ethanol was added dropwise to approximately 20 mg of Compound 1 Form B until the sample was thoroughly moistened. The wet sample was gently ground with a mortar and pestle. After granulation, the sample was dried under ambient conditions for 10 minutes. XRPD was used to assess potential crystalline form changes and crystallinity. The results are summarized in Table 15.

[1064] Table 15: Wet granulation simulation experiment

[1065] Granulation solvent XRPD water Form B: The crystallinity is not reduced. ethanol Form B: The crystallinity is not reduced.

[1066] Example 20: Preparation of Compound 1 Form B

[1067] Form B was prepared using the following procedure.

[1068] Experiment 1: About 500 mg of Compound 1 Form A was weighed into a 20 mL glass vial. Then, the mixture was stirred at 40° C. for about 5 min, and 3 mL of ACN / H 2 O (v:v=96:4) was added to the vial to obtain a suspension.

[1069] About 2 mg of Form B seed crystals were added to the above suspension.

[1070] The suspension was equilibrated under temperature cycling from 5°C to 40°C at a heating / cooling rate of 0.1°C / min for about 9 cycles.

[1071] The solid was collected by suction filtration and then dried under vacuum at 35°C for about 1 hour.

[1072] 400 mg of Form B were obtained as an off-white solid in 80% yield.

[1073] Experiment 2: About 300 mg of Compound 1 Form A was weighed into a 20 mL glass vial. The mixture was then stirred at 40° C. for about 5 min, and 3 mL of ACN / H 2 O (v:v = 96:4) was added to the vial to obtain a viscous material. (Viscous material)

[1074] About 5 mg of Form B seed crystals were added to the above viscous material. After 7 hours, it was converted into a suspension. (Viscous material to suspension)

[1075] Equilibration was performed for about 8 cycles at 300 rpm on a magnetic stirring plate with temperature cycling from 5°C to 40°C at a heating / cooling rate of 0.1°C / min (suspension).

[1076] The solid was collected by suction filtration and then dried under vacuum at 30°C for about 4 hours.

[1077] 174 mg of Form B were obtained as an off-white solid in a yield of 58%.

[1078] In other aspects, Form B of Compound 1 is obtained using the methods described herein by varying the temperature, stirring rate, or crystallization time. For example, in certain embodiments, Form B of Compound 1 is formed without temperature cycling, but rather is crystallized from solution with stirring at a temperature between about 0° C. and about 50° C., a temperature between about 20° C. and about 35° C., a temperature between about 0° C. and about 20° C., or a temperature between about 10° C. and about 30° C. In certain embodiments, the temperature is about 0° C., about 5° C., about 10° C., about 15° C., about 20° C., about 25° C., about 30° C., about 35° C., about 40° C., about 45° C., or about 50° C.

[1079] In certain embodiments, the methods described herein are modified to have a slow stirring rate, such as stirring between about 50 rpm to about 300 rpm, stirring between about 150 rpm to about 300 rpm, or stirring between about 200 rpm to about 300 rpm. In certain embodiments, stirring is about 50 rpm, about 60 rpm, about 70 rpm, about 80 rpm, about 90 rpm, about 100 rpm, about 110 rpm, about 120 rpm, about 130 rpm, about 140 rpm, about 150 rpm, about 160 rpm, about 170 rpm, about 180 rpm, about 190 rpm, about 200 rpm, about 210 rpm, about 220 rpm, about 230 rpm, about 240 rpm, about 250 rpm, about 260 rpm, about 270 rpm, about 280 rpm, about 290 rpm, or about 300 rpm.

[1080] In certain embodiments, the methods described herein are modified to have a longer crystallization period, such as a crystallization time between about 10 hours to about 30 hours, a crystallization time between about 15 hours to about 30 hours, a crystallization time between about 20 hours to about 30 hours, or a crystallization time between about 10 hours to about 25 hours. In certain embodiments, the crystallization time is about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 25 hours, about 26 hours, about 27 hours, about 28 hours, about 29 hours, about 30 hours, about 31 hours, about 32 hours, about 33 hours, about 34 hours, or about 35 hours. In other embodiments, the crystallization time does not exceed about 30 hours.

[1081] In certain aspects, Compound 1 Form B is prepared using a temperature or temperature range without temperature cycling, a slow stirring rate, and an extended crystallization time. In certain aspects, Compound 1 Form B is prepared using temperature cycling, a slow stirring rate, and an extended crystallization time.

[1082] Evaluation of the crystal form of compound 1 salt

[1083] Example 21: Slurry Equilibration

[1084] Approximately 30 mg of Compound 1 Form A and 0.5 or 1 equivalent of base were added to the solvent in a 2 mL glass vial. For experiments using NaHCO, Ca(OH), and Mg(OH), 5% by volume of water was added to improve the solubility of the counterion in the test solvent. The resulting mixture was stirred at 25°C for at least 48 hours.

[1085] The obtained suspension was centrifuged and filtered through a 0.45 μm nylon membrane filter at 14,000 rpm. After vacuum drying at 25° C. for 2 h, the solid portion was analyzed by XRPD. The results are summarized in Table 16.

[1086] Table 16: Slurry Balance

[1087]

[1088]

[1089] *According to the free form of IC, the free form is different from NH4 + The ratio is 1:0.1( Figure 37 ).

[1090] Figure 23 The XRPD pattern of Compound 1 sodium salt Form D is described. Figure 26The XRPD pattern of Compound 1 sodium salt Form E is described. Figure 17 The XRPD pattern of Compound 1 sodium salt Form F is described. Figure 29 The XRPD pattern of Compound 1 potassium salt Form G is described, and Figure 32 The XRPD pattern of Compound 1 potassium salt Form H is described.

[1091] Example 22: Antisolvent Addition

[1092] The clear solution obtained from the slurry equilibration was treated by adding anti-solvent (Table 17).

[1093] Table 17: Antisolvent Addition

[1094] counter ions Solvent / mL Antisolvent / mL Notes choline MeOH / 0.2 MTBE / 0.3 Sticky materials

[1095] Example 23: Repulping

[1096] The viscous material or clear solution obtained from the slurry equilibrium was worked up by slow evaporation under ambient conditions and addition of MTBE, toluene and ethyl acetate (EA) (Table 18).

[1097] Table 18: Repulping

[1098] counter ions MTBE Toluene Ethyl acetate choline Sticky materials choline Sticky materials Methylglucosamine Not conducted Not conducted Form A <![CDATA[Ca(OH)2]]> Not conducted Not conducted Almost amorphous

[1099] The salt forms identified in the study (Table 16) were further characterized by XRPD, DCS, TGA, KF, HPLC and 1 HNMR / IC (Table 19).

[1100] Table 19: Characterization of Salt Forms

[1101]

[1102]

[1103] Example 24: Preparation of Compound 1 Sodium Salt Form F

[1104] The sodium salt Form F was prepared using the following procedure.

[1105] 500 mg of Compound 1 Fo...

Claims

1. The isolated crystalline form B of the compound having the following structure: It is characterized by an X-ray powder diffraction (XRPD) pattern comprising at least five 2θ values ​​selected from 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

2. The isolated crystalline Form B according to claim 1, wherein the XRPD pattern comprises at least six 2θ values ​​selected from 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

3. The isolated crystalline Form B of claim 1 , wherein the XRPD pattern comprises at least seven 2θ values ​​of 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

4. The isolated crystalline Form B of claim 1 , wherein the XRPD pattern comprises at least eight 2θ values ​​selected from the group consisting of 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

5. The isolated crystalline Form B of claim 1 , wherein the XRPD pattern comprises at least nine 2θ values ​​selected from the group consisting of 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

6. The isolated crystalline Form B of claim 1 , wherein the XRPD pattern comprises at least ten 2θ values ​​selected from the group consisting of 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

7. The isolated crystalline Form B of claim 1 , wherein the XRPD pattern comprises at least eleven 2θ values ​​selected from the group consisting of 7.5±0.2°, 8.8±0.2°, 10.0±0.2°, 10.5±0.2°, 12.6±0.2°, 14.7±0.2°, 15.4±0.2°, 16.4±0.2°, 16.7±0.2°, 18.6±0.2°, 22.7±0.2°, and 25.3±0.2°.

8. The isolated crystalline Form B according to any one of claims 1 to 7, wherein the XRPD pattern comprises at least three 2θ values ​​selected from 7.5±0.1°, 8.8±0.1°, 10.0±0.1°, 10.5±0.1°, 12.6±0.1°, 14.7±0.1°, 15.4±0.1°, 16.4±0.1°, 16.7±0.1°, 18.6±0.1°, 22.7±0.1°, and 25.3±0.1°.

9. The isolated crystalline Form B according to any one of claims 1 to 8, wherein the XRPD pattern comprises a 2θ value of at least 16.4 ± 0.2°.

10. The isolated crystalline Form B according to any one of claims 1-9, wherein the XRPD pattern comprises a 2θ value of at least 10.0 ± 0.2°.

11. The isolated crystalline Form B according to any one of claims 1 to 10, wherein the XRPD pattern comprises a 2θ value of at least 15.4 ± 0.2°.

12. The isolated crystalline Form B according to any one of claims 1-11, wherein the XRPD pattern comprises a 2θ value of at least 16.7±0.2°.

13. The isolated crystalline Form B according to any one of claims 1-12, wherein the XRPD pattern comprises a 2θ value of at least 8.8±0.2°.

14. The isolated crystalline Form B according to any one of claims 1 to 13, wherein the XRPD pattern comprises a 2θ value of at least 12.6 ± 0.2°.

15. The isolated crystalline Form B according to any one of claims 1-14, wherein the XRPD pattern comprises a 2θ value of at least 14.7 ± 0.2°.

16. The isolated crystalline Form B according to any one of claims 1-15, wherein the XRPD pattern comprises a 2θ value of at least 10.5 ± 0.2°.

17. The isolated crystalline Form B according to any one of claims 1-16, wherein the XRPD pattern comprises a 2θ value of at least 25.3 ± 0.2°.

18. The isolated crystalline Form B according to any one of claims 1-17, wherein the XRPD pattern comprises a 2θ value of at least 22.7±0.2°.

19. The isolated crystalline form B according to any one of claims 1 to 18, characterized in that An XRPD pattern with characteristic 2θ values ​​of Figure 1.

20. The isolated crystalline Form B according to any one of claims 1-19, having a differential scanning calorimetry (DSC) onset endotherm of about 194±20°C.

21. The isolated crystalline Form B according to any one of claims 1-20, having a differential scanning calorimetry (DSC) onset endotherm of about 194±10°C.

22. A pharmaceutical composition comprising the isolated crystalline Form B according to any one of claims 1 to 21 in a pharmaceutically acceptable carrier.

23. A pharmaceutical composition comprising Compound 1, a solubility enhancer, a permeation enhancer, a filler, a binder / glidant, and a mucoadhesive / disintegrant, wherein Compound 1 has the following structure: or a pharmaceutically acceptable salt thereof.

24. The pharmaceutical composition according to claim 23, wherein: a. The solubility enhancer is hypromellose acetate succinate; b. The penetration enhancer is vitamin E; c. The filler is mannitol; d. The binder / glidant is cellulose; and e. The mucoadhesive is cross-linked sodium carboxymethyl cellulose.

25. A method of treating a mutant BRAF-mediated disorder, comprising administering to a patient in need thereof an effective amount of the isolated crystalline Form B according to any one of claims 1-21 or the pharmaceutical composition according to any one of claims 22-24.

26. The method of claim 25, wherein the patient is a human.

27. The method of claim 25 or 26, wherein the mutant BRAF-mediated disorder is cancer.

28. The method of claim 27, wherein the mutant BRAF-mediated cancer is melanoma.

29. The method of claim 27, wherein the mutant BRAF-mediated cancer is lung cancer.

30. The method of claim 27, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

31. The method of claim 27, wherein the mutant BRAF-mediated cancer is colorectal cancer.

32. The method of claim 27, wherein the mutant BRAF-mediated cancer is microsatellite stable colorectal cancer.

33. The method of claim 27, wherein the mutant BRAF-mediated cancer is thyroid cancer.

34. The method of claim 27, wherein the mutant BRAF-mediated cancer is ovarian cancer.

35. The method of claim 25, wherein the mutant BRAF-mediated disorder is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoastrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid carcinoma, anaplastic thyroid carcinoma, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

36. The method of any one of claims 25-35, wherein the patient is also receiving an additional active agent.

37. The method of claim 36, wherein the additional active agent is a MEK inhibitor.

38. The method of claim 37, wherein the MEK inhibitor is trametinib.

39. The method of claim 36, wherein the additional active agent is an immune checkpoint inhibitor.

40. The method of claim 39, wherein the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, cemiplimab, ipilimumab, relalizumab, atezolizumab, avelumab, and durvalumab.

41. The method of claim 36, wherein the additional active agent is cetuximab or panitumumab.

42. The isolated crystalline Form B according to any one of claims 1-21 or the pharmaceutical composition according to any one of claims 22-24 for use in the therapeutic treatment of a mutant BRAF-mediated disorder.

43. The isolated crystalline Form B of claim 42, wherein the mutant BRAF-mediated disorder is cancer.

44. The isolated crystalline Form B of claim 43, wherein the mutant BRAF-mediated cancer is melanoma.

45. The isolated crystalline Form B of claim 43, wherein the mutant BRAF-mediated cancer is lung cancer.

46. ​​The isolated crystalline Form B of claim 43, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

47. The isolated crystalline Form B of claim 43, wherein the mutant BRAF-mediated cancer is colorectal cancer.

48. The isolated crystalline Form B of claim 43, wherein the mutant BRAF-mediated cancer is microsatellite-stable colorectal cancer.

49. The isolated crystalline Form B of claim 43, wherein the mutant BRAF-mediated cancer is thyroid cancer.

50. The isolated crystalline Form B of claim 43, wherein the mutant BRAF-mediated cancer is ovarian cancer.

51. The isolated crystalline Form B of claim 42, wherein the mutant BRAF-mediated disorder is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoid astrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid carcinoma, anaplastic thyroid carcinoma, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

52. The isolated crystalline Form B according to any one of claims 1-21 or the pharmaceutical composition according to any one of claims 22-24, for use in the treatment of a mutant BRAF-mediated disorder.

53. The isolated crystalline Form B of claim 52, wherein the mutant BRAF-mediated disorder is cancer.

54. The isolated crystalline Form B of claim 53, wherein the mutant BRAF-mediated cancer is melanoma.

55. The isolated crystalline Form B of claim 53, wherein the mutant BRAF-mediated cancer is lung cancer.

56. The isolated crystalline Form B of claim 53, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

57. The isolated crystalline Form B of claim 53, wherein the mutant BRAF-mediated cancer is colorectal cancer.

58. The isolated crystalline Form B of claim 53, wherein the mutant BRAF-mediated cancer is microsatellite-stable colorectal cancer.

59. The isolated crystalline Form B of claim 53, wherein the mutant BRAF-mediated cancer is thyroid cancer.

60. The isolated crystalline Form B of claim 53, wherein the mutant BRAF-mediated cancer is ovarian cancer.

61. The isolated crystalline Form B of claim 52, wherein the mutant BRAF-mediated disorder is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoid astrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid carcinoma, anaplastic thyroid carcinoma, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

62. Use of the isolated crystalline Form B according to any one of claims 1-21 or the pharmaceutical composition according to any one of claims 22-24 in the preparation of a medicament for treating a mutant BRAF-mediated disorder.

63. The use according to claim 62, wherein the mutant BRAF-mediated disorder is cancer.

64. The use according to claim 63, wherein the mutant BRAF-mediated cancer is melanoma.

65. The use according to claim 63, wherein the mutant BRAF-mediated cancer is lung cancer.

66. The use of claim 63, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

67. The use according to claim 63, wherein the mutant BRAF-mediated cancer is colorectal cancer.

68. The use of claim 63, wherein the mutant BRAF-mediated cancer is microsatellite stable colorectal cancer.

69. The use according to claim 63, wherein the mutant BRAF-mediated cancer is thyroid cancer.

70. The use of claim 63, wherein the mutant BRAF-mediated cancer is ovarian cancer.

71. The use of claim 62, wherein the mutant BRAF-mediated disorder is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoid astrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid cancer, anaplastic thyroid cancer, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

72. The isolated crystalline form B according to any one of claims 1 to 21 for use as a therapeutically active substance.

73. A method for preparing a compound of the formula: It involves the transaminase-catalyzed reductive amination of a compound of the formula: To obtain a compound of formula A-1; where R X is selected from hydrogen and an amino protecting group.

74. The method of claim 73, wherein: The amino protecting group R X is tert-butoxycarbonyl (BOC); The first base is sodium hydroxide or potassium hydroxide; and The isopropylamine salt is isopropylamine hydrochloride.

75. A method for preparing a compound of the formula: The method comprises the following steps: (i) making the compound of the formula: reacting with a chiral acid to obtain a mixture of diastereomeric salts formed between the compound of formula A-3 and the chiral acid; (ii) crystallizing the mixture of diastereomeric salts prepared in step (i) from a first solvent to provide individually separated diastereomeric salts formed between the anion of the chiral acid and the cation of N-protected (R)-1-oxa-8-azaspiro[4.5]decan-3-ammonium of the formula: and (iii) treating the individually isolated diastereomeric salt containing a cation of formula A-4 isolated in step (ii) with a second base to liberate the compound of formula A-1; where R X is selected from hydrogen and an amino protecting group.

76. The method of claim 75, wherein: The amino protecting group R X is tert-butoxycarbonyl (BOC); The chiral acid is (R)-mandelic acid; The first solvent is acetonitrile; and The second base is sodium hydroxide or potassium hydroxide.

77. A method for preparing the compound of the formula: The method comprises reacting 2-amino-5-hydroxybenzoic acid with a compound of the formula: wherein the compound of formula A-1 is prepared in a process according to any one of claims 73 to 76; and R X is selected from hydrogen and an amino protecting group.

78. The method of claim 77, wherein: The amino protecting group R X is tert-butoxycarbonyl (BOC); the trialkyl orthoformate is triethyl orthoformate; and The second solvent is n-butanol.

79. A method for preparing a compound of the formula: It comprises reacting 2,3,6-trifluorobenzonitrile with a compound of the formula: in R X is selected from hydrogen and an amino protecting group; and The compound of formula A-5 is prepared in a process according to any one of claims 77-78.

80. The method of claim 79, wherein: The third base is potassium carbonate, and the third solvent is acetonitrile; and The amino protecting group R X It is tert-butoxycarbonyl (BOC).

81. A method for preparing a compound of the formula: It includes the following steps: (i) reacting [methyl(sulfamoyl)amino]ethane NH2SO2N(Me)Et with a compound of the formula: To obtain a compound of formula A-8: in: The compound of formula A-6 is prepared in a process according to any one of claims 79-80; R X is selected from hydrogen and an amino protecting group; and (ii) When R X When it is an amino protecting group, the amino protecting group connected to the piperidine nitrogen atom of the compound of formula A-8 is removed in the presence of the first acid.

82. The method of claim 81 , wherein: The fourth base is cesium carbonate, and the fourth solvent is N,N-dimethylacetamide; The amino protecting group R X is tert-butoxycarbonyl (BOC); and The first acid is hydrochloric acid.

83. A method for preparing a compound of the formula: It comprises reacting 2,4,5-trifluorobenzonitrile with a compound of the formula: where R Y is hydrogen or a carboxylic acid protecting group.

84. The method of claim 83, wherein: The fifth base is N,N-diisopropylethylamine, and the fifth solvent is N,N-dimethylformamide; and R Y It is tert-butyl (tBu).

85. A method for preparing a compound of the formula: It comprises reacting methylhydrazine with a compound of the formula: wherein the compound of formula B-1 is prepared in a process according to any one of claims 83-84; where R Y is hydrogen or a carboxylic acid protecting group.

86. The method of claim 85, wherein: The sixth solvent is N-methyl-2-pyrrolidone; and R Y It is tert-butyl.

87. A method for preparing a compound of the formula: It comprises reacting acrylic acid with a compound of the formula: in: The compound of formula B-3 is prepared in a process according to any one of claims 85-86; and R Y is hydrogen or a carboxylic acid protecting group.

88. The method of claim 87, wherein: The seventh solvent is an aqueous hydrochloric acid solution; and R Y It is tert-butyl.

89. A method for preparing a compound of the formula: It includes the following steps: (i) First, a metal cyanate is reacted with a compound of the formula: To obtain a reaction mixture containing a first intermediate compound of the following formula: in: The compound of formula B-4 is prepared in a process according to any one of claims 87-88; and R Y is hydrogen or a carboxylic acid protecting group; (ii) cyclizing the first intermediate compound of formula B-6 from step (i) optionally in the presence of a third acid to obtain a second intermediate compound of formula: and (iii) Then, when R Y When it is a carboxylic acid protecting group, the carboxylic acid protecting group R is removed from the second intermediate compound of formula B-7 from step (ii) in the presence of a fourth acid. Y , to obtain the compound of formula B-5.

90. The method of claim 89, wherein R Y is tert-butyl; The metal cyanate is sodium cyanate or potassium cyanate; The second acid is acetic acid; The third acid is hydrochloric acid or acetic acid; and The fourth acid is hydrochloric acid.

91. A method for preparing a compound of the formula: The method comprises the following steps: (i) First, in the presence of a sixth base and an eighth solvent, a compound of the formula: React with a carboxylic acid hydroxyl activating reagent to obtain a third intermediate compound of the following formula: The carboxylic acid hydroxyl group is converted into a more reactive group -OR Z ; wherein the compound of formula B-5 is prepared in a process according to any one of claims 89-90; and (ii) then reacting (3R)-3-[6-[2-cyano-3-[[ethyl(methyl)sulfamoyl]amino]-6-fluoro-phenoxy]-4-oxo-quinazolin-3-yl]-1-oxa-8-azaspiro[4.5]decane of the formula: reacting with the third intermediate compound of formula B-8 prepared in step (ii) to obtain a compound of formula 1, The compound of formula A-7 is prepared according to the method of any one of claims 81-82.

92. The method of claim 91, wherein (i) the sixth base is N,N-diisopropylethylamine; The eighth solvent is acetonitrile; The carboxylic acid hydroxyl activation reagent is 2-succinimidyl-1,1,3,3-tetramethyluronium tetrafluoroborate, and R Z The group is 2,5-dioxopyrrolidin-1-yl; and (ii) The seventh base is triethylamine, and the ninth solvent is N,N-dimethylformamide.

93. A method for treating brain or central nervous system (CNS) metastasis of mutant BRAF-mediated cancer comprising administering to a human patient in need thereof an effective amount of a compound of the structure or a pharmaceutically acceptable salt thereof.

94. The method of claim 93, wherein the cancer has metastasized to the brain.

95. The method of claim 93 or 94, wherein the cancer has metastasized to the CNS.

96. The method of any one of claims 93-95, wherein the mutant BRAF-mediated cancer is melanoma.

97. The method of any one of claims 93-95, wherein the mutant BRAF-mediated cancer is lung cancer.

98. The method of any one of claims 93-95, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

99. The method of any one of claims 93-95, wherein the mutant BRAF-mediated cancer is colorectal cancer.

100. The method of any one of claims 93-95, wherein the mutant BRAF-mediated cancer is microsatellite stable colorectal cancer.

101. The method of any one of claims 93-95, wherein the mutant BRAF-mediated cancer is thyroid cancer.

102. The method of any one of claims 93-95, wherein the mutant BRAF-mediated cancer is ovarian cancer.

103. The method of any one of claims 93-95, wherein the mutant BRAF-mediated cancer is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoastrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid carcinoma, anaplastic thyroid carcinoma, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

104. The method of any one of claims 93-103, wherein the patient also receives an additional active agent.

105. The method of claim 104, wherein the additional active agent is a MEK inhibitor.

106. The method of claim 105, wherein the MEK inhibitor is trametinib.

107. The method of claim 104, wherein the additional active agent is an immune checkpoint inhibitor.

108. The method of claim 107, wherein the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, cemiplimab, ipilimumab, relalizumab, atezolizumab, avelumab, and durvalumab.

109. The method of claim 104, wherein the additional active agent is cetuximab or panitumumab.

110. The method of any one of claims 93-109, wherein the mutant BRAF has a V600 mutation.

111. The method of claim 110, wherein the V600 mutation is V600E, V600K, V600R, V600D, V600M, or V600N.

112. The method of claim 110, wherein the V600 mutation is V600E.

113. The method of any one of claims 93-112, wherein the mutant BRAF has a mutation selected from the group consisting of: G469A, G469V, G469L, G469R, L597Q, and K601E.

114. The method of any one of claims 93-113, wherein the mutant BRAF has a mutation selected from the group consisting of: G466A, G466E, G466R, G466V, S467L, G469E, N581I, D594E, D594G, and D594N.

115. The method of any one of claims 93-114, wherein the mutant BRAF has a mutation selected from the group consisting of: G464I, G464R, N581T, L584F, E586K, G593D, G596C, L597R, L597S, S605I, S607F, N684T, E26A, V130M, L745L, and D284E.

116. The method of any one of claims 93-115, wherein the mutant BRAF has a splicing mutation.

117. The method of claim 116, wherein the splicing mutation is a p61-BRAF V600 splicing mutation.

118. The method of any one of claims 93-117, wherein the mutant BRAF-mediated cancer has a NRAS, MEK1, or PI3K mutation.

119. The method of claim 118, wherein the cancer has a NRAS mutation.

120. The method of claim 119, wherein the NRAS mutation is Q61K.

121. The method of claim 119, wherein the NRAS mutation is Q61R.

122. The method of claim 118, wherein the cancer has a MEK1 mutation.

123. The method of claim 118, wherein the cancer has a PI3K mutation.

124. The method of claim 123, wherein the PI3K mutation is PIK3CA H1047R or PIK3CA P449T .

125. The method of any one of claims 93-124, wherein the mutant BRAF-mediated cancer has one or more mutations that promote homodimerization or heterodimerization of the RAF protein.

126. The method of claim 125, wherein the one or more mutations promote RAF protein homodimerization.

127. The method of claim 125, wherein the one or more mutations promote RAF protein heterodimerization.

128. A compound for use in the preparation of a medicament for treating brain or central nervous system (CNS) metastasis of mutant BRAF-mediated cancer, wherein the compound has the following structure: or a pharmaceutically acceptable salt thereof.

129. The compound of claim 128, for treating brain metastases.

130. The compound of claim 128, for treating CNS metastases.

131. The compound of any one of claims 128-130, wherein the mutant BRAF-mediated cancer is melanoma.

132. The compound of any one of claims 128-130, wherein the mutant BRAF-mediated cancer is lung cancer.

133. The compound of any one of claims 128-130, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

134. The compound of any one of claims 128-130, wherein the mutant BRAF-mediated cancer is colorectal cancer.

135. The compound of any one of claims 128-130, wherein the mutant BRAF-mediated cancer is microsatellite stable colorectal cancer.

136. The compound of any one of claims 128-130, wherein the mutant BRAF-mediated cancer is thyroid cancer.

137. The compound of any one of claims 128-130, wherein the mutant BRAF-mediated cancer is ovarian cancer.

138. The compound of any one of claims 128-130, wherein the mutant BRAF-mediated cancer is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoid astrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid carcinoma, anaplastic thyroid carcinoma, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

139. The compound of any one of claims 128-130, wherein the patient is also receiving an additional active agent.

140. The compound of claim 139, wherein the additional active agent is a MEK inhibitor.

141. The compound of claim 140, wherein the MEK inhibitor is trametinib.

142. The compound of claim 139, wherein the additional active agent is an immune checkpoint inhibitor.

143. The compound of claim 142, wherein the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, cemiplimab, ipilimumab, relalizumab, atezolizumab, avelumab, and durvalumab.

144. The compound of claim 139, wherein the additional active agent is cetuximab or panitumumab.

145. The compound of any one of claims 128-144, wherein the mutant BRAF has a V600 mutation.

146. The compound of claim 145, wherein the V600 mutation is V600E, V600K, V600R, V600D, V600M, or V600N.

147. The compound of claim 145, wherein the V600 mutation is V600E.

148. The compound of any one of claims 128-147, wherein the mutant BRAF has a mutation selected from the group consisting of: G469A, G469V, G469L, G469R, L597Q, and K601E.

149. The compound of any one of claims 128-148, wherein the mutant BRAF has a mutation selected from the group consisting of: G466A, G466E, G466R, G466V, S467L, G469E, N581I, D594E, D594G, and D594N.

150. The compound of any one of claims 128-149, wherein the mutant BRAF has a mutation selected from the group consisting of: G464I, G464R, N581T, L584F, E586K, G593D, G596C, L597R, L597S, S605I, S607F, N684T, E26A, V130M, L745L, and D284E.

151. The compound of any one of claims 128-150, wherein the mutant BRAF has a splice mutation.

152. The compound of claim 151, wherein the splicing mutation is a p61-BRAF V600 splicing mutation.

153. The compound of any one of claims 128-152, wherein the mutant BRAF-mediated cancer has a NRAS, MEK1 or PI3K mutation.

154. The compound of claim 153, wherein the cancer has a NRAS mutation.

155. The compound of claim 154, wherein the NRAS mutation is Q61K.

156. The compound of claim 154, wherein the NRAS mutation is Q61R.

157. The compound of claim 153, wherein the cancer has a MEK1 mutation.

158. The compound of claim 153, wherein the cancer has a PI3K mutation.

159. The compound of claim 158, wherein the PI3K mutation is PIK3CA H1047R or PIK3CA P449T .

160. The compound of any one of claims 128-159, wherein the mutant BRAF-mediated cancer has one or more mutations that promote homodimerization or heterodimerization of the RAF protein.

161. The compound of claim 160, wherein the one or more mutations promote RAF protein homodimerization.

162. The compound of claim 160, wherein the one or more mutations promote RAF protein heterodimerization.

163. Use of a compound in the preparation of a medicament for treating brain or central nervous system (CNS) metastasis of a mutant BRAF-mediated cancer, wherein the compound has the following structure: or a pharmaceutically acceptable salt thereof.

164. Use of a compound for treating brain or central nervous system (CNS) metastasis of a mutant BRAF-mediated cancer, wherein the compound has the structure: or a pharmaceutically acceptable salt thereof.

165. The use of claim 163 or 164, wherein the cancer has metastasized to the brain.

166. The use according to any one of claims 163-165, wherein the cancer has metastasized to the CNS.

167. The use of any one of claims 163-166, wherein the mutant BRAF-mediated cancer is melanoma.

168. The use of any one of claims 163-166, wherein the mutant BRAF-mediated cancer is lung cancer.

169. The use of any one of claims 163-166, wherein the mutant BRAF-mediated cancer is non-small cell lung cancer.

170. The use of any one of claims 163-166, wherein the mutant BRAF-mediated cancer is colorectal cancer.

171. The use of any one of claims 163-166, wherein the mutant BRAF-mediated cancer is microsatellite stable colorectal cancer.

172. The use of any one of claims 163-166, wherein the mutant BRAF-mediated cancer is thyroid cancer.

173. The use of any one of claims 163-166, wherein the mutant BRAF-mediated cancer is ovarian cancer.

174. The use of any one of claims 163-166, wherein the mutant BRAF-mediated cancer is cholangiocarcinoma, Erdeheim-Chester disease, Langerhans histiocytosis, ganglioglioma, glioma, glioblastoma, hairy cell leukemia, multiple myeloma, non-small cell lung cancer, ovarian cancer, pilocytic myxoid astrocytoma, anaplastic pleomorphic xanthoastrocytoma, astrocytoma, papillary thyroid carcinoma, anaplastic thyroid carcinoma, pancreatic cancer, clear cell sarcoma of the chest, salivary gland cancer, or microsatellite stable colorectal cancer.

175. The use of any one of claims 163-174, wherein the patient also receives an additional active agent.

176. The use of claim 175, wherein the additional active agent is a MEK inhibitor.

177. The use of claim 176, wherein the MEK inhibitor is trametinib.

178. The use of claim 175, wherein the additional active agent is an immune checkpoint inhibitor.

179. The use according to claim 178, wherein the immune checkpoint inhibitor is selected from nivolumab, pembrolizumab, cemiplizumab, ipilimumab, relalizumab, atezolizumab, avelumab and durvalumab.

180. The use of claim 175, wherein the additional active agent is cetuximab or panitumumab.

181. The use of any one of claims 163-180, wherein the mutant BRAF has a V600 mutation.

182. The use according to claim 181, wherein the V600 mutation is V600E, V600K, V600R, V600D, V600M or V600N.

183. The use according to claim 181, wherein the V600 mutation is V600E.

184. The use of any one of claims 163-183, wherein the mutant BRAF has a mutation selected from the group consisting of: G469A, G469V, G469L, G469R, L597Q, and K601E.

185. The use of any one of claims 163-184, wherein the mutant BRAF has a mutation selected from the group consisting of: G466A, G466E, G466R, G466V, S467L, G469E, N581I, D594E, D594G, and D594N.

186. The use of any one of claims 163-185, wherein the mutant BRAF has a mutation selected from the group consisting of: G464I, G464R, N581T, L584F, E586K, G593D, G596C, L597R, L597S, S605I, S607F, N684T, E26A, V130M, L745L, and D284E.

187. The use according to any one of claims 163-186, wherein the mutant BRAF has a splicing mutation.

188. The use according to claim 187, wherein the splicing mutation is a p61-BRAF V600 splicing mutation.

189. The use of any one of claims 163-188, wherein the mutant BRAF-mediated cancer has a NRAS, MEK1 or PI3K mutation.

190. The use of claim 189, wherein the cancer has a NRAS mutation.

191. The use of claim 190, wherein the NRAS mutation is Q61K.

192. The use of claim 190, wherein the NRAS mutation is Q61R.

193. The use of claim 189, wherein the cancer has a MEK1 mutation.

194. The use of claim 189, wherein the cancer has a PI3K mutation.

195. The use according to claim 194, wherein the PI3K mutation is PIK3CA H1047R or PIK3CA P449T .

196. The use of any one of claims 163-195, wherein the mutant BRAF-mediated cancer has one or more mutations that promote homodimerization or heterodimerization of the RAF protein.

197. The use of claim 196, wherein the one or more mutations promote RAF protein homodimerization.

198. The use of claim 196, wherein the one or more mutations promote RAF protein heterodimerization.

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