Modified thiostrepton-inspired compounds for treatment of cancer and preparation thereof

Abstract

Disclosed herein are structures, compositions and syntheses of pharmaceutically relevant compounds inspired by thiostrepton, as well as methods of treating cancer with such compounds.

Description

[0001] MODIFIED THIOSTREPTON-INSPIRED COMPOUNDS FOR TREATMENT OF CANCER AND PREPARATION THEREOF

[0002] CROSS REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 662,826 filed on June 21, 2024, U.S. Provisional Patent Application No. 63 / 683,018, filed on August 14, 2024, and U.S. Provisional Patent Application No. 63 / 757,092, filed on February 11, 2025, each of which is incorporated by reference herein in its entirety.

[0004] BACKGROUND

[0005] Thiostrepton is a cyclic oligopeptide antibiotic that is also known by other names such as bryamycin, thiactin, alaninamide, HR4S203Y18, etc. Recent studies have shown that thiostrepton also has promising anticancer activity. There remains a need for thiostrepton derivatives having beneficial pharmacological properties.

[0006] SUMMARY

[0007] In certain aspects, the present invention provides a series of compounds having a structure according to Formula (I): or a pharmaceutically acceptable salt thereof, wherein:

[0008] R1is -CH3or -CH2CH3;

[0009] R2is -C1-C4 alkyl, -C3-C6cycloalkyl, -(C1-C3 alkylene)-C3-C6cycloalkyl, -(C1-C3 alkylene)-O-C1-C3alkyl, or -(C1-C3 alkylene)-N(R3)2; or

[0010] R1and R2are taken together with the nitrogen atom to which they are attached to form a 4-7 membered saturated heterocyclyl, optionally comprising an additional heteroatom selected from N, O and S, wherein the 4-7 membered saturated heterocyclyl is optionally substituted with 1-2 substituents independently selected from oxo, -C1-C3 alkyl, - OH, -O(C1-C3alkyl), and -C(O)-C1-C3alkyl; and

[0011] of the molecule; each R3is independently H or C1-C4 alkyl;

[0012] R4is hydrogen, C1-C4 alkyl or -C(O)C1-C4 alkyl; and

[0013] R6is C1-C4 alkyl.

[0014] In certain aspects, the present invention provides a series of compounds having a structure according to Formula (II): or a pharmaceutically acceptable salt thereof, wherein:

[0015] R1is -CH3or -CH2CH3;

[0016] R2is -C1-C4 alkyl, -C3-C6cycloalkyl, -(C1-C3 alkylene)-C3-C6cycloalkyl, -(C1-C3 alkylene)-O-C1-C3alkyl, or -(C1-C3 alkylene)-N(R3)2; or

[0017] R1and R2are taken together with the nitrogen atom to which they are attached to form a 4-7 membered saturated heterocyclyl, optionally comprising an additional heteroatom selected from N, O and S, wherein the 4-7 membered saturated heterocyclyl is optionally substituted with 1-2 substituents independently selected from oxo, -C1-C3 alkyl, - OH, -O(C1-C3alkyl), and -C(O)-C1-C3alkyl; and J wherein indicates the point of attachment of R5to the rest of the molecule; each R3is independently H or C1-C4 alkyl;

[0018] R4is hydrogen, C1-C4 alkyl or -C(0)C1-C4 alkyl; and

[0019] R6is C1-C4 alkyl.

[0020] BRIEF DESCRIPTION OF THE FIGURES

[0021] FIG. 1 is a table summarizing the results of Examples for various exemplary compounds described herein. For IC50 values, A corresponds to an IC50 of <1.39 pM; B = 1.39-2.39 pM, C - 2.39-3.39 pM, D - 3.39-4.39 pM; and E > 4.39 pM.

[0022] FIG. 2 shows the activity of Thiostrepton (TS) and 1-305 in a xenograft model of MM.

[0023] FIG. 3 shows selected PK activities of Thiostrepton (TS) and 1-305.

[0024] FIG. 4 depicts Systemic Mouse Model Experiment 1: 1-305 was injected into the peritoneal cavity (IP) lx per day to mice harboring anatomically distinct human MESO tumors engrafted subcutaneously. Tumor measurements taken every 2-3 days with calipers. No change in bodyweight observed.

[0025] FIG. 5 depicts the results of Systemic Mouse Model Experiment 2: Subcutaneous Mesothelioma tumors Treatment I.P. 3X weekly in PBS / DMSO vehicle Tumors measured in calipers.

[0026] FIG. 6a-6c show that Thiostrepton (TS) and 1-305 Reduce EMT Gene Expression. FIG. 6a shows Thiostrepton (TS) and 1-305 inhibit expression of N-cadherin, which drives EMT and cell invasion; FIG. 6b shows Thiostrepton (TS) and 1-305 inhibit expression of TGFB, which drives pro-EMT gene expression patterns; FIG. 6c shows Thiostrepton (TS) and 1-305 inhibit expression of COL5A2, high levels correlate with poor OS in Mesothelioma.

[0027] FIG. 7a- 7b show that 1-305 reduces PD-L1 Expression; PD-L1 Expression strongly correlates with poor OS in Mesothelioma. PD-1 enables tumor immune evasion (or escape); treatment with 1-305 may improve anti-tumor immune response. FIG. 7a shows that PD- LI Expression correlates with poor OS in Mesothelioma; FIG. 7b shows that Thiostrepton (TS) and 1-305 reduce PD-L1 expression.

[0028] DETAILED DESCRIPTION

[0029] Pharmaceutical Compositions

[0030] The compositions and methods described herein may be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or the compound is preferably administered as a pharmaceutical composition comprising, for example, a compound described herein and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, as a non-limiting example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are for human administration, particularly for invasive routes of administration (i.e., routes, such as injection or implantation, that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free, or substantially pyrogen-free. The excipients can be chosen, for example, to effect delayed release of an agent or to selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form such as tablet, capsule (including sprinkle capsule and gelatin capsule), granule, lyophile for reconstitution, powder, solution, syrup, suppository, injection or the like. The composition can also be present in a transdermal delivery system, e.g., a skin patch. The composition can also be present in a solution suitable for topical administration, such as a lotion, cream, or ointment.

[0031] A pharmaceutically acceptable carrier can contain physiologically acceptable agents that act, for example, to stabilize, increase solubility or to increase the absorption of a compound such as a compound described herein. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition can be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) also can be a liposome or other polymer matrix, which can have incorporated therein, for example, a compound described herein. Liposomes, for example, which comprise phospholipids or other lipids, are nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer.

[0032] The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0033] The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. 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: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer’s solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0034] A pharmaceutical composition (preparation) can be administered to a subject by any of a number of routes of administration including, for example, orally (for example, drenches as in aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); absorption through the oral mucosa (e.g., sublingually); subcutaneously; transdermally (for example as a patch applied to the skin); and topically (for example, as a cream, ointment or spray applied to the skin). The compound may also be formulated for inhalation. In certain embodiments, a compound may be simply dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions suitable for same can be found in, for example, U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, as well as in patents cited therein.

[0035] The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent to about ninety-nine percent of active ingredient, preferably from about 5 percent to about 70 percent, most preferably from about 10 percent to about 30 percent.

[0036] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound described herein, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound described herein with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0037] Formulations described herein suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), lyophile, powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil- in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and / or as mouth washes and the like, each containing a predetermined amount of a compound described herein as an active ingredient. Compositions or compounds may also be administered as a bolus, electuary or paste.

[0038] To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, such as, modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical compositions 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 sugars, as well as high molecular weight polyethylene glycols and the like.

[0039] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), 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.

[0040] The tablets, and other solid dosage forms of the pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

[0041] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophiles for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

[0042] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0043] Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0044] Dosage forms for the topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be required.

[0045] The ointments, pastes, creams and gels may contain, in addition to an active compound, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0046] Powders and sprays can contain, in addition to an active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane. Transdermal patches have the added advantage of providing controlled delivery of a compound described herein to the body. Such dosage forms can be made by dissolving or dispersing the active compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

[0047] The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

[0048] Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions described herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can 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.

[0049] These compositions may also contain adjuvants such as preservatives, 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, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin. In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0050] Injectable depot forms are made by forming microencapsulated matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly (anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissue.

[0051] For use in the methods described herein, active compounds can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.

[0052] Methods of introduction may also be provided by rechargeable or biodegradable devices. Various slow release polymeric devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including proteinaceous biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form an implant for the sustained release of a compound at a particular target site.

[0053] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0054] The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound(s) being employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound(s) employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0055] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the pharmaceutical composition or compound at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. By “therapeutically effective amount” is meant the concentration of a compound that is sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the weight, sex, age, and medical history of the subject. Other factors that influence the effective amount may include, but are not limited to, the severity of the patient’s condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being administered with the compound described herein. A larger total dose can be delivered by multiple administrations of the agent. Methods to determine efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison’s Principles of Internal Medicine 13 ed., 1814-1882, herein incorporated by reference).

[0056] In general, a suitable daily dose of an active compound used in the compositions and methods described herein will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

[0057] If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. In certain embodiments, the active compound may be administered two or three times daily. In preferred embodiments, the active compound will be administered once daily.

[0058] The patient receiving this treatment is any animal in need, including primates, in particular humans; and other mammals such as equines, cattle, swine, sheep, cats, and dogs; poultry; and pets in general.

[0059] In certain embodiments, compounds described herein may be used alone or conjointly administered with another type of therapeutic agent.

[0060] The present disclosure includes the use of pharmaceutically acceptable salts of compounds described herein in the compositions and methods described herein. In certain embodiments, contemplated salts include, but are not limited to, alkyl, dialkyl, trialkyl or tetra-alkyl ammonium salts. In certain embodiments, contemplated salts include, but are not limited to, L-arginine, benenthamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, IH-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, l-(2- hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, contemplated salts include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, contemplated salts include, but are not limited to, 1- hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2- oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, 1- ascorbic acid, 1-aspartic acid, benzenesulfonic acid, benzoic acid, (+) -camphoric acid, (+)- camphor- 10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane- 1 ,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d glucoheptonic acid, d gluconic acid, d glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, 1-malic acid, malonic acid, mandelic acid, methanesulfonic acid , naphthalene- 1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, 1- pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, 1 tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid salts.

[0061] The pharmaceutically acceptable acid addition salts can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent.

[0062] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions. Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0063] Definitions

[0064] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings that are commonly understood by those of ordinary skill in the art. Generally, nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics and protein and nucleic acid chemistry, described herein, are those well-known and commonly used in the art.

[0065] The methods and techniques of the present disclosure are generally performed, unless otherwise indicated, according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout this specification.

[0066] Chemistry terms used herein, unless otherwise defined herein, are used according to conventional usage in the art, as exemplified by “The McGraw-Hill Dictionary of Chemical Terms”, Parker S., Ed., McGraw-Hill, San Francisco, C.A. (1985).

[0067] All of the above, and any other publications, patents and published patent applications referred to in this application are specifically incorporated by reference herein. In case of conflict, the present specification, including its specific definitions, will control.

[0068] The term “agent” is used herein to denote a chemical compound (such as an organic or inorganic compound, a mixture of chemical compounds), a biological macromolecule (such as a nucleic acid, an antibody, including parts thereof as well as humanized, chimeric and human antibodies and monoclonal antibodies, a protein or portion thereof, e.g., a peptide, a lipid, a carbohydrate), or an extract made from biological materials such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues. Agents include, for example, agents whose structure is known, and those whose structure is not known. A “patient,” “subject,” or “individual” are used interchangeably and refer to either a human or a non-human animal. These terms include mammals, such as humans, primates, livestock animals (including bovines, porcines, etc.), companion animals (e.g., canines, felines, etc.) and rodents (e.g., mice and rats).

[0069] “Treating” a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results. As used herein, and as well understood in the art, “treatment” is an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0070] The term “preventing” is art-recognized, and when used in relation to a condition, such as a local recurrence (e.g., pain), a disease such as cancer, a syndrome complex such as heart failure or any other medical condition, is well understood in the art, and includes administration of a composition which reduces the frequency of, or delays the onset of, symptoms of a medical condition in a subject relative to a subject which does not receive the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving a prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable cancerous growths in a treated population versus an untreated control population, e.g., by a statistically and / or clinically significant amount.

[0071] “Administering” or “administration of’ a substance, a compound or an agent to a subject can be carried out using one of a variety of methods known to those skilled in the art. For example, a compound or an agent can be administered, intravenously, arterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (by absorption, e.g., through a skin duct). A compound or agent can also appropriately be introduced by rechargeable or biodegradable polymeric devices or other devices, e.g., patches and pumps, or formulations, which provide for the extended, slow or controlled release of the compound or agent. Administering can also be performed, for example, once, a plurality of times, and / or over one or more extended periods. Appropriate methods of administering a substance, a compound or an agent to a subject will also depend, for example, on the age and / or the physical condition of the subject and the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability and toxicity). In some embodiments, a compound or an agent is administered orally, e.g., to a subject by ingestion. In some embodiments, the orally administered compound or agent is in an extended release or slow release formulation, or administered using a device for such slow or extended release.

[0072] As used herein, the phrase “conjoint administration” refers to any form of administration of two or more different therapeutic agents such that the second agent is administered while the previously administered therapeutic agent is still effective in the body (e.g., when at least 5% of drug product is detectable systemically with industry acceptable methodology, or when the two agents are simultaneously effective in the patient, which may include synergistic effects of the two agents). For example, the different therapeutic compounds can be administered either in the same formulation or in separate formulations, either concomitantly or sequentially. In certain embodiments, the different therapeutic compounds can be administered within one hour, 12 hours, 24 hours, 36 hours, 48 hours, 72 hours, or a week of one another. Thus, an individual who receives such treatment can benefit from a combined effect of different therapeutic agents.

[0073] A “therapeutically effective amount” or a “therapeutically effective dose” of a compound or other agent described herein is an amount of a drug or an agent that, when administered to a subject will have the intended therapeutic effect. The full therapeutic effect does not necessarily occur by administration of one dose of such a drug or agent, and may occur only after administration of a series of doses (multiple consecutive doses). Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount needed for a subject will depend upon, for example, the subject’s size, health and age, and the nature and extent of the condition being treated, such as cancer.

[0074] As used herein, the terms “optional” or “optionally” mean that the subsequently described event or circumstance may occur or may not occur, and that the description includes instances where the event or circumstance occurs as well as instances in which it does not. For example, “optionally substituted alkyl” refers to the alkyl may be substituted as well as where the alkyl is not substituted. The term “modulate” as used herein includes the inhibition or suppression of a function or activity (such as cell proliferation) as well as the enhancement of a function or activity.

[0075] The phrase “pharmaceutically acceptable” is art-recognized. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0076] “Pharmaceutically acceptable salt” or “salt” is used herein to refer to an acid addition salt or a basic addition salt which is suitable for or compatible with the treatment of patients.

[0077] The term “pharmaceutically acceptable acid addition salt” as used herein means any non-toxic organic or inorganic salt of any base compounds. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric and phosphoric acids, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids that form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, benzoic, phenylacetic, cinnamic and salicylic acids, as well as sulfonic acids such as p-toluene sulfonic and methanesulfonic acids. Either the mono or di-acid salts can be formed, and such salts may exist in either a hydrated, solvated or substantially anhydrous form. In general, the acid addition salts of compounds are more soluble in water and various hydrophilic organic solvents, and generally demonstrate higher melting points in comparison to their free base forms. The selection of the appropriate salt will be known to one skilled in the art. Other non -pharmaceutically acceptable salts, e.g., oxalates, may be used, for example, in the isolation of compounds for laboratory use, or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0078] The term “pharmaceutically acceptable basic addition salt” as used herein means any non-toxic organic or inorganic base addition salt of any acid compounds or any of their intermediates. Illustrative inorganic bases which form suitable salts include lithium, sodium, potassium, calcium, magnesium, or barium hydroxide. Illustrative organic bases which form suitable salts include aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine and picoline or ammonia. The selection of the appropriate salt will be known to a person skilled in the art.

[0079] Many of the compounds useful in the methods and compositions of this disclosure have at least one stereogenic center in their structure. This stereogenic center may be present in an R or a S configuration, said R and S notation is used in correspondence with the rules described in Pure Appl. Chem. (1976), 45, 11-30. The disclosure contemplates all stereoisomeric forms such as enantiomeric and diastereoisomeric forms of the compounds, salts, prodrugs or mixtures thereof (including all possible mixtures of stereoisomers). See, e.g., WO 01 / 062726.

[0080] Some of the compounds may also exist in tautomeric forms. Such forms, although not explicitly indicated in the formulae described herein, are intended to be included within the scope of the present disclosure.

[0081] “Prodrug” or “pharmaceutically acceptable prodrug” refers to a compound that is metabolized, for example hydrolyzed or oxidized, in the host after administration to form the compound of the present disclosure. Typical examples of prodrugs include compounds that have biologically labile or cleavable (protecting) groups on a functional moiety of the active compound. Prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated to produce the active compound. Examples of prodrugs using ester or phosphoramidate as biologically labile or cleavable (protecting) groups are disclosed in U.S. Patents 6,875,751, 7,585,851, and 7,964,580, the disclosures of which are incorporated herein by reference. The prodrugs of this disclosure are metabolized to produce IPA or a salt thereof. The present disclosure includes within its scope, prodrugs of the compounds described herein. Conventional procedures for the selection and preparation of suitable prodrugs are described, for example, in “Design of Prodrugs” Ed. H. Bundgaard, Elsevier, 1985.

[0082] The phrase “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filter, diluent, excipient, solvent or encapsulating material useful for formulating a drug for medicinal or therapeutic use.

[0083] The term “linker” as used herein means any chemical functionality that “links,” or connects with chemical bonds, any two or more other chemical functionalities in a pharmaceutically relevant molecule. As a non-limiting example of the use of linkers in pharmaceutical settings, antibody-drug conjugations (ADCs) comprise a pharmaceutically active small-molecule, drug, or toxin connected to a large-molecule antibody by a linker.

[0084] Examples of common linker types include both cleavable and non-cleavable linkers. Cleavable linkers include chemical functionalities that can be cleaved in response to physiological stimuli such as chemical gradients, pH changes, or enzymatic activity. Nonlimiting examples include acid- or base-labile functional groups, pyrophosphate diester, disulfide bonds, peptides, P-glucuronides, etc. Non-cleavable linkers comprise chemical functionalities that are generally less labile to the aforementioned physiological stimuli, and non-limiting examples include certain alkyl groups and organic polymeric functionalities.

[0085] The term “reactive linker moiety” as used herein refers to a chemical structure having a terminal moiety that can react and form a covalent bond with another moiety (such as the mitochondrial targeting moiety).

[0086] The term “mitochondrial targeting peptide,” “mitochondrial targeting sequence,” “mitochondrial targeting moiety,” as used herein are art-recognized terms referring to a chemical functionality (the peptide, sequence, or moiety), which “target,” - i.e., are readily transported to and absorbed by - mitochondrial membranes (J. Zielonka, B. Kalyanaraman, et al., 2017). As used herein, mitochondrial targeting moieties may include but are not limited to the following species: berberin cation, rhodamine cation, an indolium cation, a pyridinium cation, a tetraguanidinium cation, cyanine derivatives, a guanidinium cation, a biguanidinium cation, a triphenylphosphonium cation, a triethylammonium cation, a triphenylamine, a tetraphenylethene moiety, arylphosphonium cation, an SS peptide, a mitochondrial penetrating peptide (MPP), a mitochondrial targeting sequence (MTS) peptide, a hemigramicidin S-linked nitroxide, a Dequalinium (DQA) cation, a delocalized lipophilic cation, F16 ((E)-4-(lH-indol-3-ylvinyl)-N-methylpyridinium iodide), (L- cyclohexyl alanine-D-arginine)3, a mitochondrial-targeted nanocarrier, a DDDK peptide, glycyrrhetinic acid, a-tocopheryl succinate (a-TOS), a graphene oxide nano carrier, PEG- proapoptotic peptide (KLAKLAK)2, a Dmt-D-Arg-Phe-Lys-NH2 peptide, pyruvaldehyde, N-Nonyl acridine orange, quinoline, styrl-azinium fluorophores, or 15d-PGJ2. Exemplary mitochondrial targeting moieties are listed in See. J Zielonka et al, Chem Rev 2017, 117, p 10043-10120; K L Horton et al, Chemistry & Biology 2008, 15, pp 375-382; G Battogtokh et al, Front Pharmacol 2018, 9:922; United States Patent Nos. 9,173,952 and 9,132,198, the contents of each of which are incorporated by reference herein. It is understood that substituents and substitution patterns on the compounds described herein can be selected by one of ordinary skilled person in the art to result in chemically stable compounds that can be readily synthesized by techniques known in the art, as well as those methods set forth below, from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, so long as a stable structure results.

[0087] As used herein, the term “optionally substituted” refers to the replacement of one to six hydrogen radicals in a given structure with the radical of a specified substituent including, but not limited to: hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH2-O-alkyl, -OP(O)(O-alkyl)2 or -CH2-OP(O)(O-alkyl)2. Preferably, “optionally substituted” refers to the replacement of one to four hydrogen radicals in a given structure with the substituents mentioned above. More preferably, one to three hydrogen radicals are replaced by the substituents as mentioned above. It is understood that the substituent can be further substituted.

[0088] The term “acyl” is art -recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0089] The term “acylamino” is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbylC(O)NH-.

[0090] The term “acyloxy” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0091] The term “alkoxy” refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like.

[0092] The term “alkoxyalkyl” refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

[0093] As used herein, the term “alkyl” refers to saturated aliphatic groups, including but not limited to C1-C10 straight-chain alkyl groups or C1-C10 branched-chain alkyl groups. Preferably, the “alkyl” group refers to C1-Ce straight-chain alkyl groups or C1-Ce branched- chain alkyl groups. Most preferably, the “alkyl” group refers to C1-C4 straight-chain alkyl groups or C1-C4 branched-chain alkyl groups. Examples of “alkyl” include, but are not limited to, methyl, ethyl, 1 -propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, 1 -pentyl, 2- pentyl, 3-pentyl, neo-pentyl, 1 -hexyl, 2-hexyl, 3-hexyl, 1 -heptyl, 2-heptyl, 3-heptyl, 4- heptyl, 1 -octyl, 2-octyl, 3 -octyl or 4-octyl and the like.

[0094] Moreover, the term “alkyl” as used throughout the specification, examples, and claims is intended to include both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, etc.

[0095] The term “Cx-y” or “Cx-Cy”, when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups that contain from x to y carbons in the chain. Coalkyl indicates a hydrogen where the group is in a terminal position, a bond if internal. A C1-ealkyl group, for example, contains from one to six carbon atoms in the chain.

[0096] The term “alkylamino”, as used herein, refers to an amino group substituted with at least one alkyl group.

[0097] The term “alkylthio”, as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

[0098] The term “amide”, as used herein, refers to a group wherein R9and R10each independently represent a hydrogen or hydrocarbyl group, or R9and R10taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

[0099] The terms “amine” and “amino” are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, e.g., a moiety that can be represented by wherein R9, R10, and R10’ each independently represent a hydrogen or a hydrocarbyl group, or R9and R10taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

[0100] The term “aminoalkyl”, as used herein, refers to an alkyl group substituted with an amino group. The term “aralkyl”, as used herein, refers to an alkyl group substituted with an aryl group.

[0101] The term “aryl” used alone or as part of a larger moiety as in “aralkyl”, “aralkoxy”, or “aryloxy alkyl”, refers to monocyclic and bicyclic ring systems having a total of five to fourteen ring members, wherein each ring atom is carbon, at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members. The term “aryl” may be used interchangeably with the term “aryl ring”. In certain embodiments of the present disclosure, “aryl” refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Also included within the scope of the term “aryl”, as it is used herein, is a group in which an aromatic ring is fused to one or more non-aromatic carbocyclic rings.

[0102] The term “carbamate” is art-recognized and refers to a group pio wherein R9and R10independently represent hydrogen or a hydrocarbyl group.

[0103] The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

[0104] The term “carbocycle” includes 5-7 membered monocyclic and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated and aromatic rings. Carbocycle includes bicyclic molecules in which one, two or three or more atoms are shared between the two rings. The term “fused carbocycle” refers to a bicyclic carbocycle in which each of the rings shares two adjacent atoms with the other ring. Each ring of a fused carbocycle may be selected from saturated, unsaturated and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits, is included in the definition of carbocyclic. Exemplary “carbocycles” include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5 -cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4, 5, 6, 7- tetrahydro-lH-indene and bicyclo[4.1.0]hept-3-ene. “Carbocycles” may be substituted at any one or more positions capable of bearing a hydrogen atom. The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

[0105] The term “carbonate” is art-recognized and refers to a group -OCO2-.

[0106] The term “carboxy”, as used herein, refers to a group represented by the formula -CO2H.

[0107] The term “ester”, as used herein, refers to a group -C(O)OR9wherein R9represents a hydrocarbyl group.

[0108] The term “ether”, as used herein, refers to a hydrocarbyl group linked through an oxygen to another hydrocarbyl group. Accordingly, an ether substituent of a hydrocarbyl group may be hydrocarbyl-O-. Ethers may be either symmetrical or unsymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O- heterocycle. Ethers include “alkoxyalkyl” groups, which may be represented by the general formula alkyl-O-alkyl.

[0109] The terms “halo” and “halogen” as used herein means halogen and includes chloro, fluoro, bromo, and iodo.

[0110] The terms “hetaralkyl” and “hetero aralkyl”, as used herein, refers to an alkyl group substituted with a hetaryl group.

[0111] The terms “heteroaryl” and “heteroar-”, used alone or as part of a larger moiety, e.g., “heteroaralkyl”, or “heteroaralkoxy”, refer to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 1471 electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms. The term “heteroatom” refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen. Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. The terms “heteroaryl” and “heteroar-”, as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl or heteroaryl rings such that the resulting bi- or multicyclic ring system as a whole is fully aromatic. Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, and phenoxazinyl. A heteroaryl group may be mono- or bicyclic. The term “heteroaryl” may be used interchangeably with the terms “heteroaryl ring”, “heteroaryl group”, or “heteroaromatic”, any of which terms include rings that are optionally substituted. The term “heteroaralkyl” refers to an alkyl group substituted by a heteroaryl, wherein the alkyl and heteroaryl portions independently are optionally substituted.

[0112] The term “heteroatom” as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0113] The term “heterocyclylalkyl”, as used herein, refers to an alkyl group substituted with a heterocycle group.

[0114] As used herein, the terms “heterocycle”, “heterocyclyl”, “heterocyclic radical”, and “heterocyclic ring” are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above. When used in reference to a ring atom of a heterocycle, the term “nitrogen” includes a substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or +NR (as in N-substituted pyrrolidinyl).

[0115] A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms “heterocycle”, “heterocyclyl”, “heterocyclyl ring”, “heterocyclic group”, “heterocyclic moiety”, and “heterocyclic radical”, are used interchangeably herein, and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. A heterocyclyl group may be mono- or bicyclic. The term “heterocyclylalkyl” refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.

[0116] The term “hydrocarbyl”, as used herein, refers to a group that is bonded through a carbon atom that does not have a =0 or =S substituent, and typically has at least one carbon-hydrogen bond and a primarily carbon backbone, but may optionally include heteroatoms. Thus, groups like methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered to be hydrocarbyl for the purposes of this application, but substituents such as acetyl (which has a =0 substituent on the linking carbon) and ethoxy (which is linked through oxygen, not carbon) are not. Hydrocarbyl groups include, but are not limited to aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

[0117] The term “hydroxy alkyl”, as used herein, refers to an alkyl group substituted with a hydroxy group.

[0118] The term “lower” when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups where there are ten or fewer atoms in the substituent, preferably six or fewer. A “lower alkyl”, for example, refers to an alkyl group that contains ten or fewer carbon atoms, preferably six or fewer. In certain embodiments, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are respectively lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether they appear alone or in combination with other substituents, such as in the recitations hydroxyalkyl and aralkyl (in which case, for example, the atoms within the aryl group are not counted when counting the carbon atoms in the alkyl substituent).

[0119] The terms “polycyclyl”, “polycycle”, and “polycyclic” refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls) in which two or more atoms are common to two adjoining rings, e.g., the rings are “fused rings”. Each of the rings of the polycycle can be substituted or unsubstituted. In certain embodiments, each ring of the polycycle contains from 3 to 10 atoms in the ring, preferably from 5 to 7.

[0120] The term “sulfate” is art-recognized and refers to the group -OSO3H, or a pharmaceutically acceptable salt thereof.

[0121] The term “sulfonamide” is art-recognized and refers to the group represented by the general formulae 9 wherein R9and R10independently represents hydrogen or hydrocarbyl.

[0122] The term “sulfoxide” is art-recognized and refers to the group-S(O)-.

[0123] The term “sulfonate” is art-recognized and refers to the group SO3H, or a pharmaceutically acceptable salt thereof. The term “sulfone” is art-recognized and refers to the group -S(O)2-.

[0124] The term “protecting group” is an art-recognized term referring to chemical functionalities that can modify (usually covalently) an existing labile functionality on a target molecule. This modification “protects” the labile functionality during subsequent reaction steps, and the protecting group can be removed as needed, termed “deprotection.” As a non-limiting example, the t-butyloxycarbonyl (Boc or hoc) group is commonly used to covalently modify and “protect” terminal amine groups in synthetic chemistry.

[0125] The term “substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, the heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include any substituents described herein, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.

[0126] Suitable monovalent substituents on a substitutable carbon atom of an “optionally substituted” group are independently halogen; -(CH2)o-4R°; -(CH2)o-40R°; - 0(CH2)O-4R°, -0-(CH2)O-4C(0)OR°; -(CH2)O-4CH(OR°)2; -(Clfe^SR0; -(CH2)o^Ph, which may be substituted with R°; -(CH2)O-40(CH2)O-I Ph which may be substituted with R°; -CH=CHPh, which may be substituted with R°; -(CH2)O-40(CH2)O-I -pyridyl which may be substituted with R°; -N02; -CN; -N3; -(CH2)o^N(R°)2; -(CH2)o^N(R°)C(0)R°; - N(R°)C(S)R°; -(CH2)O_4N(R°)C(0)NR02; -N(RO)C(S)NR°2; -(CH2)0-4N(RO)C(O)OR°; - N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)NR°2; -N(R°)N(R°)C(O)OR°; -(CH2)o-4C(0)R°; - C(S)R°; -(CH2)O-4C(0)OR°; -(CH2)O^C(0)SR°; -(CH2)o^C(0)OSiR°3; -(CH2)o_4OC(O)R°; -OC(0)(CH2)O^SR°, SC(S)SR°; -(CH2)O^SC(0)R°; -(CH2)O^C(0)NR°2; - C(S)NR°2; -C(S)SR°; -SC(S)SR°, -(CH2)O^OC(0)NR°2; -C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH2C(O)RO; -C(NOR°)R°; -(CH2)O^SSR°; -(CH2)O^S(0)2R°; -(CH2)O^S(0)2OR°; -(CH2)O^OS(0)2R°; -S(O)2NRO2; -(CH2)O-4S(0)R°; -N(RO)S(O)2NR°2; -N(RO)S(O)2R°; - N(OR°)R°; -C(NH)NR°2; -P(O)2RO; -P(O)RO2; -OP(O)RO2; -OP(O)(ORO)2; SiR°3; -(C1^ straight or branched alkylene)O-N(R°)2; or -(C1^ straight or branched alkylene)C(O)O- N(R°)2, wherein each R° may be substituted as defined below and is independently hydrogen, C1~6 aliphatic, -CH2Ph, -0(CH2)o-iPh, -CH2-(5-6 membered heteroaryl ring), or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0^1 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R°, taken together with their intervening atom(s), form a 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.

[0127] Suitable monovalent substituents on R° (or the ring formed by taking two independent occurrences of R° together with their intervening atoms), are independently halogen, -(CH2)o-2R*, -(haloR*), -(CH2)o-2OH, -(CH2)o-2OR*, -(CH2)o- NO2, -SiR*3, -OSiR*3, -C(O)SR* -(C1-4 straight or branched alkylene)C(O)OR*, or - SSR* wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently selected from C1-4 aliphatic, -CH2Ph, - 0(CH2)o-iPh, or a 5-7-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R° include =0 and =S.

[0128] Suitable divalent substituents on a saturated carbon atom of an “optionally substituted” group include the following: =0, =S. =NNR*2, =NNHC(0)R*, =NNHC(0)0R*, =NNHS(O)2R*, =NR*. =N0R*, -O(C(R*2))2-3O-, or -S(C(R*2))2-3S-, wherein each independent occurrence of R* is selected from hydrogen, C1~6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that are bound to vicinal substitutable carbons of an “optionally substituted” group include: -O(CR*2)2-3O-, wherein each independent occurrence of R* is selected from hydrogen, C1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0129] Suitable substituents on the aliphatic group of R* include halogen, - R*, -(haloR*), -OH, -OR*, -O(haloR’), -CN, -C(O)OH, -C(O)OR*, -NH2, -NHR*, - NR*2, or -NO2, wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C1-4 aliphatic, -CH2PI1, - 0(CH2)o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0130] Suitable substituents on a substitutable nitrogen of an “optionally substituted” group include -Rf, -NRf2, -C(O)Rf, -C(O)ORf, -C(O)C(O)Rf, -C(O)CH2C(O)Rf, - S(O)2Rf, -S(O)2NRf2, -C(S)NRf2, -C(NH)NRf2, or -N(Rt)S(O)2Rt; wherein each R' is independently hydrogen, C1-6 aliphatic which may be substituted as defined below, unsubstituted -OPh, or a substituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R' . taken together with their intervening atom(s) form an unsubstituted 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0^1 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0131] Suitable substituents on the aliphatic group and the substituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur of R' are independently halogen, -R*, -(haloR*), -OH, - OR", -O(haloR’), -CN, -C(O)OH, -C(O)OR*, -NH2, -NHR", -NR*2, or -NO2, wherein each R* is unsubstituted or where preceded by “halo” is substituted only with one or more halogens, and is independently C1-4 aliphatic, -CH2PI1, -0(CH2)o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. The term “thioalkyl”, as used herein, refers to an alkyl group substituted with a thiol group.

[0132] The term “thioester”, as used herein, refers to a group -C(O)SR9or -SC(O)R9wherein R9represents a hydrocarbyl.

[0133] The term “thioether”, as used herein, is equivalent to an ether, wherein the oxygen is replaced with a sulfur.

[0134] The term “urea” is art-recognized and may be represented by the general formula wherein R9and R10independently represent hydrogen or a hydrocarbyl.

[0135] The term “tautomer” refers to each of two or more isomers of a compound that exist together in equilibrium, and are interchanged by migration of an atom or group within the molecule, such as a hydrogen atom. Depiction in this disclosure of one tautomer within a genus or compound species is intended to encompass the compound as drawn and all of its tautomers.

[0136] In certain aspects, the present invention provides a compound having a structure according to Formula (I): or a pharmaceutically acceptable salt thereof, wherein:

[0137] R1is -CH3or -CH2CH3;

[0138] R2is -C1-C4 alkyl, -C3-C6cycloalkyl, -(C1-C3 alkylene)-C3-C6cycloalkyl, -(C1-C3 alkylene)-O-C1-C3alkyl, or -(C1-C3 alkylene)-N(R3)2; or

[0139] R1and R2are taken together with the nitrogen atom to which they are attached to form a 4-7 membered saturated heterocyclyl, optionally comprising an additional heteroatom selected from N, O and S, wherein the 4-7 membered saturated heterocyclyl is optionally substituted with 1-2 substituents independently selected from oxo, -C1-C3 alkyl, - OH, -O(C1-C3alkyl), and -C(O)-C1-C3alkyl; and

[0140] rest of the molecule; each R3is independently H or C1-C4 alkyl;

[0141] R4is hydrogen, C1-C4 alkyl or -C(O)C1-C4 alkyl; and

[0142] R6is C1-C4 alkyl.

[0143] In certain embodiments, R1is -CH3.

[0144] In certain embodiments, R2is -CH3, -CH2CH3, -CH(CH3)2, cyclopentyl, -(C1-C3 alkylene)-cyclopentyl, -CH2CH2-OCH3, or -CH2CH2N(CH3)2-

[0145] In certain such embodiments, R2is -CH3.

[0146] In other embodiments, R1and R2are taken together with the nitrogen atom to which they are attached to form a 4-6 membered heterocyclyl.

[0147] In certain such embodiments, R2represents In certain such embodiments, R5is

[0148] In certain embodiments, R4is hydrogen, -CH3 or -C(0)CH3.

[0149] In certain embodiments, the compound is selected from:

[0150]

[0151] In certain aspects, the present invention provides a compound having a structure according to Formula (II): or a pharmaceutically acceptable salt thereof, wherein:

[0152] R1is -CH3or -CH2CH3;

[0153] R2is -C1-C4 alkyl, -C3-C6cycloalkyl, -(C1-C3 alkylene)-C3-C6cycloalkyl, -(C1-C3 alkylene)-O-C1-C3alkyl, or -(C1-C3alkylene)-N(R3)2; or

[0154] R1and R2are taken together with the nitrogen atom to which they are attached to form a 4-7 membered saturated heterocyclyl, optionally comprising an additional heteroatom selected from N, O and S, wherein the 4-7 membered saturated heterocyclyl is optionally substituted with 1-2 substituents independently selected from oxo, -C1-C3 alkyl, -

[0155] OH, -O(C1-C3alkyl), and -C(O)-C1-C3alkyl; and u wherein indicates the point of attachment of R5to the rest of the molecule; each R3is independently H or C1-C4 alkyl;

[0156] R4is hydrogen, C1-C4 alkyl or -C(0)C1-C4 alkyl; and

[0157] R5is C1-C4 alkyl.

[0158] In certain such embodiments, R1is -CH3.

[0159] In certain embodiments, R2is -CH3, -CH2CH3, -CH(CH3)2, cyclopentyl, -(C1-C3 alkylene)-cyclopentyl, -CH2CH2-OCH3, or -CH2CH2N(CH3)2

[0160] In certain such embodiments, R2is -CH3.

[0161] In certain embodiments, R1and R2are taken together with the nitrogen atom to which they are attached to form a 4-7 membered heterocyclyl.

[0162] In certain embodiments, R5is hydrogen, -CH3 or -C(0)CH3. In certain embodiments, the compound is selected from: In certain embodiments, the present invention provides a pharmaceutically acceptable composition comprising any of the compounds described herein; and a pharmaceutically acceptable carrier. In certain embodiments, the composition is formulated for oral or parenteral delivery.

[0163] In certain embodiments, the present invention discloses a method of treating a cancer (e.g., solid tumor or hematological cancer) comprising administering to a subject in need thereof a therapeutically effective amount of any of the compounds described herein, or a composition of that compound. In certain embodiments, the cancer is selected from mesothelioma, lung, breast, prostate, melanoma, esophageal, leukemia, cervical, liver, colon, gastric, colorectal, glioblastoma, head and neck, pancreatic, and ovarian. In certain embodiments, the cancer is selected from mesothelioma, lung, ovarian, and breast.

[0164] EXAMPLES

[0165] General Synthetic Procedures

[0166] General Experimental Procedure 1 - Suzuki coupling and ester hydrolysis - Pd(dppf)Ch To a solution of bromide (1.05 eq.) in a mixture of dioxane and water (4:1, 0.1 M) under N2 were added the boronic acid or ester (1.0 Eq) and cesium carbonate (2.5 eq.) were added. The mixture was then degassed with N2 for a couple of minutes. Then Pd(dppf)C12 (0.15 eq.) was added, and the mixture was stirred at 80-100 °C (ext) until full conversion (overnight). The reaction was then stopped. Water was added and the aqueous phase was acidified using HC1. The resulting mixture was extracted with EtOAc twice. Combined organics were washed with brine, dried over Na2SC>4, filtered, and concentrated. The crude material was purified by automated FCC (normal phase or reverse phase). Note: if hydrolysis was not complete the crude material was submitted according to General experimental procedure 2 to provide full hydrolysis.

[0167] General Experimental Procedure 2 - Ester hydrolysis

[0168] To a solution of ester (1 eq.) in THF (0.05 M) was added Lithium hydroxide monohydrate (4 eq.) as solution in water (2M) and the mixture was stirred for 2-16 hours at room temperature. The mixture was diluted EtOAc and washed with 1 M HC1 and brine (30 mL each), dried over Na2SO4, filtered, and concentrated to provide the desired product. General Experimental Procedure 3 - HATU-mediated amide coupling

[0169] To a solution of acid (1 eq.) and amine (1 eq.) in DCM (0.05 M) were added DiPEA (3 eq.) and HATU (1.2 eq.) and the resulting mixture was stirred at room temperature for 2- 16 hours. The mixture was washed with IM HC1, water, NaHCCh and brine, dried over Na2SC>4, filtered, and concentrated. The crude material was purified by automated FCC to afford the desired product.

[0170] General Experimental Procedure 4 - Elimination of bisacetate

[0171] A solution of the bisacetate in THF / CH2CI2 (1 / 1, v / v, 0.05 M) was cooled to 0°C and DBU (4 eq.) was added. The mixture was stirred at 0°C for 2 hours. The mixture was diluted with CH2Q2 and water and HC1 (aq. IM) were added to acidify the mixture to pH=4. The mixture was extracted with CH2CI2 (3x) and the combined organic layers were washed with brine, dried over Na2SC>4, filtered, and concentrated. The crude material was purified by automated reverse phase FCC.

[0172] Analysis Methods

[0173] LCMS Method “22010199G LCMS-5 C3”

[0174] System: Agilent 1290 series with UV detector and HP 6130 MSD mass detector LCMS Method “General 3 basic”

[0175] System: Waters Acquity UPLC with PDA detector and QDA mass detector

[0176] Syntheses of Exemplary Compounds

[0177] Example 1 N,N-dimethyl-2-(2-(4-(4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][l,4]oxazin-

[0178] 7-yl)- l-oxoisoindolin-2-yl)acrylamido)acrylamide 1-305

[0179] Step 1. 2-(2-(4-(4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][l,4]oxazin-7-yl)-l- oxoisoindolin-2-yl)acrylamido)acrylic acid was prepared following General experimental procedure 1.. Methyl 2-(2-(4-bromo-l-oxoisoindolin-2-yl)acrylamido)acrylate (0.20 g, 0.49 mmol) and 4-methyl-3-oxo-3,4-dihydro-2H-benzo[b][l,4]oxazine-7-boronic acid pinacol ester (0.17 g, 0.54 mmol) gave 2-(2-(4-(4-methyl-3-oxo-3,4-dihydro-2H- benzo[b][l,4]oxazin-7-yl)-l-oxoisoindolin-2-yl)acrylamido)acrylic acid (90 mg, 0.20 mmol, 40%) as white fluffy solid.

[0180] Step 2. N,N-dimethyl-2-(2-(4-(4-methyl-3-oxo-3,4-dihydro-2H- benzo[b][l,4]oxazin-7-yl)-l-oxoisoindolin-2-yl)acrylamido)acrylamide (1-305) was prepared using the following procedure. To an ice cold suspension of 2-(2-(4-(4-methyl-3- oxo-3 ,4-dihydro-2H-benzo [b] [ 1 ,4]oxazin-7 -yl)- 1 -oxoisoindolin-2-yl)acrylamido)acrylic acid (20 mg, 45 pmol) in DMSO (1 mL) were added DiPEA (31 pL, 4 eq., 0.18 mmol) and propylphosphonic anhydride (50% wt. solution in EtOAc) (26 pL, 45 mmol). The resulting mixture was stirred for 10 minutes at 0°C. A solution dimethylamine (2.0 M solution in THF) (22 pL, 45 mmol) in DMSO (0.3 mL) was added and the resulting mixture was stirred for 4 hours at room temperature. The resulting mixture was directly purified using automated reverse phase FCC to provide N,N-dimethyl-2-(2-(4-(4-methyl-3-oxo-3,4- dihydro-2H-benzo[b][l,4]oxazin-7-yl)-l-oxoisoindolin-2-yl)acrylamido)acrylamide (1-305) (5.0 mg, 11 pmol, 24%) as a white fluffy solid. LCMS: 22010199G LCMS-5 C3 m / z - 461.2 [M+H]+. 'H NMR (400 MHz, MeOD) 57.70 (dd, 7 = 7.5. 1.2 Hz, 1H), 7.61 (dd, 7 = 7.7, 1.2 Hz, 1H), 7.53 (t, 7= 7.6 Hz, 1H), 7.23 - 7.16 (m, 2H), 7.15 - 7.12 (m, 1H), 5.65 (d, 7 = 1.5 Hz, 1H), 5.56 (d, 7= 1.5 Hz, 1H), 5.2O (d, 7 = 0.9 Hz, 1H), 4.79 (s, 2H), 4.70 (d, 7 = 0.9 Hz, 1H), 4.60 - 4.57 (m, 2H), 3.32 (s, 3H), 3.04 (s, 3H), 2.87 (s, 3H).

[0181] Example 2

[0182] The following compounds were prepared with similar methods as described in Example 1.

[0183] Example 3 2-(2-(4-(l-acetylindolin-5-yl)-l-oxoisoindolin-2-yl)acrylamido)-N,N- dimethylacrylamide 1-384

[0184] Step 1. 2-(2-(4-bromo-l-oxoisoindolin-2-yl)acrylamido)acrylic acid was prepared following General Experimental procedure 2. Methyl 2-(2-(4-bromo-l-oxoisoindolin-2- yl)acrylamido)acrylate (RSO88, 0.20 g, 0.55 mmol) gave2-(2-(4-bromo-l-oxoisoindolin-2- yl)acrylamido)acrylic acid (0.18 g, 0.52 mmol, 95%) as white solid.

[0185] Step 2. 2-(2-(4-bromo-l-oxoisoindolin-2-yl)acrylamido)-N,N-dimethylacrylamide was preprared using the following procedure. To a solution of 2-(2-(4-bromo-l- oxoisoindolin-2-yl)acrylamido)acrylic acid (0.24 g, 0.68 mmol) and DIPEA (0.48 mL, 4 eq., 2.7 mmol) in DMSO (4.0 mL) was added propylphosphonic anhydride in EtOAc (0.40 mL, 50% wt, 1 eq., 0.68 mmol). The mixture was stirred for 10 minutes followed by addition of a solution of dimethylamine in THF (0.34 mL, 2.0 molar, 1 eq, 0.683 mmol) and the resulting mixture was stirred for 1 hour. Additional propylphosphonic anhydride in EtOAc (0.40 mL, 50% wt, 1 eq., 0.68 mmol) was added and the mixture was stirred for another 45 minutes. The crude mixture was purified by automated reverse phase FCC to provide 2-(2-(4-bromo-l-oxoisoindolin-2-yl)acrylamido)-N,N-dimethylacrylamide (0.10 g, 0.26 mmol, 39 %) as a white solid.

[0186] Step 3. 2-(2-(4-(l-acetylindolin-5-yl)-l-oxoisoindolin-2-yl)acrylamido)-N,N- dimethylacrylamide 1-384 was prepared following General experimental procedure 2. 2- (2-(4-bromo-l-oxoisoindolin-2-yl)acrylamido)-N,N-dimethylacrylamide (20 mg, 52 pmol) and l-(5-(4,4,5,5-Tetramethyl-l,3,2-dioxaborolan-2-yl)indolin-l-yl)ethenone (17 mg, 1.1 eq., 58 pmol) gave 2-(2-(4-(l-acetylindolin-5-yl)-l-oxoisoindolin-2-yl)acrylamido)-N,N- dimethylacrylamide 1-384 (3.24 mg, 7.1 pmol, 13%) as a white solid. LCMS: 22010199G LCMS-5 C3 RT: 2.011 Area: 91.1 % (215 nm), 93.3 % (254 nm); m / z = 459.2 [M+H]+. 'H NMR (400 MHz, DMSO) 59.99 (s, 1H), 8.15 (d, J = 8.3 Hz, 1H), 7.75 - 7.68 (m, 2H), 7.66 - 7.60 (m, 1H), 7.51 (s, 1H), 7.42 (d, 7 = 8.1 Hz, 1H), 5.64 (d, 7 = 19.4 Hz, 2H), 5.28 (s, 1H), 4.88 (s, 2H), 4.62 (s, 1H), 4.16 (t, 7 = 8.5 Hz, 2H), 3.24 (d, 7 = 7.7 Hz, 4H), 2.96 (s, 3H), 2.82 (s, 3H), 2.20 (s, 3H).

[0187] Example 4

[0188] The following compounds were prepared with similar methods as described in

[0189] Example 3.

[0190]

[0191] Biological Studies of Exemplary Compounds

[0192] Biological Study Example 1: Cell Viability Study of Exemplary Compounds

[0193] Cell Lines were plated in 96-well plates (Corning, Kennebunk, ME, USA) at a density of 2500 cells per well. The following day, cells were treated with test compounds diluted in complete media followed by incubation for 48 h. Post-incubation cells were washed with PBS (Corning Cellgro, Manassas, VA, USA), fixed with 3.0% formaldehyde (Fisher BioReagents, Fair Lawn, NJ, USA) in PBS, and stained for 30 min with 0.1% crystal violet (Acros Organics, Fair Lawn, NJ, USA) in water. Crystal violet stain was removed, and plates were washed with H2O and allowed to dry. To quantify cell viability, plates were imaged using the Lionheart Plate reader (BioTek Instruments, Winooski, VT, USA) and / or analyzed by absorbance at 540 nm (crystal violet dye dissolved in 100% methanol) using the Synergy HTX plate reader (BioTek Instruments, Winooski, VT, USA). To determine the effective cytotoxic concentration (IC50) of test compounds, the data were plotted using a 4-parameter non-linear regression model using GraphPad Prism7 software (GraphPad Software, San Diego, CA, USA). The IC50 data were discretized into 5 groups, A, B, C, D, and E based on the IC50 values, where inclusion in group A corresponds to an IC50 of <-1.39 pM; B = 1.39-2.39 pM, C = 2.39-3.39 pM, D = 3.39-4.39 pM; and E > 4.39 pM. These results are summarized in FIG. 1. Biological Study Example 2: The activity of Thiostrepton (TS) and 1-305 in a xenograft model of MM.

[0194] Outline: 18 male SCID mice (age 10 weeks) were engrafted with 2.5 million human malignant mesothelioma cells (HMESO cell line) via intraperitoneal (IP) injection and tumors were allowed to grow for 14 days. Animals were split into 3 groups.

[0195] Groups:

[0196] 1. 5% DMSO / PBS Vehicle Control (6 mice)

[0197] 2. 50 mg / kg TS in 5% DMSO / PBS (6 mice)

[0198] 3. 50 mg / kg 1-305 in 5% DMSO / PBS (6 mice)

[0199] Following 14 days of tumor growth animals received indicated treatments or vehicle control 3x a week (M, W, F) for 3 weeks. Animal weights were monitored during treatment. On day 36 animals were sacrificed, tumors were removed, and gross anatomy was observed. Tumor burden (weight and volume) was determined at the completion of the experiment following necropsy. Tumor weight and volume were significantly reduced in the TS and 1- 305 groups. Animals in the TS group struggled to maintain body weight throughout the experiment. The result is shown in FIG. 2.

[0200] Biological Study Example 3: Testing of intraperitoneal 1-305 administration in NOD / SCID mice harboring subcutaneous human mesothelioma tumors.

[0201] 2-5xl06human mesothelioma cells (H-Meso-1 cell line) suspended in 50-100 pL of PBS are injected into the right or left flank area of the mouse. Areas of injection were shaved with clippers for the identification of palpable tumors. Once tumors are palpable, they are measured with calipers. 25-50 mg / kg of 1-305, suspended in 5-6% DMSO / PBS, was injected 3X per week in a total volume of 200 pL. Tumor burden was measured with calipers prior to dosing. The result is shown in FIG. 4 and FIG 5.

[0202] Biological Study Example 4: Metabolic stability and metabolite profiles of 1- 305, 1-380, and 1-381

[0203] Summary:

[0204] The metabolic stability and metabolite profiles of 1-305, 1-380, and 1-381 were investigated in pooled cryopreserved CD 1 -mouse, Sprague Dawley rat, New Zealand White rabbit, Gottingen minipig, Beagle dog, Cynomolgus monkey and human hepatocytes and in plasma. The incubations were performed using 1 pM initial concentration for stability determinations and with 5 pM concentration for metabolite identification and sampling at 0, 20, 40, 60, and 120 min time points. The samples were analyzed using UPLC / QE-orbitrap- MS.

[0205] 1-305

[0206] With human hepatocytes, a half-life of 41.2 min was observed, corresponding to in vitro clearance of 16.8 pL / min / mg. The half-lives and in vitro clearances in the other species were 13.1 min / 53.0 pL / min / mg in mouse, were 7.5 min / 92.2 pL / min / mg in rat, were 37.0 min / 18.7 pL / min / mg in rabbit, were 65.2 min / 10.6 pL / min / mg in minipig, were 163 min / 4.26 pL / min / mg in dog, and were 92.2 min / 7.52 pL / min / mg in monkey.

[0207] The predicted hepatic extraction ratios were 67% for human, 48% in rabbit, 36% in minipig, 46% in dog, suggesting intermediate hepatic clearance (extraction ratio 30 - 70%), 76% for mouse, 86% for rat, suggesting high hepatic extraction (extraction ration >70%) and 30% in monkey, suggesting low hepatic clearance (extraction ratio < 30%).

[0208] For 1-305, the shortest half-lives were observed in mouse and rat plasma, with halflives 42.0 min and 39.9 min, respectively, followed by dog with half-life of 71.5 min. In human plasma, the half-life was 116 min, while the half-life in rabbit was 165 min, in minipig was 110 min, and in monkey was 176 min.

[0209] In total 27 metabolites (Ml - M27) were detected for 1-305 in hepatocytes. Of these metabolites, 21 were detected in human hepatocytes. The most abundant metabolites were formed by conjugation with glutathione (M10), cysteine-glycine (M18), and cysteine (M23) and the conjugation was located to the ethene next to the aromatic structure by the MS / MS data. All three are part of the same metabolic pathway, as glutathione is first degraded to cysteine-glycine and then to cysteine, but cysteine and cysteine-cysteine can react directly as well. Two glutathione conjugates were observed, with the other (M9) having only very low abundance, and not detected in human. The MS / MS data of M9 does not clearly show, but the most likely site is the other ethene, next to the dimethylamine moiety. Additionally, double glutathione conjugation (Ml 6) was observed in low abundance (not in human), most likely both ethenes have reacted with glutathione. Cysteine conjugate (M23) behaves differently compared to others, as its abundance was high already in the 0 min sample. This suggests that 1-305 is able to react with cysteine almost immediately. This can also help explain, why the absolute parent peak area was lower in hepatocyte 0 min samples compared to buffer incubation 0 min sample - significant portion of the compound had already reacted. The abundance of metabolites other than those involving glutathione- related conjugates (Ml - M8) were low in human, with N-demethylation in dimethylamine (M5) having the highest abundance. Metabolite M8, formed by loss of C5H7NO, did not show profile of a typical metabolite, as it was observed in all samples (excluding the blank), and could be impurity, but its abundance clearly increased in incubation with monkey hepatocytes. Metabolites M5, MIO and M23 had similar abundance as in human in all other species, but Ml 8 had higher abundance in human compared to other species, particularly in mouse and rat. In animal species, glutathione (MIO), cysteine-glycine (Ml 8) and cysteine (M23) conjugates had clearly the highest abundance, but with different relative roles. Other metabolites had only minor share of metabolism. 12 metabolites were detected in mouse, 10 in rat, 12 in rabbit, 19 in minipig, 20 in dog and 25 in monkey.

[0210] The only metabolite which abundance increased during the plasma incubation was M8, formed by loss of C5H7NO form 1-305, but even that was present already in the 0 min sample. The main metabolite in hepatocytes, cysteine conjugate M23 was detected also in plasma samples, but the abundance was low and remained stable during or slowly decreased the incubation. Highest abundance of M23 was detected in rabbit, but most likely this just indicates that the used rabbit plasma had higher residual level of cysteine than other species. In addition to these metabolites detected in hepatocytes, several other metabolites were detected, in which 1-305 had conjugated with different amino acids. Similar to cysteine conjugate, the abundance of these was low and remained stable or decreased during the incubation. MS / MS data was available for the glycine conjugate (M28) and showed that the metabolite was related to 1-305 and that modification was in the same site as the in cysteine (ethene next to aromatic structure). For other amino acid conjugates, no MS / MS data was available, but it is assumed that they have similar structure. It is assumed that the conjugation is via the amine moiety of the amino acid, but this could not be confirmed by MS / MS data. Metabolite formation can’t explain the observed disappearance of the study compound in plasma incubation.

[0211] 1-380

[0212] With human hepatocytes, a half-life of 32.0 min was observed, corresponding to in vitro clearance of 21.7 pL / min / mg. The half-lives and in vitro clearances in the other species were 30.1 min / 23.0 pL / min / mg in mouse, were 7.7 min / 90.5 pL / min / mg in rat, were 53.9 min / 12.9 pL / min / mg in rabbit, were 18.0 min / 38.6 pL / min / mg in minipig, were 86.9 min / 7.98 pL / min / mg in dog, and were 38.0 min / 18.3 pL / min / mg in monkey. The predicted hepatic extraction ratios were 73% for human, and 86% in rat, suggesting high hepatic extraction (extraction ration >70%), 57% mouse, 39% in rabbit, 67% in minipig, 61% in dog, and 51% in monkey suggesting intermediate hepatic clearance (extraction ratio 30 - 70%).

[0213] For 1-380, the shortest half-lives were observed in rat and minipig plasma with halflives of 52.4 min and 68.3 min, respectively. The longest half-life (140 min) was observed in dog. In human plasma, the half-life was 87.3 min, while the half-life in mouse was 82.3 min, in rabbit was 90.8 min, and in monkey was 81.8 min.

[0214] In total 27 metabolites (Ml - M27) were detected for 1-380 in hepatocytes. Of these metabolites, 25 were detected in human hepatocytes. The most abundant metabolites were formed by conjugation with glutathione (M9), cysteine-glycine (Ml 8), and cysteine (M23) and the conjugation was located to the ethene next to the aromatic structure by the MS / MS data. All three are part of the same metabolic pathway, as glutathione is first degraded to cysteine-glycine and then to cysteine, but cysteine and cysteine-cysteine can react directly as well. Double conjugation involving glutathione, cysteine-glycine or cysteine (M14, M15, M16, M27) were also observed in low abundance, most likely both ethenes have reacted. Glutathione conjugate (M9) and cysteine conjugate (M23) behave differently compared to others, as their abundance was high already in the 0 min sample, and highest in some of the species. This suggests that 1-380 is able to react with cysteine and glutahione almost immediately. This can also help explain, why the absolute parent peak area was lower in hepatocyte 0 min samples compared to buffer incubation 0 min sample - significant portion of the compound had already reacted. The abundance of metabolites other than those involving glutathione-related conjugates (Ml - M8) were low in human, with N- demethylation in dimethylamine (M3) and hydroxylation in N-propane (M2) having the highest abundance. The N-demethylation and hydroxylation in N-propane were also combined with glutathione, cysteine-glycine or cysteine conjugations in metabolites M12, M19, M20, M24, and M25. Metabolite M5, formed by loss of C5H7NO, did not show profile of a typical metabolite, as it was observed in all samples (excluding the blank), and could be impurity. Metabolite M9 had similar or higher abundance in other species when compared to human, while the abundance of Ml 8 and M23 had lower abundance in other species, except minipig and monkey for M18, which had similar abundance as in human. The abundance of M2 was lower in other species when compared to human, but was closest in monkey. The abundance of M3 was lower in mouse, rat, and rabbit, but similar in minipig, dog and monkey. In animal species, glutathione (M9), cysteine-glycine (Ml 8) and cysteine (M23) conjugates had clearly the highest abundance, but with different relative roles. Other metabolites had only minor share of metabolism. 22 metabolites were detected in mouse, 19 in rat, 18 in rabbit, 22 in minipig, 25 in dog and 25 in monkey.

[0215] The only metabolite which abundance increased during the plasma incubation was M5, formed by loss of C5H7NO form 1-380, but even that was present already in the 0 min sample. The main metabolite in hepatocytes, cysteine conjugate M23 was detected also in plasma samples, but the abundance was low and remained stable or slowly decreased during the incubation. Highest abundance of M23 was detected in rabbit, but most likely this just indicates that the used rabbit plasma had higher residual level of cysteine than other species. In addition to these metabolites detected in hepatocytes, several other metabolites were detected, in which 1-380 had conjugated with different amino acids. Similar to cysteine conjugate, the abundance of these was low and remained stable or decreased during the incubation. MS / MS data was available for the glycine conjugate (M28) and showed that the metabolite was related to 1-380 and that modification was in the same site as the in cysteine (ethene next to aromatic structure). For other amino acid conjugates, no MS / MS data was available, but it is assumed that they have similar structure. It is assumed that the conjugation is via the amine moiety of the amino acid, but this could not be confirmed by MS / MS data. Metabolite formation can’t explain the observed disappearance of the study compound in plasma incubation.

[0216] 1-381

[0217] With human hepatocytes, a half-life of 17.7 min was observed, corresponding to in vitro clearance of 39.1 pL / min / mg. The half-lives and in vitro clearances in the other species were 5.0 min / 140 pL / min / mg in mouse, were 4.1 min / 168 pL / min / mg in rat, were 34.8 min / 19.9 pL / min / mg in rabbit, were 31.8 min / 21.8 pL / min / mg in minipig, were 55.4 min / 12.5 pL / min / mg in dog, and were 40.4 min / 17.1 pL / min / mg in monkey.

[0218] The predicted hepatic extraction ratios were 83% for human, 89% in mouse, 92% in rat, and 71% in dog, suggesting high hepatic extraction (extraction ration >70%), 50% in rabbit, 54% in minipig, and 49% in monkey, suggesting intermediate hepatic clearance (extraction ratio 30 - 70%).

[0219] For 1-381, the shortest half-lives were observed in mouse plasma with 12.8 min, followed by rat with 41.4 min. The longest half-life (233 min) was observed in minipig. In human plasma, the half-life was 147 min, while the half-life in rabbit was 168 min, in dog was 136 min, and in monkey was 198 min.

[0220] In total 24 metabolites (Ml - M24) were detected for 1-381 in hepatocytes. Of these metabolites, 17 were detected in human hepatocytes. The most abundant metabolites were formed by conjugation with glutathione (M6), cysteine-glycine (M17), and cysteine (M22) and the conjugation was located to the ethene next to the aromatic structure by the MS / MS data. All three are part of the same metabolic pathway, as glutathione is first degraded to cysteine-glycine and then to cysteine, but cysteine and cysteine-cysteine can react directly as well. In addition, double conjugation with glutathione, cysteine-glycine or cysteine or their combinations (Mi l, M12, M13, M19, M20, and M23) were observed, most likely both ethenes had reacted. Glutathione conjugate (M9) and cysteine conjugate (M23) behave differently compared to others, as their abundance was high already in the 0 min sample. This suggests that 1-381 is able to react with cysteine and glutathione almost immediately. This can also help explain, why the absolute parent peak area was lower in hepatocyte 0 min samples compared to buffer incubation 0 min sample - significant portion of the compound had already reacted. The abundance of metabolites other than those involving glutathione-related conjugates (Ml - M5) were low in human, with di-oxidation in azetidine having highest abundance of those. The di-oxidation in azetidine was found in combination with glutathione, cysteine-glycine or cysteine conjugations in metabolites M7, Ml 8 and M24. Metabolite M5, formed by loss of C5H7NO, did not show profile of a typical metabolite, as it was observed in all samples (excluding the blank), and could be also an impurity, but its abundance clearly increased in monkey hepatocytes. Metabolite M6 had similar or higher abundance in other species when compared to human, while the abundance of M17 and M22 had mostly lower abundance in other species. However, the abundances of M17 and M22 were closer to human levels in minipig and monkey. The metabolite M2 had lower in mouse and rat but was observed in similar or higher abundance in other species. In animal species, glutathione (M6), cysteine-glycine (M17), cysteine (M22), and the double glutathione conjugates (M9) had clearly the highest abundance, but with different relative roles between the species. Other metabolites had only minor share of metabolism. 14 metabolites were detected in mouse, 12 in rat, 12 in rabbit, 18 in minipig, 16 in dog and 18 in monkey.

[0221] The only metabolites which abundance increased during the plasma incubation were M25, formed by oxidation and hydrogenation in ethene next to aromatic structure and M5, formed by loss of C5H7NO form 1-381, but both were present already in the 0 min sample as well. One of the main metabolites in hepatocytes, cysteine conjugate (M22) was detected also in plasma samples, but the abundance was low and remained stable or slowly decreased during the incubation. Highest abundance of M22 was detected in rabbit, but most likely this just indicates that the used rabbit plasma had higher residual level of cysteine than other species. In addition to these metabolites, several other metabolites were detected, in which 1-381 had conjugated with different amino acids. Similar to cysteine conjugate, the abundance of these was low and remained stable or decreased during the incubation. MS / MS data was available for the glycine conjugate (M27) and lysine conjugate (M32) and showed that the metabolites were related to 1-381 and that modification was in the same site as the in cysteine (ethene next to aromatic structure). For other amino acid conjugates, no MS / MS data was available, but it is assumed that they have similar structure. It is assumed that the conjugation is via the amine moiety of the amino acid, but this could not be confirmed by MS / MS data. Metabolite formation can’t explain the observed disappearance of the study compound in plasma incubation.

[0222] Biological Study Example 4-1: Metabolic turnover in hepatocytes

[0223] The remaining LC / MS peak areas detected for 1-305 after each incubation time point with hepatocytes from each species (relative to corresponding 0 min incubation samples) are shown in Table 1. The results for 1-380 are shown in Table 2 and for 1-381 in Table 3.

[0224] 1-305

[0225] The highest metabolic turnover of 1-305 was observed with mouse and rat hepatocytes, the remaining abundances after 120 min incubation period being 2% and 4%, respectively. This was followed by rabbit, human, minipig, monkey and slowest disappearance was observed in dog. The observed disappearance was cell-dependent, as no disappearance was observed in buffer incubation.

[0226] With human hepatocytes, a half-life of 41.2 min was observed, corresponding to in vitro clearance of 16.8 pL / min / mg. The half-lives and in vitro clearances in the other species were 13.1 min / 53.0 pL / min / mg in mouse, were 7.5 min / 92.2 pL / min / mg in rat, were 37.0 min / 18.7 pL / min / mg in rabbit, were 65.2 min / 10.6 pL / min / mg in minipig, were 163 min / 4.26 pL / min / mg in dog, and were 92.2 min / 7.52 pL / min / mg in monkey.

[0227] The predicted hepatic extraction ratios were 67% for human, 48% in rabbit, 36% in minipig, 46% in dog, suggesting intermediate hepatic clearance (extraction ratio 30 - 70%), 76% for mouse, 86% for rat, suggesting high hepatic extraction (extraction ration >70%) and 30% in monkey, suggesting low hepatic clearance (extraction ratio < 30%).

[0228] 1-380

[0229] The highest metabolic turnover of 1-380 was observed with rat hepatocytes, the remaining abundances after 40 min incubation period being 3%. The disappearance was slower in other species, with 3 - 39 % remaining in the last timepoint. The observed disappearance was cell-dependent, as no disappearance was observed in buffer incubation.

[0230] With human hepatocytes, a half-life of 32.0 min was observed, corresponding to in vitro clearance of 21.7 pL / min / mg. The half-lives and in vitro clearances in the other species were 30.1 min / 23.0 pL / min / mg in mouse, were 7.7 min / 90.5 pL / min / mg in rat, were 53.9 min / 12.9 pL / min / mg in rabbit, were 18.0 min / 38.6 pL / min / mg in minipig, were 86.9 min / 7.98 pL / min / mg in dog, and were 38.0 min / 18.3 pL / min / mg in monkey.

[0231] The predicted hepatic extraction ratios were 73% for human, and 86% in rat, suggesting high hepatic extraction (extraction ration >70%), 57% mouse, 39% in rabbit, 67% in minipig, 61% in dog, and 51% in monkey suggesting intermediate hepatic clearance (extraction ratio 30 - 70%).

[0232] 1-381

[0233] The highest metabolic turnover of 1-381 was observed with mouse and rat hepatocytes, with almost complete disappearance in the 20 min timepoint, but peak area did not drop to zero in later timepoints, but remained approximately in the same level. Disappearance was slower in other species, with 12 - 29 % remaining in the last timepoint. The observed disappearance was cell-dependent, as no disappearance was observed in buffer incubation.

[0234] With human hepatocytes, a half-life of 17.7 min was observed, corresponding to in vitro clearance of 39.1 pL / min / mg. The half-lives and in vitro clearances in the other species were 5.0 min / 140 pL / min / mg in mouse, were 4.1 min / 168 pL / min / mg in rat, were 34.8 min / 19.9 pL / min / mg in rabbit, were 31.8 min / 21.8 pL / min / mg in minipig, were 55.4 min / 12.5 pL / min / mg in dog, and were 40.4 min / 17.1 pL / min / mg in monkey.

[0235] The predicted hepatic extraction ratios were 83% for human, 89% in mouse, 92% in rat, and 71% in dog, suggesting high hepatic extraction (extraction ration >70%), 50% in rabbit, 54% in minipig, and 49% in monkey, suggesting intermediate hepatic clearance (extraction ratio 30 - 70%). Table 1. The relative LC / MS peak areas for 1-305 after 0 - 120 min incubations with hepatocytes from each species. The initial substrate concentration was 1 pM and cell concentration 1 million / mL. N=2

[0236] Table 2. The relative LC / MS peak areas for 1-380 after 0 - 120 min incubations with hepatocytes from each species. The initial substrate concentration was 1 pM and cell concentration 1 million / mL. N=2 Table 3. The relative LC / MS peak areas for 1-381 after 0 - 120 min incubations with hepatocytes from each species. The initial substrate concentration was 1 pM and cell concentration 1 million / mL. N=2

[0237] Biological Study Example 4-2: Clearance calculations

[0238] The first-order rate constants k (min1) of the metabolism were obtained from the slope of time vs. In (% remaining) plot using Excel software. All time points (log-scale) were used in fitting the rate constant k (based on visual inspection of the curves).

[0239] The in vitro half-life (ti / 2) of study compound is defined as:

[0240] Intrinsic in vitro clearance was calculated as follows: where V is the volume of the incubation and M is the number of hepatocytes in the incubation.

[0241] Hepatic intrinsic clearance CL;nt,u, [1 / min] was then calculated as follows:

[0242] CLint u= ^^int.inc.uxhepatocellularity x liver weight and hepatic clearance [1 / min] was calculated as follows:

[0243] Where Q is liver flow rate, fupis fraction unbound in plasma, and B:P is blood to plasma ratio. The total clearance was then compared to total liver blood flow QH to estimate the hepatic extraction ratio.

[0244] CLH

[0245] EH~ QH

[0246] The results of the pharmacokinetic calculations for 1-305 are shown in Table 4 together with all scaling parameters used in the calculations. For 1-380, the results are shown in Table 5 and for 1-381 in

[0247] Table 6. As no information about plasma protein binding or blood to plasma (B:P)- ratio was available, the unbound fractions were considered as 1 (i.e., binding was not taken into account).

[0248] Table 4. Kinetic in vitro calculations for 1-305, based on the disappearance data in hepatocytes.

[0249] *From Ring et a / , 2011 Table 5. Kinetic in vitro calculations for 1-380, based on the disappearance data in hepatocytes.

[0250] *From Ring et a / , 2011

[0251] Table 6. Kinetic in vitro calculations for 1-381, based on the disappearance data in hepatocytes.

[0252]

[0253] *From Ring et al, 2011

[0254] Biological Study Example 4-3: Metabolic turnover in plasma

[0255] The remaining LC / MS peak areas detected for 1-305 after each incubation time point with plasma from each species (relative to corresponding 0 min incubation samples) are shown in Table 7. The results for 1-380 are shown in Table 8 and for 1-381 in Table 9. The half-lives calculated from the disappearance data in plasma is shown in Table 10.

[0256] For 1-305, the shortest half-lives were observed in mouse and rat plasma, with halflives 42.0 min and 39.9 min, respectively, followed by dog with half-life of 71.5 min. In human plasma, the half-life was 116 min, while the half-life in rabbit was 165 min, in minipig was 110 min, and in monkey was 176 min.

[0257] For 1-380, the shortest half-lives were observed in rat and minipig plasma with halflives of 52.4 min and 68.3 min, respectively. The longest half-life (140 min) was observed in dog. In human plasma, the half-life was 87.3 min, while the half-life in mouse was 82.3 min, in rabbit was 90.8 min, and in monkey was 81.8 min.

[0258] For 1-381, the shortest half-lives were observed in mouse plasma with 12.8 min, followed by rat with 41.4 min. The longest half-life (233 min) was observed in minipig. In human plasma, the half-life was 147 min, while the half-life in rabbit was 168 min, in dog was 136 min, and in monkey was 198 min. Table 7. The relative LC / MS peak areas for 1-305 after 0 - 120 min incubations with plasma from each species. The initial substrate concentration was 1 pM. N=2

[0259] Table 8. The relative LC / MS peak areas for 1-380 after 0 - 120 min incubations with plasma from each species. The initial substrate concentration was 1 pM. N=2

[0260] Table 9. The relative LC / MS peak areas for 1-381 after 0 - 120 min incubations with plasma from each species. The initial substrate concentration was 1 pM. N=2

[0261] Table 10. Calculated half-lives in plasma INCORPORATION BY REFERENCE

[0262] All publications and patents mentioned herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

[0263] EQUIVALENTS

[0264] Furthermore, the invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims are introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the invention, or aspects of the invention, is / are referred to as comprising particular elements and / or features, certain embodiments of the invention or aspects of the invention consist, or consist essentially of, such elements and / or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms “comprising” and “containing” are intended to be open and permit the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0265] The foregoing written specification is considered to be sufficient to enable one skilled in the art to practice the invention. The present invention is not to be limited in scope by examples provided, since the examples are intended as a single illustration of one aspect of the invention and other functionally equivalent embodiments are within the scope of the invention. Various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims. The advantages and objects of the invention are not necessarily encompassed by each embodiment of the invention.

Claims

We claim:

1. A compound having a structure according to Formula (I):or a pharmaceutically acceptable salt thereof, wherein:R1is -CH3or -CH2CH3;R2is -C1-C4alkyl, -C3-C6cycloalkyl, -(C1-C3 alkylene)-C3-C6cycloalkyl, -(C1-C3 alkylene)-O-C1-C3alkyl, or -(C1-C3 alkylene)-N(R3)2; orR1and R2are taken together with the nitrogen atom to which they are attached to form a 4-7 membered saturated heterocyclyl, optionally comprising an additional heteroatom selected from N, O and S, wherein the 4-7 membered saturated heterocyclyl is optionally substituted with 1-2 substituents independently selected from oxo, -C1-C3 alkyl, -OH, -O(C1-C3alkyl), and -C(O)-C1-C3alkyl; andof the molecule; each R3is independently H or C1-C4 alkyl;R4is C1-C4 alkyl, hydrogen, or -C(O)C1-C4 alkyl; and R6is C1-C4 alkyl.

2. The compound of claim 1, wherein R1is -CH3.

3. The compound of claim 1 or 2, wherein R2is -CH3, -CH2CH3, -CH(CH3)2, cyclopentyl, -(C1-C3 alkylene)-cyclopentyl, -CH2CH2-OCH3, or -CH2CH2N(CH3)2.

4. The compound of claim 3, wherein R2is -CH3.

5. The compound of claim 1, wherein R1and R2are taken together with the nitrogen atom to which they are attached to form a 4-6 membered heterocyclyl.

6. The compound of claim 5, wherein R2represents7. The compound of any one of claims 1-6, wherein R5is8. The compound of any one of claims 1-6, wherein R4is hydrogen, -CH3 or - C(O)CH3.mpound of claim 1 , wherein the compound is selected from:and 2-143.

10. A compound having a structure according to Formula (II):or a pharmaceutically acceptable salt thereof, wherein:R1is -CH3or -CH2CH3;R2is -C1-C4 alkyl, -C3-C6cycloalkyl, -(C1-C3 alkylene)-C3-C6cycloalkyl, -(C1-C3 alkylene)-O-C1-C3alkyl, or -(C1-C3 alkylene)-N(R3)2; orR1and R2are taken together with the nitrogen atom to which they are attached to form a 4-7 membered saturated heterocyclyl, optionally comprising an additional heteroatom selected from N, O and S, wherein the 4-7 membered saturated heterocyclyl is optionally substituted with 1-2 substituents independently selected from oxo, -C1-C3 alkyl, - OH, -O(C1-C3alkyl), and -C(O)-C1-C3alkyl; andof the molecule; each R3is independently H or C1-C4 alkyl;R4is hydrogen, C1-C4 alkyl or -C(O)C1-C4 alkyl; and R6is C1-C4 alkyl.

11. The compound of claim 10, wherein R1is -CH3.

12. The compound of any one of claims 10-11, wherein R2is -CH3, -CH2CH3, - CH(CH3)2, cyclopentyl, -(C1-C3 alkylene)-cyclopentyl, -CH2CH2-OCH3, or - CH2CH2N(CH3)2.

13. The compound of claim 12, wherein R2is -CH3.

14. The compound of claim 10, wherein R1and R2are taken together with the nitrogen atom to which they are attached to form a 4-7 membered heterocyclyl.

15. The compound of claim 14, wherein R2represents16. The compound of any one of claims 10-15, wherein R5is17. The compound of any one of claims 10-15, wherein R4is hydrogen, -CH3 or - C(O)CH3.

18. The compound of claim 10, wherein the compound is selected from:

19. A pharmaceutically acceptable composition comprising a compound of any preceding claim, and a pharmaceutically acceptable carrier.

20. The composition of claim 19, formulated for oral or parenteral delivery.

21. A method of treating a cancer (e.g., solid tumor or hematological cancer) comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-18, or a composition of any one of claims 19 and 20.

22. The method of claim 21, wherein the cancer is selected from mesothelioma, lung, breast, prostate, melanoma, esophageal, leukemia, cervical, liver, colon, gastric, colorectal, glioblastoma, head and neck, pancreatic, and ovarian.

23. The method of claim 22, wherein the cancer is selected from mesothelioma, lung, ovarian, and breast.

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