Pharmaceutical composition and method of use thereof

Abstract

The present invention provides methods and compositions for treating cancer through the use of compounds that form covalent bonds with kinases or pseudokinases, or pharmaceutically acceptable salts thereof, and at least one pharmaceutically acceptable excipient.

Description

TECHNICAL FIELD

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority and the benefit of U.S. Provisional Patent Application No. 63 / 585,862 filed on September 27, 2023, which is hereby incorporated by reference in its entirety into this specification for all purposes. BACKGROUND ART

[0002] Phosphatidylinositol 3-kinase (PI3K) is a family of lipid kinases that catalyze the transfer of phosphate to the 3-position of the inositol ring of phosphatidylinositol and its derivatives to produce phosphoinositol-3-phosphate (PI(3)P), phosphoinositol-3,4-bisphosphate (PI(3,4)P2), and phosphoinositol-3,4,5-trisphosphate (PI(3,4,5)P3). These kinases then act as second messengers in signaling cascades by docking with proteins containing plextrin homology. The FYVE Phox and other phospholipid-binding domains often enter various signaling complexes at the cell membrane. (Vanhaesebroeck et al., Annu. Rev. Biochem 70:535 (2001), Katso et al., Annu. Rev. Cell Dev Biol.) 17:615(2001)). PI3K is divided into three classes according to its structural characteristics and substrate specificity. Class IA PI3K is a heterodimer composed of a p110 catalytic subunit and a p85 regulatory subunit. In mammals, there are three genes encoding the p110 catalytic isoforms: PIK3CA, PIK3CB, and PIK3CD, which represent p110α, p110β, and p110δ, respectively. There are also three genes encoding the p85α (and its splicing variants p55α and p50α), p85β, and p55γ regulatory subunits: PIK3R1, PIK3R1, PIK3R2, and PIK3R3, collectively called p85. The module domain of the p85 / 55 / 50 subunit contains a Src homology (SH2) domain that binds to phosphotyrosine residues in specific sequence situations relating to the activating receptor and cytoplasmic tyrosine kinase, and is classified as Class 1A. This leads to the activation and localization of PI3K. Class IB PI3K is a heterodimer composed of a catalytic subunit p110γ and a regulatory subunit p101. p110γ is mainly expressed in leukocytes and can be directly activated by GPCRs. Class II PI3K is a monomer having only a single catalytic subunit.Class III PI3K consists of a single catalytic subunit, Vps34 (a homolog of yeast vacuolar protein sorting-deficient 34). The phospholipid products of Class I PI3K link upstream receptors to downstream cellular activities including proliferation, survival, chemotaxis, cell transport, motility, metabolism, inflammatory and allergic responses, transcription, and translation (Cantley et al., Cell 64:281 (1991), Escobedo and Williams, Nature 335:85 (1988), Fantl et al., Cell 69:413 (1992)). There is a current need for this benefit and for the binding to and regulation of PI3K for the treatment of various diseases.

[0003] This disclosure addresses these and other unmet needs. [Overview of the Initiative]

[0004] The present invention includes a pharmaceutical composition for immediate release comprising a therapeutically effective amount of a compound that forms a covalent bond with a kinase or pseudokinase or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically excipient or a method of using the excipient. In certain embodiments, the kinase is PI3 kinase (PI3K).

[0005] In another embodiment, the compound that forms a covalent bond with the kinase or pseudokinase is a compound having the following formula: [C1] (FCB)a-(L)b-(CLM)c, or a pharmaceutically acceptable salt thereof, in the formula, CLM is a covalent modality, L is a linker, a and c are each independent integers between 1 and 5. b is an integer from 0 to 5, and FCB [ka] That is the case.

[0006] In a particular embodiment, the FCB has the following structure: [ka] It has, R1 is [ka] It is selected from the group consisting of the following.

[0007] In another specific embodiment, the compound is compound I. [ka] Compound I

[0008] Compound I is disclosed in PCT application publication WO2021 / 055747 and PCT application PCT / CN2023 / 083760, both of which are incorporated herein by reference. In some embodiments, Compound I is a phosphoinositide 3-kinase (PI3K) inhibitor. In some embodiments, Compound I is an irreversible inhibitor of PI3K. In several embodiments, Compound I is an irreversible inhibitor of PI3Ka. In other embodiments, Compound I can form a covalent bond with an amino acid of PI3K. In other specific embodiments, Compound I can form a covalent bond with cysteine ​​in PI3K, for example, PI3Ka (e.g., via the Michael reaction).

[0009] The present invention also includes administering the pharmaceutical compositions described herein to a subject. In some embodiments, the method may include doing so in a subject requiring treatment for cancer, neurodegenerative disease, autoimmune disorder, or aging. In other specific embodiments, the method may include treating or inhibiting cancer in a subject. In yet another specific embodiment, the method may include treating or improving a cellular proliferation disorder in a subject, and the method includes administering to a subject requiring such treatment a therapeutically effective amount of a compound that forms a covalent bond with a kinase or pseudokinase or a pharmaceutically acceptable salt thereof, before, during, or after the subject is administered the compound that forms a covalent bond with the kinase or pseudokinase. In certain embodiments, the kinase is PI3-kinase (PI3K), preferably PI3Ka.

[0010] In another embodiment where a cell proliferation disorder is treated or improved in a subject, the cell proliferation disorder is cancer. In some embodiments, the cancer is selected from the group consisting of leukemia, brain tumors, lung (small cell and non-small cell), ovarian, prostate, breast, colon, and intestinal tissue, pancreatic tissue, liver, kidney, prostate, ovaries, lung, bone marrow, or cancer in tissue including breast tissue. In certain embodiments, the cancer is selected from the group consisting of blood cancer, colorectal cancer, ovarian cancer, breast cancer, cervical cancer, lung cancer, liver cancer, colon cancer, pancreatic cancer, lymph node cancer, colon cancer, small intestine, prostate cancer, brain cancer, bile duct cancer, gallbladder cancer, head and neck cancer, bone cancer, Ewing's sarcoma, skin cancer, kidney cancer, and heart cancer. In another embodiment, the cancer is selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, colorectal cancer, colon cancer, small intestine, and lung cancer. In some embodiments, the cancer is cancer associated with mutations in the PIK3CA gene. In another embodiment of the present invention, the subject is human. In some embodiments, the subjects have cancer with a mutation in the PIK3CA gene. [Brief explanation of the drawing]

[0011] [Figure 1] Figure 1 provides a flowchart outlining a process for manufacturing a pharmaceutical tablet of the present disclosure containing compound I. [Figure 2] Figure 2 shows the dissolution profile of Compound I tablet, 5 mg, batch 1, in 900 mL of 0.01 M HCl at 37°C, apparatus 1I (paddle), 75 rpm. [Figure 3] Figure 3 shows the dissolution profile of Compound I tablet, 50 mg, batch 1, in 900 mL of 0.01 M HCl at 37°C, apparatus 1I (paddle), 100 rpm. [Figure 4] Figure 4 shows the dissolution profile of Compound I tablet, 1 mg, batch 2, instrument 1I (paddle), 75 rpm in 900 mL of 0.01 M HCl at 37°C. [Figure 5] Figure 5 shows the dissolution profile of Compound I tablet, 5 mg, batch 2, in 900 mL of 0.01 M HCl at 37°C, apparatus 1I (paddle), 100 rpm. [Figure 6] Figure 6 shows the dissolution profile of Compound I tablet, 1 mg, batch 3, in 900 mL of 0.01 M HCl at 37°C, apparatus 1I (paddle), 75 rpm. [Figure 7] Figure 7 shows the dissolution profile of compound I, 5 mg, batch 3, in 900 mL of 0.01 M HCl at 37°C, apparatus 1I (paddle), and 100 rpm. [Figure 8] Figure 8 shows the dissolution profile of compound I tablet, 1 mg, batch 4, in 900 mL of 0.01 M HCl at 37°C, apparatus 1I (paddle), 75 rpm. [Figure 9] Figure 9 shows the dissolution profile of compound I tablets, 5 mg, batch 4, in 900 mL of 0.01 M HCl at 37°C, apparatus 1I (paddle), 100 rpm. [Modes for carrying out the invention]

[0012] definition For convenience, the specific terms used in this specification, examples, and claims are collected herein. Unless otherwise defined, all technical and scientific terms used in this disclosure have the same meaning as those commonly understood by those skilled in the art to which this disclosure belongs.

[0013] Throughout this specification, the terms “about” and / or “approximately” may be used in conjunction with numbers and / or ranges. The term “approximately” is understood to mean values ​​that are close to the listed values. Furthermore, given the definitions of terms provided herein, phrases are either less than about [a value] or greater than about [a value]. The terms “about” and “approximately” may be used interchangeably.

[0014] Throughout this specification, numerical ranges are provided for specific quantities. It should be understood that these ranges include all subranges within them. Thus, the range "50-80" includes all possible ranges within it (e.g., 51-79, 52-78, 53-77, 54-76, 55-75, 60-70, etc.). Furthermore, all values ​​within a given range may be endpoints to the ranges it encompasses (e.g., the range 50-80 includes ranges with endpoints such as 55-80, 50-75, etc.).

[0015] The terms “composition” and “formulation” may be used interchangeably throughout this specification and may mean a mixture of materials.

[0016] The terms "a" or "an" refer to one or more of the entities; for example, "PI3-kinase (PI3K) modulator" refers to one or more PI3-kinase (PI3K) modulators or at least one PI3-kinase (PI3K) modulator. In embodiments, PI3-kinase (PI3K) modulator is a PI3Ka modulator. Thus, the terms "a" (or "an"), "one or more," and "at least one" are used interchangeably herein. Furthermore, references to "inhibitors" with the indefinite article "a" or "an" do not rule out the possibility of two or more inhibitors existing unless the context explicitly requires that only one or more inhibitors exist.

[0017] As used herein, the verb “including” as used herein, in the claims, and in combination thereof is used in its non-restrictive sense, meaning that the items according to the word are included, but items not specifically mentioned are not excluded. The present invention may preferably “include,” “consist of,” or “essentially consist of” the steps, elements, and / or reagents described in the claims.

[0018] Furthermore, it should be noted that claims may be written to exclude any optional elements. Thus, such wording is intended to serve as a foresight basis for the use of such exclusive terms, such as "simply" or "only," in relation to the enumeration of elements in the claims or the use of negative limitations.

[0019] The terms “excipient,” “carrier,” and “vehicle” are used interchangeably throughout this application and refer to the substance to which the compounds of the present invention are administered. The terms “excipient,” “carrier,” and “vehicle” mean materials, compositions, or vehicles, such as liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials, that are involved in the transport or delivery of pharmaceuticals from one organ or part of the body to another organ or part of the body.

[0020] As used herein, the term “pharmaceutically acceptable” means a compound, material, composition, and / or dosage form that, within the bounds of sound medical judgment, is suitable for use in contact with human and animal tissues in a reasonable benefit-to-risk ratio without causing excessive toxicity, irritation, allergic reactions, or other problems or complications.

[0021] As used herein, the term “salt” encompasses pharmaceutically acceptable salts commonly used to form addition salts of free bases. The properties of the salt are irrelevant if it is pharmaceutically acceptable. The term “salt” also includes solvates of addition salts, such as hydrates, as well as polymorphs of addition salts. Suitable pharmaceutically acceptable acid addition salts can be prepared from inorganic or organic acids.

[0022] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting an active compound that functions as a base with an inorganic or organic acid to form a salt, such as 1-hydroxy-2-naphthic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutamic acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, ascorbic acid (L), aspartic acid (L), benzenesulfonic acid, benzoic acid, camphoric acid (+), camphor-10-sulfonic acid (+), capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, and galactamic acid. Contains lic acid, gentisic acid, glucoheptonic acid (D), gluconic acid (D), glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isethionic acid, isobutyric acid, lactic acid (DL), lactobionic acid, lauric acid, maleic acid, malic acid, (-L)malonic acid, mandelic acid, methanesulfonic acid, mucus, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, pantothenic acid, phosphoric acid, proprionic acid, pyroglutamic acid (-L), salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tartaric acid (+L), thiocyanic acid, toluenesulfonic acid (p), and undecylenic acid. Those skilled in the art will further recognize that acid addition salts can be prepared by reacting a compound with a suitable inorganic or organic acid via one of many known methods.

[0023] As used herein, the terms “disorder characterized by cell proliferation” or “condition characterized by cell proliferation” include, but are not limited to, cancer, benign and malignant tumors. Examples of cancer and tumors include, but are not limited to, cancer or tumor growth of the colorectal, breast, lung, liver, pancreas, lymph nodes, colon, prostate, brain, head and neck, skin, kidneys, blood and heart (e.g., leukemia, lymphoma, cancer).

[0024] The term “treatment” means one or more of the following: reduction, mitigation, delay, reduction, improvement, or control of at least one symptom of a condition in the subject. The term “treatment” may also mean one or more of the following: cessation, delay of onset (i.e., the period before the clinical symptoms of the condition), or reduction of the risk of developing or worsening the condition.

[0025] As used herein, the terms “inhibit” or “reduce” cell proliferation mean slowing, reducing, or, for example, stopping the amount of cell proliferation by, for example, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% compared to proliferating cells not subjected to the methods, compositions, and combinations of this application, as measured using methods known to those skilled in the art.

[0026] "Effective dose" means the amount of the formulation according to the present invention that, when administered to a patient to treat a condition, disorder, or condition, is sufficient to produce such treatment. The "effective dose" varies depending on the active ingredient, the condition, disorder, or condition being treated, its severity, and the age, weight, physical condition, and responsiveness of the mammal being treated.

[0027] "Therapeutic dose" refers to the amount of compound or therapeutic agent that, when administered to a patient to treat a disease or other undesirable medical condition, is sufficient to have a beneficial effect on that disease or condition. The therapeutic dose varies depending on the type of compound or therapeutic agent selected, the disease or condition and its severity, and the age and weight of the patient being treated. Determining the therapeutic dose of a given compound or therapeutic agent is within the ordinary art of the field and requires only routine experiments.

[0028] The term therapeutically effective, as applied to dosage or quantity, refers to the amount of compound or pharmaceutical formulation that is sufficient to produce the desired clinical benefit after administration to a patient who needs it.

[0029] As used herein, “Subject” may be a human, a non-human primate, a mammal, a rat, a mouse, a cattle, a horse, a pig, a sheep, a goat, a dog, a cat, and the like. In some embodiments, the subject is a human. In some embodiments, the subject may have cancer or be suspected of being at risk of having cancer. The terms “Subject” and “Patient” are used interchangeably throughout this application.

[0030] "Mammals" include humans, as well as domesticated animals such as laboratory animals (e.g., mice, rats, monkeys, dogs, etc.) and pets kept in captivity (e.g., cats, dogs, pigs, cows, sheep, goats, horses, rabbits), and non-domesticated animals such as wild animals.

[0031] All weight percentages referenced herein (i.e., "weight%", "wt.%", "w / w", and "%wt") are measured relative to the total weight of the pharmaceutical composition unless otherwise indicated.

[0032] As used herein, “substantially” or “substantially” refers to the complete or near-complete degree or extent of an action, characteristic, feature, state, structure, item, or result. For example, substantially enclosed object means that the object is either completely enclosed or nearly completely enclosed. The exact degree of acceptable deviation from absolute completeness may, in some cases, depend on the specific circumstances. However, generally speaking, proximity to completion is such that it has the same overall result as if absolute and complete completion had been achieved. The use of “substantially” is equally applicable when used in negative notation to refer to the complete or near-complete absence of an action, characteristic, feature, state, structure, item, or result. For example, a composition substantially free of other activators is either completely lacking in other activators or completely lacking in other activators, and its effect will be the same as if it were completely lacking in other activators. In other words, a composition substantially free of an ingredient or element or another activator may still contain such an item, as long as its measurable effect is not present.

[0033] As used herein, "D90" refers to the 90th percentile of the particle diameter, determined by sieving analysis, i.e., by the percentage of drug particles retained on a mesh screen during a sieving process with a screen of a specific mesh size. For example, if D90 = 100 μm, 90% of the drug particles remain on a 100 μm mesh screen during the sieving process. Similarly, "D80" means the 80th percentile of the particle diameter, "D70" means the 70th percentile, "D60" means the 60th percentile, "D50" means the 50th percentile, "D40" means the 40th percentile, "D30" means the 30th percentile, "D20" means the 20th percentile, and "D10" means the 10th percentile.

[0034] Compounds that form a covalent bond with PI3-kinase (PI3K) Compounds, for example, compounds having the following formula: [6] (FCB)a-(L)b-(CLM)c, Alternatively, a pharmaceutically acceptable salt thereof, and / or a subgenre thereof, or Compound I, or a salt thereof, solvate, ester, and / or prodrug, may be used in the pharmaceutical compositions disclosed herein.

[0035] In another embodiment, the compound that forms a covalent bond with the kinase or pseudokinase is a compound having the following formula: [7] (FCB)a-(L)b-(CLM)c, or a pharmaceutically acceptable salt thereof, CLM is a covalent modality, L is a linker, a and c are each independent integers between 1 and 5. b is an integer from 0 to 5, and FCB [ka] That is the case.

[0036] In a particular embodiment, the FCB has the following structure: [ka] R1 has, [ka] It is selected from the group consisting of the following.

[0037] In some embodiments, the compound that forms a covalent bond with a kinase or pseudokinase (e.g., PI3-kinase (PI3K)) is compound I. In some embodiments, the compound of formula (FCB)a-(L)b-(CLM)c is compound I.

[0038] In some embodiments, compound I is the crystalline form of compound I, or a pharmaceutically acceptable salt or solvate of compound I. In some embodiments, a pharmaceutically acceptable salt selected from the group consisting of phosphate, hydrochloride, sulfate, mesylate, benzenesulfonate, tosylate, fumarate, maleate, L-tartrate, citrate, and succinate of compound I, or their solvates,

[0039] [ka] Compound I That is the case.

[0040] In certain embodiments, compound I is a phosphate-containing salt. In some embodiments, compounds that form a covalent bond with PI3-kinase (PI3K) are called PI3K compounds.

[0041] As used herein, the term "CLM" refers to any covalent modality capable of forming a covalent bond with a biological target. The CLM may be linked to the FCB by a bond or a linker. The CLM may contain one or more chemical moieties capable of forming a covalent bond with a biological target. The chemical moieties may be electrophilic or nucleophilic groups.

[0042] As used herein, the term “biological target” refers to any target to which FCBs bind non-covalently and produce a therapeutic effect. CLMs bind covalently to biological targets. In some embodiments, the biological target is a protein. Non-limiting examples of biological targets include, but are not limited to, kinases such as phosphoinositide 3-kinase (PI3K) and pseudokinases.

[0043] The term "alkyl" refers to a saturated aliphatic radical, including linear alkyl groups, branched alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups.

[0044] In some embodiments, linear or branched alkyl groups have 30 or fewer carbon atoms in their skeleton (for example, a linear group may have C 1- C 30 The branching chain is C 3- C 30), having 20 or fewer carbon atoms, 12 or fewer carbon atoms, or 7 or fewer carbon atoms. Similarly, in some embodiments, cycloalkyls have 3 to 10 carbon atoms in the ring structure, for example, having 5, 6, or 7 carbon atoms in the ring structure. As used herein, in the examples and throughout the claims, the term “alkyl” (or “lower alkyl”) is intended to include both “unsubstituted alkyl” and “substituted alkyl,” the latter referring to an alkyl moiety having one or more substituents that substitute hydrogen at one or more carbon atoms in the hydrocarbon skeleton. Such substituents include, but are not limited to, halogens, hydroxyls, carbonyls (such as carboxyls, alkoxycarbonyls, formyl or acyls), thiocarbonyls (such as thioesters, thioacetates or thioformates), alkoxyls, phosphoryls, phosphoric acids, phosphonic acids, phosphinic acids, aminos, amides, amidines, imines, cyanos, nitros, azides, sulfhydryls, alkylthios, sulfuric acids, sulfonic acids, sulfamoyls, sulfonamidesulfonyls, heterocyclyls, aralkyls, or aromatic or heteroaromatic moieties.

[0045] Unless otherwise specified, as used herein, “lower alkyl” means an alkyl group as defined above, but having 1 to 10 carbon atoms or 1 to 6 carbon atoms in its skeletal structure. Similarly, “lower alkenyl” and “lower alkynyl” have similar chain lengths. Throughout this application, preferred alkyl groups are lower alkyl groups. In some embodiments, substituents designated as alkyl in this specification are lower alkyl groups.

[0046] Those skilled in the art will understand that a substituted portion on a hydrocarbon chain may, where appropriate, be substituted itself. For example, substituents on substituted alkyls may include halogens, hydroxyl, nitro, thiol, amino, azide, imino, amide, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl and sulfonates), and silyl groups, as well as ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, etc. Cycloalkyls may be substituted in a similar manner.

[0047] As used herein, the term “heteroalkyl” refers to a linear, branched, or cyclic carbon-containing radical or a combination thereof containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, Se, B, and S, where phosphorus and sulfur atoms are optionally oxidized, and nitrogen heteroatoms are optionally quaternized. Heteroalkyls can be substituted for alkyl groups as defined above.

[0048] The term "alkylthio" refers to an alkyl group having a sulfur radical bonded to it, as defined above. In some embodiments, the "alkylthio" portion is represented by one of -S-alkyl, -S-alkenyl, and -S-alkynyl groups. Representative alkylthio groups include methylthio and ethylthio. The term "alkylthio" also encompasses cycloalkyl groups, alkene groups, cycloalkene groups, and alkyne groups. "Arylthio" refers to an aryl group or heteroaryl group. Alkylthio groups can be substituted for alkyl groups as defined above.

[0049] The terms "alkenyl" and "alkynyl" refer to unsaturated aliphatic groups that are similar to the alkyl groups mentioned above in terms of length and possible substitutions, but each contains at least one double or triple bond.

[0050] As used herein, the term "alkoxyl" or "alkoxy" refers to an alkyl group as defined above, having an oxygen radical attached thereto. Representative alkoxyl groups include methoxy, ethoxy, propyloxy and tert-butoxy. An "ether" is two hydrocarbons covalently linked by an oxygen. Accordingly, the substituent of an alkyl that renders that alkyl an ether is or resembles an alkoxyl, and can be represented, for example, by one of -O-alkyl, -O-alkenyl and -O-alkynyl. Aroxy may be represented by -O-aryl or -O-heteroaryl, where aryl and heteroaryl are as defined below. The alkoxy and aroxy groups can be substituted as described above for alkyl.

[0051] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines, for example, moieties that can be represented by the general formula: [Chemical Formula] , wherein R9, R 10 , and R' 10 are each independently hydrogen, alkyl, alkenyl, -(CH2) m -R8, or R9 and R 10 together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure. R8 represents aryl, cycloalkyl, cycloalkenyl, heterocycle or polycycle, and m is zero or an integer in the range of 1 to 8. In some embodiments, only one of R9 or R 10 can be a carbonyl, for example, R9, R 10 and nitrogen do not together form an imide. In still other embodiments, the term "amine" does not encompass amides, for example, one of R9 and R 10 represents a carbonyl. In additional embodiments, R9 and R10 (and optionally R' 10 ) each independently represents hydrogen, alkyl or cycloalkyl, alkenyl or cycloalkenyl or alkynyl. Therefore, as used herein, the term "alkylamine" is an amine group having a substituted (as described above for alkyl) or unsubstituted alkyl bonded to it, i.e., R9 and R 10 This means that at least one of them is an alkyl group.

[0052] The term "amide" is recognized in the art as an amino-substituted carbonyl, and its general formula is: [ka] It includes parts that can be represented by R9 and R 10 It is defined as described above.

[0053] As used herein, "aryl" means C5-C 10This refers to member-based aromatic, heterocyclic, fusion aromatic, fusion heterocyclic, diaromatic, or diheterocyclic systems. In a broader sense, as used herein, "aryl" includes 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, and 10-membered monocyclic aromatic groups that may contain 0 to 4 heteroatoms, such as benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. Aryl groups having heteroatoms in their ring structure may also be called "aryl heterocyclic" or "heteroaromatic." Aromatic rings include, but are not limited to, halogens, azides, alkyls, aralkyls, alkenyls, alkynyls, cycloalkyls, hydroxyls, alkoxyls, aminos (or quaternated aminos), nitros, sulfhydryls, iminos, amides, phosphonic acids, phosphinates, carbonyls, carboxyls, silyls, ethers, alkylthios, sulfonyls, sulfonamides, ketones, aldehydes, esters, heterocycles, aromatic rings or heteroaromatic moieties, -CF3, -CN; and combinations thereof.

[0054] The term "aryl" also refers to a polycyclic ring system having two or more cyclic rings, where two or more carbons are common to two adjacent rings (i.e., fusion rings), and at least one of the rings is aromatic, and for example, the other cyclic ring or ring can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, and / or heterocyclic.Examples of heterocyclic compounds, though not limited to them, include: benzimidazolyl, benzofuranil, benzothiofuranil, benzothiophenyl, benzoxazolyl, benzoxazolinil, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinil, carbazolyl, 4aH-carbazolyl, carborinil, chromanil, chromenil, sinnolinil, decahydroquinolinil, 2H,6H-1,5,2-dithiadinyl, dihydroflo[2,3b]tetrahydrofuranil, furanil, furazanyl Imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolidinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranil, isochromanil, isoindazolyl, isoindonyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthilidinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1 ,3,4-Oxadiazolyl, Oxazolidinil, Oxazolyl, Oxadyl, Pyrimidinil, Phenanthronil, Phenadhiazinil, Phenoxatinil, Phenoxadinil, Phthalazinil, Piperadinil, Piperidinil, Piperidonil, 4-Piperidonil, Piperonil, Pteridinil, Prinnil, Pyranil, Pyrazinil, Pyrazolidinil, Pyrazolyl, Pyridadinil, Pyridoxazole, Pyridoimidazole, Pyridhiazole, Pyridinil, Pyridyl, Pyrimidinil, Pyrrolidinil Examples include pyrrolinil, 2H-pyrrolyl, pyrrolyl, quinazolinil, quinolinil, 4H-quinolidinil, quinoxalinil, quinusidinil, tetrahydrofuranil, tetrahydroisoquinolinil, tetrahydroquinolinil, tetrazolyl, 6H-1,2,5-thiadiadinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, and xanthenyl.One or more rings can be replaced with "aryls" as defined above.

[0055] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group (e.g., an aromatic group or a heteroaromatic group).

[0056] As used herein, the term “carbocyclic” refers to an aromatic or non-aromatic ring in which each atom of the ring is carbon.

[0057] "Heterocyclic" or "heterocyclic formula," as used herein, refers to a cyclic radical bonded via ring carbon or nitrogen in a monocyclic or bicyclic ring containing 3 to 10 ring atoms, preferably 5 to 6 ring atoms, and composed of 1 to 4 heteroatoms selected from the group consisting of non-peroxide oxygen, sulfur, and N(Y), with Y absent, or H, O, (C1-C 10) Alkyl, phenyl, or benzyl, optionally containing 1 to 3 double bonds, and optionally substituted with one or more substituents.Examples of heterocyclic rings, though not limited to them, include benzimidazolyl, benzofuranil, benzothiofuranil, benzothiophenyl, benzoxazolyl, benzoxazolinil, benzthiazolyl, benztriazolyl, benztetrazolyl, benzsoxazolyl, benzsothiazolyl, benzimidazolinil, carbazolyl, 4aH-carbazolyl, carborinil, chromanil, clomenil, sinnolinil, decahydroquinolinil, 2H,6H-1,5,2-dithiadinyl, dihydroflofuro[2,3-b]tetrahydrofuranil, furanil, furanil Zanil, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolidinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranil, isochromanil, isoindazolyl, isoindonyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthilidinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-Oxadiazolyl, Oxazolidinil, Oxazolyl, Oxadyl, Pyrimidinil, Phenantlidinil, Phenantronil, Phenadinil, Phenothiazinil, Phenoxatinil, Phenoxadinil, Phthalazinil, Piperadinil, Piperidinil, Piperidonil, 4-Piperidonil, Piperonil, Pteridinil, Prinnyl, Pyranil, Pyrazinil, Pyrazolidinil, Pyrazolyl, Pyrazolyl, Pyridadinil, Pyridoxazole, Pyridoimidazole, Pyridothiazol, Pyridinil, Pyridyl, Pyrimidinil, Pyrrolidinil Examples include pyrrolinil, 2H-pyrrolyl, pyrrolyl, quinazolinil, quinolinil, 4H-quinolidinil, quinoxalinil, quinusidinil, tetrahydrofuranil, tetrahydroisoquinolinil, tetrahydroquinolinil, tetrazolyl, 6H-1,2,5-thiadiadinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, and xanthenyl.The aromatic ring group, as defined above, may be optionally substituted with one or more substituents at one or more positions in, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amide, phosphonic acid, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocycle, aromatic ring or heteroaromatic moiety, -CF3 and -CN.

[0058] The term "carbonyl" is recognized in the art and has the general formula: [ka] It includes a part that can be represented by, where X is a bond, or represents oxygen or sulfur, and R 11 R' represents hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, or alkynyl. 11 X represents hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, or anyle. X is oxygen, and R 11 or R' 11 If X is not hydrogen, the formula represents an "ester". If X is oxygen, R 11 If as defined above, the part is referred to herein as a carboxyl group, and in particular R 11 When X is hydrogen, the formula represents a "carboxylic acid". 11 When X is hydrogen, the formula represents "formic acid". Generally, when the oxygen atom in the above formula is substituted with sulfur, the formula represents a "thiocarbonyl" group. When X is sulfur, R 11 or R' 11 If X is not hydrogen, the formula represents a "thioester". If X is sulfur, R 11 When X is hydrogen, the formula represents "thiocarboxylic acid". 11 If X is hydrogen, the formula represents "thioformic acid". On the other hand, if X is a bond, R 11 If X is not hydrogen, the above formula represents a "ketone" group. X is a bond, and R 11When the group is hydrogen, the above formula represents an "aldehyde" group.

[0059] As used herein, the term “monoester” refers to an analogue of a dicarboxylic acid, in which one carboxylic acid is functionalized as an ester and the other carboxylic acid is a free carboxylic acid or a salt of a carboxylic acid. Examples of monoesters include, but are not limited to, monoesters of succinic acid, glutamic acid, adipic acid, sulfite, sebacic acid, azeleic acid, oxalic acid, and maleic acid.

[0060] As used herein, the term “heteroatom” means an atom of any element other than carbon or hydrogen. Examples of heteroatoms include boron, nitrogen, oxygen, phosphorus, sulfur, and selenium. Other heteroatoms include silicon and arsenic.

[0061] As used herein, the term "nitro" means -NO2; the term "halogen" means -F, -Cl, -Br, or -I; the term "sulfhydryl" means -SH; the term "hydroxyl" means -OH; and the term "sulfonyl" means -SO2-.

[0062] As used herein, the term “substituted” refers to all permissible substituents of the compounds described herein. In its broadest sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Exemplary substituents, but not limited to, include halogens, hydroxyl groups, or any other organic group containing any number of carbon atoms, preferably 1 to 14 carbon atoms, and optionally, one or more heteroatoms such as oxygen, sulfur, or nitrogen in linear, branched, or cyclic structural formats. Typical substituents include alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, alloxy, substituted alloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amide, substituted amide, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C 3- C 20 Circular, substitution C 3- C 20 This includes cyclic, heterocyclic, substituted heterocyclic, amino acid, peptide, and polypeptide groups.

[0063] Heteroatoms such as nitrogen may have hydrogen substituents and / or any acceptable substituents of the organic compounds described herein that satisfy the valence of the heteroatom. Naturally, “substituted” or “substituted” implicitly includes the condition that such substitutions comply with the acceptable valences of the substituted atom and substituent, and that the substitutions result in stable compounds, i.e., compounds that do not undergo spontaneous transformation by reconstitution, cyclization, or removal, etc.

[0064] In broad embodiments, acceptable substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Exemplary substituents include, for example, those described herein. Acceptable substituents may be one or more identical or different substituents for a given organic compound. Heteroatoms such as nitrogen may have hydrogen substituents and / or any acceptable substituents of the organic compound described herein that satisfy the valence of the heteroatom.

[0065] In various embodiments, the substituents are selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamic acid, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydroxyl, ketone, nitro, phosphate, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone, each optionally substituted with one or more suitable substituents. In some embodiments, substitutions are selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamic acid, carboxy, cycloalkyl, ester, ether, formyl, haloalkyl, heteroaryl, heterocyclyl, ketone, phosphoric acid, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone, and alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamic acid, carboxy, cycloalkyl, ester, ether, formyl, haloalkyl, heteroaryl, heterocyclyl, ketone, phosphoric acid, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone may be further substituted with one or more suitable substituents.

[0066] Examples of substituents, though not limited to them, include halogens, azides, alkyls, aralkyls, alkenyls, alkynyls, cycloalkyls, hydroxyls, alkoxyls, aminos, nitrosulfhydryls, iminos, amides, phosphonic acids, phosphinates, carbonyls, carboxyls, silyls, ethers, alkylthios, sulfonyls, sulfonamides, ketones, aldehydes, thioketones, esters, heterocyclyls, -CNs, aryls, aryloxys, perhalalkoxys, aralkoxys, heteroaryls, heteroaryloxys, heteroarylalkyls, and hetero Examples include aralkoxy, azide, alkylthio, oxo, acylalkyl, carboxyester, carboxamide, acyloxy, aminoalkyl, alkylaminoaryl, alkylaryl, alkylaminoalkyl, alkoxyaryl, arylamino, aralkylamino, alkylsulfonyl, carboxamidealkylaryl, carboxamidearyl, hydroxyalkyl, haloalkyl, alkylaminoalkylcarboxy, aminocarboxamidealkyl, cyano, alkoxyalkyl, perhaloalkyl, and arylalkyloxyalkyl. In some embodiments, the substituent is selected from cyano, halogen, hydroxyl, and nitro.

[0067] The terms “polypeptide,” “peptide,” and “protein” generally refer to polymers of amino acid residues. As used herein, these terms also apply to amino acid polymers in which one or more amino acids are chemical analogs or modified derivatives of corresponding naturally occurring amino acids. The term “protein,” as commonly used herein, refers to a polymer of amino acids linked together by peptide bonds, forming polypeptides whose chain length is sufficient to create tertiary and / or quaternary structures. The term “protein” excludes small peptides by definition, as small peptides lack the higher-order structures necessary to be considered proteins.

[0068] As used herein, the term “substantially free” means that the compound is not present in therapeutically effective amounts when administered at the recommended dose, but may contain trace amounts of the compound that are not therapeutically effective. With respect to compositions of the present invention that are substantially free of reducing sugars such as lactose, and / or sugar alcohols such as mannitol, minimizing the amount of such reducing sugars such as lactose, and / or in which mannitol may be present, for example, less than about 10%, less than about 9%, less than about 8%, less than 7%, less than 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, less than about 0.5%, including all and partial ranges in between.

[0069] This disclosure relates to a pharmaceutical composition comprising a phosphoinositide 3 kinase (PI3K) inhibitor such as Compound I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a salt solvate thereof. [ka] (Compound I)

[0070] The present invention also includes compound I or a pharmaceutically acceptable salt thereof as a compound in any combination therapy or in use or method of treatment with additional therapeutic agents.

[0071] CLM The compounds of this disclosure may contain one or more CLMs. A CLM can be any covalent modality capable of forming a covalent bond with a biological target. A CLM may contain one or more chemical moieties, one or more chemical moieties capable of forming a covalent bond with a biological target. In certain embodiments, a CLM may contain an internal linker or spacer. The internal linker or spacer may be two parts of the CLM combined or bonded to the CLM.

[0072] In some embodiments, CLM comprises at least one substituted or unsubstituted alkyne. In some embodiments, CLM comprises at least one substituted or unsubstituted acrylamide. In some embodiments, CLM comprises at least one substituted or unsubstituted vinylsulfonamide. In some embodiments, CLM comprises at least one substituted or unsubstituted vinylsulfone. In some embodiments, CLM comprises at least one substituted or unsubstituted fumaamide. In some embodiments, CLM comprises at least one substituted or unsubstituted acrylic acid. In some embodiments, CLM comprises at least one substituted or unsubstituted isothiocyanate. In some embodiments, CLM comprises at least one substituted or unsubstituted sulfonyl fluoride. In some embodiments, CLM comprises at least one substituted or unsubstituted fluorosulfate. In some embodiments, CLM comprises at least one substituted or unsubstituted formylphenylboronic acid. In some embodiments, CLM comprises at least one substituted or unsubstituted boronic acid. In some embodiments, CLM comprises at least one activated ester. In some embodiments, CLM comprises at least one substituted or unsubstituted thioester. In some embodiments, CLM comprises at least one sulfonyl group. In some embodiments, CLM comprises at least one nitro group. In some embodiments, CLM comprises at least one substituted or unsubstituted epoxide. In some embodiments, CLM comprises at least one substituted or unsubstituted formylphenylboronic acid. In some embodiments, CLM comprises at least one substituted or unsubstituted aryl halide. In some embodiments, CLM comprises at least one substituted or unsubstituted aldehyde. In some embodiments, CLM comprises at least one substituted or unsubstituted triazine. In some embodiments, CLM comprises at least one substituted or unsubstituted cyanoacrylamide. In some embodiments, CLM comprises at least one substituted or unsubstituted chloroacetamide.

[0073] Examples of CLMs, though not limited to these, include the following: [ka] [ka] [ka] [ka] A, B, C, and D appear independently in each instance as H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C 1-6 Alkyl, N(C 1-3 Alkyl)C(O)-C 1-6 Alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkinyl, optionally substituted C 3-6 Selected from the group consisting of cycloalkyls, optionally substituted 5-10 membered heterocycles, optionally substituted aryls, and optionally substituted 5-10 membered heteroaryls, any substituent for A, B, C, and D can be independently halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, -S-CH3, or optionally substituted C 1-3 Alkyl and optionally substituted C 3-6These are 1 to 3 substituents selected from the group consisting of cycloalkyl groups. A1, A2, A3, A4, A5, and A6 are either absent in each occurrence or independently substituted with H, halogen, CF3, -OH, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C1-6 alkyl, N(C1-3 alkyl)C(O)-C1-6 alkyl, OC(O)NH2, OC(O)NH(CH3), OC(O)N(CH3)2, imidazolyl, pyrazolyl, methylimidazolyl, methylpyrazolyl, or optionally substituted. The substituents are selected from C1-6 alkyl groups, optionally substituted C2-6 alkenyl groups, optionally substituted C2-6 alkynyl groups, optionally substituted C3-6 cycloalkyl groups, optionally substituted 5-10 member heterocycles, optionally substituted aryl groups, and optionally substituted 5-10 member heteroaryl groups, and the optionally substituted substituents of A1, A2, A3, A4, A5, and A6 are independently halogens, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, and -CH2 The substituents are one to two substituents selected from the group consisting of NH2, -C(O)CH3, SH, -S-CH3, optionally substituted C1-3 alkyls, and optionally substituted C3-6 cycloalkyls, wherein the optionally substituted C1-3 alkyls and C3-6 cycloalkyls are selected from the group consisting of halogens, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, and -S-CH3, or any fragment or analog thereof.

[0074] In some embodiments, the CLM is [ka] It is selected from the group consisting of the following.

[0075] Linker The compounds in the pharmaceutical compositions of this disclosure may contain one or more arbitrary linkers that link the FCB and CLM. The linker, L, can be bonded anywhere on the FCB and CLM, as long as the efficacy of the FCB and the bonding of the CLM are not significantly affected. In some embodiments, the CLM includes an arbitrary internal linker.

[0076] In some embodiments, the linker (including the internal linker of the CLM) is a small molecule. In some embodiments, the linker (including the internal linker of the CLM) is selected, but not limited to, substituted and unsubstituted C1-C30 alkyl, substituted and unsubstituted C2-C30 alkenyl, substituted and unsubstituted C2-C30 alkynyl, substituted and unsubstituted C3-C30 cycloalkyl, substituted and unsubstituted C1-C30 heterocycloalkyl, substituted and unsubstituted C3-C30 cycloalkenyl, substituted and unsubstituted C1-C30 heterocycloalkenyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl.

[0077] In some embodiments, the linker (including the internal linker of the CLM) is C1-C10 linear alkyl, C1-C10 linear O-alkyl, C1-C10 linear substituted alkyl, C1-C10 linear substituted O-alkyl, C4-C13 branched alkyl, C4-C13 branched O-alkyl, C2-C12 linear alkenyl, C2-C12 linear O-alkenyl, C3-C12 linear substituted alkenyl, C3-C12 linear substituted O-alkenyl, polyethylene glycol, polylactic acid, polyglycolic acid, poly( The linker may be lactide-co-glycolide, polycarboxylate, polycyanoacrylate, ketone, aryl, heterocyclic, succinic acid ester, amino acid, aromatic group, ether, crown ether, urea, thiourea, amide, purine, pyrimidine, bipyridine, indole derivatives acting as crosslinkers, chelating agents, aldehydes, ketones, bisamines, bisalcohols, heterocyclic structures, aziline, disulfide, thioether, hydrazone, and combinations thereof. For example, the linker may be a C3 linear alkyl or ketone. The alkyl chain of the linker may be substituted with one or more substituents or heteroatoms. In some embodiments, the linker contains one or more atoms or groups selected from -O-, -C(=O)-, -NR, -OC(=O)-NR-, -S-, and -SS-. The linker may be selected from dicarboxylate derivatives of succinic acid, glutamic acid, or diglycolic acid.

[0078] In some embodiments, the alkyl chain of the linker may be optionally interrupted by one or more atoms or groups selected from -O-, -C(=O)-, -NR, -OC(=O)-NR-, -S-, and -SS-. The linker may be selected from dicarboxylate derivatives of succinic acid, glutamic acid, or diglycolic acid.

[0079] In some embodiments, the linker may be non-cleavable. In some embodiments, the linker may be cleavable. In some embodiments, the linker may be cleaved by an enzyme.

[0080] As an unrestricted example of a linker, [ka] [ka] These include, D1, D2, D3, D4, D5, and D6 are independently selected in each occurrence from the group consisting of N, C, O, or S. If D1-6 is N, the corresponding position is trivalent, and if D1-6 is O or S, the corresponding position is divalent. B1, B2, B3, B4, B5, and B6 are absent in each occurrence or independently H, halogen, CF3, -OH, -CH3, -NH2, -SH, -SCH3, -CN, -NO2, -CH2(NH2), -C(O)OH, -S(O)2NH2, -C(O)NH2, -C(O)CH3, NHC(O)-C1-6 alkyl, N(C1-3 alkyl)C(O)-C1-6 alkyl, OC(O)NH2, OC(O)N The optional substituents of B1, B2, B3, and B4 are independently halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2 The substituents are 1 to 3 substituents selected from the group consisting of NH2, -C(O)CH3, SH, -S-CH3, optionally substituted C1-3 alkyl, and optionally substituted C3-6 cycloalkyl, wherein the optionally substituted C1-3 alkyl and C3-6 cycloalkyl substituents are 1 to 2 substituents selected from the group consisting of halogen, OH, NH2, CH3, CF3, -CN, -NO2, -C(O)OH, -S(O)2NH2, -C(O)NH2, -CH2NH2, -C(O)CH3, SH, and -S-CH3, or any fragment or analog thereof.

[0081] In some embodiments, the linker is [ka] A selection is made from the group consisting of, and either end can be connected to the CLM. In some embodiments, [ka] The linker selected from the group consisting of the following is: [ka] It is connected to a CLM selected from the group consisting of the following.

[0082] In some embodiments, the kinase is PI3 kinase (PI3K). The therapeutic conjugate may have the structure (FCB)a-(L)b-(CLM)c, where a and c are independently integers from 1 to 5, and b is an integer from 0 to 5, and the FCB portion comprises a PI3K inhibitor or a fragment, analog, or derivative thereof.

[0083] The FCB, L (linker), and CLM parts are explained in the section above. In one non-exclusive example, the FCB is... [ka] Includes.

[0084] In some embodiments, the FCB is structured [ka] It is a compound having, and R1 is [ka] It is selected from the group consisting of the following.

[0085] Pharmaceutical composition In some embodiments, the present invention provides pharmaceutical compositions comprising a therapeutically effective amount of a compound of formula (FCB)a-(L)b-(CLM)c, and / or a subgenus thereof, or compound I, or a pharmaceutically acceptable salt, ester, solvate, and / or prodrug thereof, combined with a pharmaceutically acceptable excipient or carrier as an active ingredient.

[0086] [ka] (Compound I)

[0087] In certain embodiments, the pharmaceutical composition enables the immediate release of compound I.

[0088] In some embodiments, compound I is the crystalline form of compound I, or a pharmaceutically acceptable salt or solvate of compound I. In some embodiments, the pharmaceutically acceptable salt of compound I is selected from the group consisting of phosphates, hydrochlorides, sulfates, mesylates, benzenesulfons, tosylates, fumarates, maleates, L-tartrates, citrates, and succinates, or their solvates. In some embodiments, compound I is anhydrous or non-solvated. In another embodiment, the crystalline form is the phosphate of compound I or its solvate. In certain embodiments, the crystalline form is a hydrate. In another embodiment, the present invention may be an amorphous means of compound I.

[0089] In certain embodiments, the form of compound I in the pharmaceutical composition described herein is derived from PCT application PCT / CN2023 / 083760, which is incorporated herein by reference in its entirety.

[0090] Pharmaceutical excipients can be added to pharmaceutical compositions or formulations for a variety of purposes, including the immediate release of compounds after oral administration to a patient.

[0091] In some embodiments, the pharmaceutical compositions herein may be in the form of tablets. In some embodiments, the tablets herein have a weight range of about 20 mg to about 500 mg, or about 50 mg to about 400 mg, or about 70 mg to about 250 mg, or about 80 mg to about 150 mg. In some embodiments, the tablets herein have a weight of about 100 mg. In some embodiments, the tablets have a diameter of about 15 mm, 14 mm, 13 mm, 12 mm, 11 mm, 10 mm, 9 mm, 8 mm, 7 mm, 6 mm, 5 mm, or less than 4 mm. In some embodiments, the tablets are coated tablets. In some embodiments, the tablets have a hardness greater than about 0.5 Kp, about 0.7 Kp, about 1.0 Kp, about 1.2 Kp, about 1.5 Kp, or about 2.0 Kp when compressed using a 6 mm tablet tooling size. In some embodiments, the tablets have an abrasion degree of less than 3%, less than 2%, or less than 1%.

[0092] In some embodiments, the compound that forms a covalent bond with a kinase or pseudokinase, the compound of formula (FCB)a-(L)b-(CLM)c, and / or its subgenus, or compound I, or its pharmaceutically acceptable salts, esters, or solvates, is present in an amount ranging from about 0.1% to about 50% by weight, or 1% to about 40% by weight, or about 3% to about 20% by weight, or about 4% to about 10% by weight, based on the total weight of the composition or tablet. In some embodiments, the compound that forms a covalent bond with a kinase or pseudokinase, the compound of formula (FCB)a-(L)b-(CLM)c, and / or its subgenus, or compound I, or its pharmaceutically acceptable salts, esters, or solvates, is present in an amount of about 5% by weight.

[0093] In some embodiments, the compound that forms a covalent bond with a kinase or pseudokinase, the compound of formula (FCB)a-(L)b-(CLM)c, and / or its subgenus, or compound I, or its pharmaceutically acceptable salts, esters, or solvates, is present in an amount ranging from about 0.1 mg to about 30 mg per tablet, or about 1 mg to about 20 mg per tablet, or about 3 mg to about 7 mg per tablet, or about 5 mg to about 6 mg per tablet. In some embodiments, the compound that forms a covalent bond with a kinase or pseudokinase, the compound of formula (FCB)a-(L)b-(CLM)c, and / or its subgenus, or compound I, or its pharmaceutically acceptable salts, esters, or solvates, is present in an amount of 5 mg per tablet.

[0094] In some embodiments, suitable pharmaceutical excipients may include fillers, binders, disintegrants, lubricants, coatings, or any combination thereof.

[0095] In some embodiments, fillers include, for example, materials such as silicon dioxide, titanium dioxide, alumina, talc, kaolin, powdered cellulose, microcrystalline cellulose, urea, sodium chloride, and sugars, or combinations thereof. Any suitable sugar may be used in the compositions of the present disclosure. As used herein, “sugars” as used in the present invention includes sugar alcohols, monosaccharides, disaccharides, and oligosaccharides. Exemplary sugar alcohols include, but are not limited to, xylitol, mannitol, sorbitol, erythritol, lactitol, pentitol, and hexitol. Exemplary monosaccharides include, but are not limited to, glucose, fructose, aldoses, and ketoses. Exemplary disaccharides include, but are not limited to, sucrose, isomalt, lactose, trehalose, and maltose. Exemplary oligosaccharides include, but are not limited to, fructooligosaccharides, inulin, galactooligosaccharides, and mannanoligosaccharides. In some embodiments, the sugar is sorbitol, mannitol, or xylitol. In some embodiments, the sugar is sorbitol. In some embodiments, the sugar is sucrose.

[0096] In some embodiments, suitable pharmaceutically acceptable fillers may be selected from microcrystalline cellulose, silicified microcrystalline cellulose, mannitol EZ, pregelatinized starch, dicalcium phosphate dihydrate, lactose monohydrate, or any combination thereof.

[0097] In some embodiments, the filler used in this disclosure is in an amount ranging from about 10% to about 95% by weight, 10% to about 80% by weight, or about 20% to about 70% by weight, or about 30% to about 60% by weight, or about 40% to about 60% by weight, or about 40% to about 50% by weight, or about 42% to 46% by weight, based on the total weight of the composition or tablet. In some embodiments, the filler used herein is in an amount ranging from about 30% to about 60% by weight. In some embodiments, the filler used herein is in an amount of about 30% by weight. In some embodiments, the filler used herein is in an amount of about 50% by weight, up to about 40% by weight, based on the weight of the composition or tablet. In some embodiments, the filler used herein is in an amount ranging from about 45% to about 60% by weight, based on the total weight of the composition or tablet. In some embodiments, the filler used herein is in an amount ranging from about 70% to about 90% by weight, based on the total weight of the composition or tablet.

[0098] In some embodiments, the amount of the filler used herein is in the range of about 10 mg to about 80 mg per tablet, or about 20 mg to about 70 mg per tablet, or about 30 mg to about 60 mg per tablet, or about 40 mg to about 50 mg per tablet, or about 42 mg to about 46 mg per tablet. In some embodiments, the amount of the filler used herein is 30 mg per tablet. In some embodiments, the amount of the filler used herein is 42.5 mg per tablet.

[0099] In some embodiments, the amount of the filler used herein is in the range of about 100 mg to about 800 mg per tablet, or about 200 mg to about 700 mg per tablet, or about 300 mg to about 600 mg per tablet, or about 400 mg to about 500 mg per tablet, or about 420 mg to about 460 mg per tablet. In some embodiments, the amount of the filler used herein is 300 mg per tablet. In some embodiments, the amount of the filler used herein is about 200 mg to about 300 mg per tablet.

[0100] In certain embodiments, the filler comprises microcrystalline cellulose in the ratios and amounts described herein.

[0101] A binder can be used to impart tackiness to a tablet, thus ensuring that the tablet or tablet layer remains intact after compression. Suitable binder materials include, but are not limited to, starches (including corn starch and pregelatinized starch), gelatin, sugars (sucrose, glucose, dextrose, and lactose), polyethylene glycol, polyvinyl alcohol, waxes, natural rubber, and synthetic rubber, as well as, for example, sodium acacia alginate, polyvinylpyrrolidone, cellulosic polymers (including hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose, microcrystalline cellulose, ethylcellulose, hydroxyethylcellulose, and similar materials), Veegum, and combinations thereof. Examples of polyvinylpyrrolidone include povidone, copovidone, and crospovidone.

[0102] Solid pharmaceutical compositions that are compressed into dosage forms such as tablets may contain excipients whose function is to help bind the active ingredient and other excipients together after compression. Binders for solid pharmaceutical compositions and / or combinations include acacia, alginic acid, carbomer (e.g., carbopol), sodium carboxymethylcellulose, dextrin, ethylcellulose, gelatin, guar gum, tragacanth gum, hydrogenated vegetable oil, hydroxyethylcellulose, hydroxypropylcellulose (e.g., KLUCEL), hydroxypropylmethylcellulose (e.g., METHOCEL), liquid glucose, magnesium aluminum silicate, maltodextrin, methylcellulose, polymethacrylate, povidone (e.g., KOLLIDON, PLASDONE), pregelatinized starch, sodium alginate, and starch.

[0103] In some embodiments, suitable pharmaceutically acceptable binders may be selected from povidone K29 / 32, hydroxypropylcellulose, hydroxypropylmethylcellulose, or any combination thereof.

[0104] In some embodiments, the binder used in the present disclosure is in an amount ranging from about 0.1% to about 10% by weight, or about 1% to about 10% by weight, or about 2% to about 8% by weight, or about 2% to about 6% by weight, or about 2% to about 5% by weight, or about 3% to about 4% by weight, based on the total weight of the composition or tablet.

[0105] In some embodiments, the amount of the binder used herein is in the range of about 0.1 mg to about 10 mg per tablet, or about 1 mg to about 10 mg per tablet, or about 2 mg to about 8 mg per tablet, or about 2 mg to about 6 mg per tablet, or about 2 mg to about 5 mg per tablet, or about 3 mg to 4 mg per tablet.

[0106] In certain embodiments, the binder comprises hydroxypropyl cellulose in the ratios and amounts of the binder described herein.

[0107] Disintegrants are used to facilitate the disintegration of tablets, thereby increasing the rate of erosion relative to the rate of dissolution, and are generally starch, clay, cellulose, algin, gum, or cross-linked polymers (e.g., cross-linked polyvinylpyrrolidone). Other non-limiting examples of suitable disintegrants include, for example, lightly cross-linked polyvinylpyrrolidone, corn starch, potato starch, maize starch and modified starch, croscarmellose sodium, crospovidone, sodium starch glycolate, and combinations and mixtures thereof.

[0108] In some embodiments, the pharmaceutically acceptable disintegrant may be selected from croscarmellose sodium, crospovidone XL, or any combination thereof.

[0109] In some embodiments, the disintegrant used herein is in an amount ranging from about 0.1% to about 10% by weight, or about 1% to about 10% by weight, or about 2% to about 8% by weight, or about 2% to about 6% by weight, or about 2% to about 5% by weight, or about 3% to 4% by weight of the total weight of the composition or tablet. In some embodiments, the incorporating agent used herein is in an amount ranging from about 0.1 mg to about 10 mg per tablet, or about 1 mg to about 10 mg per tablet, or about 2 mg to about 8 mg per tablet, or about 2 mg to about 6 mg per tablet, or about 2 mg to about 5 mg per tablet, or about 3 mg to 4 mg per tablet, or about 1 mg to 2 mg per tablet, or about 8 mg to 10 mg per tablet.

[0110] In certain embodiments, the disintegrant comprises crospovidone XL in the proportions and amounts described herein.

[0111] Lubricants can be added to improve the fluidity of incompressible solid compositions and to improve the accuracy of administration. Excipients that can function as flow enhancers include, for example, colloidal silicon dioxide, magnesium trisilicate, powdered cellulose, starch, talc, and tribasic calcium phosphate.

[0112] In some embodiments, suitable pharmaceutically acceptable flow enhancers may be selected from colloidal silicon dioxide, Talc, or any combination thereof.

[0113] In some embodiments, the amount of the flow enhancer is in the range of about 0.01% to about 10% by weight, or about 0.1% to about 5% by weight, or about 0.1% to about 3% by weight, or about 0.2% to about 2% by weight, or about 0.3% to 1% by weight, or about 0.3% to 0.6% by weight, or about 1% to 5% by weight, or about 2% to 3% by weight, or about 0.4% to about 0.5% by weight, based on the total weight of the composition or tablet. In some embodiments, the amount of the flow enhancer used herein is in the range of about 0.01 mg to about 10 mg per tablet, or about 0.1 mg to about 5 mg per tablet, or about 0.1 mg to about 3 mg per tablet, or about 0.2 mg to about 2 mg per tablet, or about 0.3 mg to about 1 mg per tablet, or about 0.3 mg to about 0.6 mg per tablet, or about 0.4 mg to about 0.5 mg per tablet, or about 6 mg to about 10 mg per tablet, or about 7 mg to about 9 mg per tablet.

[0114] In certain embodiments, the flow accelerator comprises colloidal silicon dioxide in the ratio and amount of the flow accelerator described herein.

[0115] Lubricants can be used to facilitate tablet manufacturing, promote powder flow, and prevent particle capping (e.g., particle breakage) when pressure is reduced. Useful lubricants include magnesium stearate, calcium stearate, stearic acid, glyceryl behenate, talc, colloidal silicon dioxide such as Aerosil® 200, mineral oil (in PEG), hydrogenated vegetable oil (e.g., consisting of hydrogenated triglycerides and refined triglycerides of stearic acid and palmitic acid), and combinations thereof.

[0116] When dosage forms such as tablets are produced by compressing powder compositions and / or combinations, the compositions are subjected to pressure from the punch and dye. Some excipients and active ingredients tend to adhere to the surfaces of the punch and dye, which can cause pitting and other surface irregularities in the product. Lubricants can be added to the composition to reduce adhesion and facilitate the release of the product from the dye. Examples of lubricants include magnesium stearate, calcium stearate, glyceryl monostearate, glyceryl palmitostearate, hydrogenated castor oil, hydrogenated vegetable oil, mineral oil, polyethylene glycol, sodium benzoate, sodium lauryl sulfate, sodium stearyl fumarate, stearic acid, talc, and zinc stearate.

[0117] In some embodiments, suitable pharmaceutically acceptable lubricants may be selected from magnesium stearate (e.g., from a plant source), sodium steryl fumarate, glyceryl behenate, or any combination thereof.

[0118] In some embodiments, the lubricant used herein is in an amount ranging from about 0.01% to about 10% by weight of the total weight of the composition or tablet, or about 0.1% to about 5% by weight, or about 0.1% to about 3% by weight, or about 0.2% to about 2% by weight, or about 0.3% to 1% by weight, or about 0.3% to 0.6% by weight, or about 0.5% to 1.5% by weight, or about 0.7% to 1.2% by weight, or about 0.4% to 0.5% by weight.

[0119] In some embodiments, the amount of lubricant used herein is in the range of about 0.01 mg to about 10 mg per tablet, or about 0.1 mg to about 5 mg per tablet, or about 0.1 mg to about 3 mg per tablet, or about 0.2 mg to about 2 mg per tablet, or about 0.3 mg to about 1 mg per tablet, or about 1 mg to about 3 mg per tablet, or about 2 mg to about 3 mg per tablet, or about 0.3 mg to about 0.6 mg per tablet, or about 0.4 mg to 0.5 mg per tablet.

[0120] In certain embodiments, the lubricant comprises magnesium stearate in the ratios and amounts described herein.

[0121] In some embodiments, a suitable pharmaceutically acceptable coating may be selected from water-soluble polymers. In some embodiments, a suitable pharmaceutically acceptable coating may be selected from Opadry II White (PVA), Opadry II White (HPMC), or any combination thereof.

[0122] In some embodiments, the coating used herein is in an amount ranging from about 0.1% to about 10% by weight of the composition or tablet, or about 1% to about 10% by weight, or about 2% to about 8% by weight, or about 2% to about 6% by weight, or about 2% to about 5% by weight, or about 3% to 4% by weight.

[0123] In some embodiments, the coatings used herein are applied to tablets for a weight increase of about 1% by weight to about 20% by weight, based on the total weight of the tablets. In some embodiments, the coatings used herein are applied to tablets for a weight increase of about 10% by weight, or about 7% by weight, or about 5% by weight, or about 3% by weight, or about 1% by weight, based on the total weight of the tablets.

[0124] In some embodiments, the amount of coating used herein is in the range of about 0.1 mg to about 10 mg per tablet, or about 1 mg to about 10 mg per tablet, or about 2 mg to about 8 mg per tablet, or about 2 mg to about 6 mg per tablet, or about 2 mg to about 5 mg per tablet, or about 3 mg to 4 mg per tablet.

[0125] In some embodiments, the pharmaceutically active ingredients used herein are selected from microcrystalline cellulose (Avicel PH 101®), lactose monohydrate (Fast Flo®), hydroxypropyl cellulose (Klucel EXF®), croscarmellose sodium (Ac-di-sol®), colloidal silicon dioxide (Cab-O-Sil®), magnesium stearate, or any combination thereof.

[0126] In some embodiments, the pharmaceutically acceptable excipients used herein are selected from microcrystalline cellulose, crospovidone XL, colloidal silicon dioxide (Cab-O-Sil®), plant-derived magnesium stearate, hydroxypropyl cellulose, or any combination thereof.

[0127] In certain embodiments, the pharmaceutical composition comprises microcrystalline cellulose, hydroxypropyl cellulose, crospovidone XL, colloidal silicon dioxide (Cab-O-Sil®), and plant-derived magnesium stearate. In another specific embodiment, the pharmaceutical composition or formulation is a tablet. In another specific embodiment, the tablet is an oral tablet. In yet another specific embodiment, the oral tablet is an immediate-release oral tablet. In certain embodiments, the pharmaceutical composition and formulation include a tablet coating. In certain embodiments, the tablet is coated with Opadry II white.

[0128] In other embodiments, the pharmaceutical compositions and formulations may be prepared as described in the examples of this specification. In specific embodiments, the pharmaceutical compositions and formulations may be as described in Tables 2, 3, 5, 9, 12, or 15 as described herein.

[0129] In certain embodiments, the pharmaceutical composition and formulation are tablets having a specific dissolution and disintegration profile. In certain embodiments, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the drug, such as compound I, is released within about 10 minutes. In another particular embodiment, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of compound I is released within about 9 minutes, 8 minutes, 7 minutes, 6 minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes, or 1 minute.

[0130] In another specific embodiment, the dissolution of the drug in the tablet is determined using 900 mL of 0.01 M HCl at 37.0°C, Apparatus II (paddle), 75 rpm, or 100 rpm.

[0131] In another specific embodiment, the pharmaceutical composition comprises a therapeutically effective amount of a compound that forms a covalent bond with a kinase or pseudokinase, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient, the pharmaceutical composition enabling the immediate release of the compound, and the pharmaceutical composition being substantially free of impurities related to the compound. In a specific embodiment, the compound is compound I, or a pharmaceutically acceptable salt or solvated compound thereof, and the impurities are related to compound I, i.e., analogues or results of the reaction between compound I and the excipient in the composition.

[0132] In another embodiment, the pharmaceutical composition is substantially free of reducing sugars, i.e., sugars having free aldehyde or free ketone groups. In another embodiment, the pharmaceutical composition is substantially free of lactose, galactose, glucose, maltose, fructose, ribose, and / or xylose. In another embodiment, the pharmaceutical composition is substantially free of lactose. In a particular embodiment, lactose is lactose anhydride. In a particular embodiment, the composition of the present invention is substantially free of lactose isomers such as sucrose, trehalose, maltose, isomaltose, maltulose, isomaltulose, turanose, and cellobiose. In another embodiment, the composition of the present invention is substantially free of mannitol such as sorbitol, and isomers of compounds closely related to mannitol such as xylitol, erythritol, lactitol, and maltitol. In another particular embodiment, the compound is compound I or a pharmaceutically acceptable salt or solvate thereof. In a particular embodiment, compound I is in the form of a salt. In another particular embodiment, the salt is a phosphate.

[0133] The pharmaceutical formulations of this disclosure may be prepared by methods known to those skilled in the art, such as mixing, dissolving, granulation, sugar coating preparation, raising, emulsification, encapsulation, or freeze-drying processes. As described above, the pharmaceutical compositions of this disclosure may include one or more pharmaceutically acceptable carriers, such as excipients and adjuvants, that facilitate the processing of the active molecule into a preparation for pharmaceutical use.

[0134] In some embodiments, the pharmaceutical compositions of this disclosure are prepared as oral formulations. For oral administration, the compounds can be readily formulated by combining the active compound with a pharmaceutically acceptable carrier known in the art. Such carriers can formulate the pharmaceutically active ingredient for oral ingestion by a subject as tablets, pills, sugar-coated tablets, capsules, liquids, gels, syrups, slurries, suspensions, etc. The pharmaceutically active composition for oral use may be obtained as a solid excipient after the addition of a suitable adjuvant, if desired, to obtain a tablet or sugar-coated tablet core, and the resulting mixture may be optionally pulverized and processed into a granular mixture. Such oral pharmaceutical compositions may also be prepared by pulverization or melt extrusion. Suitable excipients may be fillers such as any of those disclosed herein, particularly sugars including lactose, sucrose, mannitol, or sorbitol, or cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP) preparations. Disintegrants such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or salts thereof such as sodium alginate may also be used. Wetting agents such as sodium dodecyl sulfate may be added.

[0135] In some embodiments of this disclosure, pharmaceutical compositions are prepared for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In some such embodiments, the pharmaceutical composition comprises a carrier and is formulated in an aqueous solution such as water, or a physiologically compatible buffer such as Hanks' solution, Ringer's solution, or physiological saline buffer. In some embodiments, other components are included (e.g., components that aid solubility or function as preservatives). In some embodiments, the injectable suspension is prepared using a suitable liquid carrier, suspension agent, etc. Specific pharmaceutical compositions for injection are presented in unit dosage forms, for example, in ampoules or multi-dose containers. Specific pharmaceutical compositions for injection are suspensions, solutions, or emulsions in an oily or aqueous vehicle and may contain formulations such as suspension agents, stabilizers, and / or dispersants. Specific solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils such as vegetable oils (e.g., sesame oil), and synthetic fatty acid esters such as ethyl oleate or triglycerides. The aqueous injection suspension may contain a substance that increases the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, such suspension may also contain a suitable stabilizer or agent that increases the solubility of the drug to enable the preparation of a highly concentrated solution.

[0136] Sterile injectable preparations may be prepared as sterile injectable solutions or suspensions in non-toxic, parenterally administered diluents or solvents such as 1,3-butanediol solution, or as lyophilized powders. Acceptable vehicles and solvents that may be used are water, Ringer's solution, and isotonic sodium chloride solution. Furthermore, sterile fixatives may conventionally be used as solvents or suspension media. For this purpose, any non-irritating fixative, including synthetic monoglycerides or diglycerides, may be used. In addition, fatty acids such as oleic acid may be used in the preparation of injectable preparations. Preparations for intravenous administration may include solutions in sterile isotonic aqueous buffer. If necessary, the preparation may also include solubilizers and local anesthetics to reduce pain at the injection site. Generally, the components are supplied separately or mixed together in unit dosage forms, for example, as dry lyophilized powders or anhydrous concentrates in sealed containers such as ampoules or pouches indicating the amount of the active agent. When the compound is administered by infusion, it may be dispensed into the preparation with sterile pharmaceutical-grade water, saline, or dextrose / water in an infusion bottle. If the compound is administered by injection, an ampoule of sterile water or saline solution for injection may be provided so that the components can be mixed before administration.

[0137] Suitable formulations include aqueous and non-aqueous sterile injection solutions that may contain antioxidants, buffers, bacteriostatic agents, bactericidal antibiotics, and solutes that make the formulation isotonic with the body fluids of the intended recipient, as well as aqueous and non-aqueous sterile suspensions that may contain suspending agents and thickeners. Liposome suspensions containing tissue-targeting liposomes may be present in suitable formulations.

[0138] Parenteral administration of the formulations of the present invention includes intravenous, subcutaneous, and intramuscular administration of the pharmaceutical compositions described herein. Preparations for parenteral administration include sterile solutions ready for injection, sterile, dry, soluble products ready to be mixed with a solvent immediately before use, including subcutaneous injection tablets, sterile suspensions ready for injection, sterile, dry, insoluble products ready to be mixed with a vehicle immediately before use, and sterile emulsions. The solutions may be either aqueous or non-aqueous. Pharmaceutically acceptable carriers used in parenteral preparations include aqueous vehicles, non-aqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, thickeners, emulsifiers, sequestering or chelating agents, and other pharmaceutically acceptable substances. Liposome suspensions are also suitable as pharmaceutically acceptable carriers.

[0139] In some embodiments of this disclosure, the pharmaceutical composition is formulated as a depot preparation. Such depot preparations typically have a longer action than non-depot preparations. In some embodiments, such preparations are administered parenterally (e.g., by injection or implantation). For example, subcutaneous or intramuscular depot injection of a sterile solution containing the compounds of formula (FCB)a-(L)b-(CLM)c and / or their subgenus, or compound I, or a pharmaceutically acceptable salt is an effective mode of administration. In some embodiments, the depot preparation is prepared using a suitable polymer or hydrophobic material (e.g., an emulsion in an acceptable oil) or an ion exchange resin, or as a sparingly soluble derivative, for example, a sparingly soluble salt. Thus, for example, the pharmaceutical composition may be formulated with a suitable polymer or hydrophobic material (e.g., an emulsion in an acceptable oil) or an ion exchange resin, or as a sparingly soluble derivative, for example, a sparingly soluble salt. In some embodiments, the depot preparation provides a 1-day, 7-day, or 30-day depot release profile. In some embodiments, such preparations are administered by injection on a schedule of once daily, once weekly, or once monthly.

[0140] The concentration of the pharmaceutically active compound can be adjusted so that the injection provides an effective amount to produce the desired pharmacological effect. The exact dose depends on the age, weight, and condition of the patient or animal, as is known in the art. In some embodiments, the parenteral preparation for a unit dose is packaged in an ampoule or syringe with a needle. All preparations for parenteral administration must be sterilized as is known and practiced in the art. Exemplarily, intravenous or intra-arterial infusion of a sterile aqueous solution containing compounds of formula (FCB)a-(L)b-(CLM)c and / or their subgenus, or compound I, or a pharmaceutically acceptable salt is an effective mode of administration.

[0141] In some embodiments, suitable pharmaceutically acceptable carriers may include, but are not limited to, diluents and sterile aqueous or organic solutions. Pharmaceutically acceptable carriers are well known to those skilled in the art and include, but are not limited to, aqueous and non-aqueous solutions. Pharmaceutically acceptable carriers may be aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents suitable for use in this application include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Examples of aqueous carriers suitable for use in this application include, but are not limited to, water, ethanol, alcohol solutions / aqueous solutions, glycerol, emulsions, or suspensions containing saline and a buffer medium.

[0142] Diluents may be added to the formulations described herein. Diluents can increase the proportion of solid pharmaceutical compositions and / or combinations and make pharmaceutical dosage forms containing the compositions and / or combinations easier for patients and caregivers to handle. In various embodiments, diluents for solid compositions include, for example, microcrystalline cellulose (e.g., AVICEL), finely powdered cellulose, lactose, starch, pregelatinized starch, calcium carbonate, calcium sulfate, sugar, dextrate, dextrin, dextrose, dicalcium phosphate dihydrate, tribasic calcium phosphate, kaolin, magnesium carbonate, magnesium oxide, maltodextrin, mannitol, polymethacrylate (e.g., EUDRAGIT®), potassium chloride, powdered cellulose, sodium chloride, sorbitol, and Tarc and / or mixtures of any of the above. Specific examples of microcrystalline cellulose include those marketed under the Avicel trademark (FMC Corp., Philadelphia, Pennsylvania, USA), such as Avicel® pH101, Avicel® pH102, and Avicel® pH112. Examples of lactose include lactose monohydrate, anhydrous lactose, and Pharmatose DCL21, and examples of dibasic calcium phosphate include Emcompress.

[0143] Suitable L-Ikid supports for use in this application include, but are not limited to, water (preferably a sodium carboxymethylcellulose solution partially containing additives such as cellulose derivatives), alcohols (monohydric and polyhydric alcohols, including glycols, etc.), and their derivatives, and oils (e.g., fractionated coconut oil and arachis oil).

[0144] Liquid carriers suitable for use in this application can be used in the preparation of solutions, suspensions, emulsions, syrups, elixirs, and pressurized compounds. In some embodiments, the active ingredient is dissolved or suspended in water, an organic solvent, a mixture of both, or a pharmaceutically acceptable liquid carrier such as a pharmaceutically acceptable oil or fat. In some embodiments, the liquid carrier contains other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavorings, suspending agents, thickeners, colorants, viscosity modifiers, stabilizers, or osmotic pressure modifiers.

[0145] Suitable solid carriers for use in this application include, but are not limited to, inert substances such as lactose, starch, glucose, methylcellulose, magnesium stearate, dicalcium phosphate, and mannitol. The solid carrier may further contain one or more substances that act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, flow promoters, compression aids, binders, or tablet disintegrants, and may also be encapsulants. In the case of powders, the carrier may be a finely divided solid mixed with a finely divided active compound. In the case of tablets, the active compound is mixed in an appropriate proportion with a carrier having the required compression properties and compressed to the desired shape and size. The powders and tablets may contain up to 99% of the active compound. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, polyvinylpyrrolidone, low-melt waxes, and ion-exchange resins. Tablets may be prepared by compression or molding using one or more auxiliary components as desired. Compressed tablets can be prepared by compressing the active ingredient in a free-flowing form, such as a powder or granules, with a suitable machine, optionally mixed with a binder (e.g., povidone, gelatin, hydroxypropyl methylcellulose), lubricant, inert diluent, preservative, disintegrant (e.g., sodium glycolate starch, cross-linked povidone, cross-linked sodium carboxymethylcellulose), surfactant, or dispersant. Molded tablets may be prepared by molding a mixture of powder compounds moistened with an inert liquid diluent using a suitable machine. Tablets may optionally be coated or scored, and may be formulated to provide a desired release profile by using, for example, hydroxypropyl methylcellulose in various proportions to provide delayed or controlled release of the active ingredient therein. Tablets may optionally be provided with an enteric coating to result in release into a portion of the intestine other than the stomach.

[0146] A carrier suitable for use in this application can be mixed as needed with disintegrants, diluents, granulators, lubricants, binders, etc., using conventional techniques known in the art. The carrier can also be sterilized using methods that do not react harmfully with the compound, as is generally known in the art.

[0147] The pharmaceutical formulations of this disclosure may be prepared by methods known to those skilled in the art, such as mixing, dissolving, granulation, sugar coating preparation, raising, emulsification, encapsulation, or freeze-drying processes. As described above, the pharmaceutical compositions of this disclosure may include one or more pharmaceutically acceptable carriers, such as excipients and adjuvants, that facilitate the processing of the active molecule into a preparation for pharmaceutical use.

[0148] The dissolution rate of compressed solid pharmaceutical compositions in the patient's stomach may be increased by the addition of disintegrants to the composition and / or combination. Examples of disintegrants include alginic acid, calcium carboxymethylcellulose, sodium carboxymethylcellulose (e.g., AC-DI-SOL and PRIMELLOSE), colloidal silicon dioxide, sodium croscarmellose, crospovidone (e.g., KOLLIDON and POLYPLASDONE), guar gum, magnesium silicate, methylcellulose, microcrystalline cellulose, potassium polaritrin, powdered cellulose, pregelatinized starch, sodium alginate, sodium starch glycolate (e.g., EXPLOTAB), potato starch, and starch.

[0149] Flavoring agents and flavor enhancers make the dosage form more palatable to patients. Common flavoring agents and flavor enhancers for pharmaceuticals that may be included in the compositions of the present invention include maltol, vanillin, ethyl vanillin, menthol, citric acid, fumaric acid, ethyl maltol, and tartaric acid.

[0150] Solid and liquid compositions and / or combinations may also be stained with any pharmaceutically acceptable colorants to improve their appearance and / or facilitate patient identification of products and unit dose levels.

[0151] Liquid pharmaceutical compositions and / or combinations may be prepared using excipients in which the components are dissolved or suspended in a liquid carrier such as water, vegetable oil, alcohol, polyethylene glycol, propylene glycol, or glycerin.

[0152] Liquid pharmaceutical compositions and / or combinations may contain emulsifiers for uniformly dispersing insoluble active ingredients or other excipients throughout the composition. Examples of emulsifiers that may be useful in the liquid compositions and / or combinations of the present invention include gelatin, egg yolk, casein, cholesterol, acacia, tragacanth, cartilage, pectin, methylcellulose, carbomer, cetostearyl alcohol, and cetyl alcohol.

[0153] Liquid pharmaceutical compositions and / or combinations may also contain thickeners to improve the mouthfeel of the product and / or to coat the inner lining of the gastrointestinal tract. Examples of such agents include acacia, bentonite alginate, carbomer, calcium or sodium carboxymethylcellulose, cetostearyl alcohol, methylcellulose, ethylcellulose, gelatin gum, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, maltodextrin, polyvinyl alcohol, povidone, propylene carbonate, propylene glycol alginate, sodium alginate, sodium starch glycolate, starch tragacanth, and xanthan gum.

[0154] Sweeteners such as aspartame, lactose, sorbitol, saccharin, sodium saccharin, sucrose, fructose, mannitol, and invert sugar can be added to improve the taste.

[0155] Preservatives and chelating agents such as alcohol, sodium benzoate, butylated hydroxytoluene, butylated hydroxyanisole, and ethylenediaminetetraacetic acid may be added in amounts safe for consumption to improve storage stability.

[0156] The liquid composition may also contain a buffer such as guconic acid, lactic acid, citric acid or acetic acid, sodium gucnate, sodium lactate, sodium citrate, or sodium acetate. The selection and amount of excipients used can be readily determined by a pharmaceutical scientist based on experience and consideration of standard procedures and reference work in the field.

[0157] The solid compositions of the present invention may include powders, granules, aggregates, and compacted compositions and / or combinations. Dosages include those suitable for oral, oral, rectal, parenteral (including subcutaneous, intramuscular, and intravenous), inhalation, and ophthalmic administration. The most appropriate dose in any given case depends on the nature and severity of the condition being treated, but the most preferred route of administration in the present invention is oral. Dosages are conveniently presented in unit dosage forms and can be prepared by any method well known in the pharmaceutical art.

[0158] Dosage forms include tablets, powders, capsules, suppositories, pouches, lozenges, and solid dosage forms such as liquid syrups, suspensions, aerosols, and elixirs.

[0159] The dosage form of the present invention may be a capsule containing the composition and / or combination, preferably the powder or granulated solid composition of the present invention, within either a hard or soft shell. The shell may be made from gelatin and may optionally contain plasticizers such as glycerin and sorbitol, as well as opacifying agents or colorants.

[0160] Compositions for tablet formation or capsule filling may be prepared by wet granulation. In wet granulation, some or all of the active ingredients and excipients in powder form are blended and then further mixed in the presence of a liquid, typically water, that aggregates the powder into granules. The granules are dried and then screened and / or ground to the desired particle size. The granules may be tableted, or other excipients such as lubricants and / or other additives may be added before tableting.

[0161] Tablet-forming compositions can conventionally be prepared by dry granulation. For example, a blended composition of activators and excipients can be compressed into a slag or sheet and then ground into compressed granules. The compressed granules can then be compressed into tablets.

[0162] Tablet-forming compositions may be conventionally prepared by melt granulation. For example, a blended composition of activators and excipients can be granulated into granules using a meltable binder. The granules may then be compressed into tablets.

[0163] In some embodiments, the pharmaceutical composition, such as a compressed tablet composition, comprises drug particles or drug-containing granules.

[0164] As an alternative to dry granulation, blended compositions can be directly compressed into a compressible form using direct compression technology. Direct compression produces more uniform tablets without granules. Excipients particularly well suited for direct compression tablet formation include microcrystalline cellulose, spray-dried lactose, dicalcium phosphate dihydrate, and colloidal silica. The appropriate use of these and other excipients in direct compression tablet formation is known to those skilled in the art who have experience and technique in the specific formulation challenges of direct compression tablet formation.

[0165] In some embodiments, the drug-containing granules or particles include a binder, filler, disintegrant, and / or lubricant. In some embodiments, the drug-containing granules are processed by roller compression, for example, according to the manufacturing parameters disclosed herein. In another specific embodiment, the drug-containing granules or particles may be further compressed after compression. In a particular embodiment, the drug-containing granules or particles may be compressed into a tablet. In a particular embodiment, the tablet contains compound I, or a pharmaceutically acceptable salt, ester, or solvate thereof.

[0166] In some embodiments, this disclosure is, (a) A step of mixing a mixture of compound I or a pharmaceutically acceptable salt thereof with an input material such as a binder, filler, disintegrant, and / or a first part of a lubricant, (b) Compressing or blending the blended mixture from step (a) using a roller compactor to form a compressed material, (c) Blending the roller-compressed material, i.e., the drug-containing granules or particles of step (b), with a second part of the blending material such as a disintegrant, lubricant, and / or binder, to provide a final blend. (e) A step of compressing the final blend of step (c) into a tablet, and (f) optionally coating the compressed tablet with a coating described herein, a compressed pharmaceutical tablet is provided.

[0167] In another specific embodiment, compressed pharmaceutical tablets are prepared as shown in Figure 1.

[0168] In another embodiment, the roller compression process may be modified to affect the particle size or particle size distribution of the drug-containing granules or particles before compression. In a particular embodiment, the particle size distribution of the drug-containing granules or particles may be modified to take into account a desired composition, i.e., for immediate release. In a particular embodiment, the particle size distribution is determined by sieve analysis.

[0169] In one embodiment, approximately 2% to 10% of the drug particles are retained on the sieve 20 (841 μm). In another embodiment, approximately 3% to 8% of the drug particles are retained on the sieve 20 (841 μm). In yet another embodiment, approximately 3% to 6% of the drug particles are retained on the sieve 20 (841 μm).

[0170] In one embodiment, an additional 10% to 35% of drug particles are retained on the sieve using sieve 30 (595 μm). In another embodiment, an additional 20% to 30% of drug particles are retained on the sieve using sieve 30 (595 μm). In yet another embodiment, an additional 25% to 30% of drug particles are retained on the sieve using sieve 30 (595 μm).

[0171] In one embodiment, an additional 10% to 20% of drug particles are retained on the sieve using a sieve 40 (420 μm). In another embodiment, an additional 12% to 18% of drug particles are retained on the sieve using a sieve 40 (420 μm). In yet another embodiment, an additional 13% to 16% of drug particles are retained on the sieve using a sieve 40 (420 μm).

[0172] In one embodiment, an additional 0.2% to 10% of drug particles are retained on the sieve using a 60 (250 μm) sieve. In another embodiment, an additional 1.0% to 7% of drug particles are retained on the sieve using a 60 (250 μm) sieve. In yet another embodiment, an additional 3.0% to 6.0% of drug particles are retained on the sieve using a 60 (250 μm) sieve.

[0173] In one embodiment, an additional 12% to 25% of drug particles are retained on the sieve using a sieve 100 (140 μm). In another embodiment, an additional 15% to 23% of drug particles are retained on the sieve using a sieve 100 (140 μm). In yet another embodiment, an additional 15% to 20% of drug particles are retained on the sieve using a sieve 100 (140 μm).

[0174] In one embodiment, an additional 10% to 30% of drug particles are retained on the sieve using a 230 (63 μm) sieve. In another embodiment, an additional 12% to 25% of drug particles are retained on the sieve using a 230 (63 μm) sieve. In yet another embodiment, an additional 15% to 20% of drug particles are retained on the sieve using a 230 (63 μm) sieve.

[0175] In another embodiment, the D85 value of the drug particles is approximately 50 μm to 100 μm. In yet another embodiment, the D85 value of the drug particles is approximately 60 μm to 80 μm. In yet another embodiment, the D85 value of the drug particles is approximately 63 μm.

[0176] In another embodiment, the D85 value of the drug particles is greater than approximately 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, or 100 μm.

[0177] In another embodiment, the D65 value of the drug particles is approximately 100 μm to approximately 200 μm. In another embodiment, the D65 value of the drug particles is approximately 120 μm to approximately 160 μm. In another embodiment, the D65 value of the drug particles is approximately 140 μm. In another embodiment, the D65 value of the drug particles is greater than approximately 120 μm, approximately 130 μm, approximately 140 μm, approximately 150 μm, approximately 160 μm, or approximately 180 μm.

[0178] In another embodiment, the D45 value of the drug particles is approximately 300 μm to approximately 500 μm. In another embodiment, the D45 value of the drug particles is approximately 350 μm to approximately 450 μm. In another embodiment, the D45 value of the drug particles is approximately 420 μm. In another embodiment, the D55 value of the drug particles is greater than approximately 340 μm, approximately 360 μm, approximately 380 μm, approximately 420 μm, approximately 440 μm, or approximately 480 μm. In another embodiment, the drug particles have substantially similar letters as shown in Table 1. In another embodiment, the drug particles described above are before roller compression. In another embodiment, the drug particles described above are after roller compression.

[0179] The capsule filling of the present invention may comprise any of the aforementioned blends and granules described with reference to tablet formation, provided that they are not subjected to the final tableting process.

[0180] The active ingredients and excipients may be formulated into compositions and / or combinations and dosage forms according to methods known in the art.

[0181] In some embodiments, the pharmaceutical compositions of the present disclosure are stable compositions comprising compounds of formula (FCB)a-(L)b-(CLM)c and / or their subgroups, or compound I, or pharmaceutically acceptable salts, esters, solvates, and / or prodrugs. In some embodiments of the topical pharmaceutical formulations disclosed herein, the compositions have less than 20% total impurities by HPLC after 4 weeks at 40°C / 75%RH. In some embodiments, the compositions of the present disclosure have less than 19% total impurities by HPLC after 4 weeks at 40°C / 75%RH. In some embodiments, the compositions of the present disclosure have less than 18% total impurities by HPLC after 4 weeks at 40°C / 75%RH. In some embodiments, the compositions of the present disclosure have less than 17% total impurities by HPLC after 4 weeks at 40°C / 75%RH. In some embodiments, the compositions of the present disclosure have less than 16% total impurities by HPLC after 4 weeks at 40°C / 75%RH. In some embodiments, the compositions of the disclosure have less than 15% total impurities after 4 weeks of HPLC following 40°C / 75%RH. In some embodiments, the compositions of the disclosure have less than 14% total impurities after 4 weeks of HPLC following 40°C / 75%RH. In some embodiments, the compositions of the disclosure have less than 13% total impurities after 4 weeks of HPLC following 40°C / 75%RH. In some embodiments, the compositions of the disclosure have less than 12% total impurities after 4 weeks of HPLC following 40°C / 75%RH.

[0182] therapeutic use The compounds and pharmaceutical compositions of this disclosure find use in any number of ways. For example, in some embodiments, the pharmaceutical compositions are useful in methods of modulating phosphoinositide 3-kinase (PI3K). In embodiments, the modulation of phosphoinositide 3-kinase (PI3K) activity is located in mammalian cells. In embodiments, the modulation of phosphoinositide 3-kinase (PI3K) may be for the treatment of conditions or diseases described herein, including diseases or conditions in which irreversible inhibition of PI3K provides a therapeutic benefit to subjects having the disease or condition.

[0183] In some embodiments, the Disclosure provides a method for modulating the activity of a kinase or pseudokinase, comprising administering any of the pharmaceutical compositions disclosed herein to a subject requiring such modification. In some embodiments, the method inhibits the activity of a kinase or pseudokinase.

[0184] In some embodiments, this disclosure provides a method for adjusting the activity of PI3K, which includes administering any of the pharmaceutical compositions described herein to a subject requiring such adjustment.

[0185] In some embodiments, the Disclosure provides a method for treating cancer, neurodegenerative diseases, autoimmune disorders, or aging in subjects requiring such treatment, the method comprising administering a pharmaceutical composition disclosed herein to a subject requiring such treatment. In some embodiments, the Disclosure provides a method for treating cancer, the method comprising administering a pharmaceutical composition described herein to a subject requiring such treatment. In some embodiments, the cancer is selected from the group consisting of leukemia, brain tumors, lung (small cell and non-small cell), ovarian, prostate, breast, colon, and tissue cancers including intestinal tissue, pancreatic tissue, liver, kidney, prostate, ovaries, lung, bone marrow, or breast tissue. In some embodiments, the subject has cancer having a mutation in the PIK3CA gene.

[0186] Embodiments of this disclosure provide methods for reducing, inhibiting, or improving cell proliferation in patients who require it. In some embodiments, the reduction, inhibition, or improvement in the methods disclosed herein is performed in vivo. In other embodiments, the reduction, inhibition, or improvement is performed in vitro. In embodiments, the cells of the methods disclosed herein are cancer cells. Embodiments of this disclosure provide methods for reducing or preventing tumor growth, comprising contacting tumor cells with a compound or pharmaceutical composition disclosed herein. In one embodiment, reducing or preventing tumor growth includes reducing tumor volume. In one embodiment, reducing or preventing tumor growth includes complete removal of the tumor. In one embodiment, reducing or preventing tumor growth includes stopping or halting the growth of an existing tumor. In one embodiment, reducing or preventing tumor growth includes reducing the rate of tumor growth.

[0187] The present invention also provides treatment for disorders related to cell proliferation. In one embodiment, the method involves contacting cancer and / or tumor cells with a pharmaceutical composition comprising a compound of formula (FCB)a-(L)b-(CLM)c, and / or a subgenus thereof, or compound I, or a pharmaceutically acceptable salt, ester, solvate, and / or prodrug thereof, as disclosed herein.

[0188] In some embodiments, the compound is a compound that forms a covalent bond with a kinase, or the pseudokinase is compound I. In some embodiments, the compound of formula (FCB)a-(L)b-(CLM)c is compound I. In some embodiments, compound I is the crystalline form of compound I, or a pharmaceutically acceptable salt or solvate of compound I. In some embodiments, the pharmaceutically acceptable salt of compound I is selected from the group consisting of phosphates and includes hydrochloride, sulfate, mesylate, benzenesulfonate, tosylate, fumarate, maleate, L-tartrate, citrate, succinate, or their solvates.

[0189] In another embodiment in which a cell proliferation disorder is treated or improved in a subject, the cell proliferation disorder is cancer. In a particular embodiment, the cancer is selected from the group consisting of blood cancers, colorectal cancers, ovarian cancers, breast cancers, cervical cancers, lung cancers, liver cancers, colon cancers, pancreatic cancers, lymph node cancers, colon cancers, small intestine cancers, prostate cancers, brain cancers, bile duct cancers, gallbladder cancers, head and neck cancers, bone cancers, Ewing's sarcoma, skin cancers, kidney cancers, and heart cancers. In another embodiment, the cancer is selected from the group consisting of cancers selected from the group consisting of breast cancers, ovarian cancers, pancreatic cancers, colorectal cancers, colon cancers, small intestine cancers, and lung cancers. In another embodiment of the present invention, the subject is human.

[0190] In one embodiment, the present invention provides a method for reducing or inhibiting cell proliferation and / or treating cancer, comprising the simultaneous administration to a patient in need of such treatment of a pharmaceutical composition comprising a therapeutically effective amount of at least one compound of formula (FCB)a-(L)b-(CLM)c and / or a subgenus thereof, or compound I, or a pharmaceutically acceptable salt, ester, solvate, and / or prodrug thereof. The present invention also provides a method for treating, preventing, improving and / or mitigating the progression of a disorder or condition characterized by cell proliferation in a subject. More specifically, the method of the present invention comprises the administration of an effective amount of a quinolone compound described herein in a subject to treat a disorder or condition characterized by cell proliferation.

[0191] When used herein, administration may be carried out or implemented using any of the various methods known to those skilled in the art. A pharmaceutical composition comprising a compound represented by formula (FCB)a-(L)b-(CLM)c, and / or its subgenus, or compound I, or a pharmaceutically acceptable salt, ester, solvate, and / or prodrug thereof, may be administered by conventional non-toxic and physiologically acceptable carriers or dosage forms containing carriers, for example, via subcutaneous, intravenous, parenteral, intraperitoneal, intradermal, intramuscular, topical, enteral (e.g., oral), rectal, intranasal, intrabuccal, sublingual, intravaginal, inhalation spray, drug pump, or implantable reservoir.

[0192] Furthermore, pharmaceutical compositions comprising compounds of formula (FCB)a-(L)b-(CLM)c, and / or their subgenera, or compound I, or its pharmaceutically acceptable salts, esters, solvates, and / or prodrugs, can be administered to a topical area requiring treatment. This can be achieved, for example, by local injection via surgery, topical application, percutaneous patch, injection, catheter, suppository, or implant (the implant may optionally be made of porous, non-porous, or gelatinous material), including, for example, membranes such as olfactory membranes or fibers.

[0193] The form in which the compounds of formula (FCB)a-(L)b-(CLM)c, and / or their subgenus, or compound I, or their pharmaceutically acceptable salts, esters, solvates, and / or prodrugs are administered (e.g., syrup, elixir, capsule, tablet, foam, emulsion, gel, etc.) depends in part on the route of administration. For example, for mucous membranes (e.g., oral mucosa, rectum, intestinal mucosa, bronchial mucosa), administration by administration, nasal administration, aerosol, inhalation, spray, eye drops, or suppositories may be used. The compounds of formula (FCB)a-(L)b-(CLM)c, and / or their subgenus, or compound I, or their pharmaceutically acceptable salts, esters, solvates, and / or prodrugs disclosed herein may be administered together with other bioactive agents such as anticancer agents, analgesics, anti-inflammatory agents, anesthetics, and other agents that can control one or more symptoms or causes of conditions characterized by impairment or cell proliferation.

[0194] In one embodiment, compounds of formula (FCB)a-(L)b-(CLM)c, and / or their subgenus, or compound I, or a pharmaceutically acceptable salt, ester, solvate, and / or prodrug thereof, may be administered together with a second or more therapeutic agents.

[0195] Furthermore, administration may include administering multiple doses over an appropriate period. Such administration regimens can be determined in accordance with routine practices when reviewing this disclosure.

[0196] In situations of progressively worsening disorders or conditions characterized by cell proliferation, compounds of formula (FCB)a-(L)b-(CLM)c, and / or their subgenus, or compound I, or their pharmaceutically acceptable salts, esters, solvates, and / or prodrugs, are generally administered continuously. In certain settings, administration of compounds of formula (FCB)a-(L)b-(CLM)c, and / or their subgenus, or compound I, or their pharmaceutically acceptable salts, esters, solvates, and / or prodrugs disclosed herein may be initiated before the onset of disease symptoms as part of a strategy to delay or prevent the disease. In other circumstances, the compounds of formula (FCB)a-(L)b-(CLM)c disclosed herein, and / or their subgenus, or compound I, or its pharmaceutically acceptable salts, esters, solvates, and / or prodrugs, are administered after the onset of disease symptoms as part of a strategy to delay or reverse the disease process, and / or as part of a strategy to improve cellular function and reduce symptoms.

[0197] It will be understood by those skilled in the art that the dose range depends on the specific compound of formula (FCB)a-(L)b-(CLM)c, and / or its subgenus, or compound I, or its pharmaceutically acceptable salts, esters, solvates, and / or prodrugs, and their potency. The dose range will be understood to be large enough to produce the desired effect, where neurodegeneration or other disorders and associated symptoms are improved and / or cell viability is achieved, but not so large as to cause uncontrollable adverse side effects. However, it will be understood that the specific dose level for any particular patient will depend on a variety of factors, including the age, weight, general health, sex, and diet of the individual being treated. The time and route of administration, the rate of excretion, other previously administered drugs, and the severity of the particular disease being treated will be well understood by those skilled in the art. The dosage may also be adjusted by the individual physician in case of any complications. No unacceptable toxic effects are expected when compound I disclosed herein is used in accordance with this application.

[0198] An effective amount of the compounds of formula (FCB)a-(L)b-(CLM)c disclosed herein, and / or their subgenus, or Compound I, or its pharmaceutically acceptable salts, esters, solvates, and / or prodrugs, is sufficient to produce a measurable biological response. The actual dose levels of the active ingredients of this application can be varied to administer amounts of the compounds of formula (FCB)a-(L)b-(CLM)c, and / or their subgenus, or Compound I, or its pharmaceutically acceptable salts, esters, solvates, and / or prodrugs that are effective in achieving the desired therapeutic response for a particular subject and / or application. Preferably, a minimum dose is administered, and the dose is escalated to a minimum effective dose in the absence of dose-limiting toxicity. The determination and adjustment of therapeutic effective doses, as well as the evaluation of the timing and methods for making such adjustments, are known to those skilled in the art.

[0199] Throughout this entire disclosure, various patents, patent applications, and publications are referenced. The disclosures of these patents, patent applications, and publications are incorporated herein by reference for all purposes to more fully describe the state of the art known to those skilled in the art as of the date of the present disclosure. The present disclosure shall prevail in the event of any inconsistency between the cited patents, patent applications, and publications and the present disclosure. The following examples further illustrate the present invention, but should not be construed as limiting its scope.

Examples

[0200] The present disclosure generally described herein will be more readily understood by reference to the following examples, which are included solely for the purpose of illustrating specific aspects and embodiments of the present invention and are not intended to limit the present invention.

[0201] The following examples are intended to be illustrative, and efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.), but some experimental errors and deviations should be accounted for within the range understood by those skilled in the art. Unless otherwise indicated, temperatures are in degrees Celsius. Reagents were purchased from commercial suppliers such as Sigma-Aldrich, Alfa Aesar, or TCI, and used without further purification unless otherwise indicated.

[0202] Example 1: Synthesis and characterization of methyl (5-(6-((4-(acryloylglycyl)piperazin-1-yl)methyl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazin-2-yl)-4-(trifluoromethyl)pyridin-2-yl)carbamate (Compound I) Synthesis of ethyl 1-amino-5-carbamoyl-1H-pyrrole-3-carboxylate (1).

Chemical Formula

[0203] Diethyl 1-amino-1H-pyrrole-2,4-dicarboxylate. 1002 g, 1 equivalent, 4.507 mol of diethyl 1H-pyrrole-2,4-dicarboxylate and 7.8 L of NMP were loaded into a 20 L reaction vessel. Potassium tert-butoxide (557.3 g, 1.1 equivalents, 4.967 mol) was then added, and the mixture turned pink, reaching a temperature of 37°C. The mixture was stirred until all of the potassium tert-butoxide was dissolved, and then cooled to 21°C. Subsequently, O-(4-nitrobenzoyl)hydroxylamine (839.6 g, 1.02 equivalents, 4.610 mol) was added in small amounts (exothermic reaction, reaching 36°C). The reaction mixture became a dark purple suspension. The reaction mixture was stirred overnight at 45°C (T slowly dripped, and the mixture turned orange). The sample was collected after 18 hours and diluted with ACN / water for HPLC analysis. Then, a solution of sodium dithionite (478.6 g, 0.6 equivalents, 2.749 mol) in water (2.5 L) was slowly added while maintaining the temperature below 30°C. The reaction mixture was transferred to a 50 L separatory funnel. Toluene (15 L) and water (5 L) were added to separate the phases. The aqueous phase was extracted with toluene (3 × 1 L). The combined tissue phase was washed with water (5 × 1 L), saturated sodium bicarbonate (5 × 1 L), and brine (1 L), dried over sodium sulfate, filtered, and concentrated to obtain diethyl 1-amino-1H-pyrrole-2,4-dicarboxylate (1077 g, 3.90 mol, 86%) as an orange oil (81% QNMR purity), which solidified upon standing. 1 H NMR(299MHz,DMSO-d6)δ 7.52(d,J=2.1Hz,1H),7.03(d,J=2.1Hz,1H),6.51(d,J=1.8Hz,2H),4.34-4.06(m,4H),1.38-1.14(m,6H). LCMS(ESI):Actual value 227.0[M+H] + (Calculated value 227.1[M+H]) + ).

[0204] Lithium 1-amino-4-(ethoxycarbonyl)-1H-pyrrole-2-carboxylate. A 10 L reaction vessel was packed with diethyl 1-amino-1H-pyrrole-2,4-dicarboxylate (1077 g, 1 equivalent, 3.90 mol), ethanol (3.8 L), and water (1.9 L). Lithium hydroxide monohydrate (163.9 g, 56 wt%, 0.99 equivalents, 3.831 mol) was added, and the mixture was stirred at 60°C. The transformation was monitored by LC-MS analysis. The reaction was stopped after 6 hours and cooled to room temperature. The mixture was diluted with 2 L of toluene, and the layers were separated. The aqueous phase was washed with toluene (3 × 750 ml) and concentrated in a rotary evaporator at 60°C. The resulting solid was suspended in TBME (2 L), filtered, and washed with TBME (1 L). Lithium 1-amino-4-(ethoxycarbonyl)-1H-pyrrole-2-carboxylate (853.1 g, 3.50 mol, 90%) was obtained as a pale yellow solid (83% QNMR purity). 1 H NMR(400MHz,DMSO-D6)δ 7.37(s,2H),7.09(d,J=2.2Hz,1H),6.63(d,J=2.2Hz,1H),4.12(q,J=7.1Hz,2H),1.22(t,J=7.1Hz,3H). LCMS(ESI):Actual value 199.0[M-Li+2H] + (Calculated value 199.1[M-Li+2H] + ).

[0205] Ethyl 1-amino-5-carbamoyl-1H-pyrrole-3-carboxylate (1). To a suspension of lithium 1-amino-4-(ethoxycarbonyl)-1H-pyrrole-2-carboxylate (835.8 g, 1 equivalent, 3.399 mmol) in a mixture of DMF (3.4 L) / MeTHF (8.5 mL), ammonium carbonate (2939 g, 9.00 equivalents, 30.59 mol), followed by HOBt (1041 g, 2.0 equivalents, 6.797 mol), EDCI HCl (1303 g, 2.0 equivalents, 6.797 mol), and DIPEA (2.96 L, 5 equivalents, 16.99 mol) were added. Endothermic reactions occurred upon addition of reagents. The flask was stirred at room temperature for 3 days (suspension). The conversion was monitored by LC-MS analysis. The solid was filtered, washed with MeTHF (2 L), and the filtrate was concentrated at 60°C. The crude substance (2 kg) was redissolved in MeTHF (10 L) and washed with a saturated solution of sodium bicarbonate (2 × 2 L). Three phases were formed. The upper phase (product fraction) was separated, dried over sodium sulfate, filtered, and concentrated to dryness. The resulting crude substance (908 g) was recrystallized from EtOH (1.8 L). The solid was filtered, washed with EtOH (200 mL), and dried to obtain the first harvest of ethyl 1-amino-5-carbamoyl-1H-pyrrole-3-carboxylate (67 g) as a white solid. The mother liquor was concentrated (794 g) and purified by column chromatography (2 kg silica, gradient: DCM ~ 5% MeOH). The purified substance (505 g) was recrystallized from EtOH (750 mL) and washed with EtOH (100 mL) to obtain a second harvest of ethyl 1-amino-5-carbamoyl-1H-pyrrole-3-carboxylate (68 g). The concentrated mother liquor (435 g) was further purified by column chromatography (6 kg silica, gradient: DCM ~ 5% MeOH). The product fraction was recrystallized to obtain an additional 61 g of ethyl 1-amino-5-carbamoyl-1H-pyrrole-3-carboxylate. All harvests fused to obtain ethyl 1-amino-5-carbamoyl-1H-pyrrole-3-carboxylate (1) (195.7 g, 992.4 mmol, 29%) as a white solid. 1H NMR(299MHz,DMSO-d6)δ 7.96(s,1H),7.36(d,J=2.0Hz,1H),7.31(s,1H),7.13(d,J=2.1Hz,1H),6.86(d,J=1.8Hz,2H),4.18(q,J=7.1Hz,2H),1.25(t,J=7.1Hz,3H). LCMS(ESI):Actual value 198.0[M+H] + (Calculated value 198.1[M+H]) + ).

[0206] Synthesis of 6-Amino-4-(trifluoromethyl)nicotinaldehyde [ka]

[0207] 6-amino-4-(trifluoromethyl)nicotinaldehyde (2). Equipped with a thermometer and under nitrogen, a 5 L three-necked flask was filled with 5-bromo-4-(trifluoromethyl)-2-pyridylamine (200.0 g, 1 equivalent, 829.8 mmol) and dry THF (2000 mL). The solution was cooled to -70°C. A 2.5 M solution of n-butyllithium in hexane (995.8 mL, 3.00 equivalent, 2.49 mol) was added dropwise over 90 minutes while maintaining the temperature below -60°C. The mixture was stirred at -70°C for 15 minutes. Then, DMF (160.6 mL, 2.50 equivalent, 2.074 mol) was added dropwise over 45 minutes while maintaining the temperature below -60°C. The mixture was stirred at -70°C for 30 minutes. The mixture was heated to -40°C and carefully quenched with water (74.8 mL, 5 equivalents, 4.149 mol). The resulting solution was heated to room temperature and stirred overnight. The mixture was further diluted with water / ethyl acetate (1 L / 1 L) and transferred to a separatory funnel. The organic layer was washed with water (5 × 300 mL). The combined aqueous phase was extracted with ethyl acetate (3 × 300 mL). The combined organic phase was washed with brine (300 mL), dried over sodium sulfate, filtered, and concentrated until a small amount of solvent remained and a solid was formed. The mixture was cooled to room temperature and diluted with heptane (300 mL). The mixture was filtered, the solid was washed with heptane, and dried to obtain 6-amino-4-(trifluoromethyl)nicotinaldehyde (2) (51.5 g, 271 mmol, 33%) as an orange solid. 1 H NMR(299MHz,DMSO-d6)δ 9.80(q,J=1.7Hz,1H),8.63(d,J=2.1Hz,1H),7.73(s,2H),6.84(d,J=2.0Hz,1H). LCMS(ESI): Actual value 191.0[M+H] + (Calculated value: 191.0 [M+H]) + ).

[0208] Step 1: Synthesis of 2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-oxo-3,4-dihydropyrrolo[2,1-f][1,2,4]triazine-6-carboxylate ethyl (3). [ka]

[0209] A condenser, temperature probe, and mechanical stirrer were attached to a 3 L three-necked flask, and a 1.9 L solution of 1-amino-5-carbamoyl-1H-pyrrole-3-carboxylate ethyl (1) (195.7 g, 1.00 equivalent, 962.7 mmol) and 6-amino-4-(trifluoromethyl)nicotinaldehyde (2) (192.7 g, 1.00 equivalent, 962.7 mmol) in DMSO (1.9 L) was charged. Then, cupric chloride dihydrate (213.4 g, 1.30 equivalent, 633.4 mmol) was added, and the mixture was stirred at 100 °C for 18 hours. The mixture was cooled to room temperature and poured into ice water (10 L) to induce precipitation of the product. After stirring the suspension for 30 minutes, it was filtered (Buchner filter). The pale brown filter cake was washed with water (3 × 1 L) and TBME (3 × 1 L). The solid was removed with toluene (3 × 1 L) on a rotary evaporator, and the water residue was removed. Ethyl 2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-oxo-3,4-dihydropyrrolo[2,1-f][1,2,4]triazine-6-carboxylate (3) (340.6 g, 760.0 mmol, 79%) was obtained as a solid. 1 H NMR(299MHz,DMSO-d6)δ 12.20(s,1H),8.08(d,J=1.8Hz,1H),7.23(d,J=1.8Hz,1H),7.05(s,1H),6.87(s,1H),4.26(q,J=6.9Hz,2H),1.29(t,J=7.1,3H). LCMS(ESI):Actual value 368.0[M+H] + (Calculated value 368.1[M+H]) + );366.0 [MH] - (Calculated value: 366.1 [MH]) - ).

[0210] Step 2: Synthesis of 2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazine-6-carboxylate ethyl(4). [ka]

[0211] Under nitrogen, a 5 L three-necked flask equipped with a thermometer was filled with 2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-oxo-3,4-dihydropyrrolo[2,1-f][1,2,4]triazine-6-carboxylate ethyl(3) (335.6 g, 1 equivalent, 749.2 mmol), DMF (2.5 L), and morpholine (400 mL, 6.2 equivalents, 2.580 mol). PyBOP (606.9 g, 1.55 equivalents, 1.166 mol) was then added in several portions while maintaining the temperature at approximately 15°C (exothermic). The mixture was stirred at room temperature for 48 hours. The reaction mixture was transferred to a 20 L container and water (12.5 L) was slowly added. The suspension was filtered over a Buchner filter and washed with water (3 × 5 L) and TBME (2 × 5 L). The remaining clay-like substance was redissolved in ethyl acetate under reflux. The high-temperature suspension was filtered through a glass filter containing Celite to remove copper, and the filtrate was concentrated. The remaining solid was suspended in ethyl acetate (1 L) at 60°C. The suspension was centrifuged at 3000 rpm. The resulting solution was decanted and concentrated. The residue was treated three more times with warm ethyl acetate. All organic phases were concentrated, the resulting solid was filtered, washed with TBME, and dried under vacuum at 60°C. The solid was removed with toluene (2 × 1 L) to obtain 2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazine-6-carboxylate ethyl(4) (206.5 g, 473.2 mmol, 63%) as a pale yellow solid with QNMR purity of 91%. 1 H NMR(300MHz,cdcl3)δ 8.56(s,1H),8.09(d,J=1.6Hz,1H),7.19(d,J=1.6Hz,1H),6.81(s,1H),4.88(s,2H),4. 37(q,J=7.1Hz,2H),4.08(t,J=4.8Hz,4H),3.84(t,J=4.8Hz,4H),1.39(t,J=7.1Hz,3H). LCMS(ESI):Actual value 437.2[M+H] + (Calculated value 437.2[M+H]) + ).

[0212] Step 3: Synthesis of 4-(2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazine-6-carbonyl)piperazine-1-carboxylate tert-butyl(5) [ka]

[0213] Under nitrogen, a 3 L three-necked flask was packed with 2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazine-6-carboxylate ethyl(4) (206.5 g, 91 wt%, 1 equivalent, 473.2 mmol) and THF (2 L). Then, piperazine-1-carboxylate tert-butyl (440.7 g, 5.0 equivalent, 2.366 mol) and TBD (65.87 g, 1.0 equivalent, 473.2 mmol) were added, and the solution was stirred at 65 °C for 2 days. The transformation was monitored by LC-MS and proton NMR analysis. The reaction mixture was concentrated at 50 °C to remove most of the THF. This substance was dissolved in ethyl acetate (1 L) and washed with 1 M potassium bisulfate solution (2 × 500 mL), water (3 × 500 mL, diluted with brine to enhance separation), and brine (250 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated at 50°C (foamy brown oil). The brown foam was removed with toluene (1.5 L) to remove trace amounts of ethyl acetate, and 4-(2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazine-6-carbonyl)piperazine-1-carboxylate tert-butyl(5) (382.0 g, 464 mmol, 98%) was obtained as a brown foamy substance with QNMR purity of 70%. 11H NMR (300 MHz, cdcl3) δ 8.52 (s, 1H), 7.77 (d, J=1.6 Hz, 1H), 6.98 (d, J=1.7 Hz, 1H), 6.80 (s, 1H), 4.06 (t, J=4.9 Hz, 4H), 3.82 (t, J=4.9 Hz, 4H), 3.77-3.66 (m, overlapping with THF signal, subtraction of another THF signal gives 4H), 3.48 (t, J=5.0 Hz, 4H), 1.47 (m, 27H). LCMS (ESI): Found 577.2 [M+H] + (Calculated 577.3 [M+H] + ).

[0214] Step 4: tert-Butyl 4-((2-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazin-6-yl)methyl)piperazine-1-carboxylate (6).

Chemical Formula

[0215] A 5 L three-necked flask was packed under nitrogen with tert-butyl 4-(2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazine-6-carbonyl)piperazine-1-carboxylate (5) (271.0 g, 1 equivalent, 470.0 mmol) and dry THF (2 L). The resulting solution was cooled to 0°C and a solution of TMS-Cl (107.0 nm / mL, 1.80 equivalents, 846.0 mmol) in anhydrous THF (100 mL) was added (the mixture formed a brown suspension). The mixture was further cooled to -20°C and 2.4 M LiAlH4 in THF (294.0 mL, 1.50 equivalents, 705.0 mmol) was added over 80 minutes. The reaction mixture was stirred at -20°C for 60 minutes. A 2M solution of Rochelle salt (1 L) was slowly added at -20°C to -10°C (the mixture became very thick, and a solid formed after the addition of 100 ml) (very exothermic at the start, gas generation). The solid slowly dissolved at -8°C and became easy to stir again). The mixture was warmed to room temperature overnight. The organic layer was collected, and the aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (6 kg silica, eluent: DCM in MeOH / 3.5 M NH3) to obtain 4-((2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazine-6-yl)methyl)piperazine-1-carboxylate tert-butyl(6) (172.2 g, 306.1 mmol, 65%) as an off-white, fluffy solid. 1 H NMR(299MHz,cdcl3)δ 8.53(s,1H),7.58(d,J=1.5Hz,1H),6.80(s,1H),6.67(d,J=1.6Hz,1H),4.82(s,2H),4.14-3.95(m,4H), 3.82(t,J=4.8Hz,4H),3.58(s,2H),3.44(t,J=5.3Hz,4H),2.43(t,J=5.1Hz,4H),1.45(d,J=1.4Hz,9H). LCMS(ESI): Actual value 563.3[M+H] + (Calculated value 563.3[M+H]) +); 561.2[MH] - (Calculated value 561.3 [MH]) - ).

[0216] Step 5: 4-((2-(6-((methoxycarbonyl)amino)-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrro[2,1-f][1,2,4]triazine-6-yl)methyl)piperazine-1-carboxylate tert-butyl(7). [ka]

[0217] A 3 L three-necked flask was packed with 4-((2-(6-amino-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazine-6-yl)methyl)piperazine-1-carboxylate tert-butyl(6) (172.2 g, 1 equivalent, 306.1 mmol), DCM (1.4 L), and pyridine (74 mL, 3.0 equivalents, 918.2 mmol), and the solution was cooled to 0°C. Methyl carbonochloride (26.1 mL, 1.10 equivalents, 336.7 mmol) was added to the reaction mixture over 30 minutes. The mixture was slowly warmed to room temperature and stirred overnight. HPLC analysis showed approximately 90% conversion. An additional methyl carbonochloride (2.4 mL, 0.10 equivalents, 30.6 mmol) was added dropwise over 5 minutes, and the mixture was stirred at room temperature. After 4 hours, HPLC analysis showed that complete conversion had been achieved. The reaction mixture was poured into cold water (500 ml) and transferred to a separatory funnel. The organic layer was collected, washed with water (3 × 500 ml) and brine, dried over sodium sulfate, filtered, and concentrated. The residue was removed with toluene to obtain tert-butyl 4-((2-(6-((methoxycarbonyl)amino)-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrrolo[2,1-f] [1,2,4]triazine-6-yl)methyl)piperazine-1-carboxylate (7) (174.1 g, 280.5 mmol, 92% yield) as a solid. 1H NMR(299MHz,cdcl3)δ 8.87(s,1H),8.74(s,1H),8.43(s,1H),7.59(d,J=1.5Hz,1H),6.69(d,J=1.6Hz,1H),4.05(t,J=4.8H z,4H),3.84(d,J=7.1Hz,7H),3.59(s,2H),3.45(t,J=5.1Hz,4H),2.44(t,J=4.9Hz,4H),1.45(s,9H). LCMS(ESI):Actual value 621.3[M+H] + (Calculated value 621.8 [M+H]) + );619.2 [MH] - (Calculated value 619.3 [MH]) - ).

[0218] Step 6: (5-(4-morpholino-6-(piperazine-1-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-2-yl)-4-(trifluoromethyl)pyridine-2-yl)carbamate matylate (8). [ka]

[0219] A flask was packed under nitrogen with 4-((2-(6-((methoxycarbonyl)amino)-4-(trifluoromethyl)pyridine-3-yl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazine-6-yl)methyl)piperazine-1-carboxylate tert-butyl(7) (172.1 g, 1 equivalent, 277.3 mmol) and CPME (1.0 L) (white suspension), and the mixture was cooled to 6°C. A solution of HCl in CPME (1.5 L, 3.0 mol, 16.2 equivalents, 4.50 mol) was added over 30 minutes (only slightly exothermic), and the mixture was stirred overnight at room temperature (a yellow precipitate formed immediately). HPLC analysis showed complete conversion. The mixture was filtered, and the solid was washed with 250 ml of CPME and 500 ml of TBME. The solid was transferred to a flask, but because it was very viscous (presumably hygroscopic), the solid was transferred using MeOH. The mixture was concentrated in a rotary evaporator, and the solid was dried to obtain 198 g of crude product. Before recrystallization, the starting material was removed with toluene (2 × 1 L). The crude product (191 g) was recrystallized from MeOH (300 mL). The solid was collected by filtration, washed with cold MeOH (50 mL), and dried to obtain (5-(4-morpholino-6-(piperazine-1-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-2-yl)-4-(trifluoromethyl)pyridine-2-yl)methyl dihydrochloride (8) (88.8 g, 150 mmol, yield 54%) as a solid. 1 H NMR(400MHz,DMSO-D6)δ 12.13(s,1H),10.92(s,1H),9.59(s,2H),8.73(s,1H),8.31(s,1H),8.07(s,1H),7.38(s,1 H),4.41(s,2H),4.02(t,J=4.9Hz,4H),3.80-3.69(m,8H),3.69-3.33(m,13H),3.16(s,3H). LCMS (ESI): Actual value 521.2 [M+H] + (Calculated value 521.2[M+H]) + ).

[0220] Step 7: (5-(6-((4-(acryloylglycyl)piperazin-1-yl)methyl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazin-2-yl)-4-(trifluoromethyl)pyridine-2-yl)carbamate methyl (compound 1). [ka] [ka] Acryloylglycine. 2 L of RBF was packed with glycine (200 g, 1.0 equivalent, 2.66 mol), deionized water (88 mL), and sodium hydroxide (710 mL, 30% Wt, 2.0 equivalent, 5.32 mol). After cooling the mixture to -5°C, acryloyl chloride (225 mL, 96% Wt, 1.0 equivalent, 2.66 mol) was added while maintaining the temperature at approximately 0°C. The mixture was stirred at 0°C for 1 hour. After 1 hour, the mixture was acidified to pH 2 with HCl (310 mL, 12 mol, 1.4 equivalent, 3.72 mol). The mixture was saturated with sodium sulfate (±100 g) and diluted with warm MeTHF (300 mL). The phases were separated, and the aqueous phase was extracted with MeTHF (3 × 250 mL). The combined organic layers were washed with brine (150 mL), dried over sodium sulfate, and concentrated. A solvent exchange (addition of 500 mL) was performed with approximately 500 mL of ethyl acetate. After removing 500 mL of ethyl acetate, the solvent switch was repeated two more times. The resulting slurry was stirred at room temperature for 1 hour, the solid was collected, rinsed with ethyl acetate (2 × 60 mL), dried under vacuum, and further dried on a rotary evaporator to obtain acryloyl glycine (76.00 g, 588.6 mmol, yield 22%) as a white solid. 1¹H NMR (299MHz, DMSO-d⁶) δ 12.58 (s, 1H), 8.42 (t, J=6.0Hz, 1H), 6.30 (ddd, J=17.1, 10.1, 0.8Hz, 1H), 6.10 (ddd, J=17.2, 2.3, 0.8Hz, 1H), 5.62 (ddd, J=10.1, 2.3, 0.8Hz, 1H), 3.84 (dd, J=6.0, 0.8Hz, 2H). LCMS (ESI): Measured value 130.1 [M+H] + (Calculated value 130.1 [M+H]) + ).

[0221] To a suspension of (5-(4-morpholino-6-(piperazine-1-ylmethyl)pyrrolo[2,1-f][1,2,4]triazine-2-yl)-4-(trifluoromethyl)pyridine-2-yl)carbamate methyl dihydrochloride (8) (73.2 g, 1 equivalent, 123 mmol) in THF (750 mL), DIPEA (100 mL, 4.7 equivalents, 416 mmol) was added, and the mixture was slowly dissolved, then became turbid again. Acryloylglycine (23.9 g, 1.5 equivalents, 185 mmol) was added, followed by HATU (93.8 g, 2 equivalents, 247 mmol), and the mixture (yellow) was stirred at room temperature. After 20 minutes, the mixture became a black suspension. After 1 hour, the reaction was terminated and diluted with 600 mL of ethyl acetate and then with 600 mL of saturated sodium bicarbonate solution. The organic phase was separated to obtain solutions in saturated sodium bicarbonate (600 mL) and brine (2 × 400 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated to obtain 147.2 g of crude substance as a red, viscous oil. The crude substance was dissolved in NMP (600 mL) at room temperature, and water (1.5 L) was added dropwise. The mixture was stirred at room temperature for 1 hour and then filtered. The residue was thoroughly washed with water and TBME to remove most of the NMP. The resulting solid was dissolved in DMSO (470 mL) and precipitated by adding water (900 mL). Again, the residue was thoroughly washed with water and TBME to remove most of the NMP and DMSO. After drying the materials overnight in a circulating oven at 40°C, (5-(6-((4-(acryloylglycyl)piperazin-1-yl)methyl)-4-morpholinopyrrolo[2,1-f][1,2,4]triazin-2-yl)-4-(trifluoromethyl)pyridine-2-yl)carbamate methyl (compound I) (57.7 g, 91.4 mmol, yield 74%) was obtained as a solid. 1H NMR(400MHz,DMSO-D6)δ 10.89(s,1H),8.72(s,1H),8.30(s,1H),8.20(t,J=5.5Hz,1H),7.75(d,J=1.5Hz,1 H),6.97(d,J=1.6Hz,1H),6.37(dd,J=17.1,10.2Hz,1H),6.09(dd,J=17.1,2.2Hz, 1H),5.59(dd,J=10.2,2.2Hz,1H),4.02(d,J=5.5Hz,2H),3.98(t,J=4.9Hz,4H),3. 74(d,J=4.4Hz,7H),3.57(s,2H),3.52-3.37(m,4H),2.39(dt,J=17.8,4.9Hz,4H). LCMS(ESI):Actual value 632.4[M+H] + (Calculated value 632.3 [M+H]) + );630.2 [MH] - (Calculated value 630.2 [MH]) - ).

[0222] Example 2: Transfection protocol and reading for NanoBRET screening Human embryonic kidney 293-H (HEK 293, Gibco 293-H, #11631017) cell lines were maintained in a water-saturated incubator at 37°C and 5% CO2 in Dulbecco's modified Eagle medium, high glucose, pyruvate (DMEM, Gibco, #11995065) supplemented with 10% fetal bovine serum (FBS, Gibco, #10082147) and 1× penicillin-streptomycin (100× solution, Gibco, #15140148). Cells were trypsinized using 0.05% or 0.25% trypsin-EDTA solution (trypsin-EDTA, phenol red, Gibco, #25200056 (0.25%) or #25300054). Opti-MEM medium supplemented with 10% fetal bovine serum (Opti-MEM I reduced serum medium, without phenol red, Gibco, #11058021) was used to culture cells overnight for NanoBRET reading experiments.

[0223] HEK293 cells were properly cultured before the assay. The medium was removed from the cell flask by aspiration, washed once with PBS, then aspiration, trypsinized, and the cells were dissociated from the flask. The trypsin was neutralized using growth medium, and the cells were pelleted by centrifugation at 200 × g for 5 minutes. The medium was aspirated, and the cells were resuspended in a single cell suspension using Opti-MEM I supplemented with 10% FBS. The cell density was 2 × 10⁶ in Opti-MEM I supplemented with 10% FBS in a sterile conical tube. 5 The solution was adjusted to / mL. Cells were transfected and, the following day, directly aliquoted into 96-well plates for the NanoBRET assay, thus allowing the cells to be cultured overnight in Opti-MEM. Cells were also transfected in large quantities and dispensed into 96-well plates, allowing the cells to adhere to the plates overnight, thereby enabling a wash test.

[0224] The lipid:DNA complex was prepared as follows: A 10 μg / mL solution of DNA was prepared in serum-free Opti-MEM. This solution contains carrier DNA encoding NanoLuc fused to the following biological targets in the following ratios. A serial dilution step may be ensured to accurately dilute the NanoLuc fusion DNA. In order, the following reagents, 1 mL of Opti-MEM without phenol red, were added to sterile polystyrene test tubes: 9.0 μg / mL of carrier DNA; 1.0 μg / mL of NanoLuc fusion DNA (a smaller amount for some targets). The reagents were thoroughly mixed. 30 μL of FuGENE HD was added to each mL of DNA mixture to form lipid:DNA complexes. Care was taken to avoid contact between the FuGENE HD and the plastic side of the tube, and the mixture was pipetteed directly into the liquid in the tube. The mixture was mixed 5-10 times by pipetting up and down, and incubated at room temperature for 20 minutes to form the complexes. A portion of the lipid:DNA complex (e.g., 1 mL) was divided into 2 × 10⁻⁶ 5Mix 20 parts (e.g., 20 mL) of HEK293 cells in suspension with the compound at / mL and gently mix by pipetting up and down 5 times in a sterile conical tube. Larger or smaller bulk transfections are scaled accordingly using this ratio. Dispense 100 μL of cell + lipid:DNA complex into sterile, tissue-culture-treated 96-well plates (20,000 cells / well) and incubate for at least 16 hours to express the compound. Incubate cells in a 37°C + 5% CO2 incubator for over 16 hours. Serially diluted compound I was prepared at 100 × final concentration in 100% DMSO. Stocks of serially diluted compound I were prepared in PCR plates. Add 1 μL per well of 100 serially diluted inhibitor / test compound to cells in 96-well plates transfected overnight and mix by gently tapping the plate by hand. The plates were incubated overnight in a 37°C + 5% CO2 incubator. Appropriate concentrations of the substrate mixture (500X stock) and tracer were prepared in Opti-Mem. Cells were washed by setting a plate washer in a 96-well 5X plate in pH 7.4 PBS, adding 200 μL of PBS each time. Cells were incubated at 37°C for 2 hours. 100 μL of 1X Substrate-Tracer solution was added, and the 96-well plate was gently mixed by lightly tapping. The plates were read hourly on a plate reader for the next 6 hours. Binding assay results showed that compound I resulted in over 80% inhibition of the PI3K target.

[0225] Example 3: Cell proliferation assay The purpose of this study was to investigate the effect of compound I on cell proliferation in 13 cell lines after 3 days of treatment and to determine the IC50 of compound I in each cell line. The cell lines tested included UM-UC-3 (bladder), KYSE-410 (HN / esophagus), SW1463 (rectum), Calu-1 (lung), NCI-H358 (lung), SW837 (rectum), SW756 (cervical), NCI-H2122 (lung), NCI-H1373 (lung), NCI-H1792 (lung), NCI-H23 (lung), MIA PaCa-2 (pancreas), and HC44.

[0226] Cells were harvested and maintained in appropriate culture medium. Cells were harvested during each logarithmic growth period. The cells were then resuspended and counted using a Vi cell counter (cell viability was measured by trypan blue exclusion assay). The cells were then diluted, and 90 μL of cell suspension was used in a 96-well plate according to a plate map with final cell density. Two duplicate plates were set up: one for reading on day 0 (T0) and the other cultured in an incubator for reading at the endpoint. The incubated plates were incubated overnight in a humidified incubator at 37°C and 5% CO2.

[0227] On day 0, 10 μL of culture medium was added to each well for T0 reading. Then, 50 μL of CellTiter-Glo® reagent was added to each well. The contents were then mixed on an orbital shaker for 2 minutes to promote cell lysis. The plate was then incubated at room temperature for 10 minutes to stabilize the luminescence signal. Luminescence was then recorded using an EnVision multi-label reader. Compound I, the test compound, and cisplatin, the control, were then diluted to various concentrations from a 10 mM stock solution. Compound I was diluted, and 10 μL of each 10-fold concentration of Compound I was taken from the working solution, with a concentration range from 10 μM to approximately 0.005 μM. Cisplatin was diluted to approximately 3.33 mM to approximately 150 nM. The screening plates were then returned to the incubator for the appropriate processing time (3 days).

[0228] For endpoint CTG reading, 50 μL of CellTiter-Glo® reagent was added to each well. The contents were then mixed on an orbital shaker for 2 minutes to promote cell lysis. The plate was incubated at room temperature for 10 minutes to stabilize the luminescence signal. Luminescence was then recorded using an EnVision multi-label reader. IC50 was determined for each cell line. The assay demonstrates that compound I has inhibitory effects across multiple cancer cell lines, including substantial inhibitory effects against NCI-H358 (see Table A). [Table A]

[0229] Example 4: Immediate-release tablets containing compound I Compound I was formulated into oral tablets for immediate release after administration to patients. Weighing, compression, blending, and compression were completed with Compound I as the additive drug, as shown in Figure 1. Compression tests of drug particles were also performed using different roller compression process parameters (see Table 1), and bulk density, tap density, particle size distribution by sieve analysis, Hausner ratio, and compressibility index were determined for each process defined by the respective roller compression parameters. [Table 1]

[0230] Example 5: Stability test of a formulation containing compound I. Stability tests were also conducted on various formulations using different manufacturing methods (e.g., direct blending and granulation) for immediate-release compositions formulated according to the details in Tables 2 and 3.

[0231] The stability of formulations 1 and 2 (see Tables 2 and 3) was evaluated by storing them for 4 weeks under open container conditions at 40°C / 75% relative humidity and 50°C. Samples under open container conditions, representing the worst-case scenario, were tested.

[0232] Aside from some assay variability related to sample preparation and sampling, no significant increase in degradation products was observed, indicating the stability of the formulation. [Table 2] [Table 3]

[0233] Excipient compatibility testing was also performed by mixing compound I with 18 commonly used excipients. The samples were weighed, mixed in amber vials, and stored under two stability conditions (40°C / 75%RH and 50°C) with and without caps. These were evaluated at T0, 2 weeks, and 4 weeks. The results confirm that compound I is compatible with the excipients listed in the table below. [Table 4]

[0234] Example 6: Oral tablets of compound I, 5 mg and 50 mg - Batch 1 Oral tablets of 5 mg and 50 mg were prepared as "Batch 1" by a dry granulation process. Table 5 below shows the respective formulations. [Table 5]

[0235] The tablets in Table 5 were prepared as follows: Colloidal silicon dioxide (Cab-O-Sil) and crospovidone XL, NF were combined and screened through a #20 mesh sieve. In a Gemco double cone blender (0.5 cu ft): Compound I, lactose monohydrate, NF (Fast Flo), screened colloidal silicon dioxide (Cab-O-Sil), crospovidone XL, and microcrystalline cellulose, NF (Avicel PH 102) were added. The components were blended for 10 minutes without draining. Part I magnesium stearate was screened through a #30 mesh sieve. The screened magnesium stearate was added to the blend and blended for a further 3 minutes. The initial blend was compressed using various parameters to evaluate the particle size distribution and compressibility. The compressed material was ground through a 3.15 mm coarse screen and a 1.0 mm fine screen using a granulator located below the rollers.

[0236] The weight of the granular excipients was adjusted based on the yield of the ground granules. Hydroxypropyl cellulose (Klucel EXF), colloidal silicon dioxide, NF (Cab-O-Sil), and crospovidone were screened through a #20 mesh sieve. Magnesium stearate was screened through a #30 mesh sieve. In a Gemco double cone blender (0.5 cubic feet): approximately half of the ground granules, colloidal silicon dioxide (Cab-O-Sil), crospovidone, and the screened hydroxypropyl cellulose (Klucel EXF) were added sequentially along with the remaining ground granules. The mixture was blended at 23.5 rpm for 5 minutes. The screened magnesium stearate was then added to the blend and blended further at 23.5 rpm for a further 3 minutes. Table 6 shows the particle size distribution of the ground granules and the analysis of the final blend. [Table 6]

[0237] Next, the final blend was divided into two parts for compression: one with a strength of 5 mg and the other with a strength of 50 mg. The blends were compressed using a Globe Pharma tablet press and then tested for weight (n=10), hardness (n=10), thickness (n=10), and abrasion. The data for the 5 mg and 50 mg blends are summarized in Tables 7 and 8, respectively. The batches showed uniform compression, and consistent weight, hardness, and thickness were observed throughout the batch. [Table 7] [Table 8]

[0238] Next, the tablets were loaded into an O'Hara coating pan and coated with Opadry II White (85F18422) film coating solution until a theoretical coating weight increase of 3% was achieved. The film coating solution was prepared by adding the required amount of purified water and continuing mixing for at least 45 minutes. Dissolution tests of 5 mg coated tablets of compound I were performed using 500 mL of 0.01 M HCl at 37.0°C, with Apparatus II (paddle), at 75 rpm. Dissolution data is shown in Figure 2.

[0239] Batch 1 of the drug dissolved to over 75% within 20 minutes, with complete drug release within 70 minutes. Coated tablets were tested for disintegration and completely disintegrated within 30 minutes.

[0240] Dissolution of 50 mg coated tablets from Batch 1 was performed using 900 mL of 0.01 M HCl at 37.0°C, Instrument II (paddle), and 100 rpm. Dissolution data is shown in Figure 3. The dissolution profile of the tablets (50 mg) showed complete release within 10 minutes, confirming compliance with the assay and dissolution specifications. However, chromatographic analysis identified the presence of unidentified impurities in both the 5 mg and 50 mg formulations.

[0241] Example 7: Oral tablets of compound I, 1 mg and 5 mg - Batch 2 Batch 2 was prepared to create new tablets of 1 mg and 5 mg strength. Furthermore, low-moisture grade materials were used to evaluate their impact on unknown impurities. The tablets were prepared using a common blend and the same approach as in the previous batch to obtain dose-proportional strengths of 1 mg and 5 mg. Table 9 lists the formulations. [Table 9]

[0242] Batch 2 was manufactured to prepare new tablets of 1 mg and 5 mg strength. Furthermore, low-moisture grade materials were used to evaluate their effect on unknown impurities. The tablets were prepared using the same approach as the previous batch, with a common blend to obtain dose-proportional strength.

[0243] The manufacturing process for batch 2 was as follows: Compound I, microcrystalline cellulose, NF (Avicel PH112) (part IA), and Cab-O-Sil (part I) were combined and screened through a #30 mesh stainless steel (SS) screen into a container lined with appropriately labeled double polyethylene. Half of the microcrystalline cellulose (Avicel PH112) (part IB) was added to a 0.5 cubic foot Gemco double cone blender. The previously screened material was then added. The remaining portion of the microcrystalline cellulose (Avicel PH112) (part IB) was then added. The mixture was blended for 10 minutes without engaging the agitator bar.

[0244] The mixture was then dispensed into a properly labeled double polyethylene-lined container and screened through a #30 mesh SS screen into another properly labeled double polyethylene-lined container within the same 0.5 cubic foot Gemco double cone blender. 1.1. Half of the anhydrous lactose (Part I) was added. 1.2. Crospovidone (part I) was added. 1.3. The previously screened blend was added. 1.4. The remaining portion of the anhydrous lactose (Part I) was added. 1.5. The mixture was blended for 10 minutes without using a stirring bar.

[0245] Next, magnesium stearate was weighed and screened through a #30 mesh SS screen into appropriately labeled double polyethylene-lined containers.

[0246] Approximately half of the ground granules were added to a 0.5 cubic foot Gemco double cone blender, followed by colloidal silicon dioxide, NF (Cab-O-Sil), and crospovidone, then screened hydroxypropyl cellulose (Klucel EXF), and finally the remaining ground granules.

[0247] The blend was mixed at 23.5 rpm for 5 minutes.

[0248] Next, the screened magnesium stearate was added to the blender contents. The mixture was blended for a further 3 minutes at 23.5 rpm. 25-gram composite samples were collected for both the initial blend (before preparation) and the ground granules (after preparation). Sieve analysis and density results for both the pre- and post-preparation granules are recorded in Table 10.

[0249] The initial blend was compressed using an Alexanderwerk WP 120 roller compactor at a target pressure of 60 bar. The screw feeder speed was automatically adjusted to maintain a roller gap of 2.0 mm. Subsequently, the compressed material was ground through a 3.15 mm coarse screen and a 1.0 mm fine screen using a granulator positioned below the rollers. The ground granules were then used for the final blend.

[0250] The weight of the granular excipients was adjusted based on the yield of the ground granules before weighing. Hydroxypropyl cellulose (Klucel EXF), colloidal silicon dioxide, NF (Cab-O-Sil), and crospovidone were screened through a #20 mesh screen. Magnesium stearate was screened through a #30 mesh screen.

[0251] Half of the ground granules were added to a 0.5 cubic foot Gemco double cone blender, followed by colloidal silicon dioxide, NF (Cab-O-Sil), crospovidone, and screened hydroxypropyl cellulose (Klucel EXF), and then the remaining ground granules were added. The mixture was blended at 23.5 rpm for 5 minutes. Screened magnesium stearate was then added and blended for a further 3 minutes at 23.5 rpm. [Table 10]

[0252] Next, the final blend was divided into two parts for compression: one with a strength of 1 mg and the other with a strength of 5 mg. The blend was compressed using a Globe Pharma tablet press and then tested for weight (n=10), hardness (n=10), thickness (n=10), and abrasion. The data are summarized in Table 11. The batches showed uniform compression, and consistent weight, hardness, and thickness were observed throughout the batch. [Table 11]

[0253] The tablets were loaded into an O'Hara coating pan and coated with Opadry II White (85F18422) film coating solution until a theoretical coating weight increase of 3% was achieved. The film coating solution was prepared by adding the required amount of purified water and continuing to mix for at least 45 minutes. The coating solution was mixed throughout the entire coating process.

[0254] Coated mini-tablets of compound I (1 mg) were dissolved using 900 mL of 0.01 M HCl at 37.0°C with apparatus II (paddle) at 75 rpm. Dissolution data is shown in Figure 4. Complete dissolution occurred within 10 minutes. The coated tablets were tested for disintegration and completely disintegrated within 30 minutes.

[0255] The 5 mg coated mini-tablets of Compound I were dissolved in 900 mL of 0.01 M HCl at 37.0°C using Apparatus II (paddle) at 100 rpm. The dissolution data is shown in Figure 5. The dissolution profile of Compound I 5 mg tablets showed complete release within 10 minutes, confirming compliance with the assay and dissolution specifications. However, chromatographic analysis identified the presence of unidentified impurities in both the 1 mg and 5 mg formulations.

[0256] Example 8: Oral tablets of compound I, 1 mg and 5 mg by dry granulation - Batch 3 Batch 3 was reconditioned by removing anhydrous lactose and evaluating its effect on unidentified impurities using a dry granulation method. Table 12 below shows the formulations. [Table 12]

[0257] Batch 3 was produced by screening a combination of Compound I, microcrystalline cellulose, NF (Avicel PH112) (partial IA), and Cab-O-Sil (partial I) through a #30 mesh stainless steel (SS) screen into a container lined with appropriately labeled double polyethylene, as follows:

[0258] Half of the microcrystalline cellulose (Avicel PH112) (Part IB) was added to a 0.5 cubic foot Gemco double cone blender, then the previously screened material was added, followed by the remaining portion of the microcrystalline cellulose (Avicel PH112) (Part IB), and the mixture was blended for 10 minutes without using the agitator bar.

[0259] The mixture was then dispensed into a properly labeled, double-walled polyethylene-lined container. The initial blend was screened through a #30 mesh SS screen into another properly labeled, double-walled polyethylene-lined container.

[0260] Half of the screened blend was added to 0.5 cubic feet of Gemco double cone blend, crospovidone (part I), and the remaining portion of the screened blend was added, and the mixture was then blended for 10 minutes without using a stirrer bar.

[0261] The mixture was then poured into a properly labeled double polyethylene-lined container. The first blend was then screened through a #30 mesh SS screen into another properly labeled double polyethylene-lined container, and then the weighed magnesium stearate was screened through a #30 mesh SS screen into a properly labeled double polyethylene-lined container. Half of the screened blend was added to a 0.5 cubic foot Gemco double cone blender, the screened magnesium stearate was added, and the remaining portion of the screened blend was added, and then blended for 3 minutes without using the agitator bar.

[0262] The mixture was then discharged into a properly labeled, double-walled polyethylene-lined container.

[0263] Next, the initial blend was compressed using an Alexaderwerk WP 120 roller compactor.

[0264] The blend was compressed into flake / ribbon form at a target pressure of 60 bar. The screw feeder speed was automatically adjusted to maintain a roller gap of 2.0 mm. The compressed material was then ground through a 3.15 mm coarse screen and a 1.0 mm fine screen using a granulator located below the rollers.

[0265] Approximately half of the ground granules were added to a 0.5 cubic foot Gemco double cone blender. Colloidal silicon dioxide, NF (Cab-O-Sil), and crospovidone were added, followed by screened hydroxypropyl cellulose (Klucel EXF), and then the remaining ground granules. The blender was then blended at 23.5 rpm for 5 minutes. Screened magnesium stearate was then added to the blender contents and blended for a further 3 minutes at 23.5 rpm. Sieve analysis and density results for the pre- and post-preparation granules are shown in Table 13. [Table 13]

[0266] Next, the final blend was divided into two parts for compression: one with a strength of 1 mg and the other with a strength of 5 mg. The blend was compressed using a Globe Pharma tablet press and then tested for weight (n=10), hardness (n=10), thickness (n=10), and abrasion. The data are summarized in Table 14. The batches showed uniform compression, and consistent weight, hardness, and thickness were observed throughout the batch. [Table 14]

[0267] The tablets were loaded into an O'Hara coating pan and coated with Opadry II White (85F18422) film coating solution until a theoretical coating weight increase of 3% was achieved. The film coating solution was prepared by adding the required amount of purified water and continuing to mix for at least 45 minutes. The coating solution was mixed throughout the entire coating process.

[0268] The dissolution of batch 3 was complete within 10 minutes. The coated tablets were tested for disintegration and completely disintegrated within 30 minutes.

[0269] The dissolution of 1 mg coated tablets of compound 1 was performed using 900 mL of 0.01 M HCl at 37.0°C with apparatus II (paddle) at 75 rpm. The dissolution data for 1 mg is shown in Figure 6. The dissolution data for 5 mg tablets is shown in Figure 7.

[0270] The dissolution profile of compound I batch 3 tablets (5 mg) showed complete release within 10 minutes, confirming compliance with the assay and dissolution specifications. By removing anhydrous lactose from the formulation and analyzing the impact on the resulting unknown impurity, it was observed that the impurity was no longer present. As a result, it was concluded that the unknown impurity was due to the presence of lactose.

[0271] Example 9: Compound I oral tablets 1 mg and 5 mg by wet granulation - Batch 4 This batch was reconditioned by removing anhydrous lactose and using a wet granulation process to evaluate its effect on unidentified impurities. Table 15 shows the formulations. [Table 15]

[0272] Batch 4 was prepared as follows: Raw materials were weighed into appropriate containers, including double polyethylene-lined containers and HDPE containers. Compound I and microcrystalline cellulose, NF (Avicel PH112) (Part IA) were combined and screened through a #30 mesh stainless steel screen into appropriately labeled double polyethylene-lined containers. The following materials were then placed in a 0.5 cubic foot blender: 1.1. 50% of microcrystalline cellulose (Avicel PH112) - part IB was transferred. 1.2. Screened materials from Step 2 1.3. Hydroxypropylcellulose (Klucel EXF) 1.4. Remaining portion of microcrystalline cellulose (Avicel PH112) - Part IB

[0273] Next, the ingredients were blended for 5 minutes and then discharged into a stainless steel bowl. Granulation was then initiated by manually adding water in a controlled manner. After granulation, the granules were dried in an oven set to 60°C and the loss of drying (LOD) was measured. The dried granules were then sieved through a #18 mesh and a particle size distribution (PSD) test was performed. A 25-gram composite sample of the initially screened granules was collected.

[0274] The weight of the granular excipients was adjusted based on the yield of the ground granules before weighing. Colloidal silicon dioxide, NF(Cab-O-Sil), and crospovidone were screened through a #20 mesh screen. Magnesium stearate was screened through a #30 mesh screen. Approximately 50% of the screened granules were added to a 0.5 cubic foot Gemco double cone blender. Colloidal silicon dioxide, NF(Cab-O-Sil), and crospovidone were then incorporated. The remaining screened granules were added, and the mixture was blended at 23.5 rpm for 5 minutes. The screened magnesium stearate was then added to the blender contents, and blending was continued at 23.5 rpm for a further 3 minutes. A 25-gram composite sample of the final blend was then collected. Sieve analysis and density results of the final blend granules are shown in Table 16. [Table 16]

[0275] Next, the final blend was divided into two parts for compression: one with a strength of 1 mg and the other with a strength of 5 mg. The blend was compressed using a Globe Pharma tablet press and then tested for weight (n=10), hardness (n=10), thickness (n=10), and abrasion. The data are summarized in Table 17. The batches showed uniform compression, and consistent weight, hardness, and thickness were observed throughout the batch. [Table 17]

[0276] The tablets were loaded into an O'Hara coating pan and coated with Opadry II White (85F18422) film coating solution until a theoretical coating weight increase of 3% was achieved. The film coating solution was prepared by adding the required amount of purified water and continuing to mix for at least 45 minutes. The coating solution was mixed throughout the entire coating process.

[0277] The 1 mg coated tablets of compound I were dissolved using 500 mL of 0.01 M HCl at 37.0°C with apparatus II (paddle) at 75 rpm. The dissolution data is shown in Figure 8. The 1 mg of batch 4 was completely released within 10 minutes. The coated tablets were tested for disintegration and completely disintegrated within 30 minutes.

[0278] The 5 mg coated tablets of compound I were dissolved using 500 mL of 0.01 M HCl at 37.0°C with apparatus II (paddle) at 75 rpm. The dissolution data is shown in Figure 9.

[0279] The dissolution profile of the 5 mg tablet of compound I showed complete release within 10 minutes, confirming compliance with the assay and dissolution specifications.

[0280] Example 10: A clinical study to evaluate the safety, tolerability, and efficacy of Compound I in adults with selected solid tumors. This study will be conducted in two parts: a dose-setting part to determine the maximum tolerated dose and recommended phase 2 dose (RP2D) of compound I administered orally in once-daily (QD) and twice-daily (BID) schedules as described herein; and a dose-expansion part to evaluate the safety and tolerability of compound I administered at the recommended phase 2 dose and schedule in tumor-specific cohorts.

[0281] Description of the intervention model: Part 1 (multiple dose escalation, QD or BID): Selected locally advanced, recurrent, or metastatic solid tumors with PIK3CA mutations as determined by local assessment; Part 2 (RP2D determined in Part 1): Patients with selected locally advanced, recurrent, or metastatic solid tumors with PIK3CA mutations as determined by local assessment are enrolled in the group specified in the protocol.

[0282] Compound I for use in this study can be formulated for oral use and immediate release according to tablet formulations 1 and 2 described in Examples 5-8 above.

[0283] Adults with a histologically confirmed diagnosis of colorectal cancer, gastric cancer, non-small cell lung cancer, human epidermal growth factor receptor 2 (HER2)-negative breast cancer, head and neck squamous cell carcinoma, urothelial carcinoma, or selected gastrointestinal cancers (ovarian cancer, cervical cancer, or endometrial cancer with a known PIK3CA mutation or amplification (determined by an American Association of Pathologists / Clinical Laboratory Improvement Correction [CAP / CLIA]-validated test, or an equivalently accredited diagnostic laboratory)), who meet all eligibility criteria will be enrolled in the Phase I portion of the trial.

[0284] In the dose-setting portion of the study, compound I is evaluated as a monotherapy agent at multiple oral dose levels until disease progression, unacceptable toxicity, or any other reason for designated discontinuation is met.

[0285] The final dose demonstrates that the immediate-release tablet composition containing compound I is safe and effective in treating various cancers, including colorectal cancer and breast cancer.

Claims

1. A pharmaceutical composition comprising a compound that forms a covalent bond with a kinase or pseudokinase, or a therapeutically effective amount of a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient, wherein the pharmaceutical composition enables the immediate release of the compound.

2. The pharmaceutical composition according to claim 1, wherein the kinase is PI3-kinase (PI3K).

3. The compound that forms a covalent bond with a kinase or pseudokinase is a compound having the following formula: [Chemical formula 1] (FCB)a-(L)b-(CLM)c, or a pharmaceutically acceptable salt thereof, in the formula, CLM is a covalent modality, L is the linker, a and c are each independent integers between 1 and 5. b is an integer from 0 to 5, and FCB, 【Chemistry 2】 A pharmaceutical composition according to claim 1, comprising:

4. The FCB has the following structure: 【Transformation 3】 ， In the formula, R １ but, 【Chemistry 4】 A pharmaceutical composition according to claim 3, selected from the group consisting of the following.

5. The aforementioned compound is compound I, 【Transformation 5】 Compound I The pharmaceutical composition according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof.

6. The pharmaceutical composition according to claim 5, wherein compound I is a crystalline form of compound I, or a pharmaceutically acceptable salt or solvate of compound I.

7. The pharmaceutical composition according to claim 5, wherein compound I is an amorphous form of compound I or a pharmaceutically acceptable salt thereof.

8. The pharmaceutical composition according to claim 7, wherein the pharmaceutically acceptable salt of compound I is selected from the group consisting of phosphate, hydrochloride, sulfate, mesylate, benzenesulfonate, tosylate, fumarate, maleate, L-tartrate, citrate, and succinate, or solvates thereof.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the pharmaceutical excipient is selected from fillers, binders, disintegrants, lubricants, coatings, or any combination thereof.

10. The pharmaceutical composition according to claim 9, wherein the filler is selected from microcrystalline cellulose, silicified microcrystalline cellulose, mannitol EZ, pregelatinized starch, dicalcium phosphate dihydrate, lactose monohydrate, or any combination thereof.

11. The pharmaceutical composition according to claim 9, wherein the binder is selected from povidone K29 / 32, hydroxypropylcellulose, hydroxypropylmethylcellulose, or any combination thereof.

12. The pharmaceutical composition according to claim 9, wherein the disintegrant is selected from croscarmellose sodium, crospovidone XL, sodium glycolate starch, or any combination thereof.

13. The pharmaceutical composition according to claim 9, wherein the flow promoter is selected from colloidal silicon dioxide, talc, or any combination thereof.

14. The pharmaceutical composition according to claim 9, wherein the lubricant is selected from plant-derived magnesium stearate, sodium steryl fumarate, glyceryl behenate, steric acid, or any combination thereof.

15. The pharmaceutical composition according to claim 9, wherein the coating is a water-soluble polymer selected from polyvinyl alcohol (PVA)-based or hydroxypropyl methylcellulose (HPMC)-based film coatings.

16. The pharmaceutical composition according to claim 9, wherein the coating is selected from Opadry II White (PVA), Opadry II White (HPMC), or any combination thereof.

17. The pharmaceutical composition according to any one of claims 1 to 16, wherein the compound that forms a covalent bond with a kinase or pseudokinase, or a pharmaceutically acceptable salt thereof, is present in an amount of more than 50% by weight.

18. A pharmaceutical composition according to any one of claims 1 to 17, comprising a compound that forms a covalent bond with a kinase or pseudokinase, or a pharmaceutically acceptable salt thereof, in an amount ranging from about 0.1% to about 50% by weight, or 1% to about 30% by weight, or about 1% to about 20% by weight, or about 1% to about 10% by weight, or 1% to about 4% by weight, or about 4% to about 3% by weight, or about 5% to about 6% by weight relative to the total core weight of the composition, i.e., the weight before the addition of the coating.

19. The pharmaceutical composition according to any one of claims 1 to 18, wherein the compound that forms a covalent bond with a kinase or pseudokinase, or a pharmaceutically acceptable salt thereof, is present in an amount of about 2% by weight of the total core weight of the composition.

20. The pharmaceutical composition according to any one of claims 1 to 19, wherein the amount of the filler is in the range of about 10% to about 95% by weight, or about 20% to about 90% by weight, or about 30% to about 85% by weight, or about 40% to about 80% by weight, or about 40% to about 50% by weight, or about 42% to about 46% by weight, of the total core weight of the composition.

21. The pharmaceutical composition according to any one of claims 1 to 20, wherein the amount of the filler is in the range of about 70% to about 90% by weight of the total core weight of the composition.

22. The pharmaceutical composition according to any one of claims 1 to 20, wherein the filler is in an amount of about 85% by weight of the total core weight of the composition.

23. The pharmaceutical composition according to any one of claims 1 to 20, wherein the filler is in an amount of about 86% by weight of the total core weight of the composition.

24. The pharmaceutical composition according to any one of claims 1 to 20, wherein the filler is in an amount of about 87% by weight of the total core weight of the composition.

25. The pharmaceutical composition according to any one of claims 1 to 20, wherein the filler is in an amount of about 88% by weight of the total core weight of the composition.

26. The pharmaceutical composition according to any one of claims 1 to 25, wherein the amount of the binder is in the range of about 0.1% to about 10% by weight, or about 1% to about 10% by weight, or about 2% to about 8% by weight, or about 2% to about 6% by weight, or about 2% to about 5% by weight, or about 2% to 4% by weight, or about 3% to 4% by weight, of the total core weight of the composition.

27. The pharmaceutical composition according to any one of claims 1 to 26, wherein the amount of the binder is about 3% by weight of the total core weight of the composition.

28. The pharmaceutical composition according to any one of claims 1 to 27, wherein the amount of the disintegrant is in the range of about 0.1% to about 10% by weight, or about 1% to about 10% by weight, or about 2% to about 8% by weight, or about 2% to about 6% by weight, or about 2% to about 5% by weight, or about 3% to 4% by weight, of the total core weight of the composition.

29. The pharmaceutical composition according to any one of claims 1 to 28, wherein the disintegrant is in an amount of about 3% by weight or about 3.5% by weight of the total core weight of the composition.

30. The pharmaceutical composition according to any one of claims 1 to 29, wherein the amount of the lubricant is in the range of about 0.01% to about 10% by weight of the total core weight of the composition, or about 0.1% to about 5% by weight, or about 0.1% to about 3% by weight, or about 0.2% to about 2% by weight, or about 0.3% to 1% by weight, or about 0.3% to 0.6% by weight, or about 0.4% to 0.5% by weight.

31. The pharmaceutical composition according to any one of claims 1 to 30, wherein the lubricant is in an amount of about 0.5% by weight or about 2.5% by weight of the total core weight of the composition.

32. The pharmaceutical composition according to any one of claims 1 to 31, wherein the amount of the lubricant is in the range of about 0.01% to about 10% by weight of the total core weight of the composition, or about 0.1% to about 5% by weight, or about 0.1% to about 3% by weight, or about 0.2% to about 2% by weight, or about 0.3% to 1% by weight, or about 0.3% to 0.6% by weight, or about 0.4% to 0.5% by weight.

33. The pharmaceutical composition according to any one of claims 1 to 32, wherein the lubricant is in an amount of about 0.5% by weight or about 1% by weight of the total core weight of the composition.

34. The pharmaceutical composition according to any one of claims 1 to 33, wherein the coating is in an amount ranging from about 0.1% to about 10% by weight of the composition, or from about 1% to about 10% by weight, or from about 2% to about 8% by weight, or from about 2% to about 6% by weight, or from about 2% to about 5% by weight, or from about 3% to 4% by weight.

35. The pharmaceutical composition according to any one of claims 9 to 31, wherein the coating is in an amount of about 3% by weight of the total weight of the composition.

36. The pharmaceutical composition according to claims 9 to 35, wherein the pharmaceutical composition is a solid dosage form selected from tablets, powders, capsules, suppositories, sachets, lozenges, and trozenes.

37. The pharmaceutical composition according to claims 1 to 36, wherein the pharmaceutical composition is in the form of a tablet.

38. The pharmaceutical composition according to claim 37, wherein the drug particles before compression into the tablet have a D85 value of approximately 50 μm to approximately 100 μm, approximately 60 μm to approximately 80 μm, or approximately 63 μm.

39. The pharmaceutical composition according to claim 37, wherein the drug particles before compression have a D85 value exceeding approximately 50 μm, approximately 60 μm, approximately 70 μm, approximately 80 μm, approximately 90 μm, or approximately 100 μm.

40. The pharmaceutical composition according to claim 37, wherein the drug particles before compression into the tablet have a D65 value of approximately 100 μm to approximately 200 μm, approximately 120 μm to approximately 160 μm, or approximately 140 μm.

41. The pharmaceutical composition according to claim 37, wherein the drug particles before compression into the tablet have a D65 value exceeding approximately 120 μm, approximately 130 μm, approximately 140 μm, approximately 150 μm, approximately 160 μm, or approximately 180 μm.

42. The pharmaceutical composition according to claim 37, wherein the drug particles before compression into the tablet have a D45 value of approximately 300 μm to approximately 500 μm, approximately 350 μm to approximately 450 μm, or approximately 420 μm.

43. The pharmaceutical composition according to claim 37, wherein the drug particles before compression into the tablet have a D50 value of approximately 340 μm, approximately 360 μm, approximately 380 μm, approximately 420 μm, approximately 440 μm, or approximately 480 μm.

44. A method for modulating the activity of a kinase or pseudokinase, comprising administering any pharmaceutical composition described in any one of claims 1 to 43 to a subject requiring such modification.

45. The method according to claim 44, wherein the activity of the kinase or pseudokinase is inhibited.

46. The method according to claim 44 or 45, wherein the kinase is PI3K.

47. A method for treating cancer, neurodegenerative diseases, autoimmune disorders, or aging in a subject requiring such treatment, comprising administering a pharmaceutical composition according to any one of claims 1 to 43 to the subject requiring such treatment.

48. The method according to claim 47, wherein the subject has cancer.

49. The method according to claim 47 or 48, wherein the cancer is selected from the group consisting of leukemia, brain tumors, lung (small cell and non-small cell), ovarian, prostate, breast, colon, and intracellular cancers including intestinal tissue, pancreatic tissue, liver, kidney, prostate, ovarian, lung, bone marrow, or breast tissue.

50. The method according to any one of claims 47 to 49, wherein the subject has cancer having a mutation in the PIK3CA gene.

51. The pharmaceutical composition according to any one of claims 1 to 43, wherein the pharmaceutical composition enables the immediate release of the compound, and the pharmaceutical composition is substantially lactose-free.

52. The pharmaceutical composition according to claim 51, wherein the compound is compound I.

53. The pharmaceutical composition according to claim 51 or claim 52, wherein the lactose is anhydrous lactose.

54. The pharmaceutical composition according to any one of claims 1 to 43 or 51 to 53, wherein the pharmaceutical composition enables the immediate release of the compound, and the pharmaceutical composition is substantially free of reducing sugars.

55. The pharmaceutical composition according to claim 54, wherein the reducing sugar is lactose, galactose, glucose, maltose, fructose, ribose, and / or xylose.

56. The pharmaceutical composition according to claim 54 or 55, wherein the compound is compound I.

57. The pharmaceutical composition according to any one of claims 51 to 56, wherein the pharmaceutical excipient is selected from microcrystalline cellulose, crospovidone XL, colloidal silicon dioxide (Cab-O-Sil®), hydroxypropyl cellulose, magnesium stearate, or any combination thereof.

58. The pharmaceutical composition according to any one of claims 51 to 57, wherein the tablet is an oral tablet.

59. The pharmaceutical composition according to claim 58, wherein the oral tablet is an immediate-release oral tablet.

60. The pharmaceutical composition according to any one of claims 1 to 43 or 51 to 59, wherein the composition is as described in Tables 2, 3, 5, 9, 12, or 15 as described herein.

61. The pharmaceutical composition according to claim 60, wherein at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the compound is released within about 10 minutes by dissolution studies as described herein.

62. The pharmaceutical composition according to claim 61, wherein at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of compound I is released within about 9 minutes, 8 minutes, 7 minutes, 6 minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes, or 1 minute.

63. The pharmaceutical composition according to claim 61 or 62, wherein the dissolution of the compound in the tablet is determined using 900 mL of 0.01 M HCl at 37.0°C, Apparatus II (paddle), 75 rpm, or 100 rpm.

64. The pharmaceutical composition according to any one of claims 1 to 43 or 51 to 63, wherein compound I is in the form of a salt.

65. A method for modulating the activity of a kinase or pseudokinase, comprising administering any pharmaceutical composition described in any one of claims 51 to 64 to a subject requiring such modification.

66. The method according to claim 65, wherein the activity of the kinase or pseudokinase is inhibited.

67. The method according to claim 65 or 66, wherein the kinase is PI3K.

68. A method for treating cancer, neurodegenerative diseases, autoimmune disorders, or aging in a subject requiring such treatment, comprising administering a pharmaceutical composition according to any one of claims 51 to 64 to the subject requiring such treatment.

69. The method according to claim 68, wherein the subject has cancer.

70. The method according to claim 69, wherein the cancer is selected from the group consisting of leukemia, brain tumors, lung (small cell and non-small cell), ovarian, prostate, breast, colon, and intracellular cancers including intestinal tissue, pancreatic tissue, liver, kidney, prostate, ovarian, lung, bone marrow, or breast tissue.

71. The method according to any one of claims 68 to 70, wherein the subject has cancer having a mutation in the PIK3CA gene.

Images