Tetracyclic compounds, and compositions and methods thereof

Abstract

The invention provides novel derivatives of tetracyclic compounds and thereof, and pharmaceutical compositions thereof and methods for treating diseases and disorders, such as various types of cancer.

Description

Patent Application Atty. Docket No. ENTX-038PCTTETRACYCLIC COMPOUNDS, AND COMPOSITIONS AND METHODS THEREOFPriority Claims and Related Patent Applications

[0001] This application claims the benefit of priority to U.S. Provisional Application Serial Nos. 63 / 776,785, filed March 24, 2026, and 63 / 867,160, filed August 20, 2025, the entire content of each of which is incorporated herein by reference.Technical Fields of the Invention

[0002] The invention generally relates to novel compounds and therapeutic uses thereof. More particularly, the invention provides novel derivatives of tetracyclic compounds that are shown to be potent and selective phosphoinositide 3-kinase a (PI3Ka) inhibitors. The invention also provides pharmaceutical compositions comprising compounds of the invention and methods for treating diseases and disorders associated with or related to PI3Ka activities, such as various types of cancer.Background of the Invention

[0003] Phosphoinositide 3 -kinases (PI3Ks) are a family of related intracellular signal transducer enzymes capable of phosphorylating the 3 -position hydroxyl group of the inositol ring of phosphatidylinositol (Ptdins). PI3Ks have been linked to an extraordinarily diverse group of cellular functions, including cell growth, proliferation, differentiation, motility, survival and intracellular trafficking. The PI3K signaling pathway is one of the most frequently mutated in human cancer and is also a major factor in many other diseases in humans. For examples, PI3K signaling is associated with allergic contact dermatitis, rheumatoid arthritis, osteoarthritis, inflammatory bowel disease, chronic obstructive pulmonary disorder, psoriasis, multiple sclerosis, asthma, diabetic complications and acute coronary syndrome.

[0004] The PI3K family is divided into three different classes: Class I, Class II, and Class III, based on their primary structure, regulation, and lipid substrate specificity. (Kalaany et al. 2009 Nature 458 (7239): 725-31; Leevers et al. (1999) Current Opinion in Cell Biology 11 (2): 219-25.) Class I PI3Ks (p110α, p110β, p110δ and p110γ) are activated by tyrosine kinases or G protein-coupled receptors to produce phosphatidylinositol-3,4,5-triphosphate (PIP 3).Patent Application Atty. Docket No. ENTX-038PCT Association of effectors such as PDPK1 / AKT with PIP3 activates downstream signaling pathways.

[0005] The class IA PI3K pl 10a (PI3Ka) is mutated in many human cancers. Angiogenesis has been shown to selectively require the PI3Ka isoform in the control of endothelial cell migration. Mutations in the gene encoding PI3Ka (PIK3CA) or PI3Ka up-regulation occurs in many human cancers such as ovarian cancer, cervical cancer, breast cancer, colorectal cancer, endometrial cancer, gastric carcinomas, hepatocellular carcinoma, small and non-small cell lung cancer, thyroid carcinoma, acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), and glioblastomas. In cancer, PI3Ka mutations are often hotspot point mutations in the helical or kinase domain, such as H1047R, E542K, E545K and H1047X. Graupera et al. 2008 Nature 453: 662-6; Campbell et al. 2004 Cancer Res 64, 7678-7681; Levine et al. 2005 Clin Cancer Res 11, 2875-2878; Wang et al. 2005 Hum Mutat 25, 322; Lee etal. 2005 Gynecol Oncol 97, 26-34, Bachman, et al. 2004 Cancer Biol Ther 3, 772-775; Li et al. 2006 Breast Cancer Res Treat 96, 91-95; Saal et al. 2005 Cancer Res 65, 2554-2559; Samuels and Velculescu 2004 Cell Cycle 3, 1221-1224; Samuels, et al. 2004 Science 304, 554; Velho et al 2005 Ear J Cancer 41, 1649-1654; Oda et al. 2005 Cancer Res. 65, 10669-10673; Byun et al. 2003 Int J Cancer 104, 318-327; Lee et al. 2005 Oncogene 24, 1477-1480; Tang etal. 2006 Lung Cancer 51, 181-191; Massion et al. 2004 Am J Respir Grit Care Med 170, 1088-1094; Wu etal. 2005 J Clin Endocrinol Metab 90, 4688-4693; Sujobert et al. 1997 Blood 106, 1063-1066; Hickey and Cotter 2006 J Biol Chem 281, 2441-2450; Hartmann et al. 2005 Acta Neuropathol (Berl) 109, 639-642.)

[0006] There remains an urgent and unmet need for potent and selective PI3Ka inhibitors that are safe and effective in treating diseases and conditions associated with PI3Ka, such as various types of cancer (e.g., breast cancer, ovarian cancer, colorectal cancer, lung cancer).Summary of the Invention

[0007] The invention provides novel urea derivatives of tetracyclic compounds as PI3Ka inhibitors, which are shown herein to exhibit favorable potency and selectivity profiles over known PI3Ka inhibitors. These novel compounds selectively target, bind to, inhibit and / or modulate the activity of PI3Ka. The compounds are also orally available with pharmacokinetic profiles suitable for development into an orally administered therapeutic agent for treatingPatent Application Atty. Docket No. ENTX-038PCT various diseases and disorders associated with or related to PI3Kα activities, such as various types of cancer.

[0008] In one aspect, the invention generally relates to a compound having the structural formula (I):NN(I)or a pharmaceutically acceptable form or an isotope derivative thereof,whereinX1is N or CR1;X2is N, C(O) or CR2;X3is N or CR3;each of R1, R2and R3is independently H, D, halo, CN, R, OR, C(O)NHR, C2-6 alkynyl, NRR', SO2R or SO2NR, wherein said R and C2-6 alkynyl are optionally substituted with one or more of Ra;R4is a 5- to 10-memLered monocyclic, bicyclic or bridged aryl or heteroaryl ring with 0-4 ring heteroatoms independently selected from N, O and S, optionally substituted with one or more Rc;R5is C(O)R5'or R5", whereinR5'is a 5- to 10-memLered monocyclic, bicyclic or bridged carbocyclyl, heterocyclic, aryl or heteroaryl ring with 0-4 ring heteroatoms independently selected from N, O and S, optionally substituted with one or more Rb; andR5"is a 9- or 10-memLered bicyclic or bridged heteroaryl ring with 0-4 ring heteroatoms independently selected from N, O and S, optionally substituted with one or more Rb;R6is a Ci-6 alkyl substituted with0-8 Rd;Patent Application Atty. Docket No. ENTX-038PCT an unsubstituted or substituted C3-6 carbocyclic or heterocyclic group; or an unsubstituted or substituted 5- or 6-memLered aryl or heteroaryl group; each Rais independently D, halo, OH, CN, C1-6 alkoxy, NRR' or SO2R;each of Rband Rcis independentlydeuterium, oxo, halogen, CN, NO2, OR, SR, NRR’, S(O)2R, S(0)2NRR’, S(O)R, S(O)NRR’, S(O)(NR)R, C(O)R, C(O)OR, C(O)NRR’, C(O)N(R)OR, OC(O)R, OC(O)NRR’, N(R)C(O)OR, N(R)C(O)R, N(R)C(O)NRR’, N(R)C(NR)NRR’, N(R)S(O)2NRR’ or N(R)S(O)2R; ora substituted or unsubstituted group selected from C1-6 alkyl or 4- to 6-memLered carbocyclic ring;each Rdis independently D, halogen, CN, OR, alkynyl, SR or NRR’;each of R and R’ is independently selected from H, unsubstituted or substituted C1-4 alkyl, or unsubstituted or substituted 3- to 6-memLered carbocyclic ring, or where R and R’ are attached to the same C or N atom, together form an unsubstituted or substituted 4- to 6-memLered heterocyclic ring.

[0009] In another aspect, the invention generally relates to a pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable excipient, carrier, or diluent.

[0010] In yet another aspect, the invention generally relates to a unit dosage form comprising a pharmaceutical composition disclosed herein.

[0011] In yet another aspect, the invention generally relates to a method for inhibiting cell proliferation in vitro or in vivo, comprising contacting a cell with an effective amount of a compound disclosed herein.

[0012] In yet another aspect, the invention generally relates to a method for modulating (e.g., reducing) PI3Ka activity in a cell, comprising contacting the cell with a compound disclosed herein.

[0013] In yet another aspect, the invention generally relates to a method for treating a disease or disorder mediated by PI3Ka, comprising administering to a subject in need thereof a therapeutically effective amount of a compound disclosed herein.Patent Application Atty. Docket No. ENTX-038PCT

[0014] In yet another aspect, the invention generally relates to a method for treating or reducing cancer, or a related disease or disorder, comprising administering to a subject in need thereof a therapeutically effective amount of a compound disclosed herein.

[0015] In yet another aspect, the invention generally relates to use of a compound disclosed herein, and a pharmaceutically acceptable excipient, carrier, or diluent, in preparation of a medicament for treating a disease or disorder.Definitions

[0016] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. General principles of organic chemistry, as well as specific functional moieties and reactivity, are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 2006.

[0017] As used herein, “at least” a specific value is understood to be that value and all values greater than that value.

[0018] The term “comprising”, when used to define compositions and methods, is intended to mean that the compositions and methods include the recited elements, but do not exclude other elements. The term “consisting essentially of’, when used to define compositions and methods, shall mean that the compositions and methods include the recited elements and exclude other elements of any essential significance to the compositions and methods. For example, “consisting essentially of’ refers to administration of the pharmacologically active agents expressly recited and excludes pharmacologically active agents not expressly recited. The term consisting essentially of does not exclude pharmacologically inactive or inert agents, e.g., pharmaceutically acceptable excipients, carriers or diluents. The term “consisting of’, when used to define compositions and methods, shall mean excluding trace elements of other ingredients and substantial method steps. EmLodiments defined by each of these transition terms are within the scope of this invention.

[0019] Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from context, all numerical values provided herein can be modified by the term about.Patent Application Atty. Docket No. ENTX-038PCT

[0020] In this specification and the appended claims, the singular forms "a," "an," and "the" include plural reference, unless the context clearly dictates otherwise.

[0021] As used herein, the terms “administration” of or “administering” a disclosed compound encompasses the delivery to a subject of a compound as described herein, or a prodrug or other pharmaceutically acceptable form thereof, using any suitable formulation or route of administration, as discussed herein.

[0022] As used herein, the term “co-administer” refers to the presence of two pharmacological agents in a subject’s body (e.g., in the blood) at the same time. The two pharmacological agents can be administered concurrently or sequentially.

[0023] The terms “disease”, “disorder” and “condition” are used interchangeably unless indicated otherwise.

[0024] As used herein, the terms "effective amount" or "therapeutically effective amount" refer to that amount of a compound or pharmaceutical composition described herein that is sufficient to effect the intended application including, but not limited to, disease treatment, as illustrated below.

[0025] In some emLodiments, the amount is that is sufficient to negatively modulate or inhibit the activity of PI3Ka. In some emLodiments, the amount is that effective for reduction or amelioration of a symptom to stop or reversion of progression of a disease or disorder such as cancer. In some emLodiments, the amount is that effective for detectable killing or inhibition of the growth or spread of cancer cells; the size or numLer of tumors; or other measure of the level, stage, progression or severity of the cancer.

[0026] The therapeutically effective amount can vary depending upon the intended application, or the subject and disease condition being treated, e.g., the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the weight and age of the patient, which can readily be determined by one of ordinary skill in the art. Such amount may be administered as a single dosage or according to a regimen. The term also applies to a dose that will induce a particular response in target cells, e.g., reduction of cell migration. The specific dose will vary depending on, for example, the particular compounds chosen, the species of subject and their age / existing health conditions or risk for health conditions, the dosing regimen to be followed, the severity of the disease, whether it isPatent Application Atty. Docket No. ENTX-038PCT administered in comLination with other agents, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried.

[0027] As used herein, an “inhibitor” of “PI3Ka” refers to a compound of the invention capable of negatively modulating or inhibiting all or a portion of the activity of PI3Ka.

[0028] As used herein, a “PI3Ka-associated” disease or disorder refers to diseases or disorders associated with or mediated by PI3Ka or having one or more PI3Ka mutations.Examples of PI3Ka-associated diseases or disorders include various cancer types. A PI3Ka-associated disease or disorder may also refer allergic contact dermatitis, rheumatoid arthritis, osteoarthritis, inflammatory bowel disease, chronic obstructive pulmonary disorder, psoriasis, multiple sclerosis, asthma, diabetic complications, or acute coronary syndrome.

[0029] As used herein, the term “'contacting” refers to the bringing together of indicated moieties in vitro or in vivo. For example, “contacting” a cell with a compound disclosed herein includes the administration of the compound to a subject in need thereof, as well as, for example, introducing the compound into a sample containing a cellular or purified preparation. In some emLodiments, a cell in which inhibition of PI3Ka activity is desired is contacted with an effective amount of a compound disclosed herein or pharmaceutically acceptable form thereof to negatively modulate the activity of PI3Ka. By negatively modulating the activity of PI3Ka, the methods disclosed herein are designed to inhibit undesired cellular proliferation resulting from enhanced PI3Ka activity within the cell. The cells may be contacted in a single dose or multiple doses in accordance with a particular treatment regimen to effect the desired negative modulation of PI3Ka. The abi lity of compounds to bind PI3Ka may be monitored in vitro using methods known in the art. The inhibitory activity of exemplary compounds in cells may be monitored, for example, by measuring the inhibition of PI3Ka activity using methods known in the art.

[0030] As used herein, the terms “unsubstituted or substituted” and “optionally substituted” are used interchangeably and refer to where a given chemical moiety (e.g., an alkyl group) can (but is not required to) be bonded other substituents (e.g., heteroatoms). For instance, an alkyl group that is optionally substituted can be a fully saturated alkyl chain (i.e. a pure hydrocarbon). Alternatively, the same optionally substituted alkyl group can have substituents different from hydrogen. For instance, it can, at any point along the chain be bounded to a halogen atom, a hydroxyl group, or any other substituent described herein. Thus, the term “optionally substituted” means that a given chemical moiety has the potential to contain other functionalPatent Application Atty. Docket No. ENTX-038PCT groups, but does not necessarily have any further functional groups. Suitable substituents used in the optional substitution of the described groups include, without limitation, halogen, oxo, CN, -COOH, -CH2CN, -O-C1-C6alkyl, C1-C6alkyl, -OC1-C6alkenyl, -OC1-C6alkynyl, -C1-C6alkenyl, -C1-C6alkynyl, -OH, -OP(O)(OH)2, -OC(O)C1-C6alkyl, -C(O)C1-C6alkyl, -OC(O)OC1-C6alkyl, NH2, NH(C1-C6alkyl), N(C1-C6alkyl)2, -NHC(O)C1-C6alkyl, -C(O)NHC1-C6alkyl, -S(O)2-C1-C6alkyl, -S(O)NHC1-C6alkyl, and S(O)N(C1-C6alkyl)2.

[0031] As used herein, a “pharmaceutically acceptable form” of a disclosed compound includes, but is not limited to, pharmaceutically acceptable salts, esters, hydrates, solvates, isomers, prodrugs, and isotopically labeled derivatives of disclosed compounds. In one emLodiment, a "pharmaceutically acceptable form" includes, but is not limited to, pharmaceutically acceptable salts, esters, isomers, prodrugs and isotopically labeled derivatives of disclosed compounds. In some emLodiments, a "pharmaceutically acceptable form" includes, but is not limited to, pharmaceutically acceptable salts, esters, stereoisomers, prodrugs and isotopically labeled derivatives of disclosed compounds.

[0032] In certain emLodiments, the pharmaceutically acceptable form is a pharmaceutically acceptable salt. As used herein, the term "pharmaceutically acceptable salt" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of subjects without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds provided herein include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, besylate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,Patent Application Atty. Docket No. ENTX-038PCT lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3 -phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. In some emLodiments, organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, lactic acid, trifluoracetic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

[0033] The salts can be prepared in situ during the isolation and purification of the disclosed compounds, or separately, such as by reacting the free base or free acid of a parent compound with a suitable base or acid, respectively. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(C1-4alkyl)4salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some emLodiments, the pharmaceutically acceptable base addition salt can be chosen from ammonium, potassium, sodium, calcium, and magnesium salts.

[0034] In certain emLodiments, the pharmaceutically acceptable form is a pharmaceutically acceptable ester. As used herein, the term "pharmaceutically acceptable ester" refers to esters that hydrolyze in vivo and include those that break down readily in the human body to leave the parent compound or a salt thereof. Such esters can act as a prodrug as defined herein.Pharmaceutically acceptable esters include, but are not limited to, alkyl, alkenyl, alkynyl, aryl, aralkyl, and cycloalkyl esters of acidic groups, including, but not limited to, carboxylic acids, phosphoric acids, phosphinic acids, sulfinic acids, sulfonic acids and boronic acids. Examples ofPatent Application Atty. Docket No. ENTX-038PCT esters include formates, acetates, propionates, butyrates, acrylates and ethylsuccinates. The esters can be formed with a hydroxy or carboxylic acid group of the parent compound.

[0035] In certain emLodiments, the pharmaceutically acceptable form is a “solvate” (e.g., a hydrate). As used herein, the term “solvate” refers to compounds that further include a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. The solvate can be of a disclosed compound or a pharmaceutically acceptable salt thereof. Where the solvent is water, the solvate is a "hydrate". Pharmaceutically acceptable solvates and hydrates are complexes that, for example, can include 1 to about 100, or 1 to about 10, or 1 to about 2, about 3 or about 4, solvent or water molecules. It will be understood that the term "compound" as used herein encompasses the compound and solvates of the compound, as well as mixtures thereof.

[0036] In certain emLodiments, the pharmaceutically acceptable form is a prodrug. As used herein, the term “prodrug” (or “pro-drug”) refers to compounds that are transformed in vivo to yield a disclosed compound or a pharmaceutically acceptable form of the compound. A prodrug can be inactive when administered to a subject, but is converted in vivo to an active compound, for example, by hydrolysis (e.g., hydrolysis in blood). In certain cases, a prodrug has improved physical and / or delivery properties over the parent compound. Prodrugs can increase the bioavailability of the compound when administered to a subject (e.g., by permitting enhanced absorption into the blood following oral administration) or which enhance delivery to a biological compartment of interest (e.g., the brain or lymphatic system) relative to the parent compound. Exemplary prodrugs include derivatives of a disclosed compound with enhanced aqueous solubility or active transport through the gut memLrane, relative to the parent compound.

[0037] The prodrug compound often offers advantages of solubility, tissue compatibility or delayed release in a mammalian organism (see, e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7- 9, 21-24 (Elsevier, Amsterdam). A discussion of prodrugs is provided in Higuchi, T., et al., " Pro-drugs as Novel Delivery Systems," A. C. S. Symposium Series, Vol. 14, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated in full by reference herein. Exemplary advantages of a prodrug can include, but are not limited to, its physical properties, such as enhanced water solubility for parenteral administration at physiological pHPatent Application Atty. Docket No. ENTX-038PCT compared to the parent compound, or it can enhance absorption from the digestive tract, or it can enhance drug stability for long-term storage.

[0038] As used herein, the term “pharmaceutically acceptable excipient, carrier, or diluent” refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject pharmaceutical agent from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate, magnesium stearate, and polyethylene oxide-polypropylene oxide copolymer as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0039] As used herein, the term “subject” refers to any animal (e.g., a mammal), including, but not limited to humans, non-human primates, rodents, and the like, which is to be the recipient of a particular treatment. Typically, the terms “subject” and “patient” are used interchangeably herein in reference to a human subject.

[0040] In some emLodiments, the subject has experienced and / or exhibited at least one symptom of the disease or disorder to be treated with a compound disclosed herein and / or according to a herein disclosed method. In some emLodiments, the subject has been identified or diagnosed as having a cancer having one or more PI3Ka mutations. In some emLodiments, the subject has a cancer that is positive for a PI3Ka mutation. In some emLodiments, the subject is suspected of having a PI3Ka gene-associated cancer.Patent Application Atty. Docket No. ENTX-038PCT

[0041] In some emLodiments of any of the methods or uses described herein, an assay is used to determine whether the subject has one or more PI3Ka mutations using a sample (e.g., a biological sample or a biopsy sample (e.g., a paraffin-emLedded biopsy sample) from a subject. Various techniques may be employed, for example, next generation sequencing, immunohistochemistry, fluorescence microscopy, break apart FISH analysis, Southern blotting, Western blotting, FACS analysis, Northern blotting, and PCR-based amplification (e.g., RT-PCR and quantitative real-time RT-PCR).

[0042] As used herein, the terms “treatment” or “treating” a disease or disorder refers to a method of reducing, delaying or ameliorating such a condition before or after it has occurred. Treatment may be directed at one or more effects or symptoms of a disease and / or the underlying pathology. Treatment is aimed to obtain beneficial or desired results including, but not limited to, therapeutic benefit and / or a prophylactic benefit. By therapeutic benefit is meant eradication or amelioration of the underlying disorder being treated. Also, a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the patient, notwithstanding that the patient can still be afflicted with the underlying disorder. For prophylactic benefit, the pharmaceutical compounds and / or compositions can be administered to a patient at risk of developing a particular disease, or to a patient reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease may not have been made. The treatment can be any reduction and can be, but is not limited to, the complete ablation of the disease or the symptoms of the disease. As compared with an equivalent untreated control, such reduction or degree of prevention is at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, or 100% as measured by any standard technique.

[0043] As used herein, the term "therapeutic effect" refers to a therapeutic benefit and / or a prophylactic benefit as described herein. A prophylactic effect includes delaying or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any comLination thereof.

[0044] Compounds of the present invention are, subsequent to their preparation, preferably isolated and purified to obtain a composition containing an amount by weight equal to or greaterPatent Application Atty. Docket No. ENTX-038PCT than 95% (“substantially pure”), which is then used or formulated as described herein. In certain emLodiments, the compounds of the present invention are more than 99% pure.

[0045] Solvates and polymorphs of the compounds of the invention are also contemplated herein. Solvates of the compounds of the present invention include, for example, hydrates.

[0046] As used herein, the term an “isolated” or “substantially isolated” molecule (such as a polypeptide or polynucleotide) is one that has been manipulated to exist in a higher concentration than in nature or has been removed from its native environment. For example, a subject antibody is isolated, purified, substantially isolated, or substantially purified when at least 10%, or 20%, or 40%, or 50%, or 70%, or 90% of non-subject-antibody materials with which it is associated in nature have been removed. For example, a polynucleotide or a polypeptide naturally present in a living animal is not "isolated," but the same polynucleotide or polypeptide separated from the coexisting materials of its natural state is "isolated." Further, recomLinant DNA molecules contained in a vector are considered isolated for the purposes of the present invention. Isolated RNA molecules include in vivo or in vitro RNA replication products of DNA and RNA molecules. Isolated nucleic acid molecules further include synthetically produced molecules. Additionally, vector molecules contained in recomLinant host cells are also isolated. Thus, not all “isolated” molecules need be “purified.”

[0047] As used herein, the term “purified” when used in reference to a molecule, it means that the concentration of the molecule being purified has been increased relative to molecules associated with it in its natural environment, or environment in which it was produced, found or synthesized. Naturally associated molecules include proteins, nucleic acids, lipids and sugars but generally do not include water, buffers, and reagents added to maintain the integrity or facilitate the purification of the molecule being purified. According to this definition, a substance may be 5% or more, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 98% or more, 99% or more, or 100% pure when considered relative to its contaminants.

[0048] Definitions of specific functional groups and chemical terms are described in more detail below. When a range of values is listed, it is intended to encompass each value and subrange within the range. For example, “Ci-4 alkyl” is intended to encompass, Ci, C2, C3, C4, C1-3, Ci-2, C2-4, C3-4 and C2-3 alkyl groups.Patent Application Atty. Docket No. ENTX-038PCT

[0049] As used herein, the term “aliphatic” or “aliphatic group” refers to a linear or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic.

[0050] As used herein, the term “alkyl” refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to ten carbon atoms (e.g., Ci-io alkyl). Whenever it appears herein, a numerical range such as “1 to 10” refers to each integer in the given range; e.g., “1 to 10 carbon atoms” means that the alkyl group can consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms, although the present definition also covers the occurrence of the term "alkyl" where no numerical range is designated. In some emLodiments, “alkyl” can be a Ci-6 alkyl group. In some emLodiments, alkyl groups have 1 to 10, 1 to 8, 1 to 6, or 1 to 3 carbon atoms. Representative saturated straight chain alkyls include, but are not limited to, -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl; while saturated branched alkyls include, but are not limited to, -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, 2-methylbutyl, 3-methylbutyl, 2-methylpentyl, 3 -methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3 -dimethylbutyl, and the like. The alkyl is attached to the parent molecule by a single bond. Unless stated otherwise in the specification, an alkyl group is optionally substituted by one or more of substituents which independently include: acyl, alkyl, alkenyl, alkynyl, alkoxy, alkylaryl, cycloalkyl, aralkyl, aryl, aryloxy, amino, amido, amidino, imino, azide, carbonate, carbamate, carbonyl, heteroalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, hydroxy, cyano, halo, haloalkoxy, haloalkyl, ester, ether, mercapto, thio, alkylthio, arylthio, thiocarbonyl, nitro, oxo, phosphate, phosphonate, phosphinate, silyl, sulfinyl, sulfonyl, sulfonamidyl, sulfoxyl, sulfonate, urea, -Si(Rx)3, -ORX, -SRX, -OC(O)-RX, -N(RX)2, -C(O)RX, -C(O)ORX, -OC(O)N(RX)2, -C(O)N(RX)2, -N(RX)C(O)ORX, -N(RX)C(O)RX, -N(RX)C(O)N(RX)2, -N(RX)C(NRX)N(RX)2, -N(Rx)S(O)tN(Rx)2(where t is 1 or 2), -P(=O)(RX)(RX), or -O-P(=O)(ORX)2wherein each Rxis independently hydrogen, alkyl, haloalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl, and each of these moieties can be optionally substituted as defined herein. In a non-limiting emLodiment, a substituted alkyl can be selected from fluoromethyl, difluoromethyl,Patent Application Atty. Docket No. ENTX-038PCT tri fluoromethyl, 2-fluoroethyl, 3 -fluoropropyl, hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, benzyl, and phenethyl.

[0051] Unless otherwise specifically defined, the term “aromatic” or “aryl” refers to cyclic, aromatic hydrocarbon groups that have 1 to 2 aromatic rings, including monocyclic or bicyclic groups such as phenyl, biphenyl or naphthyl. Where containing two aromatic rings (bicyclic, etc.), the aromatic rings of the aryl group may be joined at a single point (e.g., biphenyl), or fused (e.g., naphthyl). The aryl group may be optionally substituted by one or more substituents, e.g., 1 to 5 substituents, at any point of attachment. Exemplary substituents include, but are not limited to, H, halogen, -O-Ci-Ce alkyl, Ci-Ce alkyl, -Ci-Ce alkenyl, -OCi-Ce alkynyl, -Ci-Ce alkenyl, -Ci-C6alkynyl, -OH, -OP(O)(OH)2, -OC(O)C1-C6alkyl, -C(O)C1-C6alkyl, -OC(O)OC1-C6alkyl, NH2, NH(CI-C6alkyl), N(CI-C6alkyl)2, -S(O)2-Ci-C6alkyl, -S(O)NHCi-C6alkyl, and S(O)N(Ci-Ce alkyl)2. The substituents can themselves be optionally substituted. Furthermore, when containing two fused rings the aryl groups herein defined may have an unsaturated or partially saturated ring fused with a fully unsaturated ring. Exemplary ring systems of these aryl groups include indanyl, indenyl, tetrahydronaphthal enyl, and tetrahydrobenzoannulenyl.

[0052] The term “halogen” or “halo” refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

[0053] As used herein, the terms “heteroaryl” or “hetero-aromatic” refer to groups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms: having 6, 10, or 14 p electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to three heteroatoms per ring selected from the group consisting of N, O, and S. Examples of heteroaryl groups include acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, 6,7-dihydro-5H-pyrrolo[l,2- a]imidazole, furanyl, furazanyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3, 4 -oxadi zolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl,Patent Application Atty. Docket No. ENTX-038PCT pyrazolinyl, pyrazolyl, pyridazinyl, pyridoox azole, pyridoimidazole, pyridothi azole, pyridinyl, pyridyl, pyrimidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thi enoox azolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3- triazolyl, 1,2, 4-tri azolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, and xanthenyl.“Heteroaryl” also refers to bicyclic ring systems having, in addition to carbon atoms, from one to three heteroatoms per ring selected from the group consisting of N, O, and S in which one ring system may be saturated or partially saturated.

[0054] Heteroaryl groups may be substituted with 0, 1, 2, 3, or 4 substituents independently selected from alkenyl, alkoxy, alkoxyalkoxy, alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, alkyl, alkyl carbonyl, alkylcarbonylalkyl, alkylcarbonyloxy, alkylthio, alkylthioalkyl, alkynyl, carboxy, carboxyalkyl, cyano, cyanoalkyl, formyl, haloalkoxy, haloalkyl, halogen, hydroxy, hydroxyalkyl, mercapto, nitro, -NZ1Z2, and (NZiZ2)carbonyl. The term " NZ1Z2" as used herein, means two groups, Zi and Z2, which are appended to the parent molecular moiety through a nitrogen atom Z1 and Z2 are each independently selected from the group consisting of hydrogen, alkyl, alkylcarbonyl, and formyl. Representative examples of NZ1Z2 include, but are not limited to, amino, methylamino, acetylamino, and acetylmethylamino.

[0055] As used herein, the term “alkoxy” refers to an -O-alkyl radical.

[0056] As used herein, the terms “cycloalkyl” and “carbocyclyl” each refers to a monocyclic or polycyclic radical that contains only carbon and hydrogen, and can be saturated or partially unsaturated. Unless stated otherwise in the specification, the term is intended to include both substituted and unsubstituted cycloalkyl groups. Partially unsaturated cycloalkyl groups can be termed "cycloalkenyl" if the carbocycle contains at least one double bond, or "cycloalkynyl" if the carbocycle contains at least one triple bond. Cycloalkyl groups include groups having from 3 to 13 ring atoms (i.e., C3-13 cycloalkyl). Whenever it appears herein, a numerical range such as "3 to 10" refers to each integer in the given range; e.g., "3 to 13 carbon atoms" means that the cycloalkyl group can consist of 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, etc., up to and including 13 carbon atoms. The term "cycloalkyl" also includes bridged and spiro-fused cyclic structures containing no heteroatoms. The term also includes monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of ring atoms) groups. Polycyclic aryl groupsPatent Application Atty. Docket No. ENTX-038PCT include bicycles, tricycles, tetracycles, and the like. In some emLodiments, “cycloalkyl” can be a C3-8 cycloalkyl radical. In some emLodiments, “cycloalkyl” can be a C3-5 cycloalkyl radical. Illustrative examples of cycloalkyl groups include, but are not limited to the following moieties: C3-6 carbocyclyl groups include, without limitation, cyclopropyl (C3), cyclobutyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (Ce), cyclohexenyl (Ce), cyclohexadienyl (Ce) and the like. Examples of C3-7 carbocyclyl groups include norbornyl (C7). Examples of C3-8 carbocyclyl groups include the aforementioned C3-7 carbocyclyl groups as well as cycloheptyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (Cs), bicyclo[2.2.1]heptanyl, bicyclo[2.2.2]octanyl, and the like. Examples of C3-13 carbocyclyl groups include the aforementioned C3-8 carbocyclyl groups as well as octahydro-lH indenyl, decahydronaphthalenyl, spiro[4.5]decanyl and the like. Unless stated otherwise in the specification, a cycloalkyl group can be optionally substituted by one or more substituents which independently include: acyl, alkyl, alkenyl, alkynyl, alkoxy, alkylaryl, cycloalkyl, aralkyl, aryl, aryloxy, amino, amido, amidino, imino, azide, carbonate, carbamate, carbonyl, heteroalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, hydroxy, cyano, halo, haloalkoxy, haloalkyl, ester, ether, mercapto, thio, alkylthio, arylthio, thiocarbonyl, nitro, oxo, phosphate, phosphonate, phosphinate, silyl, sulfinyl, sulfonyl, sulfonamidyl, sulfoxyl, sulfonate, urea, -Si(Ra)3, -ORa, -SRa, -OC(O)-Ra, -N(Ra)2, -C(O)Ra, -C(O)ORa, -OC(O)N(Ra)2, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -N(Ra)C(O)Ra, -N(Ra)C(O)N(Ra)2, -N(Ra)C(NRa)N(Ra)2, -N(Ra)S(O)tN(Ra)2(where t is 1 or 2), -P(=O)(Ra)(Ra), or -O-P(=O)(ORa)2where each Rais independently hydrogen, alkyl, haloalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl, and each of these moieties can be optionally substituted as defined herein. The terms “cycloalkenyl" and "cycloalkynyl" mirror the above description of "cycloalkyl" wherein the prefix "alk" is replaced with "alken" or "alkyn" respectively, and the parent "alkenyl" or "alkynyl" terms are as described herein. For example, a cycloalkenyl group can have 3 to 13 ring atoms, such as 5 to 8 ring atoms. In some emLodiments, a cycloalkynyl group can have 5 to 13 ring atoms.

[0057] As used herein, the term “heterocycloalkyl” refers to a cycloalkyl radical, which have one or more skeletal chain atoms selected from an atom other than carbon, e.g., O, N, S, P or comLinations thereof. Unless stated otherwise in the specification, the term is intended to include both substituted and unsubstituted heterocycloalkyl groups. Illustrative examples ofPatent Application Atty. Docket No. ENTX-038PCT heterocycloalkyl include 2-hydroxy-aziridin-l-yl, 3-oxo-l-oxacyclobutan-2-yl, 2,2-dimethyl-tetrahydrofuran-3-yl, 3 -carboxy -morpholin-4-yl, l-cyclopropyl-4-methyl-piperazin-2-yl. 2-pyrrolinyl, 3-pyrrolinyl, dihydro-2H-pyranyl, 1,2,3,4-tetrahydropyridine, 3,4-dihydro-2H-[l,4]oxazine, etc.

[0058] As used herein, the terms “heterocycle”, “heterocyclic” or “heterocyclo” refer to fully saturated or partially unsaturated cyclic groups, for example, 3- to 8-memLered monocyclic, 7- to 12-memLered bicyclic, or 10- to 15-memLered spirocyclic or tricyclic ring systems, which have at least one heteroatom (selected from the group consisting of N, O, and S) in at least one ring, wherein 0, 1, 2 or 3 atoms of each ring may be substituted by a substituent. Each ring of the heterocyclic group containing a heteroatom may have 1, 2, 3 or 4 heteroatoms selected from nitrogen atoms, oxygen atoms and / or sulfur atoms, where the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatoms may optionally be quaternized. The heterocyclic group may be attached at any heteroatom or carbon atom of the ring or ring system. A heterocyclic group is optionally substituted. Examples of heterocyclic groups include, but not limited to, epoxy, azetidinyl, aziridinyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, pyrrolidinонyl, piperidinyl, piperazinyl, imidazolidinyl, imidazopyridinyl, thiazolidinyl, dithianyl, trithianyl, dioxolanyl, oxazolidinyl, oxazolidinonyl, decahydroquinolinyl, piperidonyl, 4-piperidinonyl, quinuclidinyl, thiomorpholinyl, thiomorpholinyl 1,1 dioxide, morpholinyl, azepanyl, oxazepanyl, azabicyclohexanyls, azabicycloheptanyl, azabicyclooctanyls, azabicyclononanyls (eg., octahydroindolizinyl), azaspiroheptanyls, di hydro- 1 H,3H,5H-oxazolo[3,4-c]oxazolyl, tetrahydro-1'H,3'H-spiro[cyclopropane-1,2'-pyrrolizine], hexahydro-1H-pyrrolizinyl, hexahydro-1H-pyrrolo[2,1-c][1,4]oxazinyl, octahydroindolizinyl, oxaazaspirononanyls, oxaazaspirooctanyls, diazaspirononanyls, oxaazabicycloheptanyls, hexahydropyrrolizinyl 4(lH)-oxide, and tetrahydro- 2H-thiopyranyl 1 -oxi de and tetrahydro-2H-thiopyranyl 1,1-dioxide.Detailed Description of the Invention

[0059] The invention is based in part on the discovery of novel urea derivatives of tetracyclic compounds as PI3Ka inhibitors. These compounds are shown herein to selectively target, bind to, inhibit and / or modulate the activity of PI3Ka. The compounds are orally available and can be used for treating various diseases and disorders associated with or related to PI3Ka activities, such as various types of cancer.Patent Application Atty. Docket No. ENTX-038PCT

[0060] In one aspect, the invention generally relates to a compound having the structural formula (I):or a pharmaceutically acceptable form or an isotope derivative thereof,whereinX1is N or CR1;X2is N, C(O) or CR2;X3is N or CR3;each of R1, R2and R3is independently H, D, halo, CN, R, OR, C(O)NHR, C2-6 alkynyl, NRR', SO2R or SO2NR, wherein said R and C2-6 alkynyl are optionally substituted with one or more of Ra;R4is a 5- to 10-memLered (e.g., 5-, 6-, 7-, 8-, 9-, 10-memLered) monocyclic, bicyclic or bridged aryl or heteroaryl ring with 0-4 (e.g., 0, 1, 2, 3 or 4) ring heteroatoms independently selected from N, O and S, optionally substituted with one or more Rc;R5is C(O)R5'or R5", whereinR5'is a 5- to 10-memLered (e.g., 5-, 6-, 7-, 8-, 9-, 10-memLered) monocyclic, bicyclic or bridged carbocyclyl, heterocyclic, aryl or heteroaryl ring with 0-4 (e.g., 0, 1, 2, 3 or 4) ring heteroatoms independently selected from N, O and S, optionally substituted with one or more Rb; andR5" is a 9- or 10-memLered bicyclic or bridged heteroaryl ring with 0-4 ring heteroatoms independently selected from N, O and S, optionally substituted with one or more Rb;R6is a C1-6 alkyl substituted with0-8 Rd;an unsubstituted or substituted C3-6 carbocyclic or heterocyclic group; orPatent Application Atty. Docket No. ENTX-038PCT an unsubstituted or substituted 5- or 6-memLered aryl or heteroaryl group; each Rais independently D, halo, OH, CN, Ci-6 alkoxy, NRR' or SO2R;each of Rband Rcis independentlydeuterium, oxo, halogen, CN, NO2, OR, SR, NRR’, S(O)2R, S(O)2NRR’, S(O)R, S(O)NRR’, S(O)(NR)R, C(O)R, C(O)OR, C(O)NRR’, C(O)N(R)OR, OC(O)R, OC(O)NRR’, N(R)C(O)OR, N(R)C(O)R, N(R)C(O)NRR’, N(R)C(NR)NRR’, N(R)S(O)2NRR’ or N(R)S(O)2R; ora substituted or unsubstituted group selected from C1-6 alkyl or 4- to 6-memLered (e.g, 4-, 5- or 6-memLered) carbocyclic ring;each Rdis independently D, halogen, CN, OR, alkynyl, SR or NRR’;each of R and R’ is independently selected from H, unsubstituted or substituted C1-4 alkyl, or unsubstituted or substituted 3- to 6-memLered (e.g., 3-, 4-, 5- or 6-memLered) carbocyclic ring, or where R and R’ are attached to the same C or N atom, together form an unsubstituted or substituted 4- to 6-memLered (e.g., 4-, 5- or 6-memLered) heterocyclic ring.

[0061] In certain emLodiments of (I), X1is N, having the structural formula (IA):

[0062] In certain emLodiments of (I), wherein X1is CR1, having the structural formula (IB):Patent Application Atty. Docket No. ENTX-038PCT

[0063] In certain emLodiments of (I)-(IB), Ring A is an aromatic ring.

[0064] In certain emLodiments of (I)-(IB), X2is N and X3is CR3.

[0065] In certain emLodiments of (I)-(IB), X2is CR2and X3is N.

[0066] In certain emLodiments of (I)-(IB), X2is CR2and X3is CR3.

[0067] In certain emLodiments of (I)-(IB), X2is C(O) and X3is N.

[0068] In certain emLodiments of (I)-(IB), R4is a 6-memLered aryl or heteroaryl ring with 0- 3 (e.g., 0, 1, 2 or 3) ringN atoms, substituted with 0-6 Rcs,

[0069] In certain emLodiments of (I)-(IB), R4is a substituted or unsubstituted phenyl, pyridyl, pyridazinyl or pyrazinyl.

[0070] In certain emLodiments of (I)-(IB), R4is a substituted or unsubstituted phenyl.

[0071] In certain emLodiments of (T)-(IB), R4is a substituted or unsubstituted pyridyl.

[0072] In certain emLodiments of (I)-(IB), R4is a substituted or unsubstituted pyridazinyl.

[0073] In certain emLodiments of (I)-(IB), R4is a substituted or unsubstituted pyrazinyl.

[0074] In certain emLodiments of (I)-(IB), R4is selected from:i iPatent Application Atty. Docket No. ENTX-038PCTF2HC

[0075] In certain emLodiments of (I)-(IB), R4is:

[0076] In certain emLodiments of (I)-(IB), the compound has the following chirality:I R4

[0077] In certain emLodiments of (I)-(IB), the compound has the following chirality:I R4

[0078] In certain emLodiments of (I)-(IB), R5is a substituted or unsubstituted 5- or 6-memLered monocyclic aryl or heteroaryl ring.

[0079] In certain emLodiments of (I)-(IB), R5is a substituted or unsubstituted 8- to 10-memLered (e.g., 8-, 9- or 10-) bicyclic carbocyclyl or heterocyclic.

[0080] In certain emLodiments of (I)-(IB), R?is a substituted or unsubstituted 8- to 10-memLered (e.g., 8-, 9- or 10-) bicyclic aryl or heteroaryl ring.

[0081] In certain emLodiments of (I)-(IB), R5is selected from:s(Rb)jPatent Application Atty. Docket No. ENTX-038PCTwherein j is 0, 1, 2, 3 or 4.

[0082] In certain emLodiments of (I)-(IB), R5is selected from:

[0083] In certain emLodiments of (I)-(IB), R5isPatent Application Atty. Docket No. ENTX-038PCT

[0084] In certain emLodiments of (I)-(IB), R5is(II)whereinY is N or CRY;RYis H or Ci-6 alkyl; andi is 0, 1, 2, 3 or 4.

[0085] In certain emLodiments of (I)-(IB), R5is:

[0086] In certain emLodiments of (II)-(IIA), Y is N.

[0087] In certain emLodiments of (II)-(IIA), Y is CH.

[0088] In certain emLodiments of (I)-(IB), R6is CR6AR6BR6C, having the structural formula (IIIA)Patent Application Atty. Docket No. ENTX-038PCT R5(IIIA)whereinR6Ais independently H, D or halogen;R6Bis H, D or halogen; andR6CisH, D, halogen, CN, OR, SR, NRR’, S(O)2R, S(O)2NRR\ S(O)R, S(O)NRR’, S(O)(NR)R or C(() NRR: oran unsubstituted or substituted C1-5 alkyl, C2-5 alkenyl, C2.5 alkynyl, C3-6 carbocyclic or heterocyclic group, or 5- or 6-memLered aryl or heteroaryl group.

[0089] In certain emLodiments of (IIIA), R6Cis CR6E)R6ER6F,whereinR6Dis H, D or halogen; andeach of R6Eand R6Fis independentlyH, D, halogen, CN, OR, SR, NRR’, S(O)2R, VOiARR. S(O)R, S(O)NRR' S(O)(NR)R or C(O)NRR’; oran unsubstituted or substituted C1-5 alkyl, C2-5 alkenyl, C2-5 alkynyl, C3-6 carbocyclic or heterocyclic group, or 5- or 6-memLered aryl or heteroaryl group.

[0090] In certain emLodiments of (TTIA), each of R6Aand R6Bis H.

[0091] In certain emLodiments of (IIIA), each of R6Aand R6Bis D.

[0092] In certain emLodiments of (IIIA), at least one of R6D, R6Eand R6Fis a halogen atom.

[0093] In certain emLodiments of (IIIA), one of R6D, R6Eand R6Fis F.

[0094] In certain emLodiments of (IIIA), two of R6D, R6Fand R6Fis F.

[0095] In certain emLodiments of (IIIA), each of R6D, R6Eand R6Fis F.

[0096] In certain emLodiments of (IIIA), at least one of R6D, R6Eand R6Fis a D.Patent Application Atty. Docket No. ENTX-038PCT

[0097] In certain emLodiments of (TTIA), one of R6D, R6Eand R6Fis D.

[0098] In certain emLodiments of (IIIA), two of R6D, R6Eand R6Fis D.

[0099] In certain emLodiments of (IIIA), each of R6D, R6Eand R6Fis D.

[0100] In certain emLodiments of (I)-(IB) and (IIIA), R6are independently selected from: CH3, CH2D, CHD2, CD3, CH2CH3, CD2CH3, CH2CD3, CD2CD3, CH2CH2F, CD2CH2F, CH2CD2F, CD2CD2F, CH2CHF2, CD2CHF2, CH2CDF2, CD2CDF2, CH2CF3, CD2CF3, CH2CN, CH2CH2N(CH3)2, CH2CH2OH, CH2CH2OCH3, CH2CH2CH2F, CH2CH2CHF2, CH2CH2CF3, propyl, isopropyl, n-butyl, isobutyl,

[0101] In certain emLodiments of (I), the compound has the structural formula (IVA):

[0102] In certain emLodiments of (I), the compound has the structural formula (IVB):Patent Application Atty. Docket No. ENTX-038PCT

[0103] In certain emLodiments of (I), the compound has the structural formula (VA):

[0104] In certain emLodiments of (I), the compound has the structural formula (VB):Patent Application Atty. Docket No. ENTX-038PCT

[0105] In certain emLodiments of (I), the compound has the structural formula (VC):

[0106] In certain emLodiments of (I), the compound has the structural formula (VD):Patent Application Atty. Docket No. ENTX-038PCT

[0107] In certain emLodiments of (I), the compound has R6is CH2R6or CD2R6, wherein R6' is a substituted or unsubstituted C1.2 alkyl.

[0108] In certain emLodiments, R6is CH3, CD3, CH2F, CHF2 or CF3.

[0109] In certain emLodiments of (I), the compound has the structural formula (VIA):(VIA)

[0110] In certain emLodiments of (I), the compound has the structural formula (VIB):Patent Application Atty. Docket No. ENTX-038PCT(VI“)

[0111] In certain emLodiments of (I), the compound has the structural formula (VIC):

[0112] In certain emLodiments of (I), the compound has the structural formula (VID):Patent Application Atty. Docket No. ENTX-038PCT(VID)

[0113] In certain emLodiments of (I), the compound has the structural formula (VIE):

[0114] In certain emLodiments of (I), the compound has the structural formula (VIF):Patent Application Atty. Docket No. ENTX-038PCT

[0115] In certain emLodiments of (I), the compound has the structural formula (VIG):(VIG)

[0116] In certain emLodiments of (I), the compound has structural formula (VIH):Patent Application Atty. Docket No. ENTX-038PCT

[0117] In certain emLodiments of (I), the compound has the structural formula (VIIA):(VIIA)

[0118] In certain emLodiments of (I), the compound has the structural formula (VIIB):Patent Application Atty. Docket No. ENTX-038PCT (VIIB)

[0119] In certain emLodiments of (I), the compound has the structural formula (VIIC):(VIIC)

[0120] In certain emLodiments of (I), the compound has the structural formula (VIID):(VIID)

[0121] In certain emLodiments of (I), the compound has the structural formula (VIIE):Patent Application Atty. Docket No. ENTX-038PCT

[0122] In certain emLodiments of (I), the compound has the structural formula (VIIF):

[0123] In certain emLodiments of (I), the compound has the structural formula (VIIG):Patent Application Atty. Docket No. ENTX-038PCT (VIIG)

[0124] In certain emLodiments of (I), the compound has the structural formula (VIIH):(VIIH)

[0125] Non-limiting examples of compounds of the invention can be found in Table 1, including pharmaceutically acceptable forms or isotope derivatives thereof.

[0126] In certain emLodiments, the compound of the invention has one or more deuterium atoms in place of hydrogen.

[0127] In another aspect, the invention generally relates to a pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable excipient, carrier, or diluent.

[0128] In certain emLodiments, the pharmaceutical composition of claim 68, being suitable for oral administration.

[0129] In yet another aspect, the invention generally relates to a unit dosage form comprising a pharmaceutical composition comprising a compound disclosed herein.

[0130] In certain emLodiments, the unit dosage form is the form of a tablet or capsule.

[0131] Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol,Patent Application Atty. Docket No. ENTX-038PCT sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[0132] The pharmaceutical compositions of the invention include those suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration. In certain emLodiments, the compound of the formulae herein is administered transdermally (e.g., using a transdermal patch). Other formulations may conveniently be presented in unit dosage form, e.g., tablets and sustained release capsules, and in liposomes, and may be prepared by any methods well known in the art of pharmacy. See, for example, Remington’s Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, PA (17th ed. 1985).

[0133] Such preparative methods include the step of bringing into association with the molecule to be administered ingredients such as the carrier that constitutes one or more accessory ingredients. In general, the compositions are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers, liposomes or finely divided solid carriers or both, and then if necessary shaping the product.

[0134] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the compounds described herein or derivatives thereof are admixed with at least one inert customary excipient (or carrier) such as sodium citrate or dicalcium phosphate or (i) fillers or extenders, as for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid, (ii) binders, as for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia, (iii) humectants, as for example, glycerol, (iv) disintegrating agents, as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate, (v) solution retarders, as for example, paraffin, (vi) absorption accelerators, as for example, quaternary ammonium compounds, (vii) wetting agents, as for example, cetyl alcohol, and glycerol monostearate, (viii) adsorbents, as for example, kaolin and bentonite, and (ix) lubricants, as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethyleneglycols, and the like. Solid dosage forms such as tablets, dragees,Patent Application Atty. Docket No. ENTX-038PCT capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and others known in the art.

[0135] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents, and emulsifiers, such as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, com germ oil, olive oil, castor oil, sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethyleneglycols, and fatty acid esters of sorbitan, or mixtures of these substances, and the like. Besides such inert diluents, the composition can also include additional agents, such as wetting, emulsifying, suspending, sweetening, flavoring, or perfuming agents.

[0136] In yet another aspect, the invention generally relates to a method for inhibiting cell proliferation in vitro or in vivo, comprising contacting a cell with an effective amount of a compound disclosed herein.

[0137] In yet another aspect, the invention generally relates to a method for modulating PI3Ka activity in a cell, comprising contacting the cell with a compound disclosed herein.

[0138] In yet another aspect, the invention generally relates to a method for reducing PI3Ka activity in a cell, comprising contacting the cell with a compound disclosed herein.

[0139] In yet another aspect, the invention generally relates to a method for treating a disease or disorder mediated by PI3Ka, comprising administering to a subject in need thereof a therapeutically effective amount of a compound disclosed herein.

[0140] In certain emLodiments, the disease or disorder is a cellular proliferative disease.

[0141] In yet another aspect, the invention generally relates to a method for treating or reducing cancer, or a related disease or disorder, comprising administering to a subject in need thereof a therapeutically effective amount of a compound disclosed herein.

[0142] In certain emLodiments, the cancer is selected from the group consisting of carcinoma, squamous carcinoma, adenocarcinoma, sarcoma, leukemia, neuroma, melanoma, and lymphoma.

[0143] Examples of cancers targeted in the present invention include, but are not particularly limited to, head and neck cancer, digestive organ cancer (esophageal cancer, stomach cancer.Patent Application Atty. Docket No. ENTX-038PCT duodenal cancer, liver cancer, biliary cancer (e.g., gallbladder and bile duct cancer), pancreatic cancer, colorectal cancer (e.g., colon cancer, and rectal cancer), etc.), lung cancer (e.g., non¬ small-cell lung cancer, small-cell lung cancer, and mesothelioma), breast cancer, genital cancer (ovarian cancer, uterine cancer (e.g., cervical cancer and endometrial cancer), etc.), urological cancer (e.g., kidney cancer, bladder cancer, prostate cancer, and testicular tumor), hematopoietic tumor (e.g., leukemia, lymphoma, malignant lymphoma, and multiple myeloma), sarcoma (e.g., osteosarcoma, and soft-tissue sarcoma), skin cancer, brain tumor, a carcinoma, squamous carcinoma, adenocarcinoma, neuroma, melanoma and the like. Examples include lung cancer, pancreatic cancer, rectal cancer, colon cancer colorectal cancer and uterine cancer. In certain emLodiments, squamous carcinoma is a cancer of uterine cervix, tarsus, conjunctiva, vagina, lung, oral cavity, skin, bladder, tongue, larynx or esophagus In one emLodiment, adenocarcinoma is a cancer of prostate, small intestine, endometrium, uterine cervix, large intestine, lung, pancreas, esophagus, rectum, uterus, stomach, breast or ovary. In certain emLodiments, tumor is rectal cancer, colon cancer, colorectal cancer, pancreatic cancer, lung cancer, breast cancer leukemia or uterine cancer.

[0144] In certain emLodiments, the cancer is selected from the group consisting of ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colorectal cancer, small and non-small cell lung cancer, endometrial cancer, appendix cancer, cholangiocarcinoma, bladder urothelial cancer, gastric carcinomas, bile duct cancer, hepatocellular carcinoma, thyroid carcinoma, and a hematologic malignancy.

[0145] In certain emLodiments, the cancer is selected from the group consisting of acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), and glioblastomas.

[0146] In certain emLodiments, the subject has a mutated class IA PI3K p110α.

[0147] In certain emLodiments, the subject has at least one of the following PI3Kα mutations: H1047R, E542K, E545K and H1047X.

[0148] In certain emLodiments, the subject does not have a PI3Ka mutant protein.

[0149] In certain emLodiments, the subject being treated is further administered one or more of chemotherapy, radiotherapy, targeted therapy, immunotherapy, and hormonal therapy.

[0150] In yet another aspect, the invention generally relates to use of a compound disclosed herein, and a pharmaceutically acceptable excipient, carrier, or diluent, in preparation of a medicament for treating a disease or disorder.Patent Application Atty. Docket No. ENTX-038PCT

[0151] In yet another aspect, the invention generally relates to use of a compound disclosed herein for treating a disease or disorder.

[0152] The amount of the active compound administered will be dependent on the subject being treated, the severity of the disorder or condition, the route of administration, the disposition of the compound and the discretion of the prescribing physician. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be used without causing any harmful side effect, with such larger doses typically divided into several smaller doses for administration throughout the day.

[0153] Any appropriate route of administration can be employed, for example, oral, intramuscular, intravenous, transdermal, subcutaneous, sublingual, parenteral, nasal, pulmonary, inhalational, buccal, intraperintoneal, rectal, intrapleural, and intrathecal administration. Most suitable means of administration for a particular patient will depend on the nature and severity of the disease or condition being treated or the nature of the therapy being used and on the nature of the active compound.

[0154] In certain preferred emLodiments, the compound is administered orally.Pharmaceutical compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, sachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion, or packed in liposomes and as a bolus, etc. Soft gelatin capsules can be useful for containing such suspensions, which may beneficially increase the rate of compound absorption.

[0155] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets optionally may be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein. Methods of formulating such slow or controlled release compositions of pharmaceutically active ingredients, such as those herein and other compounds known in the art,Patent Application Atty. Docket No. ENTX-038PCT are known in the art and described in several issued US Patents, some of which include, but are not limited to, US Patent Nos. 4,369,172; and 4,842,866, and references cited therein. Coatings can be used for delivery of compounds to the intestine (see, e.g., U. S. Patent Nos. 6,638,534, 5,217,720, and 6,569,457, 6,461,631, 6,528,080, 6,800,663, and references cited therein). A useful formulation for the compounds of this invention is the form of enteric pellets of which the enteric layer comprises hydroxypropylmethylcellulose acetate succinate.

[0156] In the case of tablets for oral use, carriers that are commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are administered orally, the active ingredient is comLined with emulsifying and suspending agents. If desired, certain sweetening and / or flavoring and / or coloring agents may be added.

[0157] Compositions suitable for topical administration include lozenges comprising the ingredients in a flavored basis, usually sucrose and acacia or tragacanth; and pastilles comprising the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and acacia.

[0158] Compositions suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example, sealed ampules and vials, and may be stored in a freeze dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.

[0159] Such injection solutions may be in the form, for example, of a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1,3 -butanediol. Among the acceptable vehicles and solvents that may be employed are mannitol, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile,Patent Application Atty. Docket No. ENTX-038PCT fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant.

[0160] Compounds of the present invention may also be administered in the form of liposomes. As is known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by mono- or multi -lamellar hydrated liquid crystals that are dispersed in an aqueous medium. Any non-toxic, physiologically acceptable and metabolizable lipid capable of forming liposomes can be used. The present compositions in liposome form can contain, in addition to a compound of the present invention, stabilizers, preservatives, excipients, and the like. The preferred lipids are the phospholipids and the phosphatidyl cholines (lecithins), both natural and synthetic. Methods to form liposomes are known in the art. See, for example, Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, N. Y. (1976), p. 33 et seq.

[0161] The pharmaceutical compositions of this invention may be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components. Such materials include, but are not limited to, cocoa butter, beeswax and polyethylene glycols.

[0162] The pharmaceutical compositions of this invention may be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and / or other solubilizing or dispersing agents known in the art.

[0163] Topical administration of the pharmaceutical compositions of this invention is especially useful when the desired treatment involves areas or organs readily accessible by topical application. For application topically to the skin, the pharmaceutical composition should be formulated with a suitable ointment containing the active components suspended or dissolvedPatent Application Atty. Docket No. ENTX-038PCT in a carrier. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical composition can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. The pharmaceutical compositions of this invention may also be topically applied to the lower intestinal tract by rectal suppository formulation or in a suitable enema formulation. Topically-transdermal patches and iontophoretic administration are also included in this invention.

[0164] Methods of treatment disclosed herein may be employed in comLination with or in addition to other therapies. In certain emLodiments, the subject being treated is further administered one or more of chemotherapy, radiotherapy, targeted therapy, immunotherapy, and hormonal therapy.

[0165] Exemplary additional therapeutically active agents include, but are not limited to, small organic molecules such as drug compounds, e.g., compounds approved by the U. S. Food and Drug Administration (FDA) as provided in the Code of Federal Regulations (CFR), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins and cells.

[0166] In certain emLodiments, a compound of the invention may be administered in comLination with endocrine therapy, e.g., agents such as letrozole, fulvestrant, tamoxifen, exemestane, or anastrozole.

[0167] In some emLodiments, a compound of the invention may be administered in comLination with a chemotherapeutic agent, e.g., docetaxel, paclitaxel, cisplatin, carboplatin, capecitabine, gemcitabine or vinorelbine. In other emLodiments, a compound of the invention may be administered in comLination with an anti-HER2 agent, e.g., trastuzumab or pertuzumab.

[0168] In certain emLodiments, the method disclosed herein is in comLination with one or more of immune check point blockade, co-signaling of T cells, and tumor targeting antibody therapies.Patent Application Atty. Docket No. ENTX-038PCT

[0169] In certain emLodiments, the method further comprises administering a chemotherapeutic agent to the subject.

[0170] In certain emLodiments, the method further comprises administering a radiotherapy to the subject. In certain emLodiments, the method further comprises administering a targeted therapy to the subject. In certain emLodiments, the method further comprises administering an immunotherapy to the subject. In certain emLodiments, the method further comprises administering hormonal therapy to the subject.

[0171] As used herein, the term "chemotherapeutic agent" refers to a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include Erlotinib (TARCEVA®, Genentech / OSI Pharm.), Bortezomib (VELCADE®, Millennium Pharm.), Fulvestrant (FASLODEX®, AstraZeneca), Sutent (SU11248, Pfizer), Letrozole (FEMARA®, Novartis), Imatinib mesylate (GLEEVEC®, Novartis), PTK787 / ZK 222584 (Novartis), Oxaliplatin (Eloxatin®, Sanofi), 5-FU (5 -fluorouracil), Leucovorin, Rapamycin (Sirolimus, RAPAMUNE®, Wyeth), Lapatinib (TYKERB®, GSK572016, Glaxo Smith Kline), Lonafarnib (SCH 66336), Sorafenib (BAY43-9006, Bayer Labs), and Gefitinib (IRESSA®, AstraZeneca), AG1478, AG1571 (SU 5271; Sugen), alkylating agents such as thiotepa and CYTOXAN® cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analog topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogs); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogs, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chloramLucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novemLichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gammall and calicheamicin omegall (Angew Chem. Inti. Ed. Engl. (1994) 33: 183-186); dynemicin, including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatinPatent Application Atty. Docket No. ENTX-038PCT chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6- diazo-5-oxo-L-norleucine, ADRIAMYCIN® (doxorubicin), morpholino-doxorubicin, cyanomorpholinodoxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esonibicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5 -fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6- mercaptopurine, thiamniprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (" Ara-C"); cyclophosphamide; thiotepa; taxoids, e.g., TAXOL® (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, N. J.), ABRAXANE® (Cremophor-free), albumin-engineered nanoparticle formulations of paclitaxel (American Pharmaceutical Partners, SchaumLerg, 111.), and TAXOTERE® (doxetaxel; Rhone-Poulenc Rorer, Antony, France); chloranmLucil; GEMZAR® (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; etoposide (VP- 16); ifosfamide; mitoxantrone; vincristine;NAVELBINE® (vinorelbine); novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine (XELODA®); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000;Patent Application Atty. Docket No. ENTX-038PCT difluoromethylomithine (DMFO); retinoids such as retinoic acid; and pharmaceutically acceptable salts, acids and derivatives of any of the above.

[0172] Examples of the second (or further) agent or therapy may include, but are not limited to, immunotherapies (e.g. PD-1 inhibitors (pemLrolizumab, nivolumab, cemiplimab), PD-L1 inhibitors (atezolizumab, avelumab, durvalumab), CTLA4 antagonist, cell signal transduction inhibitors (e.g., imatinib, gefitinib, bortezomib, erlotinib, sorafenib, sunitinib, dasatinib, vorinostat, lapatinib, temsirolimus, nilotinib, everolimus, pazopanib, trastuzumab, bevacizumab, cetuximab, ranibizumab, pegaptanib, panitumumab and the like), mitosis inhibitors (e.g., paclitaxel, vincristine, vinblastine and the like), alkylating agents (e.g., cisplatin, cyclophosphamide, chromabucil, carmustine and the like), anti-metabolites (e.g., methotrexate, 5-FU and the like), intercalating anticancer agents, (e.g., actinomycin, anthracycline, bleomycin, mitomycin-C and the like), topoisomerase inhibitors (e.g., irinotecan, topotecan, teniposide and the like), immunotherapic agents (e.g., interleukin, interferon and the like) and antihormonal agents (e.g., tamoxifen, raloxifene and the like).

[0173] Certain compounds of the present invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention.Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.

[0174] Isomeric mixtures containing any of a variety of isomer ratios may be utilized in accordance with the present invention. For example, where only two isomers are comLined, mixtures containing 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0 isomer ratios are contemplated by the present invention. Those of ordinary skill in the art will readily appreciate that analogous ratios are contemplated for more complex isomer mixtures.

[0175] If, for instance, a particular enantiomer of a compound of the present invention is desired, it may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers. Alternatively, where the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of thePatent Application Atty. Docket No. ENTX-038PCT diastereomers thus formed by fractional crystallization or chromatographic methods well known in the art, and subsequent recovery of the pure enantiomers.

[0176] Isotopically-labeled compounds are also within the scope of the present disclosure. As used herein, an "isotopically-labeled compound" refers to a presently disclosed compound including pharmaceutical salts and prodrugs thereof, each as described herein, in which one or more atoms are replaced by an atom having an atomic mass or mass numLer different from the atomic mass or mass numLer usually found in nature. Examples of isotopes that can be incorporated into compounds presently disclosed include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and chlorine, such as2H,3H,13C,14C,15N,18O,17O,31P,32P,33S,18F, and36C1, respectively.

[0177] By isotopically-labeling the presently disclosed compounds, the compounds may be useful in drug and / or substrate tissue distribution assays. Tritiated (3H) and carbon-14 (14C) labeled compounds are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (2H) can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labeled compounds presently disclosed, including pharmaceutical salts, esters, and prodrugs thereof, can be prepared by any means known in the art.

[0178] Further, substitution of normally abundant hydrogen (1H) with heavier isotopes such as deuterium can afford certain therapeutic advantages, e.g., resulting from improved absorption, distribution, metabolism and / or excretion (ADME) properties, creating drugs with improved efficacy, safety, and / or tolerability. Benefits may also be obtained from replacement of normally abundant12C with13C. (See, WO 2007 / 005643, WO 2007 / 005644, WO 2007 / 016361, and WO 2007 / 016431.)

[0179] Stereoisomers (e.g., cis and trans isomers) and all optical isomers of a presently disclosed compound (e.g., R and S enantiomers), as well as racemic, diastereomeric and other mixtures of such isomers are within the scope of the present disclosure.

[0180] Compounds of the present invention are, subsequent to their preparation, preferably isolated and purified to obtain a composition containing an amount by weight equal to or greater than 95% (“substantially pure”), which is then used or formulated as described herein. In certain emLodiments, the compounds of the present invention are more than 99% pure.Patent Application Atty. Docket No. ENTX-038PCT

[0181] Solvates and polymorphs of the compounds of the invention are also contemplated herein. Solvates of the compounds of the present invention include, for example, hydrates.

[0182] Any appropriate route of administration can be employed, for example, parenteral, intravenous, subcutaneous, intramuscular, intraventricular, intracorporeal, intraperitoneal, rectal, or oral administration. Most suitable means of administration for a particular patient will depend on the nature and severity of the disease or condition being treated or the nature of the therapy being used and on the nature of the active compound.

[0183] Compositions for parenteral injection comprise pharmaceutically-acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use.Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate. Proper fluidity may be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0184] These compositions can also contain adjuvants such as preservative, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paragen, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption, such as aluminum monostearate and gelatin.

[0185] Compounds of the present invention may also be administered in the form of liposomes. As is known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by mono- or multi-lamellar hydrated liquid crystals that are dispersed in an aqueous medium. Any non-toxic, physiologically-acceptable and metabolizable lipid capable of forming liposomes can be used. The present compositions in liposome form can contain, in addition to a compound of the present invention, stabilizers, preservatives, excipients, and the like. The preferred lipids are the phospholipids and the phosphatidyl cholines (lecithins), both natural and synthetic. Methods to form liposomes arePatent Application Atty. Docket No. ENTX-038PCT known in the art. See, for example, Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, N. Y. (1976), p. 33 et seq.

[0186] Total daily dose of the compositions of the invention to be administered to a human or other mammal host in single or divided doses may be in amounts, for example, from 0.0001 to 300 mg / kg body weight daily and more usually 1 to 300 mg / kg body weight. The dose, from 0.0001 to 300 mg / kg body, may be given twice a day.

[0187] Materials, compositions, and components disclosed herein can be used for, can be used in conjunction with, can be used in preparation for, or are products of the disclosed methods and compositions. It is understood that when comLinations, subsets, interactions, groups, etc. of these materials are disclosed that while specific reference of each various individual and collective comLinations and permutations of these compounds may not be explicitly disclosed, each is specifically contemplated and described herein. For example, if a method is disclosed and discussed and a numLer of modifications that can be made to a numLer of molecules including in the method are discussed, each and every comLination and permutation of the method, and the modifications that are possible are specifically contemplated unless specifically indicated to the contrary. Likewise, any subset or comLination of these is also specifically contemplated and disclosed. This concept applies to all aspects of this disclosure including, but not limited to, steps in methods using the disclosed compositions. Thus, if there are a variety of additional steps that can be performed, it is understood that each of these additional steps can be performed with any specific method steps or comLination of method steps of the disclosed methods, and that each such comLination or subset of comLinations is specifically contemplated and should be considered disclosed.Examples

[0188] The following examples are given for the purpose of illustrating the invention, but not for limiting the scope or spirit of the invention.

[0189] Compounds of the invention, including those specifically disclosed herein above and herein below, may be prepared as described in the following schemes. Although the present invention has been described in detail with preferred emLodiments, those of ordinary skill in the art should understand that modifications, variations, and equivalent replacements made to the present invention within the scope of the present invention belong to the protection of the present invention.Patent Application Atty. Docket No. ENTX-038PCT List of Abbreviationsaq: aqueousAc = AcetylAcO = AcetateAc2O = Acetic anhydrideAIBN = a,a'-AzoisobyronitrileAll = AllylAlloc = AllyloxycarbonylAm = Amyl (Pentyl)Ar = ArylB2Pin2 = bis(pinacolato)diboron9-BBN = 9-BorabicyclononaneBHT = tert-ButylhydroxytolueneBINAP = 2,2'-Bis(diphenylphosphino)-1,1'-binaphthylBMS = Borane-methylsulphide complexBn = BenzylBoc = tert-Butoxy carbonylBOP = Bis(2-oxo-3-oxazolidinyl)phosphineBu or n-Bu = n-Butyls-Bu or sBu = sec-Butylt-Bu or tBu = tert-ButylBuOH = ButanolBz = BenzoylBzl = BenzylCAN = Ceric ammonium nitratecataCXium A Pd G3 = mesylate [(di(l-adamantyl)-n-butylphosphine)-2-(2’-amino- l,l’biphenyl)]palladium(II)CBS = Corey-Bashki-ShibatCbz = BenzyloxycarbonylCbzCl = Benzyl chloroformate oxCod = CyclooctadienePatent Application Atty. Docket No. ENTX-038PCT Cp = CyclopentadienylCSA = Camphorsulphonic acidDABCO = 1,4-Diazabicyclo[2.2.2]octane, TriethylendiamineDAST = Diethylaminosulphur trifluoridedba = DibenzylideneacetoneDBU = 1,8-Diazabyciclo[5.4.0]undec-7-eneDCC = 1,3-DicyclohexylcarbodiimideDCM = DichloromethaneDDQ = 2,3-Dichloro-5,6-dicyano-1,4-benzoquinoneDEAD = Diethyl azodicarboxylateDHP = DihydropiranDHQD = DihydroquinidineDIBAL = Diisobutylaluminium hydrideDIBAL-H = Diisobutylaluminium hydrideDIC = DiisopropylcarbodiimideDIPEA = DiisopropylethylamineDMA = N, N-DimethylacetamideDMAC = N, N-DimethylacetamideDMAP = 4-DimethylaminopyridineDME = 1,2-DimethoxyethaneDMF = N, N-DimethylformamideDMP = Dess-Martin periodinaneDMPU = 1,3-Dimethyl-3,4,5,6-tetrahydro-2(1H)-pirimidoneDMS = DimethylsulphideDMSO = DimethylsulphoxideDPA = DiisopropylamineDPPA = Diphenylphosphoryl azideDdpb = 1,4-bis(diphenylphosphino)butaneDppe = 1,2-bis(diphenylphosphino)ethaneDppf = 1,2-bis(diphenylphosphino)ferrocenedppp = 1,3-bis(diphenylphosphino)propanePatent Application Atty. Docket No. ENTX-038PCT Dtbbpy = 4, 4’-di-tert-butyl-2,2’-dipyridylEA = Ethyl acetateEDC = 1-Ethyl-3-(3-dimethylaminopropy)carbodiimideEDCI = 1-Ethyl-3-(3-dimethylaminopropy)carbodiimide hydrochlorideEq = equivalentESI or ES = Electrospray ionizationEt = ethylEt2O = Diethyl etherEtOAc = Ethyl acetateFMOC = 9-FluorenylmethoxycarbonylFCC = Flash column chromatographyHATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphateHMDS = HexamethyldisilazaneHMPA = HexamethylphosphoramideHOAt = 7-Aza-l-hydroxybenzotriazoleHOBt= 1 -HydroxybenzotriazoleHPLC = high pressure liquid chromatographyIPA = Isopropyl alcoholIm = ImidazoleKHMDS = Potassium bis(trimethylsilyl)amideKOAc = Potassium acetateLAH = Lithium aluminium hydrideLDA = Lithium diisopropylamideLHMDS = Lithium bis(trimethylsilyl)amideMCPBA = meta-chloroperoxybenzoic acidMe = MethylMeCN = AcetonitrileMeOH = MethanolMOM = MethoxymethylMg = magnesiumPatent Application Atty. Docket No. ENTX-038PCT MS = Molecular sievesMs = MethanesulphonylMTBE = Methyl tert-butyl etherm / z = mass divided by chargeNa2SO4 = Sodium SulphateNaHMDS = Sodium bis(trimethylsilyl)amideNaCNBH3 = Sodium cyanoborohydrideNBS = N-BromosuccinimideNCS = N-ChlorosuccinimideNIS = N-IodosuccinimideNMM = N-MethylmorpholineNMO = N-Methylmorpholine-N-oxideNMP = N-MethylpyrrolidoneNMR = Nuclear magnetic resonanceNs = p-Nitrophenyl sulphonylPd(dppf)Cl2 = [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladiumPd(PPh3)4 = tetrakis(triphenylphosphine)palladiumPDC = Pyridinium dichlorochromatePCC = Pyridinium chlorochromatePE = Petroleum etherPh = PhenylPiv = Pivaloyl, 2,2-dimethylacetylPMB = p-MethoxybenzylPPA = Polyphosphoric acidPPTS = Pyridinium p-toluensulphonaten-Pr = n-PropylPr = Propyli-Pr or iPr = iso-propiloPTC = Phase transfer catalystPTS A = p-Toluenesulphonic acidPv = Pivaloyl, 2,2-dimethylacetylPatent Application Atty. Docket No. ENTX-038PCT Py = PyridineRed-Al® = Sodium bis(2-methoxyethoxy)aluminium hydrideRT = room temperatureSFC = supercritical fluid chromatographyPrep-SFC = Preparative SFCSEM = 2-(Trimethylsilyl)ethoxymethylTBAF = Tetrabutylammonium fluorideTBDMS = tert-ButyldimethylsilylTBDPS = tert-ButyldiphenylsilylTBHP = tert-ButylhydroperoxydeTBS = tert-ButyldimethylsilylTEA = TriethylamineTES = TriethylsilylTf = TrifluoromethanesulfonylTfO = TrifluoromethanesulfonateTf2O = Trifluoromethanesulfonyl anhydrideTfOH = Trifluoromethanesulfonic acidTFA = Trifluoroacetic acidTFAA = Trifluoroacetic anhydrideThexyl = 2,3-dimethyl-2-butylTHF = TetrahydrofuraneTHP = TetrahydropyranylTIPS = Triisopropyl silylTMEDA = N, N, N', N'-TetramethylethylendiamineTMG = TetramethylguanidineTMS = TrimethylsilylTol = p-ToluylTPAP = Tetra-n-propylammonium perruthenateTPS = Tripropyl silylTr = Trityl, triphenylmethylTroc = 2,2,2-TrichloroethoxycarbonylPatent Application Atty. Docket No. ENTX-038PCT Trt = Trityl, triphenylmethylTs = p-Toluenesulphonylp-TsOH = p-Toluenesulphonic acidUV = ultravioletZ = BenzyloxycarbonyTable 1. Exemplary CompoundsPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTClPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTFCl ClPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCT OMe OMePatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTPatent Application Atty. Docket No. ENTX-038PCTExemplary Synthetic ProceduresPatent Application Atty. Docket No. ENTX-038PCT

[0190] General LCMS Method:

[0191] Shimadzu LCMS2020, Reverse-phase column (Shim-Pack Scepter Cl 8, 33 x 3.0 mm, 3um), elution with A: H2O / MeCN / FA = 90 / 10 / 0.05; B: MeCN; Detection: MS, ELS, UV (100 pL split to MS with in-line UV detector); MS ionization method: Electrospray (positive and negative ion). ES-API = electrospray-atmospheric pressure ionization.

[0192] General prep-HPLC Purification Method:Instrument: Shimadzu FRC-40; Shimadzu LH-40; Shimadzu LC-8A; GX-281.Column: YMC-Triart C18, 250*20 mm, 5um; Welch Ultimate XB-C18, 250*21.2 mm, 5um. Detection wavelength: 220, 254 nM.Flow rate: 15ml / min-20ml / min.Run time: 8 min.Column temperature: 25 °C.

[0193] Chiral prep-SFC method 1:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak IH, 150x20mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 15%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle time: 15 minEluted time: 2 H

[0194] Chiral prep-SFC method 2:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak IH, 150x20mm I. D., 5pmMobile phase: A for CO2 and B for MEOHGradient: B 15%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CPatent Application Atty. Docket No. ENTX-038PCT Wavelength: 220 nmCycle time: 15 minEluted time: 2 H

[0195] Chiral prep-SFC method 3:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak IH, 250><20mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 20%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle time: 6 minInjection volume: 3.4 mLNumLer of injection needles: 5Eluted time: 2 H

[0196] Chiral prep-SFC method 4:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak C-IG, 250x30mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 40%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 8minCycle time: 6minInjection volume: 4 mLNumLer of injection needles: 6Eluted time: 2H

[0197] Chiral prep-SFC method 5:Patent Application Atty. Docket No. ENTX-038PCT Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak IA, 250 x 30 mm I. D., 5pmMobile phase: A for CO2 and B for MEOH (0.1% 7mol / L NH3 in MeOH)Gradient: B 30%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle time: lOminInjection volume: 1.5 mLNumLer of injection needles: 8Eluted time: 2H

[0198] Chiral prep-SFC method 6:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak AD, 250><30mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 20%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle time: 10 minEluted time: 2H

[0199] Chiral prep-SFC method 7:Instrument: Waters Thar 80 preparative SFCColumn: (R,R)-WHELK, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CPatent Application Atty. Docket No. ENTX-038PCT Wavelength: 220 nmCycle time: 10 minRun time: 10 minInjection volume: 3 mLNumLer of injection needles: 5Eluted time: 2 H

[0200] Chiral prep-SFC method 8:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak IB, 250x30 mm I. D., 5 gmMobile phase: A for CO2 and B for MeOH + 50% ACNGradient: B 25%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle time: 10 minEluted time: 2H

[0201] Chiral prep-SFC method 9:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IC, 250 x 20 mm I D., 5 pmMobile phase: A for CO2 and B for MeOHGradient: B 20%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 20minCycle time: 20minInjection volume: 2 mLNumLer of injection needles: 5Eluted time: 2HPatent Application Atty. Docket No. ENTX-038PCT

[0202] Chiral prep-SFC method 10:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IH, 250><30mm I. D., 5pmMobile phase: A for CO2 and B for MEOHGradient: B 25%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 8 minCycle time: 8 minInjection volume: 2 mLNumLer of injection needles: 3Eluted time: 2H

[0203] Chiral prep-SFC method 11:Instrument: SHIMADZU PREP SOLUTION SFCColumn: (R, R)- WHELK, 250x30mm I. D., 5pmMobile phase: A for CO2 and B for MEOH: CAN=1: 1Gradient: B 50%Flow rate: 60mL / minBack pressure: 100 barColumn temperature: 35°CW avelength:220nmCycle time:8minInjection volume: 3.4mLNumLer of injection needles: 3Eluted time: IH

[0204] Chiral prep-SFC method 12:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IH, 250x30mm I. D., 5pmMobile phase: A for CO2 and B for MeOHPatent Application Atty. Docket No. ENTX-038PCT Gradient: B 15%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 14 minCycle time: 14 minInjection volume: 2 mLNumLer of injection needles: 5Eluted time: 2H

[0205] Chiral prep-SFC method 13:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak AS, 250><30mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 10 minCycle time: 7 minInjection volume: 4.2 mLNumLer of injection needles: 5Eluted time: 2H

[0206] Chiral prep-SFC method 14:Instrument: Waters Thar 80 preparative SFCColumn: (R,R)-WHELK, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 40 mL / minBack pressure: 100 barPatent Application Atty. Docket No. ENTX-038PCT Column temperature: 35 °CWavelength: 220 nmCycle time: lOminRun time: 10 minInjection volume: 4 mLNumLer of injection needles: 3Eluted time: 3 H

[0207] Chiral prep-SFC method 15:Instrument: SHIMADZU PREP SOLUTION SFCColumn: (R, R)-WHELK, 250x30mm ID., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 55 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle time: 4 minInjection volume: 1.5mLNumLer of injection needles: 5Eluted time: 1 H

[0208] Chiral prep-SFC method 16:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak AS, 250x20mm I. D., 5pmMobile phase: A for CO2 and B for MEOHGradient: B 10%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle time: 10 minEluted time: 2 HPatent Application Atty. Docket No. ENTX-038PCT

[0209] Chiral prep-SFC method 17:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IH, 250><30mm I. D., 5pmMobile phase: A for CO2 and B for MEOHGradient: B 20%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 10 minCycle time: 7 minInjection volume: 2 mLNumLer of injection needles: 2Eluted time: IH

[0210] Chiral prep-SFC method 18:Instrument: Waters Thar 80 preparative SFCColumn: (R, R)-WHELK, 250×20mm I. D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 12 minRun time: 12 minInjection volume: 4.2 mLNumLer of injection needles: 5Eluted time: 2 H

[0211] Chiral prep-SFC method 19:Instrument: SHIMADZU PREP SOLUTION SFCColumn: (R, R)-WHELK, 250x20 mm ID., 5 pmPatent Application Atty. Docket No. ENTX-038PCT Mobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 10 minEluted time: 2 H

[0212] Chiral prep-SFC method 20:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak AS, 250 x 20 mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 30%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 4.5 minInjection volume: 3500 mLNumLer of injection needles: 6Eluted time: 2 H

[0213] Chiral prep-SFC method 21:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralCel OZ, 250×20mm I. D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 40%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 10 minPatent Application Atty. Docket No. ENTX-038PCT Injection volume: 2 mLNumLer of injection needles: 3Eluted time: 1 H

[0214] Chiral prep-SFC method 22:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak AD, 250><30mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 30%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 10minEluted time: 2H

[0215] Chiral prep-SFC method 23:Instrument: Waters Thar 80 preparative SFCColumn: (R, R)-WHELK, 250×20mm I. D., 5μmMobile phase: A for CO2 and B for MeOH (0.1% 7mol / L NH3 in MeOH)Gradient: B 50%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 12 minRun time: 12 minInjection volume: 3.5 mLNumLer of injection needles: 3Eluted time: 2 H

[0216] Chiral prep-SFC method 24:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralCel OX, 250×20mm I.D., 5μmPatent Application Atty. Docket No. ENTX-038PCT Mobile phase: A for CO2 and B for MeOHGradient: B 50 %Flow rate: 30 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 10 minEluted time: 2 H

[0217] Chiral prep-SFC method 25:Instrument: Waters Thar 80 preparative SFCColumn: (R, R)-WHELK, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 8 minRun time: 8 minInjection volume: 2 mLNumLer of injection needles: 5Eluted time: 2 H

[0218] Chiral prep-SFC method 26:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralCel OX, 250×20 mm I. D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 50 %Flow rate: 30 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmPatent Application Atty. Docket No. ENTX-038PCT Cycle-time: 10minEluted time: 2 H

[0219] Chiral prep-SFC method 27:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IB, 250><30mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 20%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 14minCycle-time: 14minInjection volume: 3 mLNumLer of injection needles: 5Eluted time: 2H

[0220] Chiral prep-SFC method 28:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak AD, 250×30mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 30%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 10minCycle-time: 10minInjection volume: 3 mLNumLer of injection needles: 3Eluted time: 2H

[0221] Chiral prep-SFC method 29:Patent Application Atty. Docket No. ENTX-038PCT Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak AS, 250><30mm I. D., 5 pmMobile phase: A for CO2 and B for MeOHGradient: B 20%Flow rate: 55 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 8 minInjection volume: 3.4 mLNumLer of injection needles: 2Eluted time: 1 h

[0222] Chiral prep-SFC method 30:Instrument: Waters Thar 80 preparative SFCColumn: (R,R)-WHELK, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 10minRun time: 10minInjection volume: 3 mLNumLer of injection needles: 3Eluted time: 2 H

[0223] Chiral prep-SFC method 31:Instrument: SHIMADZU PREP SOLUTION SFCColumn: (R, R)-WHELK, 250×20mm I. D., 5μmPatent Application Atty. Docket No. ENTX-038PCT Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 12minInjection volume: 3.3mLNumLer of injection needles: 4Eluted time: 2 H

[0224] Chiral prep-SFC method 32:Instrument: SHIMADZU PREP SOLUTION SFCColumn: (R, R)-WHELK, 250×20mm I. D., 5μmInjection volume: 2mLNumLer of injection needles: 3Eluted time: 1 H

[0225] Chiral prep-SFC method 33:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak AD, 250×30mm I.D., 5μmMobile phase: A for CO2 and B for EtOHGradient: B 30%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 9 minPatent Application Atty. Docket No. ENTX-038PCT Cycle-time: 79 minInjection volume: 4.2 mLNumLer of injection needles: 5Eluted time: 2 h

[0226] Chiral prep-SFC method 34:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak AD, 250><30mm I. D., 5 pmMobile phase: A for CO2 and B for MeOHGradient: B 25%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 12 minInjection volume: 3 mLNumLer of injection needles: 2Eluted time: 2 H

[0227] Chiral prep-SFC method 35:Instrument: Waters Thar 80 preparative SFCColumn: ChiralCel OX, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 25 %Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 5 minInjection volume: 3 mLNumLer of injection needles: 6Eluted time: 2 H

[0228] Chiral prep-SFC method 36:Patent Application Atty. Docket No. ENTX-038PCT Instrument: Waters Thar 80 preparative SFCColumn: (R, R)-WHELK, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 8minRun time: 8minInjection volume: 3 mLNumLer of injection needles: 3Eluted time: 2 H

[0229] Chiral prep-SFC method 37:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralCel OZ, 250×20mm I. D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 30%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 12.5 minInjection volume: 3.4 mLNumLer of injection needles: 3Eluted time: 2 H

[0230] Chiral prep-SFC method 38:Instrument: Waters Thar 80 preparative SFCColumn: (R, R)-WHELK, 250×20mm I. D., 5μmPatent Application Atty. Docket No. ENTX-038PCT Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 14 minRun time: 14 minInjection volume: 4.2 mLNumLer of injection needles: 3Eluted time: 2 H

[0231] Chiral prep-SFC method 39:Instrument: Waters Thar 80 preparative SFCColumn: ChiralCel OX, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 11 minRun time: 11 minInjection volume: 4 mLNumLer of injection needles: 2Eluted time: 2H

[0232] Chiral prep-SFC method 40:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak IG, 250×20mm I. D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 30%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CPatent Application Atty. Docket No. ENTX-038PCT Wavelength: 220 nmCycle-time: 10minEluted time: 2H

[0233] Chiral prep-SFC method 41:Instrument: Waters Thar 80 preparative SFCColumn: ChiralCel OX, 250><20mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 30 %Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 13 minRun time: 13 minInjection volume: 3 mLNumLer of injection needles: 3Eluted time: 2H

[0234] Chiral prep-SFC method 42:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak IA, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 25%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 15 minEluted time: 2 H

[0235] Chiral prep-SFC method 43:Instrument: SHIMADZU PREP SOLUTION SFCColumn: (R, R)-WHELK, 250×20mm I. D., 5μmPatent Application Atty. Docket No. ENTX-038PCT Mobile phase: A for CO2 and B for MeOHGradient: B 45%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 5.5minInjection volume: 3.2 mLNumLer of injection needles: 6Eluted time: 2 H

[0236] Chiral prep-SFC method 44:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IG, 250><20mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 30%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 9minCycle-time: 9minInjection volume: 1.5 mLNumLer of injection needles: 7Eluted time: 2H

[0237] Chiral prep-SFC method 45:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IC, 250×20mm I. D., 5μmMobile phase: A for CO2 and B for MeOH (0.1% 7mol / L NH3 in MeOH)Gradient: B 20%Flow rate: 40 mL / minBack pressure: 100 barPatent Application Atty. Docket No. ENTX-038PCT Column temperature: 35 °CWavelength: 220 nmRun time: 12minCycle-time: 12minInjection volume: 1.5 mLNumLer of injection needles: 7Eluted time: 2H

[0238] Chiral prep-SFC method 46:Instrument: AUNO LC-2000Column: ChiralPak IA, 250><20mm I. D., 5pmMobile phase: A for Hexane and B for ETOH (0.05% MEA)Gradient: B 25%Flow rate: 25 mL / minColumn temperature: 25 °CWavelength: 220 nmCycle-time: 22minRun time:25minInjection volume: 1.5 mLNumLer of injection needles: 11Eluted time: 4 H

[0239] Chiral prep-SFC method 47:Instrument: SHIMADZU PREP SOLUTION SFCColumn: (R, R)-WHELK, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 50%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 8minInjection volume: 3.4 mLPatent Application Atty. Docket No. ENTX-038PCT NumLer of injection needles: 5Eluted time: 2 h

[0240] Chiral prep-SFC method 48:Instrument: SHIMADZU PREP SOLUTION SFCColumn: (R,R)-WHELK, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 45%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 14minInjection volume: 2 mLNumLer of injection needles: 7Eluted time: 2 H

[0241] Chiral prep-SFC method 49:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPakIH, 250×30mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 22%Flow rate: 70 mL / minBack pressure: 100 barColumn temperature: 40 °CWavelength: 220 nmCycle-time: 7minInjection volume: 0.8 mLNumLer of injection needles: 12Eluted time: 2H

[0242] Chiral prep-SFC method 50:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IG, 250×20 mm I.D., 5μmPatent Application Atty. Docket No. ENTX-038PCT Mobile phase: A for CO2 and B for MeOHGradient: B 40%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 7minCycle-time: 7minInjection volume: 2 mLNumLer of injection needles: 5Eluted time: 2H

[0243] Chiral prep-SFC method 51:Instrument: SHIMADZU PREP SOLUTION SFCColumn: (R,R)-WHELK, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 37%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 12minInjection volume: 1.4 mLNumLer of injection needles: 7Eluted time: 2 H

[0244] Chiral prep-SFC method 52:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IG, 250×20mm I. D., 5μmMobile phase: A for CO2 and B for EtOHGradient: B 25%Flow rate: 40 mL / minBack pressure: 100 barPatent Application Atty. Docket No. ENTX-038PCT Column temperature: 35 °CWavelength: 220 nmRun time: 8minCycle-time: 6minInjection volume: 2 mLNumLer of injection needles: 8Eluted time: 2H

[0245] Chiral prep-SFC method 53:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IG, 250><20mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 30%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmRun time: 12minCycle-time: 12minInjection volume: 3 mLNumLer of injection needles: 6Eluted time: 2H

[0246] Chiral prep-SFC method 54:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralCel OZ, 250×20mm I. D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 40%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 40 °CWavelength: 220 nmCycle-time: 8 minPatent Application Atty. Docket No. ENTX-038PCT Injection volume: 2.5 mLNumLer of injection needles: 6Eluted time: 2 H

[0247] Chiral prep-SFC method 55:Instrument: Waters Thar 80 preparative SFCColumn: ChiralCel OZ, 250><20mm I. D., 5pmMobile phase: A for CO2 and B for MeOHGradient: B 20 %Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 11 minRun time: 11 minInjection volume: 3 mLNumLer of injection needles: 4Eluted time: 2 H

[0248] Chiral prep-SFC method 56:Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralPak IH, 250×30mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 35%Flow rate: 60 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 9 minInjection volume: 3mLNumLer of injection needles: 3Eluted time: 3H

[0249] Chiral prep-SFC method 57:Patent Application Atty. Docket No. ENTX-038PCT Instrument: SHIMADZU PREP SOLUTION SFCColumn: ChiralCel OX, 250><20mm I. D., 5 pmMobile phase: A for CO2 and B for MeOHGradient: B 35%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 40 °CWavelength: 220 nmCycle-time: 10minInjection volume: 2.5 mLNumLer of injection needles: 4Eluted time: 1 H

[0250] Chiral prep-SFC method 58:Instrument: Waters Thar 80 preparative SFCColumn: (R,R)-WHELK, 250×20mm I.D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 45%Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220 nmCycle-time: 18minRun time: 18minInjection volume: 2 mLNumLer of injection needles: 5Eluted time: 2 H

[0251] Chiral prep-SFC method 59:Instrument: Waters Thar 80 preparative SFCColumn: ChiralPak IG, 250×20mm I. D., 5μmMobile phase: A for CO2 and B for MeOHGradient: B 20 %Patent Application Atty. Docket No. ENTX-038PCT Flow rate: 40 mL / minBack pressure: 100 barColumn temperature: 35 °CWavelength: 220nmRun time: 8 minCycle-time: 8 minInjection volume: 1.5 mLNumLer of injection needles: 7Eluted time: 2HGeneral synthetic routes:

[0252] In some emLodiments, compounds of formula I are prepared according to the general procedure depicted in the Scheme I below. In some emLodiments, Step A comprises the C-N bond formation of Int-I to install R5NH. In some emLodiments, the C-N bond formation is carried out via a metal (i.e. palladium- or copper-based catalysts) catalyzed C-N coupling (i.e. R5is C(O)R5or R5) or nucleophilic aromatic substitution (i.e. R5is R5). Step B comprises the cyclization of ketone with primary amide in situ formed by the hydrolysis of CN group under basic conditions. Step B comprises the reduction of the hydroxy to form the compound of formula I. In some emLodiments, the reduction is conducted with triethylsilane (TES) under acidic condition (i.e. tri fluoroacetic acid).Scheme IR5Patent Application Atty. Docket No. ENTX-038PCT

[0253] In some emLodiments, compounds of formula I are prepared according to the general procedure depicted in the Scheme II below, by switching the sequence of C-N bond formation, cyclization and reduction. In some emLodiments, cyclization and reduction are conducted first, then followed by the C-N bond formation to install NHR5to form the compound of formula I. Scheme IICyclization Reduction KOH. ► ACN, H2O Step A Step B X1< X3C-N bond formation 'NH ^4R5NH2b 'TStepC M 6X1 / X3X2

[0254] In some emLodiments, compounds of formula I are prepared according to the general procedure depicted in the Scheme III below, i.e. X1= CR1, for R1installation by converting X (X is Cl, Br, I, OTs, or OTf, or other leaving groups or coupling partners) to different R1groups via a metal (i.e. palladium- or copper-based catalysts) catalyzed coupling ornucleophilic aromatic substitution.Scheme IIIR1installation

[0255] In some emLodiments, compounds of formula I are prepared according to the general procedure depicted in the Scheme IV below, i.e. X2= CR2, for R2installation by converting X (X is Cl, Br, I, OTs, or OTf, or other leaving groups or coupling partners) to different R2groupsPatent Application Atty. Docket No. ENTX-038PCT via a metal (i.e. palladium- or copper-based catalysts) catalyzed coupling ornucleophilic aromatic substitution.Scheme IV

[0256] In some emLodiments, compounds of formula I are prepared according to the general procedure depicted in the Scheme V below, i.e. X3= CR3, for R3installation by converting X (X is Cl, Br, I, OTs, or OTf, or other leaving groups or coupling partners) to different R3groups via a metal (i.e. palladium- or copper-based catalysts) catalyzed coupling or nucleophilic aromatic substitution.Scheme VGeneral synthetic routes to prepare Int-I:

[0257] In some emLodiments, Intermediates Int-I are prepared according to the general procedure depicted in the Scheme VI below including the introduction of different R4and R6groups, and then the construction of ring A. In some emLodiments, Step A comprises the nitration of 2,6-dibromo-4-fluorobenzene-l-carbaldehyde form 2,6-dibromo-4-fluoro-3-nitrobenzene-l-carbaldehyde. Then Step B comprises the nucleophilic addition of aldehyde with organometallic reagent bearing R4group. In some emLodiments, nucleophilic addition of aldehyde is Grignard reaction with organomagnesium compound bearing R4group. In some emLodiments, the organometallic reagent is organolithium compound bearing R4group. Step C comprises the oxidation reaction of alcohol to ketone Int-II. The oxidation conditions includePatent Application Atty. Docket No. ENTX-038PCT but are not limited to using oxidizing agents like: Pyridinium chlorochromate (PCC), Pyridinium dichromate (PDC), Jones reagent (CrO3 / H2SO4 / acetone), Dess-Martin periodinane (DMP), Collins reagent (CrCh / pyridine), the Swern oxidation (DMSO / oxalyl chloride) or other organic reactions converting alcohol to corresponding ketone. In some emLodiments, Step D comprises the SNAr reaction to displace F with amines bearing R6group to form Int-III. Then Step E comprises the cyanation via a metal-(i.e. palladium- or copper-)catalyzed reaction of the bromide with nitrile-containing reagents (i.e. CuCN, TMSCN). In some emLodiments, Step F comprises the reduction of nitro group to aniline to form key intermediate Int-IV. The common reaction conditions for nitro reduction include but are not limited to: using tin(II) chloride (SnCl2), sodium sulfide (Na2S), zinc dust in acidic solution, iron powder in acidic solution, lithium aluminum hydride (LiAlH4), sodium borohydride (NaBH4), and hydrogenation using a catalyst like palladium on carbon (Pd / C), Raney nickel, platinum oxide, or other metals or precious metals. The ring A construction from Int-IV to Int-I will be depending on the specific X1, X2and X3, which will be described in a later section.Scheme VIStep A Step Bn Step C int-u Step D Int-III Step E Br OInt-IV Int-I

[0258] In some emLodiments, intermediates Int-III are prepared from Int-II according to the general procedure depicted in the Scheme VII with additional protection and deprotection steps. Step A comprises the SNAr reaction to displace F with amines bearing protecting group PG. InPatent Application Atty. Docket No. ENTX-038PCT some emLodiments, PG is selected from the list including but not limited to benzyl type of protecting group (i.e. Bn, PMB, DMB), carbonyl type to form amide (i.e. acetyl, trifluoroacetyl, benzoyl) or carbamate (i.e. Boc, Cbz), sulfonyl type (i.e. Tosyl, SeES) or other suitable protecting groups in the art of organic chemistry. Step B comprises the deprotection of PG under suitable conditions depending on the specific PG types in the art of organic chemistry. Step C comprises the installation of R6for R6-N bond formation. In some emLodiments, it’s a N-alkylation reaction of R6bearing leaving group LG (i.e. Cl, Br, I, OMs, OTs, OTf, or other leaving groups), in which LG is displaced by the aniline NH2.Scheme VIIBr O Br O Br O O2N PG— NH2deprotectionF Step A HBrStep BH2Nlnt-llBr OR6-LG R4o6RN LG: Leaving Group = Cl, Br, I, OMs, OTs, OTfStep C H NO2Int-lllGeneral synthetic route for the construction of ring A as imidazole (X1= N, X2= CH or CR2, X3= CH or CR3):

[0259] In some emLodiments, construction of ring A as imidazole (X1= N, X2= CH or CR2and X3= CH or CR3) from Int-IV is prepared according to the general procedure depicted in the Scheme VIII below. Step A comprises the cyclization of Int-IV to guanidine with CN reagent (i.e. BrCN). Step B comprises the imidazole construction to form imidazole ring bearing with R2or R3or both. In some emLodiments, A ring is prepared by using pre-activated carbonyl or protected carbonyl compound like a-halo ketone bearing R2and R3to cyclize with the guanidine to generate the imidazole ring. In some emLodiments, A ring is prepared by using oxidative cyclization with a suitable electrophile to cyclize with the guanidine to generate the imidazole nng.Patent Application Atty. Docket No. ENTX-038PCTInt-IVHNR2lnt-1

[0260] In some emLodiments, construction of ring A as imidazole (X1= N, X2= CR2and X3= CH) Int-I is prepared according to the general procedure depicted in the Scheme IX below. Step A comprises the cyclization of guanidine motif with a-halo (Cl or Br) ester to form the lactam selectively. Step B comprises the transformation of lactam to X-(C1- or Br-)imidazole at the lactam carbonyl position with POCl3or POBn Then Step C comprises the functionalization of X (Cl or Br) into R2groups via a metal-mediated coupling reaction or S\Ar reaction (i.e. C-C bond formation, C-N bond formation or C-S bond formation) to form the compounds of formula I.Scheme IXO XBr O X= ClorBr

[0261] In some emLodiments, construction of ring A as imidazole (X1= N, X2= CH and X3= CR3) from Int-IV to Int-I is prepared according to the general procedure depicted in the Scheme X below. Step A comprises the urea formation of Int-IV dianiline with carbonylation reagents (i.e. carbonyldiimidazole (CDI), phosgene, triphosgene). Step B comprises the conversion of urea carbonyl to Cl-imidazole with chlorination reagent (i.e. POCl3, PCh, SOCI2). Step C comprises the aminoimidazole formation by replacing the Cl with protected amino acid. Step D comprises the cyclization to form amide. Then Step E comprises the chlorination ofPatent Application Atty. Docket No. ENTX-038PCT amide carbonyl by halogenation reagent (i.e. POX3, PX5, X = C, or Br). Step F comprises the R3installation by replacing the X with R3via either cross-coupling reaction or S\ Ar reactions to form Int-I.Scheme XGeneral synthetic route for the construction of ring A as triazole (X1= N, X2= N, X3= CH or CR3):

[0262] In some emLodiments, construction of ring A as triazole (X1= N, X2= N, X3= CH or CR3) Int-I is prepared according to the general procedure depicted in the Scheme XI below. Step A comprises the substitution of Cl with protected hydrazine (i.e. Boc). Followed with the deprotection of protecting group (i.e. under acidic conditions like TFA to deprotect Boc) in Step B. Then Step C comprises the cyclization of 2-hydrazine imidazole with orthoester to form Int-I with the triazole as ring A.Patent Application Atty. Docket No. ENTX-038PCT Scheme XIBr O R3C(OR)3Step Clnt-1General synthetic route for the construction of ring A as triazole (X1= N, X2= CH or CR2, X3= N):

[0263] In some emLodiments, construction of ring A as triazole (X1= N, X2= CH or CR3, X3= N) Int-I is prepared from Int-II according to the general procedure depicted in the Scheme XII below. Step A comprises the reduction of nitro group to aniline to form key intermediate Int-IV. The common reaction conditions for nitro reduction include but are not limited to: using tin(II) chloride (SnCl2), sodium sulfide (Na2S), zinc dust in acidic solution, iron powder in acidic solution, lithium aluminum hydride (LiAHH ), sodium borohydride (NaBH4), and hydrogenation using a catalyst like palladium on carbon (Pd / C), Raney nickel, platinum oxide, or other metals or precious metals. Step B comprises the hydrazine formation of aniline treated with nitrite (i.e. NaNCh) to form the diazonium salt, which is subsequently reduced using reducing agents (i.e. SnCh). Step C comprises the condensation reaction of the terminal NH2 of the hydrazine with alpha-keto acids bearing R2to form the imine. Step D comprises the cyclization reaction of the acid with aniline NH in the present of amide formation reagent (i.e. DPPA with TEA) to form the 5-memLered ring. Step E comprises the conversion of amide carbonyl to Cl using POCl3Or PCI5. Step F comprises another cyclization via a double S\Ar reactions of amines (bearing R6) to replace F and Cl to form 5-memLered ring. Step G comprises the cyanation via a metal-(i.e. palladium- or copper-)catalyzed reaction of the bromide with nitrile-Patent Application Atty. Docket No. ENTX-038PCT containing reagents (i.e. CuCN, TMSCN) to form Int-I with ring A as triazole (X1= N, X2= CH or CR2, X3= N)Scheme XIIlnt-ll Br OInt-I General synthetic route for the construction of ring A as pyrazole (X1= CH, X2= CH or CR2, X3= N):

[0264] In some emLodiments, construction of ring A as pyrazole Int-I (X1= CH, X2= CH or CR2, X3= N) is prepared from Int-IV according to the general procedure depicted in the Scheme XIII below. Step A comprises the cyclization of Int-IV with orthoacetate (i.e. trimethyl orthoacetate) to for methyl imidazole. Step B comprises the N-amination reaction of imidazole by N-amination reagents (i.e. hydroxylamines, sulfonylhydroxylamines, nitrenium ions, and other compounds). Step C comprises the cyclization to form pyrazolone using reagents like CDI. Then Step D comprises the conversion of pyrazolone carbonyl to a leaving group or coupling partner X (i.e. Cl, Br, OTs or OTf). Then Step E comprises the installation of R2by either S\. Ar reaction or coupling reaction in the art of organic chemistry.Patent Application Atty. Docket No. ENTX-038PCT Scheme XIIIN-amination Step BExemplary synthetic procedures for intermediates:(2-chloro-4-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)methanoneCl OH Br BrMg HNO3Cl H2SO4Step A Step B FCl O BrStep C F

[0265] Step A: To a solution of 2,6-dibromo-4-fluorobenzene-l-carbaldehyde (110 g, 390 mmol) in con. H SCh (550 mL) was added dropwise con. HNO3 (43.4 g, 468 mmol) at 0 °C. The reaction mixture was stirred at rt for 4 hr. The reaction mixture was poured into ice water and filtrated. The resulting solid was dissolved in EtOAc, and washed with water to remove the remaining acid, dried over Na2SO4 and concentrated in vacuo to afford 2,6-dibromo-4-fluoro-3-nitrobenzene-l-carbaldehyde (115 g, 352 mmol, 90%) as a black solid. LCMS: ESI m / z 328 [M + H]+.Patent Application Atty. Docket No. ENTX-038PCT

[0266] Step B: To a solution of 2,6-dibromo-4-fluoro-3-nitrobenzene-l-carbaldehyde (40 g, 122 mmol) in THF (400 mL) was added 2-chloro-5-fluorophenyl)magnesium chloride (734 mL, freshly made, 0.5 M in THF) at 0 °C. The mixture was stirred at rt for Ih. The reaction mixture was quenched with aqueous solution of NH4CI, extracted with EA. The organic phase was washed with brine, dried over Na2SCU and concentrated. The residue was purified by column chromatography on silica gel (eluted with EA / PE = 0-20%) to afford (2-chloro-4-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)methanol (50 g, 109mmol, 89%) as a brown oil. (crude). LCMS: ESI m / z 458 [M + H]+.

[0267] Step C: To a solution of (2-chloro-4-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)methanol (122 g, 267 mmol) in DCM (1500 mL) was added DMP (170 g, 400 mmol) at 0 °C. The reaction was stirred at rt for Ih. The reaction mixture was diluted with water, extracted with EA. The organic layer was separated, washed with brine, dried over Na2SO4and concentrated in vacuo. The residue was purified using silica gel column chromatography eluted with ethyl acetate in petroleum ether (gradient: 0-20%) to afford crude (2-chloro-4-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)methanone as a brown solid. The crude was triturated with 500 ml (EA: PE=1: 10). The suspension was filtered to get (2-chloro-4-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)methanone (88 g, 193 mmol, 72%) as a white solid. LCMS: ESI m / z 456 [M + H]+.2-amino-6-bromo-5-(2-chloro-5-fluorobenzoyl)-l-ethyl-lH-benzo[d]imidazole-4-carbonitrile

[0268] Step A: To a stirred mixture of (2-chloro-4-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)methanone (15 g, 32.9 mmol) and ethanamine (1.63 g, 36.2 mmol)Patent Application Atty. Docket No. ENTX-038PCT in tetrahydrofuran (150 mL) was added DIEA (12.8 g, 98.8 mmol). The reaction was stirred at 60 °C for 6 h. The reaction was monitored by analysis of LCMS. The reaction mixture was diluted with EtOAc, washed with brine, and organic layer was separated. The aqueous layer was extracted with EtOAc, then the comLined extracts were dried over anhydrous Na2SO4, filtered and evaporated. The crude product was triturated with PE / EA=10 / l to afford (2-chloro-4-fluorophenyl)[2,6-dibromo-4-(ethylamino)-3-nitrophenyl]methanone (15 g, 31.2 mmol, 95%) as a brown solid. LCMS: ESI m / z 481 [M + H]+.

[0269] Step B: To a solution of (2-chloro-4-fluorophenyl)[2,6-dibromo-4-(ethylamino)-3-nitrophenyl]methanone (14 g, 29.1 mmol) in N, N-dimethylacetamide (150 mL) were added copper(l+) cyanide (2.66 g, 29.7 mmol) at 90 °C. The reaction was stirred at 90 °C for 30 min. The cooled reaction was poured into EA and added H2O. The suspension was filtered on a celite and the filtrate was diluted with water, extracted with EA. The organic layer was washed with brine, dried over Na2SO4and concentrated. The residue was triturated with PE: EA=15:1 to afford 3-bromo-2-[(2-chloro-4-fluorophenyl)carbonyl]-5-(ethylamino)-6-nitrobenzene-l-carbonitrile (10 g, 23.4 mmol, 80%) as a yellow solid. LCMS: ESI m / z 428 [M+2+H]+.

[0270] Step C: To a solution of 3-bromo-2-[(2-chloro-4-fluorophenyl)carbonyl]-5-(ethylamino)-6-nitrobenzene-l-carbonitrile (10 g, 23.4 mmol) in ethanol (100 mL) and water (25 mL) was added iron(0) (6.54 g, 117 mmol) and ammonium chloride (6.27 g, 117 mmol). The reaction was stirred at 90 °C for 2 h. LCMS showed the reaction was completed. The reaction mixture was filtered, the filtrate was concentrated, diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel column chromatography eluted with ethyl EA in PE (0-30% gradient) to afford 6-amino-3-bromo-2-[(2-chloro-4-fluorophenyl)carbonyl]-5-(ethylamino)benzene-l -carbonitrile (2 g, 5.04 mmol, 22 %) as a yellow solid. LCMS (ESI): m / z 398 [M+2+H]+.

[0271] Step D: To a solution of 6-amino-3-bromo-2-[(2-chloro-4-fluorophenyl)carbonyl]-5-(ethylamino)benzene-l -carbonitrile (2.2 g, 5.55 mmol) in ethanol (25 mL) and water (5 mL) was added BrCN (2.94 g, 27.7 mmol), the reaction mixture was stirred at 100 °C for 5 h. The reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel column chromatography eluted with ethyl EA in PE (0-100% gradient) to afford 2-Patent Application Atty. Docket No. ENTX-038PCT amino-6-bromo-5-(2-chloro-5-fluorobenzoyl)-l -ethyl- lH-benzo[d]imidazole-4-carbonitrile (1.2 g, 2.85 mmol, 51%) as a brown solid. LCMS (ESI): m / z 423 [M+2+H]+.2-amino-5-bromo-6-(2-chloro-5-fluorobenzoyl)-3-((2,2-difluoroethyl)amino)benzonitriIe and 2-azanylidene-6-bromo-5-[(5-chloro-2-fluorophenyl) carbonyl]-l-(2,2-difluoroethyl)- 3H-benzo[d] imidazole-4-carbonitrile

[0272] Step A: To a solution of (2-chloro-5-fluorophenyl) (2,6-dibromo-4-fluoro-3-nitrophenyl) methanone (350.0 g, 767.0 mmol) in DMF (3000 mL) were added 2,2-difluoroethan- 1 -amine (93.5 g, 1153.0 mmol) and DIEA (199.0 g, 1537.0 mmol). The reaction mixture was stirred at 35°C for 16 h. TLC showed the reaction was completed. The solution was diluted with water and extracted with EA (4 L). The organic layer was washed with brine(4L) for four times, dried over Na2SO4 and concentrated. The residue was triturated with PE: EA=15: 1 to afford (2-chloro-5-fluorophenyl){2,6-dibromo-4-[(2,2-difluoroethyl)amino]-3-nitrophenyl (methanone (380 g, 737 mmol, 96%) as a yellow solid. LCMS: ESI m / z 517 [M + H]+. Tl NMR (400 MHz, DMSO- 6) 57.71 (dd, J= 9.6, 4.8 Hz, 1H), 7.63 - 7.56 (m, 2H), 7.51 (s, 1H), 7.02 (s, 1H), 6.15 (tt, <7 = 55.6, 3.6 Hz, 1H), 3.74 (t, J= 15.4 Hz, 2H).

[0273] Step B: To a solution of (2-chloro-5-fluorophenyl) {2,6-dibromo-4-[(2,2-difluoroethyl) amino] -3 -nitrophenyl} methanone (300.0 g, 581.0 mmol) in DMSO (500 mL) were added CuCN (53.1 g, 592.0 mmol) at 90°C. The reaction was stirred at 90°C for 30 min. LCMS showed the reaction was completed The cooled reaction was poured into EA and water was added. The suspension was filtered through celite, the filtrate was diluted with water and EA. The organic layer was separated and washed with brine, dried over Na2SO4and concentrated. The residue was triturated with PE: EA=15: 1 and IPA to afford 5-bromo-6-[(2-Patent Application Atty. Docket No. ENTX-038PCT chloro-5-fluorophenyl) carbonyl]-3-[(2,2-difluoroethyl) amino]-2-nitrobenzene-l -carbonitrile (252 g, 545 mmol, 94%) as a yellow solid.LCMS: ESI m / z 462.61 [M+H]1.

[0274] Step C: To a solution of 5-bromo-6-[(2-chloro-5-fluorophenyl) carbonyl]-3-[(2,2-difluoroethyl) amino]-2-nitrobenzene-l -carbonitrile (320.0 g, 692.0 mmol) in EtOH (2.4 L), Acetic acid (600 mL) and H2O (480 mL) was added Fe (212.0 g, 3804.0 mmol). The reaction was stirred at inner temp 75°C for 30 min. LCMS showed the reaction was completed. The reaction mixture was filtered, and the filtrate was concentrated, the residue was diluted with water and extracted with EtOAc (1500 mL*3). The organic layer was washed with brine, dried over Na2SO4and concentrated. The residue was purified using silica gel column chromatography eluted with ethyl acetate in petroleum ether (gradient: 0-50%) to give a crude product which was triturated with PE: DCM=1: 1 to afford 2-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-[(2,2-difluoroethyl) amino] benzene- 1 -carbonitrile (190.0 g, 439.2 mmol, 63.5%) as a yellow solid. LCMS: ESI m / z 432.63 [M+H]+.

[0275] Step D: To a solution of 6-amino-3-bromo-2-[(5-chloro-2-fluorophenyl) carbonyl]-5-[(2,2-difluoroethyl) amino] benzene- 1 -carbonitrile (146.8 g, 339.3 mmol), BrCN (359.4 g, 3393.2 mmol) in MeOH (1125 mL) was added H2O (375 mL), the reaction mixture was stirred at inner temp 70oC for 16 h. The reaction mixture was cooled to rt, sat. NaHCO3 (50 mL) was added, then diluted with H2O (1125 mL). After filtration, the cake was washed with MeOH: H2O=1:3 (500 mL, twice), The impure product was triturated with DCM (500 mL*2) and EA: DCM=1:1 (300 mL) and was then filtered to afford 2-azanylidene-6-bromo-5-[(5-chloro-2-fluorophenyl) carbonyl]-! -(2, 2-difluoroethyl)-3H-benzo[d] imidazole-4-carbonitrile (60.0 g, 131.1 mmol, 38.6%). LCMS: 459 [M+H]+. 1HNMR (400 MHz, DMSO-d6) 87.87 (s, 1H), 7.75 - 7.66 (m, 3H), 7.62 - 7.55 (m, 1H), 7.53 - 7.47 (m, 1H), 6.59 - 6.22 (m, 1H), 4.74 - 4.58 (m, 2H).6-bromo-2-chloro-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2,2-difluoroethyl)benzo [d] imidazole-4-carbonitriIe and 6-bromo-5-(2-chloro-5-fluorobenzoyl)-l-(2,2-difluoroethyl)-2-hydrazineyl-lH-benzo[d]imidazole-4-carbonitrilePatent Application Atty. Docket No. ENTX-038PCT Br O ClBr O Cl Br O ClNHBoc NH2

[0276] Step A: To a solution of 2-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-[(2,2-difluoroethyl)amino]benzene-l-carbonitrile (200 mg, 0.462 mmol) in THF (2 mL) was added TEA (140 mg, 1.39 mmol) and trichloromethyl [(trichloromethyl)oxy]methanoate (68.6 mg, 0.231 mmol) at 0 °C. The mixture was stirred at 20 °C for 1 hour. Themixture was diluted with H₂O and extracted with EA. The organic phase was washed with brine, dried over Na₂SO₄ and concentrated to give 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-1-(2,2-difluoroethyl)-2-oxo-3H-benzo[d]imidazole-4-carbonitrile (210 mg, 0.458 mmol, 99 %) as a red solid. LCMS: m / z 460.1 [M+H]+.

[0277] Step B: A mixture of 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2,2-difluoroethyl)-2-oxo-3H-benzo[d]imidazole-4-carbonitrile (200 mg, 0.436 mmol) in POCl3(5 mL) was stirred at 140 °C for 36 hours. The mixture was concentrated and the residue was purified by silica gel chromatography (4 g column) using 0 - 30% EtOAc / hexane to afford 6-bromo-2-chloro-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2,2-difluoroethyl)benzo[d]imidazole-4-carbonitrile (104 mg, 0.218 mmol, 50 %) as a red solid. LCMS: ESI m / z 478.1 [M + H]+.

[0278] Step C: To a solution of 6-bromo-2-chloro-5-(2-chloro-5-fluorobenzoyl)-l-(2,2-difluoroethyl)-lH-benzo[d]imidazole-4-carbonitrile (600 mg, 1.26 mmol) in DMA (8 mL) was added TEA (0.52 mL, 3.77 mmol) and tert-butyl hydrazinecarboxylate (332 mg, 2.5 mmol). The mixture was stirred at 100 °C for 2 h. The cooled mixture was diluted with water, extracted with EA. The organic phase was washed with brine, dried over Na₂SO₄ and concentrated. The residue was purified using silica gel column chromatography eluting with 0-100% ethyl acetate in petroleum ether to afford tert-butyl 2-(6-bromo-5-(2-chloro-5-fluorobenzoyl)-4-cyano-1-(2,2-Patent Application Atty. Docket No. ENTX-038PCT difluoroethyl)-1H-benzo[d]imidazol-2-yl)hydrazine-1-carboxylate (300 mg, 0.52 mmol, 41%) as a yellow solid. LCMS: ESI m / z 572 [M + H]+.

[0279] Step D: A solution of tert-butyl 2-(6-bromo-5-(2-chloro-5-fluorobenzoyl)-4-cyano-1-(2,2-difluoroethyl)-1H-benzo[d]imidazol-2-yl)hydrazine-1-carboxylate (250 mg, 0.436 mmol) in TFA (2 mL) was stirred at 25 °C for 1 h. The mixture was concentrated under vacuum to afford 6-bromo-5-(2-chloro-5-fluorobenzoyl)-1-(2,2-difluoroethyl)-2-hydrazineyl-1H-benzo[d]imidazole-4-carbonitrile (200 mg, 0.42 mmol, crude) as a yellow solid. LCMS: ESI m / z 472 [M + H]⁺.2-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-[(2-fluoroethyl)amino]benzene-1 -carbonitrileBr 0 Cl Br O Cl Br O ClStep A Step BBr O Cl Fe, AcOH EtOH, H2OStep C

[0280] Step A: To a solution of (2-chloro-5-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)m ethanone (10 g, 21.96 mmol) in dioxane (100 mL) was added DIEA (8.51 g, 65.8 mmol) and 2-fluoroethan-l -amine hydrochloride (2.40 g, 24.1 mmol) at 25 °C and the reaction mixture was stirred at 100 °C for 2 h. The cooled reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography on silica gel (EA in PE = 0 ~ 30%) to obtain (2-chloro-5-fluorophenyl){2,6-dibromo-4-[(2-fluoroethyl)amino]-3-nitrophenyl} methanone (10 g, 20.06 mmol, 91%) as an yellow solid. LCMS: 498.9, 501.9 [M+H]+.

[0281] Step B: A mixture of (2-chloro-5-fluorophenyl){2,6-dibromo-4-[(2-fluoroethyl)amino]-3 -nitrophenyl} methanone (10 g, 20.0 mmol) in DMSO (100 mL) was heated to 90 °C (inner temperature), then CuCN (1.80 g, 20.0 mmol) was added at 90 °C and thePatent Application Atty. Docket No. ENTX-038PCT suspension was stirred at 90 °C (inner temperature) for 20 min, LCMS showed the reaction was completed. The reaction mixture was cooled down to room temperature, diluted with EA and H₂O, filtered. The organic layer was separated, the aqueous layer was extracted with EA. The comLined organic layer was washed with brine, dried over Na₂SO₄, filtered and concentrated. The residue was purified by trituration (DCM: PE=1: 3) and filtered to afford 5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-[(2-fluoroethyl)amino]-2-nitrobenzene-l -carbonitrile (7.30 g, 16.42 mmol, 82%) as a yellow solid. LCMS: 445.9, 443.9 [M+H]+.

[0282] Step C: To a solution of 5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-[(2-fluoroethyl)amino]-2-nitrobenzene-l -carbonitrile (7.20 g, 16.2 mmol) in EtOH(57.6 mL), AcOH (14.4 mL) and H₂O (11.52 mL) was added Fe (4.52 g, 80.9 mmol). The reaction was stirred at inner temp 75 °C for 30 min. LCMS showed the reaction was completed. The reaction mixture was filtered, the filtrate was concentrated, diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na₂SO₄ and concentrated to afford 2-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-[(2-fluoroethyl)amino]benzene-1-carbonitrile (6 g, 14.47 mmol, 89%) as an orange solid which was used directly to the next step without further purification. LCMS: 416.0, 414.0 [M+H]+.2-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3- [(pentadeuterioethyl)aminojbenzene-l-carbonitrile

[0283] Step A: To a solution of (2-chloro-5-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)m ethanone (20 g, 43.92 mmol) in dioxane (200 mL) was added (2,4-Patent Application Atty. Docket No. ENTX-038PCT dimethoxyphenyl)methanamine (7.71 g, 46.11 mmol) and DIEA (17.0 g, 131 mmol). The reaction mixture was stirred at 100 °C under N2 for 2 h. The cooled reaction mixture was quenched with water, extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by column chromatography on silica gel to obtain (2-chloro-5-fluorophenyl)(2,6-dibromo-4-{[(2,4-dimethoxyphenyl)methyl]amino}-3-nitrophenyl)methanone (18.5 g, 30.70 mmol, 70%) as an yellow oil. LCMS: 602.9, 604.9 [M+H]+.

[0284] Step B: A mixture of (2-chloro-5-fluorophenyl)(2,6-dibromo-4-{[(2,4-dimethoxyphenyl)methyl]amino}-3-nitrophenyl)methanone (18.5 g, 30.7 mmol) in DMSO (185 mL) was heated to 90 °C (inner temperature), then CuCN (2.75 g, 30.70 mmol) was added at 90 °C and the suspension was stirred at 90 °C (inner temperature) under N2 for 20 min. LCMS showed the reaction was completed. The reaction mixture was cooled to room temperature, diluted with EA and H2O, filtered. The organic layer was separated, the aqueous layer was extracted with EA. The comLined organic layer was washed with brine, dried over Na₂SO₄, filtered and concentrated. The residue was purified by trituration (DCM: PE=1: 3.3) and filtered to afford 5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-{[(2,4-dimethoxyphenyl)methyl]amino}-2-nitrobenzene-1-carbonitrile (16 g, 24.7 mmol, 81%) as a yellow solid. LCMS: 550.0, 548.0 [M+H]+.

[0285] Step C: A solution of 5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-{[(2,4-dimethoxyphenyl)methyl]amino}-2-nitrobenzene-1-carbonitrile (15.5 g, 28.2 mmol) in DCM (150 mL) and TFA (30 mL). Then the mixture was stirred at 25 °C for 10 min. The mixture was concentrated. The residue was diluted with aqueous NaHCO3, extracted with EA. The organic phase was separated and concentrated. The residue was purified by column chromatography on silica gel to afford 3-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-2-nitrobenzene-l-carbonitrile (10 g, 25.1 mmol, 89%) as a yellow solid. LCMS: 399.9, 397.9 [M+H]+.

[0286] Step D: To a solution of 3-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-2-nitrobenzene- 1 -carbonitrile (5 g, 12.55 mmol) in DMF (50 mL) was added NaH (0.50 g, 12.5 mmol, 60% in mineral oil) at 0 °C. The mixture was stirred at 0 °C for lOmin, then 1, 1,2, 2,2-pentadeuterio-1 -iodoethane (2.02 g, 12.5 mmol) was added. The following mixture was stirred at rt for 3 h. The reaction mixture was quenched by adding sat. aq. NH4CI solution, and the mixture was extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 andPatent Application Atty. Docket No. ENTX-038PCT concentrated. The residue was purified using silica gel column chromatography eluted with ethyl acetate in petroleum ether to afford 5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-2-nitro-3-[(pentadeuterioethyl)amino]benzene-l -carbonitrile (2 g, 4.63 mmol, 37%) as a yellow solid. LCMS: 433.0, 431.0 [M+H]+.

[0287] Step E: To a solution of 5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-2-nitro-3-[(pentadeuterioethyl)amino]benzene-l -carbonitrile (2 g, 4.63 mmol) in EtOH (20 mL), AcOH (5 mL) and H₂O (4 mL) was added Fe (1.29 g, 23.1 mmol). The reaction was stirred at inner temp 75 °C for 2 h. LCMS showed the reaction was completed. The reaction mixture was filtered, the filtrate was concentrated, diluted with water and extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by column chromatography on silica gel to obtain 2-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-[(pentadeuterioethyl)amino]benzene-1-carbonitrile (1.5 g, 3.73 mmol, 81%) as an yellow solid. LCMS: 403.0, 401.0 [M+H]+.7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(ethyl-d5)-2-oxo-2,9-dihydro-3H-benzo [d] imidazo [1,2-a] imidazole-5-carbonitr ileFe / NH4Cl DIEA, THF Step A Step BPatent Application Atty. Docket No. ENTX-038PCT

[0288] Step A: To a solution of (2-chloro-5-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)m ethanone (15 g, 33 mmol) in THF (200 mL) were added ethan-d5-l -amine (3.30 g, 66 mmol) and DIEA (8.51 g, 66 mmol). The reaction was stirred at 60 °C in sealed tube under nitrogen for 18 hours. LCMS showed the reaction was 50% DP. The cooled reaction mixture was diluted water, extracted with EtOAc. The organic phase was washed with brine, dried over Na₂SO₄ and concentrated. The residue was purified using silica gel column chromatography eluted with ethyl acetate in petroleum ether (gradient: 10-20%) to afford compound (2-chloro-5-fluorophenyl)(2,6-dibromo-4-((ethyl-d5)amino)-3-nitrophenyl)methanone (6 g, 12.4 mmol, 38%) as a yellow solid. LCMS (ESI): m / z 486 [M+H]+.

[0289] Step B: To a solution of (2-chloro-5-fluorophenyl)(2,6-dibromo-4-((ethyl-d5)amino)-3-nitrophenyl)methanone (500 mg, 1.03 mmol) in EtOH (10 mL) and H₂O (10 mL) were added Fe (289 mg, 5.15 mmol) and NH₄Cl (276 mg, 5.15 mmol). The reaction was stirred at 80 °C under nitrogen for 2 hours. LCMS showed the reaction was 70% DP. The cooled reaction mixture was filtered. The filtrate was concentrated, diluted water, extracted with EtOAc. The organic phase was washed with brine, dried over Na₂SO₄ and concentrated. The residue was purified using silica gel column chromatography eluting with ethyl acetate in petroleum ether(gradient: 14-16%) to afford (3-amino-2,6-dibromo-4-((ethyl-d5)amino)phenyl)(2-chloro-5-fluorophenyl)methanone (300 mg, 0.66 mmol, 64%) as a yellow solid. LCMS (ESI): m / z 456 [M+H]+.

[0290] Step C: To a solution of {3-amino-2,6-dibromo-4-[(pentadeuterioethyl)amino]phenyl}(2-chloro-5-fluorophenyl)methanone (5 g, 10.9 mmol) in EtOH (50 mL) and H₂O (15 mL) was added Cyanogen bromide (9.3 g, 87.8 mmol). The mixture was stirred at 100 °C for 3 hours. The reaction mixture was quenched by water, extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated. The crude product was purified by flash silica chromatography, eluting with a gradient 10% to 50% EtOAc in PE. The desired fractions were evaporated to dryness to afford [2-azanylidene-4,6-dibromo-l-(pentadeuterioethyl)-3H-benzo[d]imidazol-5-yl](2-chloro-5-fluorophenyl)methanone (4 g, 76%) as a light yellow solid. LCMS (ESI): m / z 481 [M+H]+.

[0291] Step D: To a solution of [2-azanylidene-4,6-dibromo-l-(pentadeuterioethyl)-3H-benzo[d]imidazol-5-yl](2-chloro-5-fluorophenyl)methanone (2 g, 4.1 mmol) in NMP (20 mL) was added bromoacetyl bromide (3.3 g, 16.6 mmol). The mixture was stirred at 105 °C for 1Patent Application Atty. Docket No. ENTX-038PCT hour. The reaction mixture was poured into water and filtered. The filtered cake was washed with water to give the crude product. The crude product was purified by flash silica chromatography, eluting with a gradient 10% to 50% EtOAc in PE. The desired fractions were evaporated to dryness to afford a crude product (1.2 g, 48%) as a light yellow solid, which will be used directly in the next step. LCMS (ESI): m / z 601 [M+H]+.

[0292] Step E: To a solution of the crude product (1.2 g, 1.99 mmol) obtained in the last step D in DMF (40 mL) was added K2CO3 (0.41 g, 2.99 mmol). The mixture was stirred at 60 °C for 1.5 hours. The reaction mixture was quenched by water, extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated. The crude product was purified by flash silica chromatography, eluting with a gradient 0% to 100% EtOAc in PE. The desired fractions were evaporated to dryness to afford 5,7-dibromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(pentadeuterioethyl)-3,9-dihydro-2H-benzo[4,5]imidazo[2,3-b]imidazol -2-one (400 mg, 39%) as a light yellow solid. LCMS (ESI): m / z 521 [M+H]+.

[0293] Step F: To a solution of 5,7-dibromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(pentadeuterioethyl)-3,9-dihydro-2H-benzo[4,5]imidazo[2,3-b]imidazol-2-one (400 mg, 0.77 mmol) in DMSO (5 mL) was added Copper(I) cyanide (103 mg, 1.15 mmol). The mixture was stirred at 130 °C for 2 hours. The reaction mixture was filtered and the filtrate was concentrated to give the residue. The reaction mixture was quenched by water, extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated. The crude product was purified by flash silica chromatography, eluting with a gradient 10% to 90% EtOAc in PE to afford 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-2-oxo-9-(pentadeuterioethyl)-3,9-dihydro-2H-benzo[4,5]imidazo[2,3-b]imidazole-5-carbonitrile (25 mg, 7%) as a white solid. LCMS (ESI): m / z 466 [M+H]+.6-bromo-5-(2-chloro-5-fluorobenzoyl)-2-hydrazineyl-l-propyl-lH-benzo[d]imidazole-4-carbonitrilePatent Application Atty. Docket No. ENTX-038PCTCuCN DMSOStep B

[0294] Step A: To a solution of (2-chloro-5-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)m ethanone (15 g, 32.9 mmol) in THF (150 mL) was added propan- 1 -amine (2.04 g, 34.6 mmol) and DIEA (8.5 g, 65.9 mmol). The mixture was stirred at 60 °C for 5 h. The cooled mixture was quenched with water, extracted with EtOAc. The organic layer was washed with water and brine, dried over anhydrous Na₂SO₄, filtered and concentrated afford (2-chloro-5-fluorophenyl)[2,6-dibromo-3-nitro-4-(propylamino)phenyl]methanone (15 g, 92%). LCMS (ESI): m / z 493 [M+H]+.

[0295] Step B: To a solution of (2-chloro-5-fluorophenyl)[2,6-dibromo-3-nitro-4-(propylamino)phenyl]methanone (13.7 g, 27.7 mmol) in DMSO (130 mL) was added CuCN (2.47 g, 27.7 mmol). The mixture was stirred at 90 °C for 0.5 h. The cooled mixture was quenched with water, extracted with EtOAc. The organic layer was washed with water and brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by silica gel chromatography, eluted with a gradient of 0-50% ethyl acetate in petroleum ether to afford 3-bromo-2-[(2-chloro-5-fluorophenyl)carbonyl]-6-nitro-5-(propylamino)benzene-1-carbonitrile (11 g, 90%). LCMS (ESI): m / z 440 [M+H]+.

[0296] Step C: To a solution of 3-bromo-2-[(2-chloro-5-fluorophenyl)carbonyl]-6-nitro-5-(propylamino)benzene-l -carbonitrile (8.4 g, 19.1 mmol) in EtOH (60 mL) and H2O (12 mL) was added Fe (1.06 g, 19.1 mmol) and AcOH (15 mL). The mixture was stirred at 80°C for 1 h. ThePatent Application Atty. Docket No. ENTX-038PCT cooled mixture was quenched with water, extracted with EtOAc. The organic layer was washed with water and brine, dried over anhydrous Na₂SO₄, filtered and concentrated to afford 6-amino-3-bromo-2-[(2-chloro-5-fluorophenyl)carbonyl]-5-(propylamino)benzene-1-carbonitrile (7 g, 89%). LCMS (ESI): m / z 410 [M+H]+.

[0297] Step D: To a solution of 6-amino-3-bromo-2-[(2-chloro-5-fluorophenyl)carbonyl]-5-(propylamino)benzene-l -carbonitrile (3 g, 7.31 mmol) in THF (30 mL) was added trichloromethyl [(trichloromethyl)oxy]methanoate (1.52 g, 5.11 mmol) at 0 °C. The mixture was stirred at 20 °C overnight. The cooled mixture was quenched with water, extracted with DCM. The organic layer was washed with water and brine, dried over anhydrous Na₂SO₄, filtered and concentrated to afford 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-2-oxo-1-propyl-3H-benzo[d]imidazole-4-carbonitrile (2.8 g, 87 %). LCMS (ESI): m / z 436 [M+H]+.

[0298] Step E: To a solution of 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-2-oxo-l-propyl-3H-benzo[d]imidazole-4-carbonitrile (2.8 g, 6.41 mmol) in POCl₃ (20 mL) was added DIEA (4.14 g, 32.1 mmol). The mixture was stirred at 105°C for 4 h. Then the mixture extracted with EtOAc. The organic layer was washed with NaHCO₃ and brine, dried over anhydrous Na₂SO₄, filtered and concentrated. The residue was purified by silica gel flash chromatography, eluted with a gradient of 0-50% ethyl acetate in petroleum ether to afford 6-bromo-2-chloro-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-propylbenzo[d]imidazole-4-carbonitrile (400 mg, 13%). LCMS (ESI): m / z 627 [M+H]+.

[0299] Step F: To a solution of 6-bromo-2-chloro-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-propylbenzo[d]imidazole-4-carbonitrile (400 mg, 0.88 mmol) in DMA (12 mL) was added TEA (266 mg, 2.64 mmol) and tert-butyl hydrazinecarboxylate (232 mg, 1.76 mmol). The mixture was stirred at 100 °C for 2 h. The cooled mixture was quenched with water, extracted with EtOAc. The organic layer was washed with water and brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by silica gel chromatography, eluted with a gradient of 0-30% ethyl acetate in petroleum ether to afford tert-butyl 2-(6-bromo-5-(2-chloro-5-fluorobenzoyl)-4-cy ano- 1 -propyl- lH-benzo[d]imidazol -2 -yl)hydrazine-l -carboxylate (300 mg, 61%). LCMS (ESI): m / z 550 [M+H]+.

[0300] Step G: A solution of tert-butyl 2-(6-bromo-5-(2-chloro-5-fluorobenzoyl)-4-cyano-l-propyl-lH-benzo[d]imidazol-2-yl)hydrazine-l-carboxylate (300 mg, 0.54 mmol) in TFA (3 mL) was stirred at 20°C for 0.5 h. The mixture was concentrated to afford 6-bromo-5-(2-chloro-5-Patent Application Atty. Docket No. ENTX-038PCT fluorobenzoyl)-2-hydrazineyl-l -propyl- lH-benzo[d]imidazole-4-carbonitrile (150 mg, 61%). LCMS (ESI): m / z 450 [M+H]+.[dioxo(2,4,6-trimethylphenyl)-X6-sulfanyl]oxidanamine

[0301] Step A: TFA (10 mL) was cooled to ice temperature, then 2-methylpropan-2-yl ({[dioxo(2,4,6-trimethylphenyl)-λ6-sulfanyl]oxy}amino)methanoate (1 g, 3.17 mmol) was added. The reaction mixture was stirred at 0 °C for 1.5 h and then poured onto crushed ice. The resulting mixture was filtered. The filter cake was washed with water and dried under vacuum to obtain [dioxo(2,4,6-trimethylphenyl)-λ6-sulfanyl]oxidanamine (600 mg, 2.79 mmol, 87%) as a white solid. This material was taken though to the next step without any purification.6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazin-4-amineHATU, NH4Cl HCl, NaNO2DIEA, DMF DMF Step A Step BPOCl₃ HCl / EA 140 °C, 5 h 80 °CStep E Step C

[0302] Step A: To a solution of 2-amino-5-fluoro-3-(trifluoromethyl)benzoic acid (2 g, 8.9mmol) in DMF (20 mL) was added NH₄Cl (0.58 g, 10.7 mmol), DIEA (3.48 g, 26.9 mmol) and HATU (5.11 g, 13.4 mmol). The reaction mixture was stirred at 25 °C for 1 h. The mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-50%Patent Application Atty. Docket No. ENTX-038PCT ethyl acetate in petroleum ether to afford 2-amino-5-fluoro-3-(trifluoromethyl)benzamide (2 g, 8.1 mmol, 90%) as a white solid. LCMS: ESI m / z 223 [M + H]+.

[0303] Step B: To a solution of 2-amino-5-fluoro-3-(trifluoromethyl)benzamide (2 g, 9 mmol) in DMF (30 mL) was added HC1 (2M, 15 mL, 30 mmol) and sodium nitrite (0.75 g, 10.8 mmol). The mixture was stirred at 25 °C for 1 h. The mixture was diluted with EA and washed with NaHCO3and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-50% ethyl acetate in petroleum ether to afford 6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazin-4(3H)-one (1 g, 4.29 mmol, 47%) as a white solid. LCMS: ESI m / z 234 [M + H]+.

[0304] Step C: A solution of 6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazin-4(3H)-one (200 mg, 0.86 mmol) in POCl3(4 mL, 42.8 mmol) was stirred at 140 °C for 5h. The mixture was diluted with EA and washed with NaHCO3and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-30% ethyl acetate in petroleum ether to afford 4-chloro-6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazine (60 mg, 0.24 mmol, 27%) as a yellow solid. LCMS: ESI m / z 252 [M + H]+.

[0305] Step D: To a solution of 4-chloro-6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazine (460 mg, 1.8 mmol) in 2,2,2-Trifluoroethanol (6 mL) was added DMBNH2 (611 mg, 3.6 mmol). The mixture was stirred at 25 °C for 1 h. The mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-50% ethyl acetate in petroleum ether to afford 4-{[(2,4-dimethoxyphenyl)methyl]amino}-6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazine (600 mg, 1.6 mmol, 85%) as a yellow solid. LCMS: ESI m / z 383 [M + H]+.

[0306] Step E: A solution of 4-{[(2,4-dimethoxyphenyl)methyl]amino}-6-fluoro-8-(trifhjoromethyl)benzo[d][l,2,3]triazine (600 mg, 1.6 mmol) in HC1 (4 M in EA, 10 mL, 40 mmol) was added to 20 mL sealed tube. The mixture was stirred at 80 °C for 2 h. The cooled mixture was quenched with NaHCO3, diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-50% ethyl acetate in petroleum ether to afford 6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazin-4-amine (120 mg, 0.52 mmol, 32%) as a yellow solid. LCMS: ESI m / z 233 [M + H]+.Patent Application Atty. Docket No. ENTX-038PCT Exemplary synthetic proceduresExample 1: N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-[(dimethylamino)methyl]-l-oxo-2,3-dihydro-lH-imidazo[l',2':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide

[0307] Step A: To a solution of 2-azanylidene-6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2,2-difluoroethyl)-3H-benzo[d]imidazole-4-carbonitrile (200 mg, 0.437 mmol) in DMA (2 mL) was added 1,3 -dibrom opropan-2-one (471.70 mg, 2.185 mmol). The reaction mixture was stirred at 120 °C in a sealed tube for 2h. The cooled reaction mixture was diluted with EtOAc and water. The organic phase was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by columnPatent Application Atty. Docket No. ENTX-038PCT chromatography on silica gel (MeOH in DCM = 0 ~ 3%) to obtain 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(2,2-difluoroethyl)-2-[(dimethylamino)methyl]benzo[4,5]imidazo[l,2-a]imidazole-5-carbonitrile (50 mg, 0.084 mmol, 19%) as a yellow solid. LCMS: 539.0, 537.0 [M+H]+.

[0308] Step B: To a solution of 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(2,2-difluoroethyl)-2-[(dimethylamino)methyl]benzo[4,5]imidazo[l,2-a]imidazole-5-carbonitrile (50 mg, 0.084 mmol), 5-fluoro-3-(trifluoromethyl)benzene-l-carboxamide (20.76 mg, 0.100 mmol), CS2CO3 (54.43 mg, 0.167 mmol), XANT PHOS (9.67 mg, 0.017 mmol) in dioxane (2 mL) was added Pd2(dba)3 (7.65 mg, 0.008 mmol). The reaction mixture was stirred at 100 °C under N2 for 2 h. The cooled reaction mixture was quenched with water, extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel (MeOH in DCM = 0 ~ 3%) to obtain N-{6-[(2-chloro-5-fluorophenyl)carbonyl]-5-cyano-9-(2,2-difluoroethyl)-2-[(dimethylamino)methyl]benzo[4,5]imidazo[l,2-a]imidazol-7-yl}-5-fluoro-3-(trifluoromethyl)benzamide (34 mg, 0.051 mmol, 61%) as an yellow solid. LCMS: 664.1 [M+H]+.

[0309] Step C: To a solution ofN-{6-[(2-chloro-5-fluorophenyl)carbonyl]-5-cyano-9-(2,2-difluoroethyl)-2-[(dimethylamino)methyl]benzo[4,5]imidazo[l,2-a]imidazol-7-yl}-5-fluoro-3-(trifluoromethyl)benzamide (34 mg, 0.051 mmol) in ACN (2 mL) and H2O (1 mL) was added KOH (14.3 mg, 0.256 mmol). The reaction was stirred at 25 °C for 1 h. The reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated to afford {N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-[(dimethylamino)methyl]-3-hydroxy-l -oxo-2, 3-dihydro-lH-imidazo[l',2': l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (30 mg, 0.035 mmol, 69%) as a yellow solid. LCMS (ESI) m / z: 682.1 [M+H]+

[0310] Step D: To a solution ofN-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-[(dimethylamino)methyl]-3-hydroxy-l-oxo-2,3-dihydro-lH-imidazo[l',2':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (30 mg, 0.035 mmol) in 2,2,2-trifluoroacetic acid (1 mL) was added triethylsilane (20.4 mg, 0.176 mmol). The reaction mixture was stirred at 70 °C for 10 min. The cooled reaction mixture was concentrated. The residue was purified by prep-HPLC to obtain N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-Patent Application Atty. Docket No. ENTX-038PCT difluoroethyl)-8-[(dimethylamino)methyl]-l-oxo-2,3-dihydro-lH-imidazo[l',2':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (15.2 mg, 0.023 mmol, 65%) as a white solid. LCMS: 666.1 [M+H]+. 'H NMR (400 MHz, DMSO-d6) 8 10.61 (s, 1H), 9.78 (s, 1H), 9.43 (s, 1H), 8.19 (s, 1H), 8.02 -7.92 (m, 1H), 7.87 - 7.65 (m, 3H), 7.38 - 7.26 (m, 1H), 7.17 - 7.07 (m, 1H), 6.74 - 6.36 (m, 2H), 6.17 (brs, 1H), 4.94 - 4.62 (m, 3H), 4.44 - 4.33 (m, 2H), 2.85 -2.80 (m, 6H).Example 2: N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide, Example 6: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 7: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCT

[0311] Step A: To a solution of 6-bromo-5-(2-chloro-5-fluorobenzoyl)-l-(2,2-difluoroethyl)-2-hydrazineyl-lH-benzo[d]imidazole-4-carbonitrile (200 mg, 0.42 mmol) in dioxane (10 mL) was added triethoxymethane (627 mg, 4.2 mmol). The mixture was stirred at 100 °C for 2 h. The cooled mixture was diluted with water, extracted with EA. The organic phase was washed with brine, dried over Na2SO4and concentrated. The residue was purified using silica gel column chromatography eluting with 0-100% ethyl acetate in petroleum ether to afford 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-9H-benzo[4,5]imidazo[2,l-c][l,2,4]triazole-5-carbonitrile (60 mg, 0.12 mmol, 29%) as a yellow solid. LCMS: ESI m / z 482 [M + H]+.

[0312] Step B: To a solution of 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-9H-benzo[4,5]imidazo[2,l-c][l,2,4]triazole-5-carbonitrile (60 mg, 0.12 mmol) in dioxane (5 mL) was added 3-fluoro-5-(trifluoromethyl)benzamide (31 mg, 0.15 mmol), Cs2CO3 (81 mg, 0.25 mmol), Xantphos (15 mg, 0.025 mmol) and Pd2(dba)3(12 mg, 0.012 mmol). The mixture wasPatent Application Atty. Docket No. ENTX-038PCT stirred at 100 °C under N2 for 3 h. The cooled mixture was diluted with water, extracted with EA. The organic phase was washed with brine, dried over Na2SO4and concentrated. The residue was purified using silica gel column chromatography eluting with 0-100% ethyl acetate in petroleum ether to afford N-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-9H-benzo[4,5]imidazo[2,l-c][l,2,4]triazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (60 mg, 0.099 mmol, 79%) as a yellow solid. LCMS: ESI m / z 609 [M + H]+.

[0313] Step C: To a solution ofN-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-9H-benzo[4,5]imidazo[2,l-c][l,2,4]triazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (50 mg, 0.082 mmol) in ACN (3 mL) and H2O (0.5 mL) was added KOH (46 mg, 0.82 mmol). The mixture was stirred at 25 °C for 1 h. The cooled mixture was diluted with water, extracted with EA. The organic phase was washed with brine, dried over Na2SO4 and concentrated. The residue was purified using silica gel column chromatography eluting with 0-50% ethyl acetate in petroleum ether to afford N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (30 mg, 0.048 mmol, 58%) as a yellow solid. LCMS: ESI m / z 627 [M + H]+.

[0314] Step D: To a solution of N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (30 mg, 0.048 mmol) in TFA (2 mL) was added Et3SiH (1 mL, 6.26 mmol). The mixture was stirred at 75 °C for 1 h. The cooled mixture was concentrated under vacuum. The residue was purified using pre-HPLC to afford N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydro- [1,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (10.3 mg, 0.017 mmol, 35%) as a white solid. LCMS: ESI m / z 611 [M + H]+. ’H NMR (400 MHz, DMSO-d6) 5 10.65 (s, 1H), 9.49 (s, 1H), 8.96 (s, 1H), 7.96 (d, J= 8.4 Hz, 1H), 7.84 - 7.65 (m, 3H), 7.36 - 7.26 (m, 1H), 7.16 - 7.03 (m, 1H), 6.84 - 5.88 (m, 3H), 4.97 - 4.58 (m, 2H). N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydro- [1,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (130 mg, 0.21 mmol) was purified by chiral prep-SFC separation method lc(Step E) to afford two isomers. Both were acidified with HC1 (1 M), concentrated under vacuum respectively and then purified by prep-HPLC to afford (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-Patent Application Atty. Docket No. ENTX-038PCT oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (40.2 mg, 0.066 mmol, 62%) as a white solid. LCMS: ESI m / z 611 [M + H]+. 'H NMR (400 MHz, DMSO-d6) 8 10.71 (s, 1H), 9.51 (s, 1H), 9.07 (s, 1H), 7.96 (d, J = 8.4 Hz, 1H), 7.86 - 7.65 (m, 3H), 7.36 - 7.26 (m, 1H), 7.15 - 7.06 (m, 1H), 6.84 - 5.90 (m, 3H), 4.93 -4.68 (m, 2H). And (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (28.8 mg, 0.047 mmol, 44%) as a white solid. LCMS: ESI m / z 611 [M + H]+. ’H NMR (400 MHz, DMSO-d6) 8 10.69 (s, 1H), 9.51 (s, 1H), 9.05 (s, 1H), 7.96 (d, J = 8.4 Hz, 1H), 7.84 - 7.68 (m, 3H), 7.36 - 7.26 (m, 1H), 7.15 - 7.06 (m, 1H), 6.80 - 5.85 (m, 3H), 4.93 -4.68 (m, 2H).Example 3: N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methyl-l-oxo-2,3-dihydro-lH-imidazo[l',2':l,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5- (trifluoromethyl)benzamideKOH, MeCN,H2O, rt, 1hStep C

[0315] Step A. To a stirred solution of 2-azanylidene-6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2,2-difluoroethyl)-3H-benzo[d]imidazole-4-carbonitrile (600 mg, 1.31 mmol) in DMA (8 mL) was added l-bromopropan-2-one (359 mg, 2.62 mmol) at rt. After stirred at 100 °C overnight, the mixture was quenched with ice-water and extracted with EtOAc (10 mL*3). The comLined organic phase was washed with brine, dried with Na2SO4, filtered. ThePatent Application Atty. Docket No. ENTX-038PCT residue was purified by chromatography (silica gel, 0- 80 %, EtOAc in PE) to give 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(2,2-difluoroethyl)-2-methylbenzo[4,5]imidazo[l,2-a]imidazole-5-carbonitrile (400 mg, 0.81 mmol, 61%) as a brown solid. LCMS: m / z 496 [M+H]+.

[0316] Step B. To a stirred mixture of 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(2,2-difluoroethyl)-2-methylbenzo[4,5]imidazo[l,2-a]imidazole-5-carbonitrile (120 mg, 0.24 mmol) in dioxane (8 mL) was added 5-fluoro-3-(trifluoromethyl)benzene-l-carboxamide (75 mg, 0.36 mmol), Pd2(dba)3(44 mg, 0.05 mmol), XANT PHOS (56 mg, 0.09mmol) and CS2CO3 (197 mg, 0.61 mmol) at rt. After stirred at 100 °C under N2 for Ih, the cooled mixture was quenched with ice-water and extracted with EtOAc (15 mL*3). The comLined organic phase was washed with brine, dried with Na2SO4, filtered and concentrated. The residue was purified by chromatography (silica gel, 0-8 %, MeOH in DCM) to give N-{6-[(2-chloro-5-fluorophenyl)carbonyl]-5-cyano-9-(2,2-difluoroethyl)-2-methylbenzo[4,5]imidazo[l,2-a]imidazol-7-yl}-3-fluoro-5-(trifluoromethyl)benzamide (100 mg, 0.16 mmol, 66 %) as a yellow solid. LCMS: m / z 622 [M+H]+.

[0317] Step C. To a stirred solution ofN-{6-[(2-chloro-5-fluorophenyl)carbonyl]-5-cyano-9-(2,2-difluoroethyl)-2-methylbenzo[4,5]imidazo[l,2-a]imidazol-7-yl}-3-fluoro-5-(trifluoromethyl)benzamide (100 mg, 0.16 mmol) in ACN (3 mL) / H2O (1 mL) was added KOH (90 mg, 1.6 mmol) at rt. After stirred at rt for 1 h, the mixture was quenched with ice-water and extracted with EtOAc (10 mL*3). The comLined organic phase was washed with brine, dried with Na2SO4, filtered and concentrated to give crude N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-8-methyl-l-oxo-2,3-dihydro-lH-imidazo[l',2':l,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (90 mg, 0.14 mmol, 87%) as a brown solid. LCMS: m / z 640 [M+H]+.

[0318] Step D. To a stirred mixture of N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3 -hydroxy-8-methyl- 1 -oxo-2,3 -dihydro- lH-imidazo[ 1 ',2': 1,2]imidazo[5,4-e]isoindol-4-yl]-3 -fluoro-5-(trifluoromethyl)benzamide (90 mg, 0.14 mmol) in TFA (2 mL) was added Et3SiH (0.5 mL) at rt. After stirred at 50 °C for Ih, the cooled mixture was concentrated. The residue was purified by prep-HPLC (C18, 20 ~ 90 % MeCN in H2O with 0.1 % FA) to give N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methyl-l-oxo-2,3-dihydro-lH-imidazo[r,2':l,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (33 mg,Patent Application Atty. Docket No. ENTX-038PCT 0.05 mmol, 37%) as a white solid. LCMS: m / z 624 [M+H]+. ’H NMR (400 MHz, DMSO-d6) 5 10.62 (s, 1H), 9.43 (s, 1H), 7.97 (d, J= 8.4 Hz, 1H), 7.86 (s, 1H), 7.82 - 7.76 (m, 2H), 7.72 (s, 1H), 7.34 - 7.29 (m, 1H), 7.15 - 7.09 (m, 1H), 6.70- 6.44 (m, 1H), 6.16 (brs, 1H), 4.89 - 4.75 (m, 2H), 2.38 (s, 3H).Example 4: N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]>midazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide

[0319] Step A: To a solution of 6-bromo-5-(2-chloro-5-fluorobenzoyl)-l-(2,2-difluoroethyl)-2-hydrazineyl-lH-benzo[d]imidazole-4-carbonitrile (80 mg, 0.17 mmol) in dioxane (10 mL) was added 1,1,1 -tri ethoxy ethane (275 mg, 1.7 mmol) and TFA (38 mg, 0.34 mmol). The mixture was stirred at 110 °C overnight. The cooled mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-30% ethyl acetate in petroleum ether to afford 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-3-methyl-9H-benzo[4,5]imidazo[2,l-Patent Application Atty. Docket No. ENTX-038PCT c][l,2,4]triazole-5-carbonitrile (20 mg, 0.04 mmol, 24%) as a yellow solid. LCMS: ESI m / z 496 [M + H]+.

[0320] Step B: To a solution of 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-3-methyl-9H-benzo[4,5]imidazo[2,l-c][l,2,4]triazole-5-carbonitrile (20 mg, 0.04 mmol) in dioxane (5 mL) was added 3-fluoro-5-(trifluoromethyl)benzamide (13 mg, 0.063 mmol), CS2CO3 (27 mg, 0.083 mmol), Xantphos (5 mg, 0.009 mmol) and Pd2(dba)3(4 mg, 0.004 mmol). The mixture was stirred at 100 °C under N2 for 2 h. The cooled mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-10% methanol in dichloroform to afford N-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-3-methyl-9H-benzo[4,5]imidazo[2,l-c][l,2,4]triazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (20 mg, 0.032 mmol, 79%) as a yellow solid. LCMS: ESI m / z 623 [M + H]+.

[0321] Step C: To a solution of N-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-3-methyl-9H-benzo[4,5]imidazo[2,l-c][l,2,4]triazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (20 mg, 0.032 mmol) in ACN (5 mL) and H2O (1 mL) was added KOH (18 mg, 0.32 mmol). The mixture was stirred at 25 °C for 1 h. The mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-10% methanol in dichloroform to afford N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-9-methyl- 1-oxo- 1,2,3, 6-tetrahydro-[ 1,2, 4]triazolo[3', 4':2,3]imidazo[4, 5-e]isoindol -4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (15 mg, 0.023 mmol, 72%) as a yellow solid. LCMS: ESI m / z 641 [M + H]+.

[0322] Step D: To a solution ofN-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-9-methyl-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (15 mg, 0.023 mmol) in TFA (2 mL) was added Et3SiH (27 mg, 0.23 mmol). The mixture was stirred at 75 °C for 1 h. The cooled mixture was concentrated under vacuum. The residue was purified using pre-HPLC to afford N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (5.2 mg, 0.008 mmol, 35%) as a white solid. LCMS: ESI m / z 625 [M + H]+. ’H NMR (400 MHz,Patent Application Atty. Docket No. ENTX-038PCT DMSO-d6) 8 10.57 (s, 1H), 9.41 (s, 1H), 7.96 (d, J= 7.6 Hz, 1H), 7.86- 7.63 (m, 3H), 7.35 -7.24 (m, 1H), 7.16 - 7.02 (m, 1H), 6.87 - 5.80 (m, 3H), 4.92 - 4.47 (m, 2H), 3.01 (s, 3H).Example 5: N-(9-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methyl-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideNCSDMF. 0 °CStep A

[0323] Step A: To a solution ofN-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methyl-l-oxo-2,3-dihydro-lH-imidazo[r,2':l,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (20 mg, 0.032 mmol) in DMF (3 mL) was added NCS (7 mg, 0.048 mmol). The reaction mixture was stirred at 0 °C for 0.5h. The cooled reaction mixture was diluted with H₂O, extracted with EA. The organic phase was washed with brine, dried by Na₂SO₄, filtered and concentrated. The residue was purified using prep-HPLC (0.1% NH3. H2O as additive) to afford N-[9-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methyl-l-oxo-2,3-dihydro-lH-imidazo[T,2':l,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (1.8 mg, 0.003 mmol, 8%) as a white solid. LCMS: ESI m / z 658.3 [M+H]+. 'H NMR (400 MHz, DMSO-d6) 8 10.65 (s, 1H), 9.26 (s, 1H), 8.02 (d, J= 8.2 Hz, 1H), 7.91 (s, 1H), 7.88 - 7.77 (m, 2H), 7.39 - 7.28 (m, 2H), 7.21 - 7.10 (m, 1H), 6.77 - 6.42 (m, 2H), 6.13 (brs, 1H), 4.89 -4.68 (m, 2H), 2.34 (s, 3H).Example 8: N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-2,3-dihydro-lH-pyrazolo [1 ',5': 1,2] imidazo[5,4-e] isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCT

[0324] Step A: To a solution of 2-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-[(2,2-difluoroethyl)amino]benzene-l -carbonitrile (600 mg, 1.39 mmol) in dioxane (10 mL) was added 1,1,1 -tri ethoxy ethane (2250 mg, 13.87 mmol) and 2,2,2-trifluoroacetic acid (0.011 mL, 0.14 mmol). The reaction was stirred at 110 °C for 16 h. The reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated. The residue was purified by column chromatography on silica gel (EA in PE = 0 ~ 50%) to obtain 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2,2-difluoroethyl)-2-methylbenzo[d]imidazole-4-carbonitrile (400 mg, 0.88 mmol, 63%) as an yellow oil. LCMS: 457.0, 455.0 [M+H]+.

[0325] Step B: To a solution of 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2,2-difluoroethyl)-2-methylbenzo[d]imidazole-4-carbonitrile (400 mg, 0.88 mmol) in DCM (4 mL) was added [dioxo(2,4,6-trimethylphenyl)-X6-sulfanyl]oxidanamine (283 mg, 1.31 mmol). The reaction was stirred at rt for 16 h and then filtered. The filter cake was washed with DCM and petroleum ether to give l-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-7-cyano-3-Patent Application Atty. Docket No. ENTX-038PCT (2,2-difluoroethyl)-2-methylbenzo[d]imidazol-l-ium (300 mg, 0.508 mmol, 58%) as a white solid. LCMS: 473.0, 471.0, [M+H]+.

[0326] Step C: A solution of l-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-7-cyano-3-(2,2-difluoroethyl)-2-methylbenzo[d]imidazol-l-ium (200 mg, 0.34 mmol) in DMF-DMA (2 mL) was stirred at 100 °C for 1 h. The cooled reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4and concentrated. The residue was purified by column chromatography on silica gel (EA in PE = 0 ~ 45%) to obtain 6-bromo-7-[(2-chloro-5-fluorophenyl)carbonyl]-4-(2,2-difluoroethyl)benzo[4,5]imidazo[2,l-e]pyrazole-8-carbonitrile (30 mg, 0.062 mmol, 18%) as an yellow oil. LCMS: LCMS: 482.0, 480.0, [M+H]+.

[0327] Step D: To a solution of 6-bromo-7-[(2-chloro-5-fluorophenyl)carbonyl]-4-(2,2-difluoroethyl)benzo[4,5]imidazo[2,l-e]pyrazole-8-carbonitrile (30 mg, 0.062 mmol), 5-fluoro-3-(trifluoromethyl)benzene-l-carboxamide (15 mg, 0.075 mmol), CS2CO3 (40 mg, 0.12 mmol), XANT PHOS (7 mg, 0.012 mmol) in dioxane (3 mL) was added Pd2(dba)3(5 mg, 0.006 mmol). The reaction mixture was stirred at 100 °C under N2 for 2 h. The cooled reaction mixture was quenched with water, extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel (MeOH in DCM = 0 ~ 3%) to obtain N-{7-[(2-chloro-5-fluorophenyl)carbonyl]-8-cyano-4-(2,2-difluoroethyl)benzo[4,5]imidazo[2,l-e]pyrazol-6-yl}-5-fluoro-3-(trifluoromethyl)benzamide (30 mg, 0.049 mmol, 79%) as an yellow solid. LCMS: 607.1 [M+H]+.

[0328] Step E: To a solution of N-{7-[(2-chloro-5-fluorophenyl)carbonyl]-8-cyano-4-(2,2-difluoroethyl)benzo[4,5]imidazo[2,l-e]pyrazol-6-yl}-5-fluoro-3-(trifluoromethyl)benzamide (30 mg, 0.049 mmol) in ACN (2 mL) and H₂O (0.5 mL) was added KOH (14 mg, 0.25 mmol). The reaction was stirred at RT for 0.5 h. The reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated to afford N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-2,3-dihydro-lH-pyrazolo[l',5': l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (30 mg, 0.048 mmol, 97%) as a yellow solid. LCMS: 625.1 [M+H]+.

[0329] Step F: To a solution ofN-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-2,3-dihydro-lH-pyrazolo[T,5':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-Patent Application Atty. Docket No. ENTX-038PCT (trifluoromethyl)benzamide (30 mg, 0.048 mmol) in 2,2,2-trifluoroacetic acid (2 mL) was added triethylsilane (28 mg, 0.24 mmol). The reaction mixture was stirred at 90 °C for 3 hr. The cooled reaction mixture was concentrated. The residue was purified by prep-HPLC to obtain N-[3-(2-chl oro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l -oxo-2, 3-dihy dro-lH-pyrazolo[l',5': l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (7.3 mg, 0.012 mmol, 25%) as a white solid LCMS: 609.1 [M+H]+. ‘HNMR (400 MHz, DMSO-d6) 5 10.52 (s, 1H), 9.16 (s, 1H), 7.97 - 7.92 (m, 1H), 7.87 -7.84 (m, 1H), 7.82 - 7.70 (m, 3H), 7.33 -7.26 (m, 1H), 7.14 - 7.03 (m, 1H), 6.68 - 6.35 (m, 2H), 6.22 - 6.01 (m, 2H), 4.92 - 4.65 (m, 2H).Example 9: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l -oxo-1,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 10: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide

[0330] Step A: N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (180 mg, 0.28 mmol) was purified with chiral prep-SFC method 2 to afford two isomers, which were acided with HC1(1 M) and concentrated under vacuum to afford (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (65.5 mg, 0.105 mmol, 72%) as a white solid. LCMS: ESI m / z 625 [M + H]+. ’H NMR (400 MHz, DMSO-d6) 5 10.77 (s, 1H), 9.55 (s, 1H), 8.02 - 7.90 (m, 2H), 7.88 - 7.79 (m, 1H), 7.76 (s,Patent Application Atty. Docket No. ENTX-038PCT 1H), 7.34 -7.28 (m, 1H), 7.16 - 7.05 (m, 1H), 6.82- 6.08 (m, 3H), 4.96 -4.73 (m, 2H), 3.17 (s, 3H). And (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5- (trifluoromethyl)benzamide (70 mg, 0.11 mmol, 77%) as a white solid. LCMS: ESI m / z 625 [M + H]+. 'HNMR (400 MHz, DMSO-d6) 5 10.74 (s, 1H), 9.54 (s, 1H), 8.03 - 7.92 (m, 2H), 7.87 -7.78 (m, 1H), 7.75 (s, 1H), 7.36 - 7.27 (m, 1H), 7.16 - 7.04 (m, 1H), 6.77 - 5.98 (m, 3H), 4.96 -4.72 (m, 2H), 3.16 (s, 3H).Example 11: 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((6-fluoro-8- (trifluoromethyl)benzo[d][l,2,3]triazin-4-yl)amino)-3,6-dihydro- [l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-one

[0331] Step A: To a solution of 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-9H-benzo[4,5]imidazo[2,l-c][l,2,4]triazole-5-carbonitrile (100 mg, 0.207 mmol) in ACN (5 mL) and H2O (1 mL) was added KOH (58 mg, 1.04 mmol). The mixture was stirred at 25 °C for 0.5 h. The mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-10% methanol in dichloroform to afford 4-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-3,6-dihydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-one (80 mg, 0.16 mmol, 77%) as a yellow solid. LCMS: ESI m / z 500 [M + H]+.Patent Application Atty. Docket No. ENTX-038PCT

[0332] Step B: To a solution of 4-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-3,6-dihydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-one (80 mg, 0.16 mmol) in TFA (3 mL) was added Et3SiH (185 mg, 1.6 mmol). The mixture was stirred at 70 °C for 1 h. The mixture was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-10% methanol in di chloroform to afford 4-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-3,6-dihydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-one (70 mg, 0.14 mmol, 90%) as a white solid. LCMS: ESI m / z 484 [M + H]+.

[0333] Step C: To a solution of 4-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-3,6-dihydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-one (30 mg, 0.062 mmol) in dioxane (6 mL) was added 6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazin-4-amine (21 mg, 0.093 mmol), Xantphos (7 mg, 0.012 mmol), CS2CO3 (40 mg, 0.12 mmol) and Pd (dba)3 (6 mg, 0.006 mmol). The mixture was stirred at 120 °C under N2 for 2 h. The cooled mixture was diluted with EA and washed with H2O. The organic layer was concentrated under vacuum. The residue was purified using pre-HPLC and then pre-TLC to afford 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((6-fluoro-8-(trifluoromethyl)benzo[d][l, 2, 3]tri azin-4-yl)amino)-3,6-dihydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-one (2.5 mg, 0.004 mmol, 6%) as a white solid. LCMS: ESI m / z 636 [M + H]+. 'H NMR (400 MHz, DMSO-d6) 8 10.49 (s, 1H), 9.47 (s, 1H), 9.00 (s, 1H), 8.52 (d, J= 8.0 Hz, 1H), 8.45 - 8.27 (m, 1H), 7.92 (s, 1H), 6.97 - 6.79 (m, 2H), 6.79 - 6.72 (m, 1H), 6.71- 6.42 (m, 1H), 6.20 - 5.96 (m, 1H), 4.89 -4.74 (m, 2H).Example 12: 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazin-4-yl)amino)-9-methyl-3,6-dihydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-onePatent Application Atty. Docket No. ENTX-038PCTExample 12

[0334] Step A: To a solution of 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-3-methyl-9H-benzo[4,5]imidazo[2,l-c][l,2,4]triazole-5-carbonitrile (170 mg, 0.34 mmol) in ACN (15 mL) and H₂O (3 mL) was added KOH (192 mg, 3.42 mmol). The mixture was stirred at 25 °C for 1 h. The mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-10% methanol in dichloroform to afford 4-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-9-methyl-3,6-dihydro- [1,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-1(2H)-one (140 mg, 0.27 mmol, 79%) as a yellow solid. LCMS: ESI m / z 514 [M + H]+.

[0335] Step B: To a solution of 4-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-9-methyl-3,6-dihydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-one (140 mg, 0.27 mmol) in TFA (4 mL) was added Et3SiH (2 mL, 12.5 mmol). The mixture was stirred at 70 °C for 1 h. The mixture was diluted with EA and washed with saturated NaHCO3and H2O. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-10% methanol in di chloroform to afford 4-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-3,6-dihydro- [1.2.4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-one (100 mg, 0.2 mmol, 73%) as a white solid. LCMS: ESI m / z 498 [M + H]+.Patent Application Atty. Docket No. ENTX-038PCT

[0336] Step C: To a solution of 4-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-3,6-dihydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-one (20 mg, 0.04 mmol) in dioxane (4 mL) was added 6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazin-4-amine (18 mg, 0.08 mmol), Xantphos (5 mg, 0.008 mmol), CS2CO3 (26 mg, 0.08 mmol) and Pdc(dba)3 (4 mg, 0.004 mmol). The mixture was stirred at 120 °C under N2 for 3 h. The mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using prep-HPLC to afford 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((6-fluoro-8-(trifluoromethyl)benzo[d][l,2,3]triazin-4-yl)amino)-9-methyl-3,6-dihydro-[l,2,4]triazolo[3',4':2,3]imidazo[4,5-e]isoindol-l(2H)-one (1 mg, 0.002 mmol, 3%) as a white solid. LCMS: ESI m / z 650 [M + H]+. 'H NMR (400 MHz, CD3OD) δ 8.58 – 8.49 (m, 1H), 8.34 - 8.24 (m, 1H), 8.10 - 7.90 (m, 2H), 7.65 - 7.54 (m, 1H), 7.50 - 7.37 (m, 1H), 7.17 -7.01 (m, 1H), 6.61 - 6.18 (m, 1H), 4.77 - 4.72 (m, 2H), 3.20 (s, 3H).Example 13: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-1-oxo-1,2,3,6-tetrahydro-[1,2,4]triazolo[5',1':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 14: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-Patent Application Atty. Docket No. ENTX-038PCT (trifluoromethyl)benzamideFe / NH4CI NaNO2, SnCI2EtOH, H2O HCI, 0 - rt step A step BDPPA, TEA POCI3HCI, water Tol, 100 °C 120 °C, 48h step C step D step ECuCN K2CO3, DMA NMP step GKOHPd2(dba)3, xantphos MeCN, H2O Cs2CO3, dioxane step H step IExample 14 Example 13Patent Application Atty. Docket No. ENTX-038PCT

[0337] Step A: To a solution of (2-chloro-5-fluorophenyl)(2,6-dibromo-4-fluoro-3-nitrophenyl)m ethanone (5 g, 11.0 mmol) in EtOH (50 mL) and H2O (10 mL) was added Fe (3.07 g, 54.9 mmol) and NH4CI (2.94 g, 54.9 mmol)). The reaction was stirred at 80 °C for 2 h. The reaction mixture was filtrated, concentrated, diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4and concentrated. The residue was triturated with PE / EA=5 / 1 to afford (3-amino-2,6-dibromo-4-fluorophenyl)(2-chloro-5-fluorophenyl)methanone (3.7 g, 8.69 mmol, 79%) as a yellow solid. LCMS: 425.8 [M+H]+.

[0338] Step B: To a solution of (3-amino-2,6-dibromo-4-fluorophenyl)(2-chloro-5-fluorophenyl)methanone (4 g, 9.40 mmol) in concentrated hydrochloric acid (40 mL) was added dropwise a solution of sodium nitrite (0.71 g, 10.34 mmol) in water (5 mL) at -5° C (with a rate of the dropwise addition to keep the internal temperature below 5° C). After 0.5 h at -5° C, the above-mentioned solution was added dropwise into a solution of stannous chloride dihydrate (5.30 g, 23.50 mmol) in concentrate HC1 solution ( 30 mL) at -15°C. After the dropwise addition completed, the mixture was allowed to warm to 35° C and stirred for 12 h. The mixture was cooled to 0 °C, adjusted to pH=9 with concentrated ammonia aqueous solution. The aqueous layer was extracted with EtOAc. The organic layers were washed with water, dried over Na2SO4, filtered and evaporated to give the residue which was purified by column chromatography to provide the title compound (2-chloro-5-fluorophenyl)(2,6-dibromo-3-diazanyl-4-fluorophenyl)methanone (1.4 g, 3.18 mmol, 34%) yellow solid. LCMS: 440.9 [M+H]+.

[0339] Step C: To a solution of hydrogen chloride (2 mL) in H2O (30 mL) was added (2-chloro-4-fluorophenyl)(2,6-dibromo-3-diazanyl-4-fluorophenyl)methanone (2 g, 4.54 mmol) and formylmethanoic acid (670 mg, 4.54 mmol, 50% w / w). The mixture was stirred at room temperature for 5 h. The mixture was extracted with ethyl acetate. The organic phase was washed with a saturated brine, and dried on anhydrous sodium sulfate. The organic phase was separated and concentrated. The residue was purified by column chromatography on silica gel (eluted with MeOH / DCM = 0-10%) to afford [(lE)-2-{2,4-dibromo-3-[(2-chloro-4-fluorophenyl)carbonyl]-6-fluorophenyl}diazanylidene]acetic acid (2 g, 4.03 mmol, 89%) as a white solid. LCMS: 494.9 [M-H]+.

[0340] Step D: To a solution of [(lE)-2-{2,4-dibromo-3-[(2-chloro-4-fluorophenyl)carbonyl]-6-fluorophenyl}diazanylidene]acetic acid (2.25 g, 4.53 mmol) in toluene (25 mL) was added DPPA (1.87 g, 6.80 mmol) and TEA (1.38 g, 13.6 mmol). The reactionPatent Application Atty. Docket No. ENTX-038PCT mixture was stirred at 100 °C for 2 h. The cooled mixture was diluted with H₂O, extracted with EA. The organic phase was separated and concentrated. The residue was purified by column chromatography on silica gel (eluted with MeOH / DCM = 0-10%) to afford 2-{2,4-dibromo-3-[(2-chloro-4-fluorophenyl)carbonyl]-6-fluorophenyl}-4H,2H,3H-l,2,4-triazol-3-one (1.07 g, 2.17 mmol, 48%) as abrown solid. LCMS: 493.9 [M+H]+.

[0341] Step E: A solution of2-{2,4-dibromo-3-[(2-chloro-4-fluorophenyl)carbonyl]-6-fluorophenyl}-4H,2H,3H-l,2,4-triazol-3-one (1.6 g, 3.24 mmol) in POCl₃ (20 mL) was stirred at 120 °C for 24 h. The mixture was concentrated. The residue was diluted with aqueous NaHCO3, extracted with EA. The organic phase was separated and concentrated. The residue was purified by column chromatography on silica gel (eluted with MeOH / DCM = 0-10%) to afford (2-chloro-4-fluorophenyl)[2,6-dibromo-3-(5-chloro-l,2,4-triazol-l-yl)-4-fluorophenyl]methanone (600 mg, 1.17 mmol, 36%) as abrown oil. LCMS: 511.8 [M+H]+.

[0342] Step F: A solution of (2-chloro-4-fluorophenyl)[2,6-dibromo-3-(5-chloro-l,2,4-triazol-l-yl)-4-fluorophenyl]methanone (600 mg, 1.17 mmol) in DMF (20 mL) was added K₂CO₃ (486 mg, 3.52 mmol) and 2,2-difluoroethan- 1 -amine (4.75 g, 58.6 mmol). The mixture was stirred at 100 °C for 2 h. The reaction mixture was diluted with H₂O, extracted with EA. The organic was concentrated to give a residue. The residue was purified by column (eluted with EA: PE=50%) to give (2-chloro-4-fluorophenyl)[6,8-dibromo-4-(2,2-difluoroethyl)benzo[4,5]imidazo[2,3-e][l,2,4]triazol-7-yl]methanone (450 mg, 0.839 mmol, 72%) as a brown solid. LCMS: 536.9 [M+H]+.

[0343] Step G: To a solution of (2-chloro-4-fluorophenyl)[6,8-dibromo-4-(2,2-difluoroethyl)benzo[4,5]imidazo[2,3-e][l,2,4]triazol-7-yl]methanone (590 mg, 1.10 mmol) in NMP (6 mL) was added CuCN (118 mg, 1.32 mmol). The mixture was stirred at 140 °C under N2 for 2 h. The cooled mixture was diluted with water and extracted with EA twice. The comLined organic layer was washed with water, brine, dried and concentrated. The residue was purified by column (eluted with EA: PE=50%) to get a crude. The crude was purified by pre-HPLC to give 6-bromo-7-[(2-chloro-4-fluorophenyl)carbonyl]-4-(2,2-difluoroethyl)benzo[4,5]imidazo[2,3-e][l,2,4]triazole-8-carbonitrile (110 mg, 0.228 mmol, 21%) as a brown solid. LCMS: 484.0 [M+H]+.

[0344] Step H: A solution of 6-bromo-7-[(2-chloro-4-fluorophenyl)carbonyl]-4-(2,2-difluoroethyl)benzo[4,5]imidazo[2,3-e][l,2,4]triazole-8-carbonitrile (115 mg, 0.238 mmol), 5-Patent Application Atty. Docket No. ENTX-038PCT fluoro-3-(trifluoromethyl)benzene-l-carboxamide (51.8 mg, 0.250 mmol), Pd₂(dba)₃ (21.8 mg, 24 μmol), xantphos (27.5 mg, 48 μmol), Cs₂CO₃ (232 mg, 0.715 mmol) in dioxane (6 mL) was stirred at 100 °C for 2h. The reaction mixture was concentrated in vacuo. The residue was purified using silica gel column chromatography eluted with MeOH in DCM (gradient:0%~10%) to afford N-{7-[(2-chloro-4-fluorophenyl)carbonyl]-8-cyano-4-(2,2-difluoroethyl)benzo[4,5]imidazo[2,3-e][l,2,4]triazol-6-yl}-3-fluoro-5-(trifluoromethyl)benzamide (110 mg, 0.181 mmol, 76%) as a brown solid. LCMS: 609 [M+H]+.

[0345] Step I: To a solution ofN-{7-[(2-chloro-4-fluorophenyl)carbonyl]-8-cyano-4-(2,2-difluoroethyl)benzo[4,5]imidazo[2,3-e][l,2,4]triazol-6-yl}-3-fluoro-5-(trifluoromethyl)benzamide (100 mg, 0.164 mmol) in ACN (2 mL) and H₂O (0.2 mL) was added KOH (46.1 mg, 0.821 mmol). The reaction mixture was stirred at rt for 1 h. The mixture was diluted with aqueous H₂O, extracted with EA. The organic phase was separated and concentrated to afford N-[3-(2-chloro-4-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-2,3-dihydro-lH-[l,2,4]triazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (100 mg, 0.160 mmol, 97%) as a brown solid. LCMS: 627.0 [M+H]+.

[0346] Step J: To a solution of N-[3-(2-chloro-4-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-2,3-dihydro-lH-[l,2,4]triazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (110 mg, 0.175 mmol) in TFA (3 mL) was added TES (0.3 mL). The reaction mixture was stirred at 60 °C for 30 min. Then the reaction mixture was concentrated to give a residue. The residue was purified by pre-HPLC to give N-[3-(2-chloro-4-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-2,3-dihydro-lH-[l,2,4]triazolo[5',r:2,3]imidazo[4,5-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (45 mg, 74 μmol, 42%) as a white solid. LCMS: 611.1 M+H]+. It was purified by chiral prep-SFC method 3 to give (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[5',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (14.4 mg, 24 μmol, 13%, ee=99) as a white solid. LCMS: 611.1 [M+H]+. ’H NMR (400 MHz, CD3OD) 58.18 (s, 1H), 7.91 (s, 1H), 7.72 - 7.65 (m, 3H), 7.29 - 7.25 (m, 1H), 7.03 - 6.98 (m, 1H), 6.55 - 6.28 (m, 3H), 4.85 - 4.78 (m, 2H). And (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[5',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (14.2 mg, 23 μmol, 13%) as a white solid. LCMS: 611.1 [M+H]+. ’H NMR (400 MHz, CD3OD)Patent Application Atty. Docket No. ENTX-038PCT 88.17 (s, 1H), 7.91 (s, 1H), 7.72 - 7.65 (m, 3H), 7.29 - 7.25 (m, 1H), 7.03 - 6.98 (m, 1H), 6.42 (m, 3H), 4.85 - 4.78 (m, 2H).Example 15: N-[(3S)-3-(2-chloro-5-fluorophenyl)-6-(2-fluoroethyl)-l-oxo-2,3-dihydro-lH-[l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide Example 16: N-[(3R)-3-(2-chloro-5-fluorophenyl)-6-(2-fluoroethyl)-l-oxo-2,3-dihydro-lH-[l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamideTriphosgene, TEA THFStep APatent Application Atty. Docket No. ENTX-038PCT

[0347] Step A: To a solution of 2-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-[(2-fluoroethyl)amino]benzene-l -carbonitrile (6 g, 14.47 mmol) and TEA (6.03 mL, 43.4 mmol) in THF (60 mL) was added trichloromethyl [(trichloromethyl)oxy]methanoate (2.15 g, 7.24 mmol) at 0 °C. The reaction was stirred at 25 °C for 1 h. LCMS showed the reaction was completed. The reaction mixture was added to ice water and extracted with EA. The organic layer was washed with brine, dried over Na₂SO₄ and concentrated and the residue was purified by flash chromatography to afford 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2-fluoroethyl)-2-oxo-3H-benzo[d]imidazole-4-carbonitrile (4.9 g, 11.1 mmol, 77%) as a yellow solid. LCMS: 442.0, 440.0 [M+H]+.

[0348] Step B: A solution of 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2-fluoroethyl)-2-oxo-3H-benzo[d]imidazole-4-carbonitrile (4 g, 9.08 mmol) in POCl₃ (40 mL), was added DIEA (2.35 g, 18.16 mmol). Then the mixture was stirred at 120 °C for 16 h. The mixture was concentrated. The residue was diluted with aqueous NaHCO3, extracted with EA. The organic phase was separated and concentrated. The residue was purified by column chromatography on silica gel to afford 6-bromo-2-chloro-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2-fluoroethyl)benzo[d]imidazole-4-carbonitrile (2.74 g, 5.97 mmol, 66%) as a brown oil. LCMS: 459.9, 457.9 [M+H]+.

[0349] Step C: To a solution of 6-bromo-2-chloro-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2-fluoroethyl)benzo[d]imidazole-4-carbonitrile (670 mg, 1.46 mmol) in DMA (7 mL) was added 2-methylpropan-2-yl diazanylmethanoate (385 mg, 2.92 mmol) and triethylamine (0.61 mL, 4.37 mmol) at 25 °C. The reaction was stirred at 100 °C for 16 h. LCMS showed the reaction was completed. The reaction mixture was added to water and extracted with EA. The organic layer was washed with brine, dried over Na₂SO₄ and concentrated and the residue was purified by flash chromatography to afford 2-methylpropan-2-yl (2-{6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-4-cyano-1-(2-fluoroethyl)benzo[d]imidazol-2-yl}diazanyl)methanoate (550 mg, 0.99 mmol, 68%) as a yellow solid. LCMS: 556.0, 554.0 [M+H]+.

[0350] Step D: A solution of 2-methylpropan-2-yl (2-{6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-4-cyano-l-(2-fluoroethyl)benzo[d]imidazol-2-yl}diazanyl)methanoate (450 mg, 0.81 mmol) in TFA (5 mL) was stirred at 25 °C for 10 min. The mixture was concentrated. The residue was diluted with aqueous NaHCO3, extracted with EA. The organic phase was separated and concentrated. The residue was purified by column chromatography onPatent Application Atty. Docket No. ENTX-038PCT silica gel to afford 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-2-diazanyl-l-(2-fluoroethyl)benzo[d]imidazole-4-carbonitrile (220 mg, 0.484 mmol, 60%) as a yellow solid. LCMS: 456.0, 454.0 [M+H]+.

[0351] Step E: To a solution of 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-2-diazanyl-l-(2-fluoroethyl)benzo[d]imidazole-4-carbonitrile (220 mg, 0.48 mmol) in dioxane (5 mL) was added triethoxymethane (717 mg, 4.84 mmol) and TFA (0.004 mL, 0.048 mmol) at 25 °C. The reaction was stirred at 110 °C for 1 h. LCMS showed the reaction was completed. The reaction mixture was added to ice water and extracted with EA. The organic layer was washed with brine, dried over Na₂SO₄ and concentrated and the residue was purified by flash chromatography to afford 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(2-fluoroethyl)benzo[4,5]imidazo[2,l-c][l,2,4]triazole-5-carbonitrile (200 mg, 0.430 mmol, 88.9%) as a yellow solid. LCMS: 466.0, 464.0 [M+H]+.

[0352] Step F: To a solution of 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(2-fluoroethyl)benzo[4,5]imidazo[2,l-c][l,2,4]triazole-5-carbonitrile (200 mg, 0.430 mmol), 3-fluoro-5-(trifluoromethyl)benzene-l-carboxamide (98 mg, 0.473 mmol), XANT PHOS (49.81 mg, 0.086 mmol), CS2CO3 (280 mg, 0.861 mmol) in dioxane (5 mL) wasadded Pd2(dba)3(39 mg, 0.043 mmol). The reaction mixture was stirred at 100 °C under N2 for 2 h. The cooled reaction mixture was quenched with water, extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel (MeOH in DCM = 0 ~ 5%) to obtain N-[3-(2-chloro-5-fluorophenyl)-6-(2-fluoroethyl)-3-hydroxy-l-oxo-2,3-dihydro-lH-[l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (210 mg, 0.35 mmol, 80%) as an yellow solid. LCMS:609.1 [M+H]+.

[0353] Step G: To a solution ofN-[3-(2-chloro-5-fluorophenyl)-6-(2-fluoroethyl)-3-hydroxy-l-oxo-2,3-dihydro-lH-[l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (210 mg, 0.345 mmol) in TFA (4 mL) was added tri ethyl si lane (1 mL). The reaction mixture was stirred at 70 °C for 30 min. Then the reaction mixture was concentrated to give a residue. The residue was purified by pre-HPLC to give N-[3-(2-chloro-5-fluorophenyl)-6-(2-fluoroethyl)- 1 -oxo-2, 3 -dihydro- lH-[ 1,2,4]triazolo[4',3 ': 1,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (170 mg, 0.287 mmol, 83%) as a white solid. LCMS: 593.1 [M+H]+.Patent Application Atty. Docket No. ENTX-038PCT

[0354] Step H: N-[3-(2-chloro-5-fluorophenyl)-6-(2-fluoroethyl)-l -oxo-2, 3-dihydro-lH-[l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (200 mg, 0.337 mmol) was purified by chiral prep-SFC method 4 to afford N-[(3S)-3-(2-chloro-5-fluorophenyl)-6-(2-fluoroethyl)- 1 -oxo-2, 3 -dihydro- lH-[ 1,2,4]triazolo[4',3 ': 1,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (83.1 mg, 0.140 mmol, 42%) as a white solid. LCMS: ESI m / z 593.1 [M + H]+. 'H NMR (400 MHz, DMSO-d6) 8 10.63 (s, 1H), 9.47 (s, 1H), 8.96 (s, 1H), 7.96 (d, J= 8.0 Hz, 1H), 7.77 (d, J= 12.6 Hz, 2H), 7.71 (s, 1H), 7.35 - 7.27 (m, 1H), 7.16 - 7.06 (m, 1H), 6.16 (s, 2H), 4.99 - 4.95 (m, 2H), 4.75 - 4.48 (m, 2H). And N-[(3 R)-3 -(2-chloro-5 -fluorophenyl)-6-(2 -fluoroethyl)- 1 -oxo-2, 3 -dihydro- 1 H-[l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-3-fluoro-5-(trifluoromethyl)benzamide (80.9 mg, 0.136 mmol, 40%) as a white solid. LCMS: ESI m / z 593.1 [M + H]+. 'HNMR (400 MHz, DMSO-d6) 8 10.63 (s, 1H), 9.47 (s, 1H), 8.96 (s, 1H), 7.96 (d, J= 8.0 Hz, 1H), 7.77 (d, J = 12.6 Hz, 2H), 7.71 (s, 1H), 7.35 - 7.27 (m, 1H), 7.16- 7.06 (m, 1H), 6.16 (s, 2H), 4.99-4.95 (m, 2H), 4.75 - 4.48 (m, 2H).Example 17: N-[(3S)-3-(2-chloro-5-fluorophenyl)-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH-[l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamideExample 18: N-[(3R)-3-(2-chloro-5-fluorophenyl)-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-1H-[1,2,4]triazolo[4',3':1,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCTPd₂(dba)₃ xantphos Cs₂CO₃ Step F

[0355] Step A: To a solution of 2-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-3-[(pentadeuterioethyl)amino]benzene-l -carbonitrile (1.5 g, 3.73 mmol) and TEA (1.6 mL, 11.2 mmol) in THF (20 mL) was added trichloromethyl [(trichloromethyl)oxy]methanoate (0.55 g, 1.87 mmol) at inner temp -20 °C. The reaction was stirred at inner temp -20 °C for 10 min and stirred at inner temp 0 °C for additional 30 min. LCMS showed the reaction was completed. The reaction mixture was added to ice water and extracted with EA. The organic layer was washed with brine, dried over Na₂SO₄ and concentrated and the residue was purified by flash chromatography to afford 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-2-oxo-l-(pentadeuterioethyl)-3H-benzo[d]imidazole-4-carbonitrile (1.3 g, 3.04 mmol, 81%) as a yellow solid. LCMS: 429.0, 427.0 [M+H]+.

[0356] Step B: A solution of 6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-2-oxo-l-(pentadeuterioethyl)-3H-benzo[d]imidazole-4-carbonitrile (1 g, 2.34 mmol) in POCl3(10 mL), was added DIEA (0.60 g, 4.68 mmol). Then the mixture was stirred at 120 °C for 16 h. The mixture was concentrated. The residue was diluted with aqueous NaHCO3, extracted with EA. The organic phase was separated and concentrated. The residue was purified by column chromatography on silica gel to afford 6-bromo-2-chloro-5-[(2-chloro-5-fluorophenyl)carbonyl]-Patent Application Atty. Docket No. ENTX-038PCT l-(pentadeuterioethyl)benzo[d]imidazole-4-carbonitrile (500 mg, 1.12 mmol, 48%) as a yellow solid. LCMS: 447.0, 445.0 [M+H]+.

[0357] Step C: To a solution of 6-bromo-2-chloro-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(pentadeuterioethyl)benzo[d]imidazole-4-carbonitrile (500 mg, 1.12 mmol) in DMA(10 mL) was added 2-methylpropan-2-yl diazanylmethanoate (296 mg, 2.24 mmol) and TEA (0.5 mL, 3.36 mmol) at 25 °C. The reaction was stirred at 100 °C for 1 h. LCMS showed the reaction was completed. The reaction mixture was added to ice water and extracted with EA. The organic layer was washed with brine, dried over Na2SO4and concentrated and the residue was purified by flash chromatography to afford 2-methylpropan-2-yl (2-{6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-4-cyano-l-(2-fluoroethyl)benzo[d]imidazol-2-yl}diazanyl)methanoate (550 mg, 0.99 mmol, 68%) as ayellow solid. LCMS: 543.1, 541.1 [M+H]+.

[0358] Step D: A solution of 2-methylpropan-2-yl {2-[(2E)-6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-4-cyano-l-(pentadeuterioethyl)-2,3-dihydro-lH-benzo[d]imidazol-2-ylidene]diazanyl}methanoate (500 mg, 0.923 mmol) in TFA (5 mL). Then themixture was stirred at 25 °C for 10 min. The mixture was concentrated. The residue was diluted with aqueous NaHCO3, extracted with EA. The organic phase was separated and concentrated. The residue was purified by column chromatography on silica gel to afford (2E)-6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-2-hydrazinylidene-l-(pentadeuterioethyl)-2,3-dihydro-lH-benzo[d]imidazole-4-carbonitrile (300 mg, 0.68 mmol, 74%) as ayellow solid. LCMS: 443.1, 441.1 [M+H]+.

[0359] Step E: To a solution of (2E)-6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-2-hydrazinylidene-l-(pentadeuterioethyl)-2,3-dihydro-lH-benzo[d]imidazole-4-carbonitrile (300 mg, 0.679 mmol) in dioxane (5 mL) was added triethoxymethane (1006 mg, 6.79mmol) and TFA (0.005 mL, 0.068 mmol) at 25 °C. The reaction was stirred at 110 °C for 1 h. LCMS showed the reaction was completed. The reaction mixture was added to ice water and extracted with EA. The organic layer was washed with brine, dried over Na2SO4and concentrated and the residue was purified by flash chromatography to afford 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(pentadeuterioethyl)benzo[4,5]imidazo[2,l-c][l,2,4]triazole-5-carbonitrile (180 mg, 0.399 mmol, 59%) as a yellow solid. LCMS: 453.1, 451.1 [M+H]+.

[0360] Step F: To a solution of 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(pentadeuterioethyl)benzo[4,5]imidazo[2,l-c][l,2,4]triazole-5-carbonitrile (180 mg, 0.399Patent Application Atty. Docket No. ENTX-038PCT mmol), 5-fluoro-3-(trifluoromethyl)benzene-l -carboxamide (99 mg, 0.48 mmol), XANT PHOS (46.12 mg, 0.080 mmol), CS2CO3 (259 mg, 0.80 mmol) in dioxane (5 mL) was added Pd₂(dba)₃ (36 mg, 0.040 mmol). The reaction mixture was stirred at 100 °C under N2 for 2 h. The cooled reaction mixture was quenched with water, extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel (MeOH in DCM = 0 ~ 6%) to obtain N-[3-(2-chloro-5-fluorophenyl)-3-hydroxy-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH- [l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (120 mg, 0.201 mmol, 51%) as an yellow solid. LCMS: 496.1 [M+H]+.

[0361] Step G: To a solution ofN-[3-(2-chloro-5-fluorophenyl)-3-hydroxy-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH-[l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (120 mg, 0.201 mmol) in TFA (2 mL) wasadded triethylsilane (0.5 mL, 0.201 mmol). The reaction mixture was stirred at 70 °C for 30 min. Then the reaction mixture was concentrated to give a residue. The residue was purified by pre-HPLC to give N-[3-(2-chloro-5-fluorophenyl)-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH- [l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (70 mg, 0.121 mmol, 60%) as a white solid. LCMS: 580.1 [M+H]+.

[0362] Step H: N-[3-(2-chloro-5-fluorophenyl)-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH-[l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (70 mg, 0.121 mmol) was purified by chiral prep-SFC method 5 to afford N-[(3S)-3-(2-chloro-5-fluorophenyl)-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH- [l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (19 mg, 0.033 mmol, 27%) as a white solid. LCMS: ESI m / z 580.1 [M + H]+. 'H NMR (400 MHz, DMSO-d6) 8 10.60 (s, 1H), 9.46 (s, 1H), 8.93 (s, 1H), 8.00 - 7.92 (m, 1H), 7.82 - 7.63 (m, 3H), 7.37 - 7.28 (m, 1H), 7.10 (s, 1H), 6.75 - 5.91 (m, 2H). AndN-[(3R)-3-(2-chloro-5-fluorophenyl)-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH- [l,2,4]triazolo[4',3':l,2]imidazo[5,4-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (19.4 mg, 0.033 mmol, 28%) as a white solid.

[0363] LCMS: ESI m / z 580.1 [M + H]+. 'HNMR (400 MHz, DMSO-d6) 8 10.60 (s, 1H), 9.46 (s, 1H), 8.93 (s, 1H), 8.00 - 7.92 (m, 1H), 7.82 - 7.63 (m, 3H), 7.37 - 7.28 (m, 1H), 7.10 (s, 1H), 6.75 - 5.91 (m, 2H).Patent Application Atty. Docket No. ENTX-038PCT Example 19: N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo-l, 2,3,6-tetrahydroimidazo [2 ’, 1 ':2,3] imidazo[4,5-e] isoindol-4-yl)-3-fluoro-5- (trifluoromethyl)benzamideExample 19

[0364] Step A: To a solution of 6-bromo-5-(2-chloro-5-fluorobenzoyl)-l-(2,2-difluoroethyl)- 2-imino-2, 3 -dihydro- lH-benzo[d]imidazole-4-carbonitrile (300 mg, 0.66 mmol) in DMA (3 mL) were added 2 -bromo- 1,1 -dimethoxypropane (600 mg, 3.28 mmol). The reaction was stirred at 140 °C in sealed tube under nitrogen for 2 hours. The cooled reaction mixture was diluted water, extracted with EtOAc. The organic phase was washed with brine, dried over Na2SO4and concentrated. The residue was purified by silica gel flash chromatography, eluted with a gradient of 45 - 55% ethyl acetate in petroleum ether to afford 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-3-methyl-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (27 mg, 8%). LCMS (ESI): m / z 495, 497 [M+H]+.

[0365] Step B: To a solution of 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)- 3-methyl-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (14 mg, 0.03 mmol) in dioxane (5 mL) were added 3-fluoro-5-(trifluoromethyl)benzene-l-carboxamide (8.78 mg, 0.04 mmol), Cs₂CO₃ (3 g, 9.18 mmol), XantPhos (3.27 mg, 0.01 mmol) and Pd₂(dba)₃ (2.59 mg). The reaction was stirred at 100 °C under nitrogen for 2 hours. The cooled reaction mixture was diluted water, extracted with EtOAc. The organic phase was washed with brine, dried over Na2SO4 and concentrated. The residue was purified by silica gel flash chromatography, elutedPatent Application Atty. Docket No. ENTX-038PCT with a gradient of 60 - 70% ethyl acetate in petroleum ether to afford to afford N-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-3-methyl-9H-benzo[d]imidazo[l,2-a]imidazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (10 mg, 57%). LCMS (ESI): m / z 622 [M+H]+.

[0366] Step C: To a solution of N-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-3-methyl-9H-benzo[d]imidazo[l,2-a]imidazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (20 mg, 0.03 mmol) in ACN (1 mL) and H2O (3 mL) were added KOH (9.02 mg, 0.16 mmol). The reaction was stirred at room temperature under nitrogen for 1 hour. LCMS showed the reaction was 80% DP. The reaction mixture was diluted water, extracted with EtOAc. The organic phase was washed with brine, dried over Na2SO4 and concentrated afford N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-9-methyl-l-oxo-1, 2,3, 6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (15 mg, 73%). LCMS (ESI): m / z 640 [M+H]+.

[0367] Step D: To a solution ofN-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-9-methyl-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (15 mg, 0.02 mmol) in TFA (2 mL) were added Et3SiH (8.18 mg, 0.07 mmol). The reaction was stirred at 70 °C under nitrogen for 30 minutes. The cool reaction mixture was concentrated. The residue was purified by prep-HPLC to afford N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl- 1 -oxo- 1,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (2.4 mg, 16%). LCMS (ESI): m / z 624 [M+H]+. ‘HNMR (400 MHz, CD3OD) 88.03 (s, 1H), 7.75 - 7.69 (m, 2H), 7.67(d, 8.4 Hz, 1H), 7.30 - 7.25 (m, 2H), 7.06 - 6.97 (m, 1H), 6.62 - 6.31 (m, 2H), 4.96 -4.90 (m, 1H), 4.89 - 4.87 (m, 1H), 4.83 - 4.81 (m, 1H), 2.96 (d, J= 1.2 Hz, 3H).Example 20: N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l, 2,3,6-tetrahydroimidazo [2 ’, 1 ':2,3] imidazo[4,5-e] isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCT

[0368] Step A: To a mixture of 6-bromo-5-(2-chloro-5-fluorobenzoyl)-l-(2,2-difluoroethyl)-2-imino-2,3-dihydro-lH-benzo[d]imidazole-4-carbonitrile (700 mg, 1.53 mmol) in DMA (10 mL) was added 2-bromoacetyl bromide (461 mg, 2.31 mmol). The resulting mixture was stirred at 120 °C overnight. The cooled reaction mixture was diluted with EtOAc, washed with water and brine, dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified by silica gel flash chromatography eluted with 20 - 100% ethyl acetate in petroleum ether to afford 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-2-oxo-2,9-dihydro-3H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (280 mg, 37%). LCMS (ESI): m / z 497 / 499 [M+H]+.

[0369] Step B: A solution of 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-2-oxo-2, 9-dihydro-3H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (280 mg, 0.56 mmol) in POCl3(5 mL) was stirred at 80 °C for 3 hours. The cooled reaction mixture was concentrated, and the residue was diluted with EtOAc, washed with water and brine, dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified by silica gel flash chromatography eluted with 0 - 50% ethyl acetate in petroleum ether to afford 7-bromo-2-chloro-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (130 mg, 45%). LCMS (ESI): m / z 515 / 517 / 519 [M+H]+.Patent Application Atty. Docket No. ENTX-038PCT

[0370] Step C: To a mixture of 7-bromo-2-chloro-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (70 mg, 0.14 mol) in 1,4-dioxane (3 mL) were added 3-fluoro-5-(trifluoromethyl)benzamide (42 mg, 0.20 mol), XantPhos (16 mg, 0.03 mol), Pd2(dba)3 (12 mg, 0.01 mol) and CS2CO3 (133 mg, 0.4 mol). The reaction resulting mixture was purged with nitrogen three times, then stirred at 100 °C under N2 for 2 h. The cooled mixture was filtered and the filter cake was washed with EtOAc. The filtrate was concentrated and the residue was purified by silica gel flash chromatography, eluted with a gradient of 0 -100 % ethyl acetate in petroleum ether to afford N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (40 mg, 45%). LCMS (ESI): m / z 660 / 662 [M+H]+.

[0371] Step D: To a mixture of N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hy droxy-1 -oxo- 1,2,3, 6-tetrahydroimidazo[2', T:2, 3]imidazo[4, 5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (25 mg, 0.04 mmol) in TFA (2 mL) were added TES (0.5 mL) at 0 °C. The resulting mixture was stirred at 40 °C for 50 mins. The reaction mixture was concentrated and the residue was purified by prep-HPLC to afford N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (3 mg, 12%). LCMS (ESI): m / z 644 [M+H]+. 'H NVIR (400 MHz, DMSO-d6) 5 10.59 (s, 1H), 9.40 (s, 1H), 7.94 (d, J= 8.4 Hz, 1H), 7.88 (s, 1H), 7.81 - 7.69 (m, 3H), 7.30 (dd, J= 8.8, 5.2 Hz, 1H), 7.13 - 7.08 (m, 1H), 6.66 - 6.38 (m, 1H), 6.14 (brs, 1H), 4.86 - 4.66 (m, 2H).Example 21: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo- 1.2.3.6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 22: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo- 1.2.3.6-tetrahydroimidazo [2',1': 2,3] imidazo [4,5-e] isoindol-4-yl)-3-fluoro-5-Patent Application Atty. Docket No. ENTX-038PCT (trifluoromethyl)benzamide

[0372] Step A: N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (150 mg, 0.24 mmol) was prepared by chiral prep-SFC method 6 to afford (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo-l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (65.2 mg, 0.1 mmol, 43%) LCMS (ESI): m / z 424 [M+H]+. 1HNMR (400 MHz, DMSO-d6) 5 10.51 (s, 1H), 9.26 (s, 1H), 7.95 (d,. / = 8.4 Hz, 1H), 7.77 (d,.7= 10.0 Hz, 2H), 7.73 (s, 1H), 7.34 - 7.25 (m, 1H), 7.14 -7.05 (m, 1H), 6.83 (d, J= 1.2 Hz, 1H), 6.68 - 5.97 (m, 2H), 4.86 - 4.56 (m, 2H), 2.80 (s, 3H). And (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l-oxo- 1,2,3, 6-tetrahydroimidazo[2',r:2, 3]imidazo[4, 5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (37 mg, 0.06 mmol, 25%). 'HNMR (400 MHz, DMSO-d6) 6 10.50 (s, 1H), 9.26 (s, 1H), 7.95 (d, J= 8.4 Hz, 1H), 7.77 (d, J= 10.8 Hz, 2H), 7.73 (s, 1H), 7.34 - 7.26 (m, 1H), 7.14 - 7.05 (m, 1H), 6.83 (d, J= 1.2 Hz, 1H), 6.72 - 5.96 (m, 2H), 4.85 - 4.58 (m, 2H), 2.80 (s, 3H).Example 23: N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l,8-dioxo-l,2,3,9-tetrahydropyrazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-Patent Application Atty. Docket No. ENTX-038PCT (trifluoromethyl)benzamideExample 23

[0373] Step A: To a solution of l-amino-5-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-7-cyano-3-(2,2-difluoroethyl)-2-methylbenzo[d]imidazol-l-ium (300 mg, 0.51 mmol) in dioxane (5 mL) was added TEA (0.14 mL, 1.02 mmol) and di(imidazol-l-yl)methanone (164 mg, 1.02 mmol). The reaction was stirred at 50 °C for 1 h. The reaction mixture was purified by prep-HPLC to afford 6-bromo-7-[(2-chloro-5-fluorophenyl)carbonyl]-4-(2,2-difluoroethyl)-2-oxo-lH-benzo[4,5]imidazo[2,3-e]pyrazole-8-carbonitrile (15 mg, 0.03 mmol, 6%). LCMS (ESI): m / z 497, 499 [M+H]+.

[0374] Step B: To a solution of 6-bromo-7-[(2-chloro-5-fluorophenyl)carbonyl]-4-(2,2-difluoroethyl)-2-oxo-lH-benzo[4,5]imidazo[2,3-e]pyrazole-8-carbonitrile (15 mg, 0.03 mmol) in dioxane (3 mL), was added 3-fluoro-5-(trifluoromethyl)benzene-l -carboxamide (9.36 mg, 0.05 mmol), XANT PHOS (3.49 mg, 0.01 mmol), Cs2CO3(19.64 mg, 0.06mmol) and Pd2(dba)3 (2.76 mg, 0.00 mmol). The reaction mixture was stirred at 100 °C under N2for 2 h. The cooled reaction mixture was quenched with water, extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous Na2SC>4, filtered and concentrated. The residue was purified by column chromatography on silica gel (MeOH in DCM = 0 ~ 3%) to obtain N-{7-[(2-chloro-5-fhiorophenyl)carbonyl]-8-cyano-4-(2,2-difluoroethyl)-2-oxo-lH-Patent Application Atty. Docket No. ENTX-038PCT benzo[4,5]imidazo[2,3-e]pyrazol-6-yl]-5-fluoro-3-(trifluoromethyl)benzamide (15 mg, 0.02 mmol, 80%). LCMS (ESI): m / z 624 [M+H]+.

[0375] Step C: To a solution ofN-{7-[(2-chloro-5-fluorophenyl)carbonyl]-8-cyano-4-(2,2-difluoroethyl)-2-oxo-lH-benzo[4,5]imidazo[2,3-e]pyrazol-6-yl}-5-fluoro-3-(trifluoromethyl)benzamide (15 mg, 0.02 mmol) in ACN (2 mL) and H2O (0.5 mL) was added KOH (6.75 mg, 0.12 mmol). The reaction was purified by prep-HPLC to afford N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l,8-dioxo-l,2,3,9-tetrahydropyrazolo[5', T:2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (15 mg, 0.02 mmol, 78%). LCMS (ESI): m / z 642 [M+H]+.

[0376] Step D: To a solution ofN-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l,8-dioxo-l,2,3,9-tetrahydropyrazolo[5', T:2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (15 mg, 0.02 mmol) in TFA (1 mL) was added tri ethyl silane (0.3 mL, 0.02 mmol). The reaction mixture was stirred at 60 °C for 30 min. The cooled reaction mixture was concentrated. The residue was purified by prep-HPLC to obtain N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l,8-dioxo-l,2,3,9-tetrahydropyrazolo[5', T:2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (2 mg, 0.00 mmol, 17.09%). LCMS (ESI): m / z 626 [M+H]+. 'H NMR (400 MHz, CD3OD) 57.75 - 7.59 (m, 4H), 7.29 - 7.21 (m, 1H), 7.05 - 6.93 (m, 1H), 6.76 - 6.09 (m, 3H), 4.61 (t, J= 16.2 Hz, 2H).Example 24: N-(3-(2-chloro-5-fluorophenyl)-8-cyano-6-(2,2-difluoroethyl)-l-oxo-l, 2,3,6-tetrahydroimidazo [2 ', 1 ': 2,3] imidazo[4,5-e] isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 34: N-(8-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l, 2,3,6-tetrahydroimidazo [2 ’, 1 ':2,3] imidazo[4,5-e] isoindol-4-yl)-3-fluoro-5-Patent Application Atty. Docket No. ENTX-038PCT (trifluoromethyl)benzamideTFA, TES Pd(PPh3)4, Zn(CN)2DMA, 110 °C, O / N Step C Step DExample 34 Example 24

[0377] Step A: To a solution of N-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-2-oxo-2,9-dihydro-3H-benzo[d]imidazo[l,2-a]imidazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (80 mg, 0.13 mmol) in MeCN (3 mL) was added POBr3 (367.61 mg, 1.28 mmol). The mixture was stirred at 80 °C for 5 hours. The reaction mixture was concentrated. The crude product was purified by flash silica chromatography, eluting with a gradient 10% to 60% EtOAc in PE. The desired fractions were evaporated to dryness to afford N-(2-bromo-6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (60 mg, 68%). LCMS (ESI): m / z 686 [M+H]+.

[0378] Step B: To a solution of N-(2-bromo-6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (60 mg, 0.09 mmol) in MeCN (3 mL) and H2O (1 mL) was added KOH (4.90 mg, 0.09 mmol). The mixture was stirred at room temperature for 0.5 hour. The reaction mixture was quenched by water, extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated. The crude product was purified by flash silica chromatography, eluting with aPatent Application Atty. Docket No. ENTX-038PCT gradient 1% to 5% MeOH in DCM. The desired fractions were evaporated to dryness to afford N-(8-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (60 mg, 97%). LCMS (ESI): m / z 704 [M+H]+.

[0379] Step C: To a solution ofN-(8-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzarnide (60 mg, 0.09 mmol) in TFA (2 mL) was added triethylsilane (0.2 mL, 0.13 mmol). The mixture was stirred at 50 °C for 0.5 hour. The reaction mixture was concentrated. The crude product was purified by flash silica chromatography, eluting with a gradient 10% to 90% EtOAc in PE. The desired fractions were evaporated to dryness to afford N-(8-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (20 mg, 34%). LCMS (ESI): m / z 688 [M+H]+.

[0380] StepD: To a solution of N-(8-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l -oxo- 1,2,3, 6-tetrahydroimidazo[2', T:2, 3]imidazo[4, 5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (20 mg, 0.03 mmol) in DMA (2 mL) was added Pd(PPh3)4 (3.36 mg, 0.01 mmol) and Zn(CN)2 (34.0 mg, 0.29 mmol). The mixture was stirred at 110°C for 16 hours. The reaction mixture was quenched by water, extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated. The residue was purified by pre-TLC (70% EtOAc in PE) to afford N-(3-(2-chloro-5-fluorophenyl)-8-cyano-6-(2,2-difluoroethyl)-l-oxo- 1,2,3, 6-tetrahydroimidazo[2', T:2, 3]imidazo[4, 5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (2.8 mg, 15%).1HNMR(400 MHz, CD3OD) 58.64 (s, 1H), 7.78 (s, 1H), 7.7 - 7.64 (m, 3H), 7.28 (dd, J= 8.8, 5.0 Hz, 1H), 7.03 - 7.00 (m, 1H), 6.39 (t, J= 54.6 Hz, 2H), 4.73 (t, J= 15.2 Hz, 2H). LCMS (ESI): m / z 635 [M+H]+.Example 25: (R)-N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo- 1.2.3.6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 26: (S)-N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo- 1.2.3.6-tetrahydroimidazo[2',l':2,3]iniidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCTExample 25 Example 26

[0381] Step A: N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (50 mg, 0.08 mmol) was prepared by chiral prep-SFC method 7 to afford (R)-N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (20 mg, 0.06 mmol, 40%). LCMS (ESI): m / z 644 [M+H]+. 'H NMR (400 MHz, DMSO-d6) 8 10.60 (s, 1H), 9.41 (s, 1H), 7.95 (d, J= 8.4 Hz, 1H), 7.89 (s, 1H), 7.82 - 7.74 (m, 2H), 7.71 (s, 1H), 7.31 (dd, J= 8.8, 5.2 Hz, 1H), 7.13 - 7.08 (m, 1H), 6.67 - 6.39 (m, 1H), 6.13 (brs, 1H), 4.87 -4.68 (m, 2H). And (S)-N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo- 1,2,3, 6-tetrahydroimidazo[2',l':2, 3]imidazo[4, 5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (20 mg, 0.06 mmol, 40%). LCMS (ESI): m / z 644 [M+H]+. 'HNMR (400 MHz, DMSO-d6)8 10.59 (s, 1H), 9.41 (s, 1H), 7.95 (d, J= 8.4 Hz, 1H), 7.89 (s, 1H), 7.82 - 7.74 (m, 2H), 7.71 (s, 1H), 7.31 (dd, J= 8.8, 5.2 Hz, 1H), 7.13 - 7.08 (m, 1H), 6.67 - 6.39 (m, 1H), 6.15 (brs, 1H), 4.86 - 4.68 (m, 2H).Example 27: N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l,8-dioxo-2,3-dihydro-lH-pyrazolo [5',1 ': 2,3] imidazo [4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCT

[0382] Step A: To a solution of 6-bromo-7-[(2-chloro-5-fluorophenyl)carbonyl]-4-(2,2-difluoroethyl)-2-oxo-lH-benzo[4,5]imidazo[2,3-e]pyrazole-8-carbonitrile (140 mg, 0.28 mmol) in DMF (5 mL) was added NaH (22.51 mg, 0.56 mmol, 60% in mineral) at 0 °C. The reaction was stirred at 0 °C for 10 min. Then was added iodomethane (0.05 mL, 0.56 mmol) and the reaction mixture was stirred at rt for 1 h. After 1 h, the reaction mixture was quenched by adding sat. aq. NH4CI solution, extracted with EA, comLined all organic phases, washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel to give 6-bromo-7-[(2-chloro-5-fluorophenyl)carbonyl]-4-(2,2-difluoroethyl)-l-methyl-2-oxobenzo[4,5]imidazo[2,3-e]pyrazole-8-carbonitrile (50 mg, 0.10 mmol, 35%) and 6-bromo-7-[(2-chloro-5-fluorophenyl)carbonyl]-4-Patent Application Atty. Docket No. ENTX-038PCT (2,2-difluoroethyl)-2-methoxybenzo[4,5]imidazo[2,3-e]pyrazole-8-carbonitrile (20 mg, 0.04 mmol, 14%). LCMS (ESI): m / z 511, 513 [M+H]+.

[0383] Step B: To a solution of 6-bromo-7-[(2-chloro-5-fluorophenyl)carbonyl]-4-(2,2-difluoroethyl)-l-methyl-2-oxobenzo[4,5]imidazo[2,3-e]pyrazole-8-carbonitrile (50 mg, 0.10 mmol) in dioxane (5 mL), was added 3-fluoro-5-(trifluoromethyl)benzene-l -carboxamide (30.36 mg, 0.15 mmol), XANT PHOS (11.31 mg, 0.02 mmol), Cs2CO3(63.68 mg, 0.20mmol) and Pd2(dba)3(8.95 mg, 0.01 mmol). The reaction mixture was stirred at 100 °C under N2for 2 h. The cooled reaction mixture was quenched with water, extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous Na2SC>4, filtered and concentrated. The residue was purified by column chromatography on silica gel (MeOH in DCM = 0 ~ 3%) to obtain N-{7-[(2-chloro-5-fluorophenyl)carbonyl]-8-cyano-4-(2,2-difluoroethyl)-l-methyl-2-oxobenzo[4,5]imidazo[2,3-e]pyrazol-6-yl}-5-fluoro-3-(trifluoromethyl)benzamide (50 mg, 0.08 mmol, 80%) as an yellow solid. LCMS (ESI): m / z 638 [M+H]+.

[0384] Step C: To a solution of N-{7-[(2-chloro-5-fluorophenyl)carbonyl]-8-cyano-4-(2,2-difluoroethyl)-l-methyl-2-oxobenzo[4,5]imidazo[2,3-e]pyrazol-6-yl}-5-fluoro-3-(trifluoromethyl)benzamide (50 mg, 0.08 mmol) in ACN (3 mL) and H2O (1 mL) was added KOH (21.99 mg, 0.39 mmol). The reaction was stirred at RT for 0.5 h. The reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated to afford N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hy droxy-9-methyl-l, 8-di oxo-2, 3-dihydro-lH-pyrazolo[5', T:2,3]imidazo[4, 5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (50 mg, 0.08 mmol, 97%) as a yellow solid. LCMS (ESI): m / z 656 [M+H]+.

[0385] Step D: To a solution of N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-9-methyl-l, 8-di oxo-2, 3-dihydro- lH-pyrazolo[5',l':2, 3]imidazo[4, 5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (50 mg, 0.08 mmol) in TFA (2 mL) wasadded triethylsilane (0.5 mL, 0.08 mmol). The reaction mixture was stirred at 60 °C for 30 min. The cooled reaction mixture was concentrated. The residue was purified by prep-HPLC to obtain N-[3 -(2-chl oro-5 -fhjorophenyl)-6-(2,2-difluoroethyl)-9-methyl- 1, 8-di oxo-2, 3 -dihydro-lH-pyrazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (34 mg, 0.05 mmol, 70%). LCMS (ESI): m / z 640 [M+H]+.Patent Application Atty. Docket No. ENTX-038PCT! HNMR (400 MHz, DMSO-d6) 8 10.45 (s, 1H), 9.32 (s, 1H), 7.95 (d, J= 8.4 Hz, 1H), 7.82 -7.62 (m, 3H), 7.34 - 7.27 (m, 1H), 7.20 - 7.02 (m, 1H), 6.65 (s, 1H), 6.50 (t, J= 54.0 Hz, 1H), 6.05 (s, 1H), 5.04 (s, 1H), 4.79 -4.48 (m, 2H), 3.66 (s, 3H).Example 28: N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methoxy-l-oxo-2,3-dihydro-lH-pyrazolo [5',1 ': 2,3] imidazo [4,5-e]isoindol-4-yl]-5-fluoro-3- (trifluoromethyl)benzamideStep B

[0386] Step A: To a solution of 6-bromo-7-[(2-chloro-5-fluorophenyl)carbonyl]-4-(2,2-difluoroethyl)-2-methoxybenzo[4,5]imidazo[2,3-e]pyrazole-8-carbonitrile (20 mg, 0.04 mmol) in dioxane (3 mL), was added 3-fluoro-5-(trifluoromethyl)benzene-l -carboxamide (12.14 mg, 0.06 mmol), XANT PHOS (4.52 mg, 0.01 mmol), Cs2CO3(25.47 mg, 0.08mmol) and Pd2(dba)3(3.58 mg, 0.00 mmol). The reaction mixture was stirred at 100 °C under N2for 2 h. The cooled reaction mixture was quenched with water, extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous Na2SC>4, filtered and concentrated. The residue was purified by column chromatography on silica gel to obtain N-{7-[(2-chloro-5-fluorophenyl)carbonyl]-8-cyano-4-(2,2-difluoroethyl)-2-methoxybenzo[4,5]imidazo[2,3-Patent Application Atty. Docket No. ENTX-038PCT e]pyrazol-6-yl]-5-fluoro-3-(trifluoromethyl)benzamide (20 mg, 0.03 mmol, 80%). LCMS (ESI): m / z 638 [M+H]+.

[0387] Step B: To a solution of N-{7-[(2-chloro-5-fluorophenyl)carbonyl]-8-cyano-4-(2,2-difluoroethyl)-2-methoxybenzo[4,5]imidazo[2,3-e]pyrazol-6-yl}-5-fluoro-3-(trifluoromethyl)benzamide (20 mg, 0.03 mmol) in ACN (3 mL) and H2O (0.5 mL) was added KOH (8.80 mg, 0.16 mmol). The reaction was stirred at RT for 0.5 h. The reaction mixture was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated to afford N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-8-methoxy-l-oxo-2,3-dihydro-lH-pyrazolo[5', T:2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (20 mg, 0.03 mmol, 97%). LCMS (ESI): m / z 656 [M+H]+.

[0388] Step C: To a solution ofN-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-8-methoxy-l -oxo-2, 3-dihy dro-lH-pyrazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (20 mg, 0.03 mmol) in TFA (2 mL) wasadded Triethylsilane (0.5 mL, 0.03 mmol). The reaction mixture was stirred at 60 °C for 30 min. The cooled reaction mixture was concentrated. The residue was purified by prep-HPLC to obtain N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methoxy-l -oxo-2, 3-dihy dro-lH-pyrazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (12.7 mg, 0.02 mmol, 65%). LCMS (ESI): m / z 640 [M+H]+. 'HNMR (400 MHz, DMSO-d6) 5 10.45 (s, 1H), 9.10 (s, 1H), 7.95 (d, J= 8.4 Hz, 1H)„ 7.79 - 7.68 (m, 3H), 7.33 - 7.26 (m, 1H), 7.14 - 7.02 (m, 1H), 6.50 ((t, J= 54.0 Hz, 1H), 6.08 (s, 1H), 5.60 (s, 1H), 4.84 - 4.61 (m, 2H), 3.95 (s, 3H).Example 29: N-[(3S)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l,8-dioxo-2,3-dihydro-lH-pyrazolo [5 ’, 1 ': 2,3] imidazo [4,5-e] isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamideExample 30: N-[(3R)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l,8-dioxo-2,3-dihydro-lH-pyrazolo [5',1 ': 2,3] imidazo [4,5-e] isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCTExample 29

[0389] Step A: N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l,8-dioxo- 2,3-dihydro-lH-pyrazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (32 mg, 0.05 mmol) was separated by chiral prep-SFC method 8 to give N-[(3 S)-3-(2-chl oro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l,8-di oxo-2, 3-dihydro-lH-pyrazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (13.5 mg, 0.02 mmol, 42%) as a white solid. LCMS: ESI m / z 640 [M + H]+.1H NMR (400 MHz, CD3OD) 5 7.77 - 7.60 (m, 4H), 7.29 - 7.21 (m, 1H), 7.03 - 6.96 (m, 1H), 6.85 - 6.15 (m, 3H), 5.16 (s, 1H), 4.65 -4.50 (m, 2H), 3.86 (s, 3H). AndN-[(3R)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-methyl-l,8-dioxo-2,3-dihydro-lH-pyrazolo[5',r:2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (13.7 mg, 0.02 mmol, 42%) as a white solid. LCMS: ESI m / z 640 [M + H]+. ’H NMR (400 MHz, CD3OD) 67.75 - 7.57 (m, 4H), 7.29 - 7.20 (m, 1H), 7.06 - 6.93 (m, 1H), 6.80 - 5.95 (m, 3H), 5.16 (s, 1H), 4.67 - 4.51 (m, 2H), 3.86 (s, 3H).Example 31: N-[(3R)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methoxy-l-oxo- 2,3-dihydro-lH-pyrazolo [5',1 ': 2,3] imidazo [4,5-e] isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamideExample 32: N-[(3S)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methoxy-l-oxo- 2,3-dihydro-lH-pyrazolo [5',1 ': 2,3] imidazo [4,5-e] isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCT

[0390] Step A: N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methoxy-l-oxo-2,3-dihydro-lH-pyrazolo[5',r:2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3- (trifluoromethyl)benzamide (10.5 mg, 0.02 mmol) was separated by chiral prep-SFC method 9 to give N-[(3R)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methoxy-l-oxo-2,3-dihydro-lH-pyrazolo[5',r:2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (4.3 mg, 0.01 mmol, 41%) as a white solid. LCMS: ESI m / z 640[M + H]+. NMR (400 MHz, CD3OD) 8 7.74 - 7.62 (m, 4H), 7.28 - 7.23 (m, 1H), 7.02 - 6.95 (m, 1H), 6.78 - 6.08 (m, 3H), 5.55 (s, 1H), 4.62 (t, J= 16.0 Hz, 2H), 4.05 (s, 3H). And N-[(3S)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-methoxy-l-oxo-2,3-dihydro-lH-pyrazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (4 mg, 0.01 mmol, 38%) as a white solid. LCMS: ESI m / z 640 [M + H]+. 'H NMR (400 MHz, CD3OD) 87.74 - 7.57 (m, 4H), 7.29 - 7.21 (m, 1H), 7.05 - 6.93 (m, 1H), 6.76 -6.13 (m, 3H), 5.54 (s, 1H), 4.61 (t, J= 16.0 Hz, 2H), 4.04 (s, 3H). Example 33: N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-ethynyl-l-oxo-l,2,3,6-tetrahydroimidazo [2 ', 1 ': 2,3] imidazo[4,5-e] isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCT

[0391] Step A: To a solution ofN-(8-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (50 mg, 0.07 mmol) in dioxane (5 mL) was added tributyl(ethynyl)-X4-stannane (68.62 mg, 0.22 mmol), Bis(tri-tert-butylphosphine)palladium(0) (3.71 mg, 0.01 mmol) and CsF (22.05 mg, 0.15 mmol). The mixture was stirred at 80 °C for 1 hour. The reaction mixture was concentrated. The crude product was purified by flash silica chromatography, eluting with a gradient of 10% to 70% EtOAc in PE to give the residue. The residue was purified by prep-HPLC to give N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-ethynyl-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (4.5 mg, 10%) as a white solid. LCMS(ESI): m / z 634 [M+H]+.1H NMR (400 MHz, DMSO-d6) 5 10.60 (s, 1H), 9.40 (s, 1H), 8.10 (s, 1H), 7.96 (d, J= 6.4 Hz, 1H), 7.76 (d, J= 9.0 Hz, 2H), 7.71 (s, 1H), 7.35 - 7.27 (m, 1H), 7.11 (s, 1H), 6.54 (t, J = 54.2 Hz, 2H), 4.81 - 4.75 (m, 2H), 4.24 (s, 1H).Example 35: 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindole-8-carboxylic acidPatent Application Atty. Docket No. ENTX-038PCTXantPhos, Cs2CO3Pd2(dba)3, dioxanestep BKOH TES, TFA MeOH, H2O step C step D

[0392] Step A: To a solution of 2-azanylidene-6-bromo-5-[(2-chloro-4-fluorophenyl)carbonyl]-l-(2,2-difluoroethyl)-3H-benzo[d]imidazole-4-carbonitrile (400 mg, 0.87 mmol) in N, N-dimethylacetamide (4 mL) was added ethyl 3-bromo-2-oxopropanoate (170 mg, 0.87 mmol). The reaction was stirred at 100 °C for 5 hs. LCMS showed the reaction was completed. The reaction mixture was added to ice water and extracted with EA. The organic layer was washed with brine, dried over Na2SO4and concentrated and the residue was purified by flash chromatography to afford ethyl 7-bromo-6-[(2-chloro-4-fluorophenyl)carbonyl]-5-Patent Application Atty. Docket No. ENTX-038PCT cyano-9-(2,2-difluoroethyl)benzo[4,5]imidazo[2,3-b]imidazole-2-carboxylate (230 mg, 0.42 mmol, 48%) as a brown solid. LCMS (ESI): m / z 553; 555 [M+H]+.

[0393] Step B: A solution of ethyl 7-bromo-6-[(2-chloro-4-fluorophenyl)carbonyl]-5-cyano-9-(2,2-difluoroethyl)benzo[4,5]imidazo[2,3-b]imidazole-2-carboxylate (200 mg, 0.36 mmol), 3-fluoro-5-(trifluoromethyl)benzene-1-carboxamide (74.8 mg, 0.36 mmol), Pd2dba3 (33.1 mg, 0.04 mmol), Xantphos (41.8 mg, 0.07 mmol), CS2CO3 (352 mg, 1.08 mmol) in dioxane (10 mL) was stirred at 90 °C under N2 for 1 h. LCMS showed the reaction was completed. The reaction mixture was concentrated, diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4and concentrated in vacuo. The residue was purified by silica gel column chromatography eluted with ethyl EA in PE (0-50% gradient) to afford ethyl 6-[(2-chloro-4-fluorophenyl)carbonyl]-5-cyano-9-(2,2-difluoroethyl)-7-({[5-fhioro-3-(trifluoromethyl)phenyl]carbonyl}amino)benzo[4,5]imidazo[2,3-b]imidazole-2-carboxylate (100 mg, 0.15 mmol, 41%) as a brown solid. LCMS (ESI): m / z 680 [M+H]+.

[0394] Step C: To a solution of ethyl 6-[(2-chloro-4-fluorophenyl)carbonyl]-5-cyano-9-(2,2-difluoroethyl)-7-({[5-fluoro-3-(trifluoromethyl)phenyl]carbonyl}amino)benzo[4,5]imidazo[2,3-b]imidazole-2-carboxylate (20 mg, 0.03 mmol) in methanol (1 mL) and water (0.2 mL) was added potassium hydroxide (8 mg, 0.15 mmol). The mixture was stirred at rt for 16 h. LCMS showed the reaction was completed. The reaction mixture was concentrated, diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo to get the 3-(2-chloro-4-fluorophenyl)-6-(2,2-difluoroethyl)-4-({[5-fluoro-3 -(trifluoromethyl)phenyl]carbonyl } amino)-3 -hydroxy- 1 -oxo-2,3 -dihydro- 1 H-imidazo[2',3':2,3]imidazo[4,5-e]isoindole-8-carboxylic acid (15 mg, 0.02 mmol, 76%) as a crude. LCMS (ESI): m / z 670 [M+H]+.

[0395] Step D: A solution of 3-(2-chloro-4-fluorophenyl)-6-(2,2-difluoroethyl)-4-({[5-fluoro-3-(tri fluoromethyl)phenyl]carbonyl}amino)-3-hydroxy-l -oxo-2, 3-dihy dro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindole-8-carboxylic acid (15 mg, 0.02 mmol) in TFA (0.5 mL) was added TES (0.1 mL). The mixture was stirred at 70 °C for 16 h. LCMS showed the reaction was completed. The reaction mixture was concentrated, and purified by prep-HPLC to afford 3-(2-chloro-4-fluorophenyl)-6-(2,2-difluoroethyl)-4-({[5-fluoro-3-(trifluoromethyl)phenyl]carbonyl}amino)-l-oxo-2,3-dihydro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindole-8-carboxylic acid (3.2 mg, 0.00 mmol, 22%) as a white solid. LCMS(ESI): m / z 654Patent Application Atty. Docket No. ENTX-038PCT [M+H]+. ‘HNMR (400 MHz, DMSO-d6) 8 10.64 (s, 1H), 9.47 (s, 1H), 8.47 (s, 1H), 7.97 (d, J= 8.2 Hz, 1H), 7.80 (s, 1H), 7.78 (d, J= 9.4 Hz, 1H), 7.72 (s, 1H), 7.32 (dd, J= 8.8, 5.2 Hz, 1H), 7.11 (t, J = 6.8 Hz, 1H), 6.56 (t, J= 54.2 Hz, 2H), 6.17 (s, 1H), 4.94 - 4.65 (m, 2H).Example 36: (S)-N-(3-(2-chloro-5-fluorophenyl)-8-cyano-6-(2,2-difluoroethyl)-l-oxo-l, 2,3,6-tetrahydroimidazo [2 ’, 1 ': 2,3] imidazo[4,5-e] isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 37: (R)-N-(3-(2-chloro-5-fluorophenyl)-8-cyano-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l ': 2,3] imidazo [4,5-e] isoindol-4-yl)-3-fluoro-5- (trifluoromethyl)benzamideExample 37 Example 36

[0396] Step A: To a solution of N-(8-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (60 mg, 0.0871 mmol) in DMA (5 mL) was added Pd(PPh3)4 (67.11 mg, 0.06 mmol) and ZINC CYANIDE (68.19 mg, 0.58 mmol). The mixture was stirred at 115 °C for 2 hours under microwave. The cooled reaction mixture was quenched by water, extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4and concentrated. The residue was purified by pre-TLC (70% EtOAc in PE) to afford the crude product (11 mg, 20%). The crude product was purified by chiral prep-SFC method 10 to give (S)-N-(3-(2-chloro-5-fluorophenyl)-8-cyano-6-(2,2-difluoroethyl)- 1 -oxo- 1,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (3 mg, 3%) as a white solid. 'HNMR (400 MHz, CD3OD) 88.63 (s, 1H), 7.78 (s, 1H), 7.70 -7.61 (m, 3H), 7.28 (dd, J = 8.8, 5.0 Hz, 1H), 7.05 - 6.96 (m, 1H), 6.39 (t, J = 54.4 Hz, 1H), 4.73 (t, J= 15.2 Hz, 2H).Patent Application Atty. Docket No. ENTX-038PCT

[0397] And (S)-N-(3-(2-chloro-5-fluorophenyl)-8-cyano-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (3 mg, 3%) as a white solid. LCMS (ESI): m / z 635 [M+H]+. 'H NMR (400 MHz, CD3OD) 5 8.64 (s, 1H), 7.78 (s, 1H), 7.70 - 7.61 (m, 3H), 7.28 (dd, J= 8.8, 5.0 Hz, 1H), 7.04 - 6.96 (m, 1H), 6.39 (t, J= 54.6 Hz, 1H), 4.73 (t, J= 15.2 Hz, 2H).Example 38: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-8-(prop-l-yn-l-yl)-l,2,3,6-tetrahydroimidazo |2',1 ': 2,3 J imidazo [4,5-eJ isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 39: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyI)-l-oxo-8-(prop-l-yn-l-yl)-l,2,3,6-tetrahydroimidazo[2',l':2,3]iniidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide

[0398] Step A: To a mixture of N-(8-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-Patent Application Atty. Docket No. ENTX-038PCT (trifluoromethyl)benzamide (20 mg, 0.03 mmol), caesium fluoride (8.82 mg, 0.06 mmol) and Bis(tri-tert-butylphosphine)palladium(0) (1.48 mg, 0.001 mmol) in dioxane (2 mL) was added tributyl(prop-l-ynyl)-X4-stannane (19.1 mg, 0.06 mmol). The reaction mixture was purged with nitrogen three times, then heated to 80 °C for 1 hour under nitrogen. The reaction was monitored by analysis of LCMS. After cooling to room temperature, the reaction mixture was diluted with EtOAc, washed with brine, and the organic layer was separated. The aqueous layer was extracted with EtOAc, then the comLined extracts were dried over anhydrous Na2SO4, filtered and evaporated. The residue was purified prep-HPLC (Cl 8, acetonitrile / H2O, 0.1% trifluoroacetic acid modifier, 40% to 95% gradient, 15 min run) to afford N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-8-(prop-l-yn-l-yl)-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (7 mg, 37%). LCMS (ESI): m / z 648.4 [M+H]+.

[0399] Step B: N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-8-(prop-l-yn-l-yl)-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (7 mg, 0.01 mmol) was purified by chiral prep-SFC method 11 to give (R)-N-(3 -(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)- 1 -oxo-8-(prop- 1 -yn- 1 -yl)- 1,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (1.6 mg, 22%). LCMS (ESI): m / z 648.4 [M+H]+. *HNMR (400 MHz, DMSO-d6) 8 10.58 (s, 1H), 9.36 (s, 1H), 7.97 - 7.93 (m, 2H), 7.78 - 7.70 (m, 3H), 7.32 - 7.28 (m, 1H), 7.12 - 7.07 (m, 1H), 6.65 - 6.41 (m, 1H), 6.14 (brs, 1H), 4.79 - 4.65 (m, 2H), 2.08 (s, 3H). And (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-8-(prop-l-yn-l-yl)-l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (1.0 mg, 14%). ‘HNMR (400 MHz, DMSO-d6) 8 10.58 (s, 1H), 9.36 (s, 1H), 7.98 - 7.93 (m, 2H), 7.78 - 7.70 (m, 3H), 7.34 - 7.28 (m, 1H), 7.13 - 7.07 (m, 1H), 6.66 - 6.40 (m, 1H), 6.14 (brs, 1H), 4.81 - 4.64 (m, 2H), 2.08 (s, 3H).Example 40: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l, 2,3,6-tetrahydroimidazo [2 ’, 1 ': 2,3] imidazo[4,5-e] isoindol-4-yl-8,9-d2)-3-fluoro-5-(trifluoromethyl)benzamideExample 41: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo [2 *, 1 ': 2,3] imidazo[4,5-e] isoindol-4-yl-8,9-d2)-3-fluoro-5-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCTKOH, MeCN, TFA, Et3SiH, H2O, RT, 1h 60 °C, 1hPd2(dba)3, xantphos, Cs2CO3,dioxane, 100°C, 2hStep C Step D Step E

[0400] Step A: To a solution of 2-azanylidene-6-bromo-5-[(2-chloro-5-fluorophenyl)carbonyl]-l-(2,2-difluoroethyl)-3H-benzo[d]imidazole-4-carbonitrile (500 mg, 1.09 mmol) in DMA (5 mL) was added l,2-dibromo-l,l,2,2-tetradeuterioethane (1.88 g, 9.83 mmol). The mixture was stirred at 140 °C for ON. The reaction mixture was diluted water, extracted with EA. The organic phase was washed with brine, dried over Na₂SO₄ and concentrated. The residue was purified using silica gel column chromatography eluted with ethyl acetate in petroleum ether (gradient:©- 100%) to afford 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-Patent Application Atty. Docket No. ENTX-038PCT 2,2,3,3-tetradeuterio-9-(2,2-difluoroethyl)-3,9-dihydro-2H-benzo[4,5]imidazo[l,2-a]imidazole-5-carbonitrile (100 mg, 0.21 mmol, 19%) as a yellow solid. LCMS: (ESI) m / z 489.0 [M+H]+.

[0401] Step B: To a solution of 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-2,2,3,3-tetradeuterio-9-(2,2-difluoroethyl)-3,9-dihydro-2H-benzo[4,5]imidazo[l,2-a]imidazole-5-carbonitrile (100 mg, 0.21 mmol) in toluene (2 mL) was added MnO2(357 mg, 4.10 mmol). The mixture was stirred at 100 °C O / N with sealed tube. The reaction mixture was diluted water, extracted with EA. The organic phase was washed with brine, dried over Na2SO4and concentrated. The residue was purified using silica gel column chromatography eluted with ethyl acetate in petroleum ether (gradient:0-50%) to afford 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-2,3-dideuterio-9-(2,2-difluoroethyl)-3,9-dihydro-2H-benzo[4,5]imidazo[l,2-a]imidazole-5-carbonitrile (80 mg, 0.16 mmol, 80%) as a yellow solid. LCMS: (ESI) m / z 485.1 [M+H]+.

[0402] Step C: To a solution of 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-2,3-dideuterio-9-(2,2-difluoroethyl)benzo[4,5]imidazo[l,2-a]imidazole-5-carbonitrile (80 mg, 0.17 mmol) in dioxane (5 mL) were added 3-fluoro-5-(trifluoromethyl)benzene-1-carboxamide (51.4 mg, 0.25 mmol), Pd2(dba)3(15.2 mg, 0.02 mmol), XANT PHOS (19.1 mg, 0.03 mmol) and CS2CO3 (162 mg, 0.50 mmol). The mixture was stirred at 100 °C under N2 atmosphere for 2h. The cooled reaction mixture was diluted water, extracted with EA. The organic phase was washed with brine, dried over Na2SO4and concentrated. The residue was purified using silica gel column chromatography eluted with methanol in dichloromethane (gradient: 0-10%) to afford N-{6-[(2-chloro-5-fluorophenyl)carbonyl]-5-cyano-2,3-dideuterio-9-(2,2-difluoroethyl)benzo[4,5]imidazo[l,2-a]imidazol-7-yl}-5-fluoro-3-(trifluoromethyl)benzamide (80 mg, 0.13 mmol, 79%) as a yellow solid. LCMS: (ESI) m / z 610.2 [M+H]+.

[0403] Step D: To a solution ofN-{6-[(2-chloro-5-fluorophenyl)carbonyl]-5-cyano-2,3-dideuterio-9-(2,2-difluoroethyl)benzo[4,5]imidazo[l,2-a]imidazol-7-yl}-5-fluoro-3-(trifluoromethyl)benzamide (80 mg, 0.13 mmol) in ACN (3 mL) and H2O (1.00 mL) was added KOH (14.7 mg, 0.26 mmol). The mixture was stirred at RT for Ih. The reaction mixture was diluted water, extracted with EA. The organic phase was washed with brine, dried over Na2SO4 and concentrated to afford N-[3-(2-chloro-5-fluorophenyl)-8,9-dideuterio-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-2,3-dihydro-lH-imidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-Patent Application Atty. Docket No. ENTX-038PCT (trifluoromethyl)benzamide (70 mg, 0.11 mmol, 85%) as a yellow solid. LCMS: (ESI) m / z 628.2 [M+H]+.

[0404] Step E: To a solution of N-[3-(2-chloro-5-fluorophenyl)-8,9-dideuterio-6-(2,2-difluoroethyl)-3-hy droxy-1 -oxo-2, 3-dihy dro-lH-imidazo[2',l':2, 3]imidazo[4, 5-e]isoindol -4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (70 mg, 0.11 mmol) in TFA (2.00 mL) was added Et3SiH (0.2 mL). The mixture was stirred at 60 °C for Ih. The reaction mixture was concentrated. The residue was purified by prep-HPLC to afford N-[3-(2-chloro-5-fluorophenyl)-8,9-dideuterio-6-(2, 2-difluoroethyl)-l -oxo-2, 3-dihy dro-lH-imidazo[2',l':2, 3]imidazo[4, 5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (30 mg, 0.05 mmol, 44%) as a white solid. LCMS: (ESI) m / z 612.1 [M+H]+.

[0405] Step F: N-[3-(2-chloro-5-fluorophenyl)-8,9-dideuterio-6-(2,2-difluoroethyl)-l-oxo- 2, 3-dihy dro-lH-imidazo[2', l':2,3]imidazo[4, 5-e]isoindol -4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (30 mg, 0.05 mmol) was purified with chiral prep-SFC method 12 to afford ( S)-N -(3 -(2-chl oro-5 -fluoropheny l)-6-(2,2-difluoroethy 1 )- 1 -oxo- 1,2,3, 6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl-8,9-d2)-3-fluoro-5-(trifluoromethyl)benzamide (9.6 mg ) as a white solid.JH NMR (400 MHz, DMSO-d6) 8 10.59 (s, IH), 9.36 (s, IH), 7.99 - 7.90 (m, IH), 7.85 - 7.66 (m, 3H), 7.36 - 7.24 (m, IH), 7.15 - 7.04 (m, IH), 6.88 - 5.81 (m, 3H), 4.88 - 4.61 (m, 2H). And (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl-8,9-d2)-3-fluoro-5-(trifluoromethyl)benzamide (11.7 mg ) as a white solid. ’H NMR (400 MHz, DMSO-d6) 8 10.57 (s, IH), 9.36 (s, IH), 7.98 - 7.91 (m, IH), 7.82 - 7.69 (m, 3H), 7.37 - 7.28 (m, IH), 7.15 - 7.06 (m, IH), 6.80 - 5.78 (m, 3H), 4.87 - 4.64 (m, 2H).Example 42: (S)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e] isoindole-8-carboxamideExample 43: (R)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-Patent Application Atty. Docket No. ENTX-038PCT e]isoindole-8-carboxamideExample 42 Example 43

[0406] Step A: To a stirred mixture of 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindole-8-carboxylic acid (50 mg, 0.08 mmol) in DMF (2 mL), were added NH4CI (40.9 mg, 0.76 mmol), DIEA (29.6 mg, 0.23 mmol) and HATU (43.6 mg, 0.11 mmol). The mixture was stirred at room temperature for 1 hour. The reaction was monitored by analysis of LC-MS. The reaction mixture was diluted with EtOAc, washed with brine, and organic layer was separated. The aqueous layer was extracted with EtOAc, then the comLined extracts were dried over anhydrous Na2SO4, filtered and evaporated. The residue was purified Prep-HPLC (C18, acetonitrile / H2O, 0.1% trifluoroacetic acid modifier, 30% to 95% gradient, 15 min run) to afford 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindole-8-carboxamide (22 mg, 44%). LCMS: ESI m / z 653.4 [M+H]+.Patent Application Atty. Docket No. ENTX-038PCT

[0407] Step B: 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-l-oxo-l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindole-8-carboxamide (22 mg, 0.03 mmol) was purified by chiral Prep-SFC method 13 to give (S)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindole-8-carboxamide (2.9 mg, 13%) 'H NMR (400 MHz, DMSO-d6) 8 10.60 (s, 1H), 9.42 (s, 1H), 8.36 (s, 1H), 7.97 - 7.93 (m, 1H), 7.82 - 7.75 (m, 2H), 7.72 (s, 1H), 7.45 (s, 1H), 7.33 - 7.29 (m, 2H), 7.13 - 7.08 (m, 1H), 6.70 -6.46 (m, 2H), 6.16 (brs, 1H), 4.82 - 4.69 (m, 2H). And (R)-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindole-8-carboxamide (2.9 mg, 13%). LCMS: ESI m / z 653.4 [M+H]+. ’H NMR (400 MHz, DMSO-d6) 8 10.62 (s, 1H), 9.44 (s, 1H), 8.36 (s, 1H), 7.96 (d, J= 8.0 Hz, 1H), 7.80 (s, 1H), 7.79 - 7.75 (m, 1H), 7.72 (s, 1H), 7.46 (s, 1H), 7.35 - 7.29 (m, 2H), 7.15 - 7.08 (m, 1H), 6.80 - 6.37 (m, 2H), 6.18 (brs, 1H), 4.87 - 4.68 (m, 2H).Example 44: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-ethynyl-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 45: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-ethynyl-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideprep-SFCStep A

[0408] Step A: N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-ethynyl-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5- (trifluoromethyl)benzamide (5 mg, 0.01 mmol) was purified with chiral prep-SFC method 14 to afford (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-ethynyl-l-oxo-l,2,3,6-Patent Application Atty. Docket No. ENTX-038PCT tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (4 mg, 0.01 mmol, 40%) as a white solid. LCMS: ESI m / z 634 [M + H]+.JH NMR (400 MHz, CD3OD) 5 8.16 (s, 1H), 7.76 - 7.63 (m, 4H), 7.31 - 7.24 (m, 1H), 7.03 - 6.97 (m, 1H), 6.78 - 6.04 (m, 3H), 4.73 - 4.63 (m, 2H), 3.66 (s, 1H). And (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-ethynyl- 1-oxo- 1,2,3, 6-tetrahydroimidazo[2',l':2, 3]imidazo[4, 5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (3.9 mg, 0.01 mmol, 39%) as a white solid. LCMS: ESI m / z 634 [M + H]+.1H NMR (400 MHz, CD3OD) δ 8.16 (s, 1H), 7.75 - 7.63 (m, 4H), 7.31 - 7.24 (m, 1H), 7.04 - 6.97 (m, 1H), 6.81 - 6.02 (m, 3H), 4.73 - 4.64 (m, 2H), 3.66 (s, 1H).Example 46: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-8-(trifluoromethyl)-l,2,3?6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 47: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-8-(trifluoromethyl)-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideBr O ClKOHMeCN, H2O Step CPatent Application Atty. Docket No. ENTX-038PCTCF3Example 47

[0409] Step A: To a solution of 6-bromo-5-(2-chloro-5-fluorobenzoyl)-l-(2,2-difluoroethyl)-2-imino-2, 3 -dihydro- lH-benzo[d]imidazole-4-carbonitrile (2 g, 4.37 mmol) in DMA (20 mL) was added 3 -bromo- 1,1,1 -trifl uoropropan-2-one (1.25 g, 6.56 mmol). The mixture was stirred at 120 °C for 2 hours. The cooled mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-30% ethyl acetate in petroleum ether to afford 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-2-(trifluoromethyl)-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (2 g, 3.64 mmol, 83%) as a yellow solid. LCMS: ESI m / z 549, 551 [M + H]+.

[0410] Step B: To a solution of 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-2-(trifluoromethyl)-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (500 mg, 0.91 mmol) in dioxane (30 mL) was added 3-fluoro-5-(trifluoromethyl)benzamide (282.62 mg, 1.36 mmol), XantPhos (105 mg, 0.18 mmol), CS2CO3 (889 mg, 2.73 mmol) and Tris(dibenzylideneacetone)dipalladium (83 mg, 0.09 mmol). The mixture was stirred at 100 °C under nitrogen for 2 hours. The cooled mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-30% ethyl acetate in petroleum ether to afford N-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-2-(trifluoromethyl)-9H-benzo[d]imidazo[l,2-a]imidazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (600 mg, 0.89 mmol, 97%) as a yellow solid. LCMS: ESI m / z 676 [M + H]+.

[0411] Step C: To a solution ofN-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-2-(trifluoromethyl)-9H-benzo[d]imidazo[l,2-a]imidazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (600 mg, 0.89 mmol) in ACN (10 mL) and H2O (2 mL) was addedPatent Application Atty. Docket No. ENTX-038PCT KOH (249 mg, 4.44 mmol). The mixture was stirred at 25 °C for 1 hour. The mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-10% methanol in dichloromethane to afford N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-1-oxo-8-(trifluoromethyl)-1,2,3,6-tetrahydroimidazo[2',1':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (500 mg, 0.72 mmol, 81%) as a yellow solid. LCMS: ESI m / z 694 [M + H]+.

[0412] Step D: To a solution of N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-1-oxo-8-(trifluoromethyl)-1,2,3,6-tetrahydroimidazo[2',1':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (500 mg, 0.72 mmol) in TFA (10 mL) was added Et3SiH (418 mg, 3.60 mmol). The mixture was stirred at 60 °C for 1 hour. The mixture was concentrated under vacuum. The residue was purified using prep-HPLC to afford N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-8-(trifluoromethyl)-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (150 mg, 0.22 mmol, 30%) as a white solid. LCMS: ESI m / z 678 [M + H]+.

[0413] Step E: N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-1-oxo-8-(trifluoromethyl)-1,2,3,6-tetrahydroimidazo[2',1':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (5 mg, 0.07 mmol) was purified with chiral prep-SFC method 15 to afford (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-8-(trifluoromethyl)-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (14.9 mg, 0.02 mmol, 60%) as a white solid. LCMS: ESI m / z 678 [M + H]+. 'HNMR (400 MHz, DMSO-d6) 5 10.64 (s, 1H), 9.48 (s, 1H), 8.36 - 8.28 (m, 1H), 7.96 (d, J= 8.4 Hz, 1H), 7.83 (s, 1H), 7.77 (d, J= 8.4 Hz, 1H), 7.71 (s, 1H), 7.35 - 7.27 (m, 1H), 7.16 - 7.08 (m, 1H), 6.77 - 5.75 (m, 3H), 4.94 - 4.68 (m, 2H). And (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-8-(trifluoromethyl)-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (15.1 mg, 0.02 mmol, 60%) as a white solid. LCMS: ESI m / z 678 [M + H]+.JH NMR (400 MHz, DMSO-d6) 5 10.64 (s, 1H), 9.48 (s, 1H), 8.34 - 8.32 (m, 1H), 7.96 (d, J= 8.4 Hz, 1H), 7.83 (s, 1H), 7.77 (d, J= 8.4 Hz, 1H), 7.71 (s, 1H), 7.35 - 7.27 (m, 1H), 7.16 - 7.05 (m, 1H), 6.74 - 6.01 (m, 3H), 4.93 - 4.70 (m, 2H).Patent Application Atty. Docket No. ENTX-038PCT Example 48: (S)-N-(3-(2-chloro-5-fluorophenyl)-l -oxo-6-(2,2,2-trifluoroethyl)-l,2,3,6-tetrahydroimidazo [2 ’, 1 ':2,3] imidazo[4,5-e] isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 49: (R)-N-(3-(2-chloro-5-fluorophenyl)-l-oxo-6-(2,2,2-trifluoroethyl)-l,2,3,6-tetrahydroimidazo [2 ’, 1 ': 2,3] imidazo[4,5-e] isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideBr O Cl " O CuCN DMA, 100 °C DMSO, 130 °C Step A Step BXantPhos, CS2CO3, Pd2(dba)3, dioxane, Step CTFA, TES70 °CStep EPatent Application Atty. Docket No. ENTX-038PCTExample 48 Example 49

[0414] Step A: To a solution of (2-chloro-5-fluorophenyl)(4,6-dibromo-2-imino-l -(2,2,2-trifluoroethyl)-2,3-dihydro-lH-benzo[d]imidazol-5-yl)methanone (3 g, 5.67 mmol) in DMA (25 mL) was added 2-bromo- 1,1 -dimethoxy ethane (1.44 g, 8.5 mmol). The mixture was stirred at 100 °C for 2 h. The cooled mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-30% ethyl acetate in petroleum ether to afford (2-chloro-5-fluorophenyl)(5,7-dibromo-9-(2,2,2-trifluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazol-6-yl)methanone (1 g, 1.81 mmol, 32%) as a yellow solid. LCMS: ESI m / z 552, 554, 556 [M + H]+.

[0415] Step B: To a solution of (2-chloro-5-fluorophenyl)(5,7-dibromo-9-(2,2,2-trifluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazol-6-yl)methanone (800 mg, 1.45 mmol) in DMSO (10 mL) was added CuCN (261 mg, 2.90 mmol). The mixture was stirred at 130 °C for 1 h. The cooled mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-50% ethyl acetate in petroleum ether to afford 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2,2-trifluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (90 mg, 0.181 mmol, 12%) as a yellow solid. LCMS: ESI m / z 498.9, 500.9 [M + H]+.

[0416] Step C: A solution of 7-bromo-6-(2-chloro-5-fluorobenzoyl)-9-(2,2,2-trifluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (90 mg, 0.181 mmol), 5-fluoro-3-(trifluoromethyl)benzene-l -carboxamide (45 mg, 0.217 mmol), Pd2(dba)3 (17 mg, 0.019 mmol), XantPhos (21 mg, 0.036 mmol), Cs2CO3(118 mg, 0.362 mmol) in dioxane (5 mL) was stirred at 100 °C for 2 h. The cooled mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel columnPatent Application Atty. Docket No. ENTX-038PCT chromatography eluting with 0-100% ethyl acetate in petroleum ether to afford N-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2,2-trifluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (80 mg, 0.128 mmol, 71%) as a yellow solid. LCMS: ESI m / z 626.0 [M + H]+.

[0417] Step D: To a solution of N-(6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2,2-trifluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazol-7-yl)-3-fluoro-5-(trifluoromethyl)benzamide (80 mg, 0.128 mmol) in MeCN (2 mL) and H2O (0.2 mL) was added KOH (40 mg, 0.714 mmol). The reaction mixture was stirred at rt for 30 min. The mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum to give N-(3-(2-chloro-5-fluorophenyl)-3 -hydroxy- 1 -oxo-6-(2,2,2-trifluoroethyl)- 1,2, 3, 6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (55 mg, 0.086 mmol, 67%) as a yellow solid. LCMS: ESI m / z 644.0 [M + H]+.

[0418] Step E: To a solution ofN-(3-(2-chloro-5-fluorophenyl)-3-hydroxy-l-oxo-6-(2,2,2-tri fluoroethyl)-!, 2, 3, 6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (55 mg, 0.086 mmol) in TFA (2 mL) was added Et3SiH (0.5 mL). The reaction mixture was stirred at 70 °C for 1 h. The reaction mixture was concentrated to give a residue. The residue was purified by prep-HPLC to give N-(3-(2-chloro-5-fluorophenyl)-l-oxo-6-(2,2,2-trifluoroethyl)-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (35 mg, 0.056 mmol, 65%) as a white solid. LCMS: ESI m / z 628.0 [M + H]+.

[0419] Step F: N-(3-(2-chloro-5-fluorophenyl)-l-oxo-6-(2,2,2-trifluoroethyl)-l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (35 mg, 0.056 mmol) was purified with chiral prep-SFC method 16 to afford (S)-N-(3-(2-chloro-5-fluorophenyl)-l-oxo-6-(2,2,2-trifluoroethyl)-l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fhioro-5-(trifhioromethyl)benzamide (11 mg, 0.018 mmol, 31%) as a white solid. 'H NMR (400 MHz, CD3OD) 68.05 (s, 1H), 7.84 - 7.60 (m, 4H), 7.33 - 7.24 (m, 1H), 7.19 (s, 1H), 7.03 - 7.00 (m, 1H), 6.65 (brs, 1H), 6.38 (brs, 1H), 5.18 - 5.00 (m, 2H). and (R)-N-(3-(2-chloro-5-fluorophenyl)-l-oxo-6-(2,2,2-trifluoroethyl)-l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (14 mg, 0.022 mmol, 40%) as a white solid. H NMR (400 MHz, CD3OD) 58.05 (s, 1H), 7.80 -Patent Application Atty. Docket No. ENTX-038PCT 7.60 (m, 4H), 7.32 - 7.24 (m, 1H), 7.19 (s, 1H), 7.03 - 7.00 (m, 1H), 6.65 (brs, 1H), 6.36 (brs, 1H), 5.17 - 4.99 (m, 2H).Example 50: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo [2 ', 1 ': 2,3] imidazo[4,5-e] isoindol-4-yl-8-d)-3-fluoro-5-(trifluoromethyl)benzamideExample 51: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3?6-tetrahydroimidazo [2 ', 1 *:2,3] imidazo[4,5-e] isoindol-4-yl-8-d)-3-fluoro-5-(trifluoromethyl)benzamideD2, PtQ>:SFCCD3ODStep AF6example 50 example 51

[0420] Step A: To a solution of N-(8-bromo-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (12 mg, 0.017 mmol) in CD3OD (1 mL) was added PtO2(4.1 mg, 0.017 mmol) at room temperature under nitrogen and the mixture was stirred under a D2 balloon at room temperature for 0.5 hr. The reaction was monitored by analysis of LCMS. The mixture was filtered through celite and the filtrate was concentrated to dryness. The residue was purified by prep-HPLC and then by chiral prep-SFC method 17 to afford: (S)-N-(3-(2-chloro-5-Patent Application Atty. Docket No. ENTX-038PCT fluorophenyl)-6-(2,2-difluoroethyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl-8-d)-3-fluoro-5-(trifluoromethyl)benzamide (1 mg, 4%), LCMS (ESI): m / z 611 [M+H]+.1HNMR(400 MHz, CD3OD) 88.04 (s, 1H), 7.77 - 7.63 (m, 4H), 7.27 (dd, J= 8.8, 5.0 Hz, 1H), 7.07 - 6.96 (m, 1H), 6.37 (t, J= 54.6 Hz, 2H), 4.71 (t, J= 15.0 Hz, 2H). And (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)- 1 -oxo- 1,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl-8-d)-3-fluoro-5-(trifluoromethyl)benzamide (1 mg, 4%), LCMS (ESI): m / z 611 [M+H]+. *H NMR (400 MHz, CD3OD) 8 8.03 (s, 1H), 7.75 - 7.60 (m, 4H), 7.26 (dd, J= 8.8, 5.0 Hz, 1H), 7.06 - 6.95 (m, 1H), 6.36 (t, J= 54.6 Hz, 2H), 4.71 (t, J= 15.0 Hz, 2H).Example 52: N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-(hydroxymethyl)-l-oxo-l,2,3?6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideBr O ClDMA, 140 °C Pd2(dba)3, xantphos,dioxane, 100 °Cstep BTES, TFA, 70 °CStep DPatent Application Atty. Docket No. ENTX-038PCT

[0421] Step A: To a solution of 2-azanylidene-6-bromo-5-[(2-chloro-4-fluorophenyl)carbonyl]-l-(2,2-difluoroethyl)-3H-benzo[d]imidazole-4-carbonitrile (3 g, 6.56 mmol) in N, N-dimethylacetamide (30 mL) was added ethyl 2-chloro-2-formylacetate (2.96 g, 19.7 mmol). The reaction was stirred at 140 °C for 2 h. LCMS showed the reaction was completed. The cooled reaction mixture was added to ice water and extracted with EA. The organic layer was washed with brine, dried over Na₂SO₄ and concentrated and the residue was purified by flash chromatography to afford ethyl 7-bromo-6-[(2-chloro-4-fluorophenyl)carbonyl]-5-cyano-9-(2,2-difluoroethyl)benzo[4,5]imidazo[2,3-b]imidazole-3-carboxylate (1 g, 1.81 mmol, 28%) as a brown solid. LCMS (ESI): m / z 553 [M+H]+.

[0422] Step B: A solution of ethyl 7-bromo-6-[(2-chloro-4-fluorophenyl)carbonyl]-5-cyano-9-(2,2-difluoroethyl)benzo[4,5]imidazo[2,3-b]imidazole-3-carboxylate (1.2 g, 2.17 mmol), 3-fluoro-5-(trifluoromethyl)benzene-l-carboxamide (0.45 g, 2.17 mmol), Pd2dba3 (0.20 g, 0.22 mmol), Xantphos (0.25 g, 0.43 mmol), CS2CO3 (2.11 g, 6.50 mmol) in dioxane (60 mL) was stirred at 90 °C for 2 h. LCMS showed the reaction was completed. The reaction mixture was concentrated, diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4and concentrated in vacuo. The residue was purified by silica gel column chromatography eluted with ethyl EA in PE (0-100% gradient) to afford ethyl 6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-7-(3-fluoro-5-(trifluoromethyl)benzamido)-9H-benzo[d]imidazo[l,2-a]imidazole-3-carboxylate (0.8 g, 1.18 mmol, 54%) as a brown solid. LCMS (ESI): m / z 680 [M+H]+.

[0423] Step C: To a solution of ethyl 6-(2-chloro-5-fluorobenzoyl)-5-cyano-9-(2,2-difluoroethyl)-7-(3-fluoro-5-(trifluoromethyl)benzamido)-9H-benzo[d]imidazo[l,2-a]imidazole-3 -carboxylate (800 mg, 1.18 mmol) in methanol (10 mL) and water (1 mL) was added potassiumPatent Application Atty. Docket No. ENTX-038PCT hydroxide (330 mg, 5.88 mmol). The mixture was stirred at rt for 16 h. LCMS showed the reaction was complete. The reaction mixture was concentrated, diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4and concentrated in vacuo to get ethyl 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-3-hydroxy-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindole-9-carboxylate (700 mg, 1.00 mmol, 85%) as a crude. LCMS (ESI): m / z 698 [M+H]+.

[0424] Step D: A solution of ethyl 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(tri fluoromethyl)benzamido)-3 -hydroxy- 1 -oxo-1, 2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindole-9-carboxylate (700 mg, 1.00 mmol) in TFA (10 mL) was added TES (1 mL). The mixture was stirred at 70 °C for 16 h. LCMS showed the reaction was completed. The cooled reaction mixture was concentrated, and purified by silica gel column chromatography eluted with ethyl EA in PE (0-100% gradient) to afford to afford ethyl 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindole-9-carboxylate (420 mg, 0.62 mmol, 64%) as a brown solid. LCMS (ESI): m / z 698 [M+H]+.

[0425] Step E: To a solution of ethyl 3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-(3-fluoro-5-(trifluoromethyl)benzamido)-l-oxo-l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindole-9-carboxylate (420 mg, 0.62 mmol ) in THF (2 mL) was added LiAlH4(93 mg, 2.46 mmol). The mixture was stirred at 0°C for 16 h. LCMS showed the reaction was completed. The mixture was then re-cooled to 0° C. and water (0.1 mL), 15% sodium hydroxide solution (O.lmL) and water (0.3 mL) were carefully added. The mixture was diluted with THF, filtered and the filtrate was concentrated in vacuo and purified by pre-HPLC to afford N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-9-(hydroxymethyl)- 1 -oxo- 1,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (260 mg, 0.41 mmol, 66%) as a white solid. LCMS (ESI): m / z 640 [M+H]+.1H NMR (400 MHz, DMSO-d6) 8 10.55 (s, 1H), 9.68 (s, 1H), 7.96 (d, J= 8.4 Hz, 1H), 7.85 (s, 1H), 7.78 (d, J= 9.2 Hz, 1H), 7.74 (s, 1H), 7.33 - 7.29 (m, 1H), 7.13 - 7.10 (m, 1H), 7.07 (s, 1H), 6.54 (t, J= 54.4 Hz, 2H), 6.12 (s, 1H), 5.37 (t, J= 6.0 Hz, 1H), 4.99 (d, J= 6.6 Hz, 2H), 4.84 - 4.70 (m, 2H).Example 53: (R)-8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((6-fluoro-8-(trifluoromethyl)quinazolin-4-yl)amino)-3,6-dihydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-l(2H)-onePatent Application Atty. Docket No. ENTX-038PCT Example 54: (S)-8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((6-fluoro-8-(trifluoromethyl)quinazolin-4-yl)amino)-3,6-dihydroimidazo[2',l':2,3]imidazo[4,5-elisoindol-l(2H)-oneNHPd2dba3, XantPhos CS2CO3, dioxane Step AExample 53 Example 54Patent Application Atty. Docket No. ENTX-038PCT

[0426] Step A: To a solution of 7-bromo-2-chloro-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (1.4 g, 2.7 mmol) in dioxane (80 mL) was added diphenylmethanimine (0.74 g, 4.07 mmol), Xantphos (0.31 g, 0.54 mmol), CS2CO3 (2.65 g, 8.14 mmol) and Pd2(dba)3(0.25 g, 0.27 mmol). The mixture was stirred at 100 °C under nitrogen for 12 hours. The cooled mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-30% ethyl acetate in petroleum ether to afford 2-chloro-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-7-((diphenylmethylene)amino)-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (1.3 g, 2.11 mmol, 77%) as a yellow solid. ESI m / z 616 [M + H]+.

[0427] Step B: To a solution of 2-chloro-6-(2-chloro-5-fluorobenzoyl)-9-(2,2-difluoroethyl)-7-((diphenylmethylene)amino)-9H-benzo[d]imidazo[l,2-a]imidazole-5-carbonitrile (1.2 g, 1.95 mmol) in ACN (4 mL) and H2O (20 mL) was added KOH (0.55 g, 9.73 mmol). The mixture was stirred at 25 °C for 1 hour. The mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-10% methanol in dichloroform to afford 8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((diphenylmethylene)amino)-3-hydroxy-3,6-dihydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-l(2H)-one (1.2 g, 1.95 mmol, 99%) as a yellow solid. LCMS: ESI m / z 634 [M + H]+.

[0428] Step C: To a solution of 8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((diphenylmethylene)amino)-3-hydroxy-3,6-dihydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-l(2H)-one (1.2 g, 1.95 mmol) in ACN (20 mL) was added HC1 (2N, 5 mL, 10.00 mmol). The mixture was stirred at 25 °C for 1 hours. The mixture was diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-30% ethyl acetate in petroleum ether to afford 4-amino-8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-3,6-dihydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-l(2H)-one (800 mg, 1.70 mmol, 87%) as a yellow solid. LCMS: ESI m / z 470 [M + H]+.

[0429] Step D: To a solution of 4-amino-8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-3,6-dihydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-l(2H)-one (700 mg, 1.49 mmol) in TFA (5 mL) was added Et3SiH (1 mL, 6.02 mmol). The mixture was stirredPatent Application Atty. Docket No. ENTX-038PCT at 40 °C for 10 minutes. The cooled mixture was concentrated under vacuum. The residue was purified using silica gel column chromatography eluting with 0-30% ethyl acetate in petroleum ether to afford 4-amino-8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3,6-dihydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-l(2H)-one (400 mg, 0.88 mmol, 59%) as a yellow solid. LCMS: ESI m / z 454 [M + H]+.

[0430] Step E: To a solution of 4-chloro-6-fluoro-8-(trifluoromethyl)quinazoline (200 mg, 0.80 mmol) in 1,1 -dideuteri o-l-(deuteriooxidanyl)-2, 2, 2-trifluoroethane (5 mL) was added 4-amino-8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3,6-dihydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-l(2H)-one (100 mg, 0.22 mmol). The mixture was stirred at 100 °C for 2 hours. The cooled mixture was concentrated under vacuum. The residue was purified using pre-HPLC to afford 8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((6-fluoro-8-(trifluoromethyl)quinazolin-4-yl)amino)-3,6-dihydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-l(2H)-one (30 mg, 0.04 mmol, 20%) as a yellow solid. LCMS: ESI m / z 668 [M + H]+.

[0431] Step F: 8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((6-fluoro-8-(trifluoromethyl)quinazolin-4-yl)amino)-3,6-dihydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-l(2H)-one (30 mg, 0.04 mmol) was purified with chiral prep-SFC method 18 to afford (R)-8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((6-fluoro-8-(trifluoromethyl)quinazolin-4-yl)amino)-3,6-dihydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-l(2H)-one (8.0 mg, 0.01 mmol, 53%) as a white solid. LCMS: ESI m / z 668 [M + H]+. ’H NMR (400 MHz, DMSO-d6) 8 10.09 (s, 1H), 9.36 (s, 1H), 8.46 (s, 1H), 8.27 - 8.15 (m, 2H), 7.93 -7.78 (m, 2H), 7.06 - 6.83 (m, 2H), 6.69 - 6.37 (m, 2H), 6.19 - 5.89 (m, 1H), 4.86 - 4.70 (m, 2H). And (S)-8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-4-((6-fluoro-8-(trifluoromethyl)quinazolin-4-yl)amino)-3,6-dihydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-l(2H)-one (9.5 mg, 0.01 mmol, 63%) as a white solid. LCMS: ESI m / z 668 [M + H]+. 'H NMR (400 MHz, DMSO-d6) 8 10.08 (s, 1H), 9.36 (s, 1H), 8.47 (s, 1H), 8.31 - 8.15 (m, 2H), 7.93 -7.76 (m, 2H), 7.05 - 6.81 (m, 2H), 6.68 - 6.34 (m, 2H), 6.21 - 5.93 (m, 1H), 4.85 - 4.70 (m, 2H).Example 55: (R)-N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(ethyl-d5)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCT Example: 56 (S)-N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(ethyl-d5)-l-oxo-l, 2,3,6-tetrahydroimidazo [2 ’, 1 ':2,3] imidazo[4,5-e] isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideKOHStep CSFCStep ECl Cl Example 55 Example 56

[0432] Step A: To a solution of 7-bromo-6-[(2-chloro-5-fluorophenyl)carbonyl]-2-oxo-9- (pentadeuterioethyl)-3,9-dihydro-2H-benzo[4,5]imidazo[2,3-b]imidazole-5-carbonitrile (25 mg, 0.05 mmol) in POCl3(1 mL) was added DIEA (13.8 mg, 0.11 mmol). The mixture was stirred at 80 °C for 2 hours. The reaction mixture was concentrated. The crude product was purified by pre-TLC (50% EtOAc in PE) to afford 7-bromo-2-chloro-6-[(2-chloro-5-fluorophenyl)carbonyl]-Patent Application Atty. Docket No. ENTX-038PCT 9-(pentadeuterioethyl)benzo[4,5]imidazo[2,3-b]imidazole-5-carbonitrile (10 mg, 39%) as a white solid. LCMS(ESI): m / z 484 [M+H]+.

[0433] Step B: To a solution of 7-bromo-2-chloro-6-[(2-chloro-5-fluorophenyl)carbonyl]-9-(pentadeuterioethyl)benzo[4,5]imidazo[2,3-b]imidazole-5-carbonitrile (10 mg, 0.02 mmol) in dioxane (1 mL) was added 3-fluoro-5-(trifluoromethyl)benzene-l-carboxamide (6.40 mg, 0.03 mmol), Pd2(dba)3 (1.89 mg, 0.00 mmol), XANT PHOS (2.39 mg, 0.00 mmol) and CS2CO3 (16.79 mg, 0.05 mmol). The mixture was stirred at 100 °C under N2 for 2 hours. The cooled reaction mixture was quenched by water, extracted with EtOAc. The organic layer was washed with brine, dried over Na₂SO₄ and concentrated. The crude product was purified by prep-TLC (60% EtOAc in PE) to afford N-{2-chloro-6-[(2-chloro-5-fluorophenyl)carbonyl]-5-cyano-9-(pentadeuterioethyl)benzo[4,5]imidazo[2,3-b]imidazol-7-yl)-5-fluoro-3-(trifluoromethyl)benzamide (10 mg, 0.02 mmol, 80%) as a white solid. LCMS (ESI): m / z 611 [M+H]+.

[0434] Step C: To a solution of N-{2-chloro-6-[(2-chloro-5-fluorophenyl)carbonyl]-5-cyano-9-(pentadeuterioethyl)benzo[4,5]imidazo[2,3-b]imidazol-7-yl]-5-fluoro-3-(trifluoromethyl)benzamide (10 mg, 0.02 mmol) in MeCN (1 mL) and H2O (0.5 mL) was added KOH (4.59 mg, 0.08 mmol). The mixture was stirred at room temperature for 0.5 hour. The reaction mixture was quenched by water, extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated. The crude product was purified by prep-TLC (80% EtOAc in PE) to afford N-[8-chloro-3-(2-chloro-5-fluorophenyl)-3-hydroxy-l -oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (10 mg, 97%) as a light white solid. LCMS(ESI): m / z 629 [M+H]+.

[0435] Step D: To a solution of N-[8-chloro-3-(2-chloro-5-fluorophenyl)-3-hydroxy-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (6 mg, 0.01 mmol) in TFA (1 mL) was added triethylsilane (0.4 mL). The mixture was stirred at 40 °C for 0.5 hour. The reaction mixture was poured into sodium bicarbonate aqueous solution and extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4 and concentrated. The crude product was purified by prep-TLC (50% EtOAc in PE) to give N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(ethyl-d5)-l-oxo-l,2,3,6-tetrahydroimidazo[2', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (6 mg, 80%). LCMS(ESI): m / z 613 [M+H]+.Patent Application Atty. Docket No. ENTX-038PCT

[0436] Step E: N-(8-chloro-3-(2-chloro-5-fluorophenyl)-6-(ethyl-d5)-l-oxo-l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (6 mg) was purified by chiral prep-SFC method 19 to give N-[(3R)-8-chloro-3-(2-chloro-5-fluorophenyl)-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (2.4 mg, 41%) as a white solid. ’HNMR (400 MHz, CD3OD) 87.91 (s, 1H), 7.71 - 7.65 (m, 4H), 7.27 (dd, J= 8.8, 5.0 Hz, 1H), 7.03 -6.89 (m, 1H), 6.34 (s, 1H). and N-[(3S)-8-chloro-3-(2-chloro-5-fluorophenyl)-l-oxo-6-(pentadeuterioethyl)-2,3-dihydro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (1.3 mg, 22%) as a white solid. 'H NMR (400 MHz, CD3OD) 87.92 (s, 1H), 7.71 - 7.65 (m, 4H)„ 7.27 (dd, J= 8.8, 5.0 Hz, 1H), 7.03 - 6.89 (m, 1H), 6.34 (s, 1H). LCMS(ESI): m / z 613 [M+H]+Example 57: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-(methylsulfonyl)-l-oxo-1,2,3,6-tetrahydroimidazo [2',1 ':2,3 Jim idazo [4,5-e] isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamideExample 58: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-(methylsulfonyl)-l-oxo-1,2,3,6-tetrahydroimidazo [2*, 1 *:2,3]imidazo [4,5-e] isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCTSFCStep C

[0437] Step A: To a solution ofN-[8-chloro-3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-3-hydroxy-l-oxo-2,3-dihydro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (50 mg, 0.08 mmol) in ACN (1 mL) were added NaI (56.8 mg, 0.38 mmol) and TMSCl (41.1 mg, 0.38 mmol). The mixture was stirred at RT for ON. The reaction mixture was diluted water, extracted with EA. The organic phase was washed with brine, dried over Na₂SO₄ and concentrated. The residue was purified using silica gel column chromatography eluted with ethyl acetate in petroleum ether (gradient: 0-50%) to afford N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-iodo-l-oxo-2,3-dihydro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (40 mg, 0.05 mmol, 72%) as a yellow solid. LCMS: (ESI) m / z 736.1 [M+H]+.

[0438] Step B: To a solution ofN-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-iodo-l-oxo-2,3-dihydro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (40 mg, 0.05 mmol) in DMSO (3 mL) were added sodium methanesulfmate (6.66 mg, 0.07 mmol), Cui (1.04 mg, 0.01 mmol) and L-proline (2.50 mg, 0.02 mmol). The mixture was stirred at 140 °C with Microwave for Ih. The cooled reactionPatent Application Atty. Docket No. ENTX-038PCT mixture was purified by prep-HPLC to afford N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-(methyldioxo-X6-sulfanyl)-l-oxo-2,3-dihydro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (7 mg, 0.01 mmol, 19%) as a white solid. LCMS: (ESI) m / z 688.1 [M+H]+.

[0439] Step C: N-[3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-(methyldioxo-X6-sulfanyl)-l-oxo-2,3-dihydro-lH-imidazo[2',3':2,3]imidazo[4,5-e]isoindol-4-yl]-5-fluoro-3-(trifluoromethyl)benzamide (7 mg, 0.01 mmol) was purified with chiral prep-SFC method 20 to afford (S)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-(methylsulfonyl)-l-oxo- l,2,3,6-tetrahydroimidazo[2',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (3.0 mg ) as a white solid.NMR (400 MHz, DMSO-d6) 8 10.67 (s, 1H), 9.52 (s, 1H), 8.43 (s, 1H), 7.96 (d, J= 8.0 Hz, 1H), 7.86 (s, 1H), 7.80 - 7.70 (m, 2H), 7.37 - 7.29 (m, 1H), 7.15 - 7.08 (m, 1H), 6.72 - 6.06 (m, 3H), 4.96 - 4.72 (m, 2H), 3.28 (s, 3H). And (R)-N-(3-(2-chloro-5-fluorophenyl)-6-(2,2-difluoroethyl)-8-(methylsulfonyl)-l-oxo-l,2,3,6-tetrahydroimidazo[2',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (2.5 mg ) as a white solid. ‘HNMR (400 MHz, DMSO-d6) 8 10.68 (s, 1H), 9.51 (s, 1H), 8.43 (s, 1H), 7.96 (d, J= 8.0 Hz, 1H), 7.86 (s, 1H), 7.82 - 7.70 (m, 2H), 7.37 - 7.29 (m, 1H), 7.15 - 7.08 (m, 1H), 6.72 - 6.06 (m, 3H), 4.97 - 4.68 (m, 2H), 3.27 (s, 3H).Example 59: (R)-N-(3-(2-chloro-5-fluorophenyl)-6-ethyl-l-oxo-l,2,3,6-tetrahydro- [1.2.4]triazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide Example 60: (S)-N-(3-(2-chloro-5-fluorophenyl)-6-ethyl-l-oxo-l,2,3>6-tetrahydro- [1.2.4]triazolo[5',r:2,3]imidazo[4,5-elisoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamidePatent Application Atty. Docket No. ENTX-038PCTExample 59 Example 60

[0440] Step A: A solution of (2-chloro-5-fluorophenyl)(2,6-dibromo-3-(5-chloro-lH-l,2,4-triazol-l-yl)-4-fluorophenyl)methanone (600 mg, 1.17 mmol) in DMA (10 mL) was added K2CO3 (1.62 g, 11.7 mmol) and Ethylamine hydrochloride (478 mg, 5.86 mmol). The mixture was stirred at 120 °C for 16 hrs. The cooled reaction mixture was diluted with H2O and extracted with EtOAc. The comLined organic layers were dried over anhydrous Na2SO4, filtered and the filtrate was concentrated to dryness. The residue was purified by silica gel flash chromatography, eluted with a gradient of 0 - 40% EtOAc in PE to afford (2-chloro-5-fluorophenyl)(6,8-dibromo-4-ethyl-4H-benzo[4,5]imidazo[l,2-b][l,2,4]triazol-7-yl)methanone (350 mg, 71%) as a brown solid. LCMS: (ESI) m / z 501 [M+H]+.

[0441] Step B: To a solution of (2-chloro-5-fluorophenyl)(6,8-dibromo-4-ethyl-4H-benzo[4,5]imidazo[l,2-b][l,2,4]triazol-7-yl)methanone (350 mg, 0.70 mmol) in DMSO (4 mL) was added CuCN (75.2 mg, 0.84 mmol). The mixture was stirred at 140 °C under N2 for 2 hrs. The cooled mixture was diluted with water and filtered through celite. The filtrate was extracted with EtOAc. The comLined organic layers were washed with water, brine, filtered and the filtrate was concentrated to dryness. The residue was purified by silica gel flash chromatography, eluted with a gradient of 0 - 40% EtOAc in PE to afford 6-bromo-7-(2-chloro-Patent Application Atty. Docket No. ENTX-038PCT 5-fluorobenzoyl)-4-ethyl-4H-benzo[4,5]imidazo[l,2-b][l,2,4]triazole-8-carbonitrile (60 mg, 19%) as a brown solid. LCMS: (ESI) m / z 446 / 448 [M+H]+.

[0442] Step C: To a mixture of 6-bromo-7-(2-chloro-5-fluorobenzoyl)-4-ethyl-4H-benzo[4,5]imidazo[l,2-b][l,2,4]triazole-8-carbonitrile (60 mg, 0.13 mmol), 3-fluoro-5-(trifluoromethyl)benzamide (27.82 mg, 0.13 mmol), CS2CO3 (87.5 mg, 0.27mmol) and Xantphos (15.5 mg, 0.03 mmol) in dioxane (3 mL) was added Pd2(dba)3(12.3 mg, 0.01 mmol). The mixture was stirred at 100 °C under N2 for 2 hrs. The cooled reaction was monitored by analysis of LCMS. The mixture was filtered through celite and the filtrate was diluted with water. The mixture was extracted with EtOAc. The comLined organic layers were dried over anhydrous Na2SO4, filtered and concentrated to dryness to afford N-(7-(2-chloro-5-fluorobenzoyl)-8-cyano-4-ethyl-4H-benzo[4,5]imidazo[l,2-b][l,2,4]triazol-6-yl)-3-fluoro-5-(trifluoromethyl)benzamide (60 mg, 78%) as a yellow solid. LCMS: (ESI) m / z 573 [M+H]+.

[0443] Step D: To a solution of N-(7-(2-chloro-5-fluorobenzoyl)-8-cyano-4-ethyl-4H-benzo[4,5]imidazo[l,2-b][l,2,4]triazol-6-yl)-3-fluoro-5-(trifluoromethyl)benzamide (60 mg, 0.10 mmol) in CH3CN (2 mL) and H2O (0.2 mL) was added KOH (29.3 mg, 0.52 mmol) at room temperature and the mixture was stirred at room temperature for 1 hour. The reaction was monitored by analysis of LCMS. The mixture was diluted with water and extracted with EtOAc. The comLined organic layers were dried over anhydrous Na2SO4, filtered and the filtrate was concentrated to dryness. The residue was purified by prep-TLC (DCM: MeOH= 10: 1) to afford N-(3-(2-chloro-5-fluorophenyl)-6-ethyl-3-hydroxy-l -oxo-1,2,3, 6-tetrahydro-[l,2,4]triazolo[5', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (30 mg, 48%). LCMS: (ESI) m / z m / z 591 [M+H]+.

[0444] Step E: To a solution ofN-(3-(2-chloro-5-fluorophenyl)-6-ethyl-3-hydroxy-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[5', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (30 mg, 0.05 mmol) in TFA (1.5 mL) was added Et3SiH (0.3 mL, 1.88 mmol) at room temperature and the mixture was stirred at 60 °C for 0.5hour. The reaction was monitored by analysis of LCMS. The mixture was concentrated to dryness under reduced pressure. The residue was purified by prep-HPLC to afford N-(3-(2-chloro-5-fluorophenyl)-6-ethyl-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[5', T:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (18 mg, 65%). LCMS: (ESI) m / z 575 [M+H]+.Patent Application Atty. Docket No. ENTX-038PCT

[0445] Step F: N-(3-(2-chloro-5-fluorophenyl)-6-ethyl-l-oxo-l,2,3,6-tetrahydro- [1.2.4]triazolo[5',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (18 mg, 65%) was purified by chiral prep-SFC method 21 to give (R)-N-(3-(2-chloro-5-fluorophenyl)-6-ethyl-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[5',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (3.3 mg, 12%) as a white solid. ’H NVIR (400 MHz, DMSO-d6) 5 10.55 (s, 1H), 9.25 (s, 1H), 8.18 (s, 1H), 8.01 - 7.88 (m, 2H), 7.81 - 7.69 (m, 2H), 7.30 (dd, J= 8.8, 5.0 Hz, 1H), 7.13 - 7.02 (m, 1H), 6.80 - 5.96 (m, 2H), 4.46 - 4.28 (m, 2H), 1.46 (t, J= 7.2 Hz, 3H). And (S)-N-(3-(2-chloro-5-fluorophenyl)-6-ethyl-l-oxo-l,2,3,6-tetrahydro-[l,2,4]triazolo[5',r:2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide (4.7 mg, 16%) as a white solid. ’H NMR. (400 MHz, DMSO-d6) 8 10.55 (s, 1H), 9.25 (s, 1H), 8.18 (s, 1H), 8.01 - 7.88 (m, 2H), 7.84 - 7.68 (m, 2H), 7.30 (dd, J = 8.8, 5.0 Hz, 1H), 7.13 - 7.02 (m, 1H), 6.76 - 5.72 (m, 2H), 4.44 - 4.28 (m, 2H), 1.46 (t, J= 7.2 Hz, 3H).Example 61: (S)-N-(3-(2-chloro-5-fluorophenyl)-l-oxo-6-propyl-l,2,3>6-tetrahydro- [1.2.4]triazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamide Example 62: (R)-N-(3-(2-chloro-5-fluorophenyl)-l-oxo-6-propyl-l,2,3,6-tetrahydro- [l,2,4]triazolo[5',l':2,3]imidazo[4,5-e]isoindol-4-yl)-3-fluoro-5-(trifluoromethyl)benzamidePd2(dba)3, xantphosCs2CO3, dioxane Step CPatent Application Atty. Docket No. ENTX-038PCT

[0446] Step A: To a solution of (2-chloro-5-fluorophenyl)(2,6-dibromo-3-(5-chloro-lH-l,2,4-triazol-l-yl)-4-fluorophenyl)methanone (400 mg, 0.78 mmol) in DMA (8 mL) was added propan- 1 -amine (230 mg, 3.91 mmol) and K2CO3 (539 mg, 3.91 mmol). The mixture was stirred at 120 °C for 2 h. The cooled mixture was quenched with water, extracted with EtOAc. The organic layer was washed with water and brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purifie...

Claims

1. Patent Application Atty. Docket No. ENTX-038PCT What is claimed is:CLAIMS1. A compound having the structural formula (I):(I)or a pharmaceutically acceptable form or an isotope derivative thereof,whereinX1is N or CR1;X2is N, C(O) or CR2;X3is N or CR3;each of R1, R2and R3is independently H, D, halo, CN, R, OR, C(O)NHR, C2-6 alkynyl, NRR', SO2R or SO2NR, wherein said R and C2-6 alkynyl are optionally substituted with one or more of Ra;R4is a 5- to 10-memLered monocyclic, bicyclic or bridged aryl or heteroaryl ring with 0-4 ring heteroatoms independently selected from N, O and S, optionally substituted with one or more Re;R5is C(O)R3or R5", whereinR5'is a 5- to 10-memLered monocyclic, bicyclic or bridged carbocyclyl, heterocyclic, aryl or heteroaryl ring with 0-4 ring heteroatoms independently selected from N, O and S, optionally substituted with one or more Rb; andR5"is a 9- or 10-memLered bicyclic or bridged heteroaryl ring with 0-4 ring heteroatoms independently selected from N, O and S, optionally substituted with one or more Rb;R6is a C1-6 alkyl substituted withPatent Application Atty. Docket No. ENTX-038PCT 0-8 Rd;an unsubstituted or substituted C3-6 carbocyclic or heterocyclic group; or an unsubstituted or substituted 5- or 6-memLered aryl or heteroaryl group; each Rais independently D, halo, OH, CN, C1-6 alkoxy, NRR' or SO2R; each of Rband Rcis independentlydeuterium, oxo, halogen, CN, NO2, OR, SR, NRR’, S(O)2R, S(O)2NRR’, S(O)R, S(O)NRR’, S(O)(NR)R, C(O)R, C(O)OR, C(O)NRR’, C(O)N(R)OR, OC(O)R, OC(O)NRR’, N(R)C(O)OR, N(R)C(O)R, N(R)C(O)NRR’, N(R)C(NR)NRR’, N(R)S(O)2NRR’ or N(R)S(O)2R; ora substituted or unsubstituted group selected from C1-6 alkyl or 4- to 6- memLered carbocyclic ring;each Rdis independently D, halogen, CN, OR, alkynyl, SR or NRR’; each of R and R’ is independently selected from H, unsubstituted or substituted C1-4 alkyl, or unsubstituted or substituted 3- to 6-memLered carbocyclic ring, or where R and R’ are attached to the same C or N atom, together form an unsubstituted or substituted 4- to 6-memLered heterocyclic ring.

2. The compound of claim 1, wherein X1is N, having the structural formula (IA):R5NHr4r vNX2(IA)3. The compound of claim 1, wherein X1is CR1, having the structural formula (IB):Patent Application Atty. Docket No. ENTX-038PCT4. The compound of claim 2 or 3, wherein Ring A is an aromatic ring.

5. The compound of claim 2 or 3, wherein X2is N and X3is CR3.

6. The compound of claim 2 or 3, wherein X2is CR2and X3is N.

7. The compound of claim 2 or 3, wherein X2is CR2and X3is CR3.

8. The compound of claim 2 or 3, wherein X2is C(O) and X3is N.

9. The compound of any one of claims 1-8, wherein R4is a 6-memLered aryl or heteroaryl ring with 0-3 ring N atoms, substituted with 0-6 Res,10. The compound of claim 9, wherein R4is a substituted or unsubstituted phenyl, pyridyl, pyridazinyl or pyrazinyl.

11. The compound of claim 10, wherein R4is a substituted or unsubstituted phenyl.

12. The compound of claim 10, wherein R4is a substituted or unsubstituted pyridyl.

13. The compound of claim 10, wherein R4is a substituted or unsubstituted pyridazinyl.

14. The compound of claim 10, wherein R4is a substituted or unsubstituted pyrazinyl.

15. The compound of any one of claims 1-11, wherein R4is selected from:Patent Application Atty. Docket No. ENTX-038PCT16. The compound of claim 15, wherein R4is:i17. The compound of any one of claims 1-16, having the following chirality:i °18. The compound of any one of claims 1-16, having the following chirality:I R4O19. The compound of any one of claims 1-18, wherein R5' is a substituted or unsubstituted 5- or 6-memLered monocyclic aryl or heteroaryl ring.

20. The compound of any one of claims 1-18, wherein R5is a substituted or unsubstituted 8- to 10-memLered bicyclic carbocyclyl or heterocyclic.

21. The compound of any one of claims 1-18, wherein R5is a substituted or unsubstituted 8- to 10-memLered bicyclic aryl or heteroaryl ring.

22. The compound of any one of claims 1-18, wherein R5'is selected from:Patent Application Atty. Docket No. ENTX-038PCTwherein j is 0, 1, 2, 3 or 4.

23. The compound of any one of claims 1-18, wherein R5’ is selected from:

24. The compound of any one of claims 1-18, wherein R5isPatent Application Atty. Docket No. ENTX-038PCT25. The compound of any one of claims 1-18, wherein R5is(II)whereinY is N or CRY;RYis H or Ci-6 alkyl; andis 0, 1, 2, 3 or 4.

26. The compound of claim 25, wherein R5is:(IIA)27. The compound of claim 25 or 26, wherein Y is N.

28. The compound of claim 25 or 26, wherein Y is CH.

29. The compound of any one of claims 1-28, wherein R6is CR6AR6B]6C, having the structural formula (IIIA)Patent Application Atty. Docket No. ENTX-038PCTR6^R6BZR6C(IIIAwhereinR6Ais independently H, D or halogen;R6Bis H, D or halogen; andR6CisH, D, halogen, CN, OR, SR, NRR’, S(O)2R, S(O)2NRR', S(O)R, S(O)NRR' S(O)(NR)R or C(O)NRR’; oran unsubstituted or substituted C1-5 alkyl, C2-5 alkenyl, C2-5 alkynyl, C3-6 carbocyclic or heterocyclic group, or 5- or 6-memLered aryl or heteroaryl group.

30. The compound of claim 29, wherein R6Cis CR6DR6ER6F,whereinR6Dis H, D or halogen; andeach of R6Eand R6Fis independentlyH, D, halogen, CN, OR, SR, NRR’, S(O)2R, S(O)2RR’, S(O)R, S(O)NRR’, S(O)(NR)R or C(O)NRR’; oran unsubstituted or substituted C1-5 alkyl, C2-5 alkenyl, C2-5 alkynyl, C3-6 carbocyclic or heterocyclic group, or 5- or 6-memLered aryl or heteroaryl group.

31. The compound of claim 29 or 30, wherein each of R6Aand R6Bis H.

32. The compound of claim 29 or 30, wherein each of R6Aand R6Bis D.

33. The compound of any one of claims 30-32, wherein at least one of R6D, R6Eand R6Fis a halogen atom.

34. The compound of claim 33, wherein one of R6D, R6Eand R6Fis F.

35. The compound of claim 33, wherein two of R6D, R6Eand R6Fis F.

36. The compound of claim 33, wherein each of R6D, R6Eand R6Fis F.

37. The compound of any one of claims 30-32, wherein at least one of R6D, R6Eand R6Eis aPatent Application Atty. Docket No. ENTX-038PCT D.

38. The compound of claim 33, wherein one of R6D, R6Eand R6Fis D.

39. The compound of claim 33, wherein two of R6D, R6Eand R6Fis D.

40. The compound of claim 33, wherein each of R6D, R6Eand R6Eis D.

41. The compound of any one of claims 1-28, wherein R6are independently selected from:CH3, CH2D, CHD2, CD3, CH2CH3, CD2CH3, CH2CD3, CD2CD3, CH2CH2F, CD2CH2F, CH2CD2F, CD2CD2F, CH2CHF2, CD2CHF2, CH2CDF2, CD2CDF2, CH2CF3, CD2CF3, CH2CN, CH2CH2N(CH3)2, CH2CH2OH, CH2CH2OCH3, CH2CH2CH2F, CH2CH2CHF2, CH2CH2CF3, propyl, isopropyl, n-butyl, isobutyl,42. The compound of claim 1, having the structural formula (IVA):(IVA)43. The compound of claim 1, having the structural formula (IVB):Patent Application Atty. Docket No. ENTX-038PCT(IVB)44. The compound of claim 1, having the structural formula (VA):(VA)45. The compound of claim 1, having the structural formula (VB):Patent Application Atty. Docket No. ENTX-038PCT46. The compound of claim 1, having the structural formula (VC):<7X2(VC)47. The compound of claim 1, having the structural formula (VD):

48. The compound of claim 1, wherein R6is CH2R6or CD2R6, wherein R6is a substituted or unsubstituted C1.2 alkyl.

49. The compound of claim 48, wherein R6' is CH3, CD3, CH2F, CHF2, CF3.

50. The compound of claim 48 or 49, having the structural formula (VIA):Patent Application Atty. Docket No. ENTX-038PCT51. The compound of claim 48 or 49, having the structural formula (VIB):(VI“)52. The compound of claim 48 or 49, having the structural formula (VIC):Patent Application Atty. Docket No. ENTX-038PCT 53. The compound of claim 48 or 49, having the structural formula (VID):(VID)54. The compound of claim 48 or 49, having the structural formula (VIE):

55. The compound of claim 48 or 49, having the structural formula (VIF):Patent Application Atty. Docket No. ENTX-038PCT 56. The compound of claim 48 or 49, having the structural formula (VIG):

57. The compound of claim 48 or 49, having the structural formula (VIH):

58. The compound of claim 48 or 49, having the structural formula (VIIA):(VIIA)Patent Application Atty. Docket No. ENTX-038PCT 59. The compound of claim 48 or 49, having the structural formula (VIIB):(VIIB)60. The compound of claim 48 or 49, having the structural formula (VIIC):(VIIC)61. The compound of claim 48 or 49, having the structural formula (VIID):

62. The compound of claim 48 or 49, having the structural formula (VIIE):Patent Application Atty. Docket No. ENTX-038PCT63. The compound of claim 48 or 49, having the structural formula (VIIF):

64. The compound of claim 48 or 49, having the structural formula (VIIG):

65. The compound of claim 48 or 49, having the structural formula (VIIH):Patent Application Atty. Docket No. ENTX-038PCT66. A compound selected from Table 1, or a pharmaceutically acceptable form or an isotope derivative thereof.

67. The compound of any of claims 1-66, having one or more deuterium atoms in place of hydrogen.

68. A pharmaceutical composition comprising a compound according to any one of claims 1- 67 and a pharmaceutically acceptable excipient, carrier, or diluent.

69. The pharmaceutical composition of claim 68, being suitable for oral administration.

70. A unit dosage form comprising a pharmaceutical composition according to claim 68 or 6971. The unit dosage form of claim 70, being in the form of a tablet or capsule.

72. A method for inhibiting cellular proliferation in vitro or in vivo, comprising contacting a cell with an effective amount of a compound according to any one of claims 1-67.

73. A method for modulating phosphoinositide 3-kinase α (PI3Kα) activity in a cell, comprising contacting the cell with a compound according to any one of claims 1-67.

74. A method for reducing phosphoinositide 3-kinase α (PI3Kα) activity in a cell, comprising contacting the cell with a compound according to any one of claims 1-67.

75. A method for treating a disease or disorder mediated by phosphoinositide 3-kinase α (PI3Kα), comprising administering to a subject in need thereof a therapeutically effective amount of the compound according to any one of claims 1-67.

76. The method of claim 75, wherein the disease or disorder is a cellular proliferative disease.

77. A method for treating or reducing cancer, or a related disease or disorder, comprising administering to a subject in need thereof a therapeutically effective amount of thePatent Application Atty. Docket No. ENTX-038PCT compound according to any one of claims 1-67.

78. The method of claim 77, wherein the cancer is selected from the group consisting of carcinoma, squamous carcinoma, adenocarcinoma, sarcoma, leukemia, neuroma, melanoma, and lymphoma.

79. The method of claim 77 or 78, wherein the cancer is selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, colorectal cancer, lung cancer, endometrial cancer, appendix cancer, cholangiocarcinoma, bladder urothelial cancer, gastric cancer, bile duct cancer, and a hematologic malignancy.

80. The method of any one of claims 77-79, wherein the subject has a mutated class IA PI3K p110α.

81. The method of any one of claims 77-80, wherein the subject has at least one of the following PI3Kα mutations: H1047R, E542K, E545K and H1047X.

82. The method of any one of claims 77-81, wherein the subject being treated is further administered one or more of chemotherapy, radiotherapy, targeted therapy, immunotherapy, and hormonal therapy.

83. Use of the compound according to any one of claims 1-67, and a pharmaceutically acceptable excipient, carrier, or diluent, in preparation of a medicament for treating a disease or disorder.

84. The use of claim 83, wherein the disease or disorder is a cellular proliferative disease.

85. The use of claim 84, wherein the disease or disorder is cancer.

86. The use of claim 85, wherein the cancer is selected from the group consisting of carcinoma, squamous carcinoma, adenocarcinoma, sarcoma, leukemia, neuroma, melanoma, and lymphoma.

87. The use of claim 85, wherein the cancer is selected from the group consisting of ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colorectal cancer, small and non-small cell lung cancer, endometrial cancer, appendix cancer, cholangiocarcinoma, bladder urothelial cancer, gastric carcinomas, bile duct cancer, hepatocellular carcinoma, thyroid carcinoma, and a hematologic malignancy.

88. The use of claim 87, wherein the cancer is selected from the group consisting of acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), andglioblastomas.

Images