Thiostrepton-based compounds for treatment of cancer and preparation thereof

Abstract

Disclosed herein are structures, compositions, and synthesis of thiostrepton-based pharmaceutically related compounds, as well as methods of treating cancer with such compounds.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 310,400 filed on February 15, 2022 and U.S. Provisional Patent Application No. 63 / 341,238 filed on May 12, 2022, each of which is hereby incorporated by reference in its entirety. Background Art

[0003] Thiostrepton is a cyclic oligopeptide antibiotic, also known by other names, such as Bryamycin, Thiactin, Alaninamide, HR4S203Y18, etc. Recent studies have shown that Thiostrepton also has promising anticancer activity. There is still a need for Thiostrepton derivatives with beneficial pharmacological properties. Summary of the invention

[0004] In certain embodiments, the present invention provides a series of compounds having the structure of Formula (IA):

[0005]

[0006] or a pharmaceutically acceptable salt thereof, wherein:

[0007] R 1 is -NH2, -NH(CH3), -O-CH3 or -NH-CH2-C(O)-NH2;

[0008] R 2 is -H, -CH3, =CH2 or =CH(alkyl);

[0009] R 3 is -H, -CH3, =CH2 or =CH(alkyl);

[0010] R 5 is -C(O)-R1 or -CN;

[0011] Ring A is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0012] Ring B is absent or present, and when present is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0013] R 4 is hydrogen, a protecting group, -C(O)-CH3, -L' or -LY;

[0014] L', when present, is a reactive linker moiety;

[0015] L, when present, is a linker moiety;

[0016] Y, when present, is the mitochondrial targeting moiety;

[0017] Each are independently a single bond or a double bond; and

[0018] Any hydrogen atoms are optionally replaced with deuterium.

[0019] In certain embodiments, the present invention also provides a series of compounds having the structure of formula (IB):

[0020]

[0021] or a pharmaceutically acceptable salt thereof, wherein:

[0022] R 1 is -NH2 or -O-CH3;

[0023] R 2 is CH2 or CH(alkyl);

[0024] R 3 is CH2 or CH(alkyl);

[0025] Ring A is heteroaryl, cycloalkyl or heterocyclyl;

[0026] Ring B is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0027] R 4 is hydrogen, a protecting group, -C(O)-CH3, -L' or -LY;

[0028] L', when present, is a reactive linker moiety;

[0029] L, when present, is a linker moiety;

[0030] Y, when present, is a mitochondrial targeting moiety; and

[0031] Any hydrogen atoms are optionally replaced with deuterium.

[0032] Also provided herein are methods of treating cancer comprising administering any of the pharmaceutical compositions described herein to a subject in need thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 : Cell viability assay data in malignant mesothelioma (MM) cells (HMESO cell line) treated with compound (1)-HCl or (5)-HCl. N = 2 technical replicates and 2 biological replicates performed by 2 different laboratory technicians. All compounds were dissolved in 100% DMSO. (5)-HCl precipitates at concentrations of approximately 20-80 μM when added to complete tissue culture medium.

[0034] FIG. 2A : View of HMESO mesothelioma cells treated with (5)-HCl (40× magnification).

[0035] FIG. 2B : View of HMESO mesothelioma cells treated with (5)-HCl (20× magnification).

[0036] Figure 3 : Immunoblot of PRX3 from HMESO mesothelioma cells treated with (1) or (5).

[0037] Figure 4 : Bar graph of PRX3 (PRX3-X-PRX3):monomer ratios of HMESO mesothelioma cells treated with (1) or (5).

[0038] Figure 5 is a table summarizing the results of Examples 42-44 for various compounds described herein. NT indicates "not tested". For EC 50 Values, "A" represents an EC less than or equal to 5.0 μM 50 , "B" represents an EC of 5.1 μM to 10.0 μM 50 , "C" represents an EC of 10.1 μM to 50.0 μM 50 , and "D" represents an EC greater than 50.0 μM 50 . DETAILED DESCRIPTION

[0039] Pharmaceutical composition

[0040] The compositions and methods described herein can be used to treat individuals in need. 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 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 non-limiting examples, 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 a preferred embodiment, when such pharmaceutical compositions are used for administration to humans, especially for invasive administration routes (i.e., routes to avoid transport or diffusion through epithelial barriers, such as injection or implantation), the aqueous solution is pyrogen-free, or substantially pyrogen-free. Excipients can be selected, for example, to achieve delayed release of the agent or selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in unit dosage form, such as tablets, capsules (including spray capsules and gelatin capsules), granules, lyophilized agents for reconstitution, powders, solutions, syrups, suppositories, injections or similar dosage forms. The composition may also be presented in a transdermal delivery system, such as a skin patch. The composition may also be presented in a solution suitable for topical administration, such as a lotion, cream or ointment.

[0041] A pharmaceutically acceptable carrier may contain a physiologically acceptable agent that acts, for example, to stabilize a compound (such as a compound described herein), increase the solubility of the compound, or increase the absorption of the compound. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran; 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 formulation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) may also be a liposome or other polymer matrix, into which a compound such as described herein may be incorporated. For example, liposomes comprising phospholipids or other lipids are non-toxic, physiologically acceptable and metabolizable carriers that are relatively simple to manufacture and administer.

[0042] The phrase "pharmaceutically acceptable" is used 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.

[0043] As used herein, the phrase "pharmaceutically acceptable carrier" 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 that can be used 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 carboxymethylcellulose, 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, oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffered saline; and (21) other non-toxic compatible substances used in pharmaceutical preparations.

[0044] The pharmaceutical composition (formulation) can be administered to a subject by any of a number of routes of administration, including, for example, oral (e.g., lavage, tablets, capsules (including spray capsules and gelatin capsules) in aqueous or non-aqueous solutions or suspensions, boluses, powders, granules, pastes applied to the tongue); absorption through the oral mucosa (e.g., sublingually); subcutaneously; transdermally (e.g., as a patch applied to the skin); and topically (e.g., as a cream, ointment or spray applied to the skin). The compound can also be formulated for inhalation. In certain embodiments, the compound can simply be dissolved or suspended in sterile water. Details of suitable routes of administration and compositions suitable for such routes of administration can be found, for example, in U.S. Patent Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896 and the patents cited therein.

[0045] The preparation can be conveniently presented in unit dosage form and can be prepared by any method well-known in the pharmaceutical technology. The amount of active ingredient that can be combined with carrier materials to produce a single dosage form will vary depending on the host treated, the specific mode of administration. The amount of active ingredient that can be combined with carrier materials to produce a single dosage form will generally be the amount of the compound that produces the therapeutic effect. In general, in 100%, this amount will be in the range of about 1% to about 99% active ingredient, preferably about 5% to about 70%, most preferably about 10% to about 30%.

[0046] The method for preparing these preparations or compositions includes the step of bringing into association the active compound (such as the compounds described herein) with a carrier and optionally one or more auxiliary ingredients. In general, the preparations are prepared by uniformly and intimately bringing into association the compounds described herein with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

[0047] Formulations described herein suitable for oral administration can be in the form of capsules (including spray capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), lyophilizates, powders, granules, or as a solution or 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 a pastille (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and / or as a mouthwash, etc., each containing a predetermined amount of a compound described herein as the active ingredient. The composition or compound can also be administered as a bolus, elixir or paste.

[0048] To prepare solid dosage forms for oral administration (capsules (including spray capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), 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 starch, lactose, sucrose, glucose, mannitol and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrants such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) solution retardants such as paraffin; (6) absorption promoters such as quaternary ammonium compounds; (7) wetting agents such as cetyl alcohol and glyceryl monostearate; (8) absorbents such as kaolin and bentonite; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, 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 spray capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also contain a buffer. Solid compositions of a similar type may also be used as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols.

[0049] Tablets can be made by compression or molding, optionally with one or more auxiliary ingredients. Compressed tablets can be prepared using a binder (e.g., gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), a surfactant or a dispersant. Molded tablets can be made by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0050] Tablets and other solid dosage forms of pharmaceutical compositions, such as dragees, capsules (including spray capsules and gelatin capsules), pills and granules, can be optionally scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in pharmaceutical formulation technology. The dosage form can also be formulated to provide a slow or controlled release of the active ingredient therein, for example, using different proportions of hydroxypropyl methylcellulose to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. The dosage form can be sterilized by, for example, filtering through a bacterial retention filter, or by immediately incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water or some other sterile injectable medium before use. These compositions can also optionally contain an opacifier and can have a composition that releases the active ingredient only or preferentially in a certain part of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. When appropriate, the active ingredient can also be in a microencapsulated form together with one or more of the above-mentioned excipients.

[0051] Liquid dosage forms that can be used for oral administration include pharmaceutically acceptable emulsions, lyophilized agents for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage form may contain an inert diluent commonly used in the art, such as water or other solvents, cyclodextrins and their derivatives; solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oil (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuran methanol, polyethylene glycol and sorbitan fatty acid esters, and mixtures thereof.

[0052] 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.

[0053] 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.

[0054] Dosage forms for 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 any preservatives, buffers, or propellants that may be required.

[0055] Ointments, pastes, creams and gels may contain, in addition to the active compounds, excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0056] In addition to the active compound, powders and sprays can contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of these substances. In addition, sprays can contain conventional propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0057] Transdermal patches have the additional advantage of controlled delivery of the compounds described herein to the body. Such dosage forms can be made by dissolving or dispersing the active compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound through the skin. The rate of this flux can be controlled by providing a rate-controlling membrane or dispersing the compound in a polymer matrix or gel.

[0058] As used herein, the phrases "parenteral administration" and "parenteral administration" refer to modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise a combination of one or more active compounds with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions or sterile powders that can be reconstituted into sterile injectable solutions or dispersions prior to use, and the pharmaceutical compositions may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0059] Examples of suitable aqueous and nonaqueous carriers that can be used in the pharmaceutical compositions described herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) 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 maintaining the required particle size in the case of dispersions, and by the use of surfactants.

[0060] These compositions may also contain adjuvants, such as preservatives, wetting agents, emulsifiers and dispersants. By including various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc., protection against microbial action can be ensured. It may also be necessary to include isotonic agents, such as sugar, sodium chloride, etc. in the composition. In addition, delayed absorption of injectable pharmaceutical forms may be achieved by including delayed absorption agents, such as aluminum monostearate and gelatin.

[0061] In some cases, in order to prolong the drug effect, it is necessary to slow down the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material with poor water solubility. The absorption rate of the drug depends on its dissolution rate, which in turn may depend on the crystal size and crystalline form. Alternatively, delayed absorption of a drug form administered parenterally can be achieved by dissolving or suspending the drug in an oil vehicle.

[0062] By forming a microencapsulation matrix of the subject compound in a biodegradable polymer, such as polylactide-polyglycolide, an injectable storage form is made. Depending on the ratio of the drug to the polymer and the properties of the specific polymer used, the drug release rate can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Storage injectable preparations are also prepared by encapsulating the drug in a liposome or microemulsion compatible with body tissues.

[0063] For use in the methods described herein, the active compounds may be administered per se or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably 0.5% to 90%) of the active ingredient in combination with a pharmaceutically acceptable carrier.

[0064] The introduction method can also be provided by a rechargeable or biodegradable device. Various slow-release polymer devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including protein biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable polymers and non-degradable polymers, can be used to form implants for sustained release of compounds at specific target sites.

[0065] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without causing toxicity to the patient.

[0066] The selected dosage level will depend upon a variety of factors, including the activity of the specific compound or combination of compounds employed, or the esters, salts or amides thereof; the route of administration; the time of administration; the rate of excretion of the specific compound employed; the duration of the treatment; other drugs, compounds and / or materials used in combination with the specific compound 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.

[0067] A physician or veterinarian of ordinary skill in the art can easily determine and prescribe the therapeutically effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian may start the dosage of the pharmaceutical composition or compound at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. "Therapeutically effective amount" means a concentration of the compound sufficient to induce the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the subject's weight, sex, age, and medical history. Other factors affecting the effective amount may include, but are not limited to, the severity of the patient's illness, the condition being treated, the stability of the compound, and, if necessary, another type of therapeutic agent administered with the compound described herein. A larger total dose may be delivered by multiple administrations of the agent. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13th edition, 1814-1882, incorporated herein by reference).

[0068] In general, a suitable daily dose of the active compound used in the compositions and methods described herein will be that amount which is the lowest dose of the compound effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above.

[0069] If necessary, the effective daily dose of the active compound can be administered as one, two, three, four, five, six or more sub-doses, which are administered independently at appropriate time intervals throughout the day, optionally in unit dosage form. In certain embodiments, the active compound can be administered twice or three times a day. In a preferred embodiment, the active compound will be administered once a day.

[0070] The patient receiving such treatment is any animal in need thereof, including primates, particularly humans; and other mammals, such as horses, cattle, pigs, sheep, cats, and dogs; poultry; and pets in general.

[0071] In certain embodiments, the compounds described herein may be used alone or in combination with another type of therapeutic agent.

[0072] The present disclosure includes the use of pharmaceutically acceptable salts of compounds described herein in compositions and methods described herein. In certain embodiments, expected salts include but are not limited to alkyl, dialkyl, trialkyl or tetraalkyl ammonium salts. In certain embodiments, expected salts include but are not limited to L-arginine, benthamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucosamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine and zinc salts. In certain embodiments, expected salts include but are not limited to Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, the 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, l-ascorbic acid, l-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, caprylic acid (decanoic acid), caprylic acid (caproic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid (cyclamic acid), acid), dodecyl sulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptanoic 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, l-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, l-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, l-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenate.

[0073] Pharmaceutically acceptable acid addition salts may also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates may also be prepared. The source of such solvates may be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or foreign to such solvent.

[0074] 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.

[0075] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfite, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0076] definition

[0077] Unless otherwise defined herein, the scientific and technical terms used in this application shall have the meanings commonly understood by those of ordinary skill in the art. In general, the nomenclature and techniques associated with the 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.

[0078] The methods and techniques of the present disclosure are generally performed 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 the present specification unless otherwise indicated.

[0079] Unless otherwise defined herein, chemical terms used 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, CA (1985).

[0080] All of the above, as well as any other publications, patents, and published patent applications mentioned in this application, are specifically incorporated herein by reference. In the event of a conflict, the present specification, including its specific definitions, will control.

[0081] The term "agent" is used herein to refer to a compound (such as an organic or inorganic compound, a mixture of compounds), a biopolymer (such as a nucleic acid, an antibody, including a portion thereof and a humanized antibody, a chimeric antibody and a human antibody and a monoclonal antibody, a protein or a portion thereof, such as a peptide, a lipid, a carbohydrate), or an extract made from a biological material, such as a bacterial, plant, fungal or animal (particularly mammalian) cell or tissue. Agents include, for example, agents of known structure and those of unknown structure.

[0082] "Patient", "subject" or "individual" are used interchangeably and refer to human or non-human animals. These terms include mammals, such as humans, primates, livestock animals (including cattle, pigs, etc.), companion animals (e.g., dogs, cats, etc.) and rodents (e.g., mice and rats).

[0083] "Treatment" of a disease or patient refers to taking steps to obtain beneficial or desired results, including clinical results. As used herein and as is well understood in the art, "treatment" is a method for obtaining beneficial or desired results (including clinical results). Beneficial or desired clinical results may include, but are not limited to, alleviating or improving one or more symptoms or diseases, alleviating the extent of the disease, stabilizing (i.e., not worsening) the disease state, preventing disease transmission, delaying or slowing disease progression, improving or alleviating the disease state, and alleviating (partially or completely), whether detectable or undetectable. "Treatment" may also mean extending the survival period compared to the expected survival period without treatment.

[0084] The term "prevention" is art-recognized and is well understood in the art when used in connection with a condition such as local recurrence (e.g., pain), a disease such as cancer, a syndromic complex such as heart failure, or any other medical condition, and includes administering a composition that reduces the frequency of symptoms of the medical condition or delays the onset of symptoms in a subject relative to a subject not receiving the composition. Thus, cancer prevention includes, for example, reducing the number of detectable cancerous growths in a patient population receiving prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable cancerous growths in a treated population compared to an untreated control population, e.g., by a statistically and / or clinically significant amount.

[0085] The "administration" or "administration" of a substance, compound or agent to a subject can be performed using one of a variety of methods known to those skilled in the art. For example, the compound or agent can be administered intravenously, intraarterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinal, intracerebrally, and transdermally (by absorption, such as a transdermal catheter). The compound or agent can also be suitably introduced by a rechargeable or biodegradable polymer device or other device (such as a patch and pump) or formulation that provides extended, slow or controlled release of the compound or agent. It can also be administered, for example, once, multiple times and / or over one or more extended periods of time.

[0086] The appropriate method of administering a substance, compound or agent to a subject will also depend on, for example, the age and / or physical condition of the subject and the chemical and biological properties (e.g., solubility, digestibility, bioavailability, stability and toxicity) of the compound or agent. In some embodiments, the compound or agent is orally administered to the subject, for example, by ingestion. In some embodiments, the orally administered compound or agent is in an extended release or slow release formulation, or is administered using a device for such slow or extended release.

[0087] As used herein, the phrase "combined 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 the drug product can be detected systemically using industry-accepted methods, or when the two agents are effective for the patient at the same time, this can include a synergistic effect of the two agents). For example, different therapeutic compounds can be administered concomitantly or sequentially in the same formulation or in separate formulations. In certain embodiments, different therapeutic compounds can be administered within one hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, or a week of each other. Therefore, individuals receiving such treatment can benefit from the combined effects of different therapeutic agents.

[0088] A "therapeutically effective amount" or "therapeutically effective dose" of a compound or other agent described herein is an amount of the drug or agent that will have the desired therapeutic effect when administered to a subject. The full therapeutic effect does not necessarily occur by administering one dose of such a drug or agent, and may occur only after administering a series of doses (multiple consecutive doses). Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount required for a subject will depend, for example, on the subject's size, health, and age, as well as the nature and extent of the disorder (such as cancer) being treated.

[0089] As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where the event or circumstance does not occur. For example, "optionally substituted alkyl" means that the alkyl group may be substituted as well as instances where the alkyl group is not substituted.

[0090] As used herein, the term "modulate" includes inhibiting or suppressing a function or activity (such as cell proliferation) as well as enhancing a function or activity.

[0091] The term "pharmaceutically acceptable" is art-recognized. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and / or dosage forms that are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio, within the scope of sound medical judgment.

[0092] "Pharmaceutically acceptable salt" or "salt" is used herein to refer to acid addition salts or base addition salts that are suitable or compatible with patient treatment.

[0093] As used herein, the term "pharmaceutically acceptable acid addition salt" refers to any non-toxic organic or inorganic salt of any basic compound. Illustrative inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid, and metal salts, such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids that form suitable salts include monocarboxylic acids, dicarboxylic acids and tricarboxylic acids, such as glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, benzoic acid, phenylacetic acid, cinnamic acid and salicylic acid, and sulfonic acids, such as p-toluenesulfonic acid and methanesulfonic acid. Monoacids or diacids can be formed, and such salts can exist in hydrated, solvated and substantially anhydrous forms. In general, the acid addition salts of the compound are more soluble in water and various hydrophilic organic solvents, and generally exhibit higher melting points compared to their free base forms. The selection of appropriate salts will be known to those skilled in the art. Other non-pharmaceutically acceptable salts, such as oxalates, may be useful, for example, in isolating the compound for laboratory use or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0094] As used herein, the term "pharmaceutically acceptable base addition salt" means any non-toxic organic or inorganic base addition salt of any acid compound or any intermediate thereof. Illustrative inorganic bases that form suitable salts include lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide or barium hydroxide. Illustrative organic bases that form suitable salts include aliphatic, alicyclic or aromatic organic amines such as methylamine, picoline and picoline or ammonia. The selection of appropriate salts will be known to those skilled in the art.

[0095] Many compounds that can be used in the methods and compositions of the present disclosure have at least one stereogenic center in their structure. This stereogenic center can exist in the R or S configuration, and the R and S symbols are used to correspond to the rules described in Pure Appl. Chem. (1976), 45, 11-30. The present disclosure covers all stereoisomeric forms of compounds, salts, prodrugs or mixtures thereof (including all possible stereoisomer mixtures), such as enantiomeric and diastereomeric forms. See, for example, WO 01 / 062726.

[0096] Some compounds may also exist in tautomeric forms. Such forms, although not explicitly indicated in the formulas described herein, are intended to be included within the scope of the present disclosure.

[0097] "Prodrug" or "pharmaceutically acceptable prodrug" refers to a compound that is metabolized, e.g., hydrolyzed or oxidized, in a host to form a compound of the present disclosure after administration. Typical examples of prodrugs include compounds having biologically labile or cleavable (protecting) groups on a functional portion of an 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 an active compound. Examples of prodrugs using esters or phosphoramidates 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 the present 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.

[0098] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filter, diluent, excipient, solvent or encapsulating material, which can be used to formulate a medicament for medical or therapeutic use.

[0099] As used herein, the term "linker" means any chemical functional group that "links" or chemically connects any two or more other chemical functional groups in a pharmaceutically relevant molecule. As a non-limiting example of the use of a linker in a pharmaceutical context, an antibody-drug conjugate (ADC) comprises a pharmaceutically active small molecule, drug or toxin connected to a macromolecular antibody by a linker.

[0100] Examples of common linker types include cleavable and non-cleavable linkers. Cleavable linkers include chemical functional groups that can be cleaved in response to physiological stimuli, such as chemical gradients, pH changes, or enzymatic activity. Non-limiting examples include acid or base labile functional groups, pyrophosphate diesters, disulfide bonds, peptides, β-glucuronides, etc. Non-cleavable linkers include chemical functional groups that are generally less stable to the aforementioned physiological stimuli, and non-limiting examples include certain alkyl and organic polymeric functional groups.

[0101] As used herein, the term "reactive linker moiety" refers to a chemical structure having a terminal portion that can react with another moiety (such as a mitochondrial targeting moiety) and form a covalent bond.

[0102] As used herein, the terms "mitochondrial targeting peptide", "mitochondrial targeting sequence", "mitochondrial targeting moiety" are art-recognized terms that refer to a chemical functional group (peptide, sequence or moiety) that is "targeted" to the mitochondrial membrane - that is, it is easily transported to and taken up by the mitochondrial membrane (J. Zielonka, B. Kalyanaraman et al., 2017). As used herein, the mitochondrial targeting moiety can include, but is not limited to, the following types: berberine cation, rhodamine cation, indolium cation, pyridinium cation, tetraguanidinium cation, cyanine derivative, guanidinium cation, biguanidinium cation, triphenylphosphonium cation, triethylammonium cation, triphenylamine, tetraphenylethylene moiety, arylphosphonium cation, SS peptide, mitochondrial penetrating peptide (MPP), mitochondrial targeting sequence (MTS) peptide, hemigramicidin S-linked nitroxide, dequalinium (DQA) cation, non-localized Lipophilic cation, F16 ((E) -4- (1H-indol-3-ylvinyl) -N-methylpyridinium iodide), (L-cyclohexylalanine-D-arginine) 3, mitochondrial targeting nanocarrier, DDDK peptide, glycyrrhetinic acid, α-tocopheryl succinate (α-TOS), graphene oxide nanocarrier, PEG-pro-apoptotic peptide (KLAKLAK) 2, Dmt-D-Arg-Phe-Lys-NH2 peptide, methylglyoxal, N-nonylacridine orange, quinoline, styryl-azinium fluorophore or 15d-PGJ2. Exemplary mitochondrial targeting moieties are listed in reference. J Zielonka et al., Chem Rev 2017, 117, pp. 10043-10120; KL Horton et al., Chemistry & Biology 2008, 15, pp. 375-382; G Battogtokh et al., Front Pharmacol 2018, 9:922; U.S. Patent Nos. 9,173,952 and 9,132,198, the contents of each of which are incorporated herein by reference.

[0103] It will be appreciated that substituents and substitution patterns on the compounds described herein can be selected by one of ordinary skill in the art to produce chemically stable compounds that can be readily synthesized from readily available starting materials by techniques known in the art and those described below. If a substituent is itself substituted with more than one group, it will be appreciated that these multiple groups may be on the same carbon or on different carbons, as long as a stable structure results.

[0104] As used herein, the term "optionally substituted" means that one to six hydrogen radicals in a given structure are replaced by the group of a specified substituent, including but not limited to: hydroxy, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silicon, 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" means that one to four hydrogen radicals in a given structure are replaced by the substituents mentioned above. More preferably, one to three hydrogen radicals are replaced by the substituents as mentioned above. It should be understood that the substituents may be further substituted.

[0105] The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0106] 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 hydrocarbyl C(O)NH-.

[0107] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbyl C(O)O-, preferably alkyl C(O)O-.

[0108] 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.

[0109] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

[0110] As used herein, the term "alkyl" refers to a saturated aliphatic group, including but not limited to C1-C 10 Straight chain alkyl or C1-C 10 Branched alkyl. Preferably, the "alkyl" group refers to a C1-C6 straight chain alkyl or a C1-C6 branched chain alkyl. More preferably, the "alkyl" group refers to a C1-C4 straight chain alkyl or a C1-C4 branched chain alkyl. 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, neopentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl or 4-octyl, etc.

[0111] Furthermore, the term "alkyl" as used throughout the specification, examples and claims is intended to include unsubstituted and substituted alkyl groups, the latter referring to alkyl moieties having substituents replacing hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl and the like.

[0112] When used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, the term "C x-y ” or “C x -C y " is intended to include groups containing from x to y carbons in the chain. C0 alkyl indicates that the group has hydrogen in a terminal position, or a bond if internal. For example, C 1-6 Alkyl groups contain from one to six carbon atoms in the chain.

[0113] As used herein, the term "alkylamino" refers to an amino group substituted with at least one alkyl group.

[0114] As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

[0115] As used herein, the term "amide" refers to the group

[0116]

[0117] Where R 9 and R 10 Each independently represents hydrogen or a hydrocarbon group, or R 9 and R 10 Together with the N atom to which they are attached they form a heterocyclic ring having 4 to 8 atoms in the ring structure.

[0118] The terms "amine" and "amino" are art-recognized and refer to unsubstituted and substituted amines and salts thereof, for example, the moiety represented by

[0119]

[0120] Where R 9 , R 10 and R 10 ' each independently represents hydrogen or a hydrocarbon group, or R 9 and R 10 Together with the N atom to which they are attached they form a heterocyclic ring having 4 to 8 atoms in the ring structure.

[0121] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0122] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0123] The term "aryl", used alone or as part of a larger moiety such as "aralkyl", "aralkyloxy", or "aryloxyalkyl", 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" is used interchangeably with the term "aryl ring". In certain embodiments of the present disclosure, "aryl" refers to an aromatic ring system, including but not limited to phenyl, biphenyl, naphthyl, anthracenyl, etc., which may bear one or more substituents. As used herein, groups in which an aromatic ring is fused to one or more non-aromatic carbocyclic rings are also included within the scope of the term "aryl".

[0124] The term "carbamate" is art-recognized and refers to the group

[0125]

[0126] Where R 9 and R 10 independently represent hydrogen or a hydrocarbon group.

[0127] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0128] The term "carbocycle" includes 5-7 membered monocycles and 8-12 membered bicyclic rings. Each ring of the bicyclic carbocycle can be selected from saturated, unsaturated and aromatic rings. Carbocycles include bicyclic molecules, wherein one, two or three or more atoms are shared between the two rings. The term "fused carbocycle" refers to a bicyclic carbocycle, wherein each of the rings shares two adjacent atoms with another ring. Each ring of the fused carbocycle can be selected from saturated, unsaturated and aromatic rings. In an exemplary embodiment, an aromatic ring, such as a phenyl ring, can be fused with a saturated or unsaturated ring, such as cyclohexane, cyclopentane or cyclohexene. Where the valence number permits, any combination of saturated, unsaturated and aromatic bicyclic rings is included in the definition of carbocycle. 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 decahydronaphthalene, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. "Carbocycle" may be substituted at any position or positions capable of bearing a hydrogen atom.

[0129] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0130] The term "carbonate" is art-recognized and refers to the group -OCO2-.

[0131] As used herein, the term "carboxyl" refers to a group represented by the formula -CO2H.

[0132] As used herein, the term "ester" refers to the group -C(O)OR 9 , where R 9 It represents a hydrocarbon group.

[0133] As used herein, the term "ether" refers to a hydrocarbon group connected to another hydrocarbon group via an oxygen. Thus, an ether substituent of a hydrocarbon group may be hydrocarbon-O-. An ether may be symmetrical or asymmetrical. 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.

[0134] As used herein, the terms "halo" and "halogen" mean halogen and include chlorine, fluorine, bromine and iodine.

[0135] As used herein, the term "hetaralkyl" or "heteroaralkyl" refers to an alkyl group substituted with a heteroaryl group.

[0136] The terms "heteroaryl" and "heteroar-", used alone or as part of a larger moiety such as "heteroaralkyl" or "heteroaralkoxy", refer to groups having 5 to 10 ring atoms, preferably 5, 6 or 9 ring atoms; having 6, 10 or 14 π electrons shared in a cyclic array; and having one to five heteroatoms in addition to carbon atoms. The term "heteroatom" refers to nitrogen, oxygen or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of basic nitrogen. Heteroaryl includes, but is not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl and pteridinyl. As used herein, the terms "heteroaryl" and "heteroaromatic-" also include groups in which a heteroaromatic ring is fused to one or more aryl or heteroaryl rings so that the resulting bicyclic or polycyclic system as a whole is fully aromatic. Non-limiting examples include indolyl, isoindolyl, benzothiophenyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl and phenoxazinyl. The heteroaryl group can be a monocyclic or bicyclic ring. The term "heteroaryl" is used interchangeably with the terms "heteroaryl ring", "heteroaryl" or "heteroaromatic", any of which includes optionally substituted rings. The term "heteroaralkyl" refers to an aryl group substituted with a heteroaryl group, wherein the alkyl and heteroaryl moieties are independently and optionally substituted.

[0137] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen and sulfur.

[0138] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group.

[0139] As used herein, the terms "heterocycle", "heterocyclyl", "heterocyclic group" 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 saturated or partially unsaturated and has one or more, preferably one to four, heteroatoms as defined above in addition to carbon atoms. The term "nitrogen" when used to refer to the ring atoms of a heterocycle includes 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).

[0140] Heterocycle can be connected to its side group at any heteroatom or carbon atom, which will produce a stable structure and any one of the ring atoms can be optionally substituted. The example of such saturated or partially unsaturated heterocyclic group includes but is not limited to tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolyl, tetrahydroisoquinolyl, decahydroquinolyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolane, diazepanyl, oxazepanyl, thiazopanyl, morpholinyl and quinuclidine. The terms "heterocycle", "heterocyclic radical", "heterocyclic ring", "heterocyclic group", "heterocyclic moiety" and "heterocyclic group" are used interchangeably, and also include groups in which heterocyclic rings are fused with one or more aryls, heteroaryls or alicyclic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl or tetrahydroquinolyl. Heterocyclic radical can be monocyclic or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocycloalkyl portions independently are optionally substituted.

[0141] As used herein, the term "hydrocarbyl" refers to a group bonded via a carbon atom that does not have a =O or =S substituent, and typically has at least one carbon-hydrogen bond and a main carbon backbone, but may optionally include heteroatoms. Therefore, for the purposes of this application, groups such as methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered hydrocarbyl, but substituents such as acetyl (which has a =O substituent on the connecting carbon) and ethoxy (which is connected via oxygen rather than carbon) are not hydrocarbyl. Hydrocarbyl includes, but is not limited to, aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

[0142] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group.

[0143] When used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy, the term "low" is intended to include groups in which the substituent has ten or fewer atoms, preferably six or fewer atoms. For example, "low alkyl" refers to an alkyl group containing ten or fewer carbon atoms, preferably six or fewer carbon atoms. In certain embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy substituents defined herein are respectively low acyl, low acyloxy, low alkyl, low alkenyl, low alkynyl or low alkoxy, whether they occur alone or in combination with other substituents, such as in the narration of hydroxyalkyl and aralkyl (in this case, for example, when calculating the carbon atoms in the alkyl substituent, the atoms in the aryl are not calculated).

[0144] The terms "polycyclic", "polycyclic" and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl) wherein two or more atoms are common to two adjacent rings, e.g., the rings are "fused rings". Each ring of the polycyclic ring may be substituted or unsubstituted. In certain embodiments, each ring of the polycyclic ring contains 3 to 10 atoms, preferably 5 to 7, in the ring.

[0145] The term "sulfate" is art-recognized and refers to the group -OSO3H or a pharmaceutically acceptable salt thereof.

[0146] The term "sulfonamide" is art-recognized and refers to a group represented by the general formula

[0147]

[0148] Where R 9 and R 10 independently represent hydrogen or a hydrocarbon group.

[0149] The term "sulfoxide" is art-recognized and refers to the group -S(O)-.

[0150] The term "sulfonate" is art-recognized and refers to the group SO3H or a pharmaceutically acceptable salt thereof.

[0151] The term "sulfone" is art-recognized and refers to the group -S(O)2-.

[0152] The term "protecting group" is an art-recognized term that refers to a chemical functional group that can modify (usually covalently modify) an existing unstable functional group on a target molecule. This modification "protects" the unstable functional group in subsequent reaction steps, and the protecting group can be removed as needed, which is called "deprotection." As a non-limiting example, the tert-butyloxycarbonyl (Boc or boc) group is commonly used to covalently modify and "protect" terminal amino groups in synthetic chemistry.

[0153] The term "substituted" refers to the part of the substituent having a hydrogen on one or more carbons of the replacement main chain. It should be understood that "replacement" or "replaced by ... " includes the following implicit conditions: such substitution is consistent with the allowed valence of the substituted atom and the substituent, and the substitution produces a stable compound, for example, it will not spontaneously undergo transformations such as rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad sense, permissible substituents include non-cyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. For appropriate organic compounds, permissible substituents can be one or more and are the same or different. For the purposes of this disclosure, heteroatoms such as nitrogen can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that meet the heteroatom valence. Substituents may include any of the substituents described herein, for example, halogen, hydroxy, carbonyl (such as carboxyl, alkoxycarbonyl, formyl or acyl), thiocarbonyl (such as thioester, thioacetate or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl or aromatic or heteroaromatic moieties. Those skilled in the art will appreciate that the moieties substituted on the hydrocarbon chain may themselves be substituted where appropriate.

[0154] Suitable monovalent substituents on the substitutable carbon atoms of an "optionally substituted" group are independently halogen; -(CH2) 0-4 R°; -(CH2) 0-4 OR°; -O(CH2) 0-4 R o 、-O-(CH2) 0-4 C(O)OR°; -(CH2) 0-4 CH(OR°)2; -(CH2) 0- 4SR°; -(CH2) 0-4 Ph, which may be substituted by R°; -(CH2) 0-4 O(CH2) 0-1 Ph, which may be substituted by R°; -CH=CHPh, which may be substituted by R°; -(CH2) 0-4 O(CH2) 0-1 -pyridyl, which may be substituted by R°; -NO2; -CN; -N3; ​​-(CH2) 0-4 N(R°)2; -(CH2) 0-4 N(R°)C(O)R°;-N(R°)C(S)R°;-(CH2) 0-4N(R°)C(O)NR°2;-N(R°)C(S)NR°2;-(CH2) 0-4 N(R°)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) 0-4 C(O)R°;-C(S)R°;-(CH2) 0-4 C(O)OR°; -(CH2) 0-4 C(O)SR°; -(CH2) 0-4 C(O)OSiR°3; -(CH2) 0-4 OC(O)R°; -OC(O)(CH2) 0-4 SR°, SC(S)SR°; -(CH2) 0-4 SC(O)R°; -(CH2) 0-4 C(O)NR°2; -C(S)NR°2; -C(S)SR°; -SC(S)SR°, -(CH2) 0-4 OC(O)NR°2; -C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH2C(O)R°; -C(NOR°)R°; -(CH2) 0-4 SSR°; -(CH2) 0-4 S(O)2R°; -(CH2) 0-4 S(O)2OR°; -(CH2) 0-4 OS(O)2R°;-S(O)2NR°2;-(CH2) 0-4 S(O)R°; -N(R°)S(O)2NR°2; -N(R°)S(O)2R°; -N(OR°)R°; -C(NH)NR°2; -P(O)2R°; -P(O)R°2; -OP(O)R°2; -OP(O)(OR°)2; SiR°3; -(C 1-4 Straight or branched alkylene)ON(R°)2; or -(C 1-4 linear or branched alkylene) C(O)ON(R°)2, wherein each R° may be substituted as defined below and is independently hydrogen, C 1-6 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, -CH2-(5-6 membered heteroaryl ring) or a 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definition, two independent occurrences of R° together with their intervening atoms form a 3-12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which heteroatoms may be substituted as defined below.

[0155] Suitable monovalent substituents on R° (or two independent occurrences of R° together with their intervening atoms to form a ring) are independently halogen, -(CH2) 0-2 R ● 、-(halogen R ● )、-(CH2) 0-2 OH, -(CH2) 0-2 OR ● 、-(CH2) 0-2 CH(OR ● )2; -O(halogen R ● )、-CN、-N3、-(CH2) 0-2 C(O)R ● 、-(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR ● 、-(CH2) 0-2 SR ● 、-(CH2) 0-2 SH, -(CH2) 0-2 NH2, -(CH2) 0-2 NHR ● 、-(CH2) 0-2 NR ● 2. -NO2, -SiR ● 3. -OSiR ● 3. -C(O)SR ● 、-(C 1-4 Straight or branched alkylene)C(O)OR ● or -SSR ● , where each R ● unsubstituted or, when preceded by "halo", substituted only by one or more halogens, and independently selected from C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph 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.

[0156] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: =0, =S, =NNR * 2. =NNHC(O)R * 、=NNHC(O)OR * 、=NNHS(O)2R * , =NR * , =NOR * 、-O(C(R *2)) 2-3 O-or-S(C(R * 2)) 2-3 S-, where each independent occurrence of R * is selected from hydrogen, C which may be substituted as defined below 1-6 aliphatic or unsubstituted 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents bonded to the ortho-substitutable carbon of an "optionally substituted" group include: -O(CR * 2) 2-3 O-, where each independent occurrence of R * is selected from hydrogen, C which may be substituted as defined below 1-6 Aliphatic or unsubstituted 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0157] R * Suitable substituents on the aliphatic group include halogen, -R ● 、-(halogen R ● ), -OH, -OR ● 、-O(halogen R ● )、-CN、-C(O)OH、-C(O)OR ● 、-NH2、-NHR ● 、-NR ● 2 or -NO2, where each R ● unsubstituted or, when preceded by "halo", substituted only by one or more halogens, and independently C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph or a 5-7 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0158] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include or Each of these are independently hydrogen, C which may be substituted as defined below 1-6 aliphatic, unsubstituted -OPh or substituted 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or, notwithstanding the above definition, two independent occurrences of Together with their intervening atoms they form an unsubstituted 3-12 membered saturated, partially unsaturated or aromatic monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0159] Suitable substituents on the aliphatic group and the substituted 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur are independently halogen, -R ● 、-(halogen R ● ), -OH, -OR ● 、-O(halogen R ● )、-CN、-C(O)OH、-C(O)OR ● 、-NH2、-NHR ● 、-NR ● 2 or -NO2, where each R ● unsubstituted or, when preceded by "halo", substituted only by one or more halogens, and independently C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph or a 5-6 membered saturated, partially unsaturated or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0160] As used herein, the term "sulfanyl" refers to an alkyl group substituted with a thiol group.

[0161] As used herein, the term "thioester" refers to the group -C(O)SR 9 or -SC(O)R 9 , where R 9 It represents a hydrocarbon group.

[0162] As used herein, the term "thioether" is equivalent to an ether in which the oxygen is replaced by sulfur.

[0163] The term "urea" is art-recognized and may be represented by the general formula

[0164]

[0165] Where R 9 and R 10 independently represent hydrogen or a hydrocarbon group.

[0166] The term "tautomer" refers to each of two or more isomers of a compound that exist together in equilibrium and are interchangeable by the migration of atoms or groups (such as hydrogen atoms) within the molecule. Exemplary tautomers of the present disclosure include, but are not limited to The depiction of one tautomer within a genus or compound class in this disclosure is intended to encompass the drawn compound and all tautomers thereof. Specifically, for the above examples, any of those structures is also disclosed for a series of 2n different classes. where n is the number of tautomeric sites on the molecule.

[0167] For purposes of the present disclosure, any of the embodiments described herein are applicable to any of the general formulas described herein, provided that interdependencies exist where appropriate.

[0168] In certain embodiments, the present invention provides a series of compounds having the structure of Formula (IA):

[0169]

[0170] or a pharmaceutically acceptable salt thereof, wherein:

[0171] R 1 is -NH2, -NH(CH3), -O-CH3 or -NH-CH2-C(O)-NH2;

[0172] R 2 is -H, -CH3, =CH2 or =CH(alkyl);

[0173] R 3 is -H, -CH3, =CH2 or =CH(alkyl);

[0174] R 5 is -C(O)-R1 or -CN;

[0175] Ring A is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0176] Ring B is absent or present, and when present is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0177] R 4 is hydrogen, a protecting group, -C(O)-CH3, -L' or -LY;

[0178] L', when present, is a reactive linker moiety;

[0179] L, when present, is a linker moiety;

[0180] Y, when present, is the mitochondrial targeting moiety;

[0181] Each are independently a single bond or a double bond; and

[0182] Any hydrogen atoms are optionally replaced with deuterium.

[0183] In certain embodiments, the present invention provides a series of compounds having the structure of formula (2-1):

[0184]

[0185] or a pharmaceutically acceptable salt thereof, wherein:

[0186] R 1 is -NH2, -NH(CH3), -O-CH3 or -NH-CH2-C(O)-NH2;

[0187] R 2 is -H, -CH3, =CH2 or =CH(alkyl);

[0188] R 3 is -H, -CH3, =CH2 or =CH(alkyl);

[0189] R 5 is -C(O)-R1 or -CN;

[0190] Ring A is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0191] Ring B is absent or present, and when present is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0192] R 4 is hydrogen, a protecting group, -C(O)-CH3, -L' or -LY;

[0193] L', when present, is a reactive linker moiety comprising an alkynyl group;

[0194] L, when present, is a linker moiety;

[0195] Y, when present, is the mitochondrial targeting moiety;

[0196] Each are independently a single bond or a double bond;

[0197] Any hydrogen atoms are optionally replaced by deuterium;

[0198] The prerequisite is

[0199] If Ring A is selected from unsubstituted thiazolyl, oxazolyl, cyclopentyl and bicyclo[2.1.1]hexane and Ring B-NHR 4 yes but

[0200] a) R 4 is -L'-Y or L; or

[0201] b) R 2 and R 3 Not simultaneously =CH2 or not simultaneously =CH(alkyl).

[0202] In certain embodiments, L is T is selected from alkyl and (-CH2CH2-O-) v -; v is an integer selected from 3-9; and T is bonded to Y.

[0203] In certain embodiments, the present invention also provides a series of compounds having the structure of formula (2-2):

[0204]

[0205] or a pharmaceutically acceptable salt thereof, wherein:

[0206] R 1 is -NH2 or -O-CH3;

[0207] R 2 is =CH2 or CH(alkyl);

[0208] R 3 is =CH2 or CH(alkyl);

[0209] Ring A is heteroaryl, cycloalkyl or heterocyclyl;

[0210] Ring B is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0211] R 4 is hydrogen, a protecting group, -C(O)-CH3, -L' or -LY;

[0212] L', when present, is a reactive linker moiety comprising an alkynyl group;

[0213] L, when present, is a linker moiety;

[0214] Y, when present, is a mitochondrial targeting moiety; and

[0215] Any hydrogen atoms are optionally replaced by deuterium;

[0216] The prerequisite is

[0217] If Ring A is selected from unsubstituted thiazolyl, oxazolyl, cyclopentyl and bicyclo[2.1.1]hexane and Ring B-NHR 4 yes but

[0218] a) R 4 is -L'-Y or L; or

[0219] b) R 2 and R 3 Not simultaneously =CH2 or not simultaneously =CH(alkyl).

[0220] In certain embodiments, L is T is selected from alkyl and (-CH2CH2-O-) v -; v is an integer selected from 3-9; and T is bonded to Y.

[0221] In certain embodiments, the present invention also provides a series of compounds having the structure of formula (2-3):

[0222]

[0223] or a pharmaceutically acceptable salt thereof, wherein:

[0224] R 1 is -NH2 or -O-CH3;

[0225] R 2 is =CH2 or =CH(alkyl);

[0226] R 3 is =CH2 or =CH(alkyl);

[0227] Ring A is heteroaryl, cycloalkyl or heterocyclyl;

[0228] Ring B is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0229] R 4 is hydrogen, a protecting group or -C(O)-CH3; and

[0230] Any hydrogen atoms are optionally replaced by deuterium;

[0231] The prerequisite is

[0232] If Ring A is selected from unsubstituted thiazolyl, oxazolyl, cyclopentyl and bicyclo[2.1.1]hexane and Ring B-NHR 4 yes but

[0233] R 2 and R 3 Not simultaneously =CH2 or not simultaneously =CH(alkyl).

[0234] In certain embodiments, the present invention provides a series of compounds having the structure of formula (2-4):

[0235]

[0236] or a pharmaceutically acceptable salt thereof, wherein:

[0237] R 1 is -NH2, -NH(CH3), -O-CH3 or -NH-CH2-C(O)-NH2;

[0238] R 2 is -H, -CH3, =CH2 or =CH(alkyl);

[0239] R 3 is -H, -CH3, =CH2 or =CH(alkyl);

[0240] R 5 is -C(O)-R1 or -CN;

[0241] Ring A is selected from Where * represents the bond to ring B and ** and *** represent -NH-R 4 ;

[0242] Ring B is absent or present, and when present is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0243] R 4 is hydrogen, a protecting group, -C(O)-CH3, -L' or -LY;

[0244] L', when present, is a reactive linker moiety comprising an alkynyl group;

[0245] L, when present, is a linker moiety;

[0246] Y, when present, is the mitochondrial targeting moiety;

[0247] Each are independently a single bond or a double bond; and

[0248] Any hydrogen atoms are optionally replaced with deuterium.

[0249] In certain embodiments, L is T is selected from alkyl and (-CH2CH2-O-) v -; v is an integer selected from 3-9; and T is bonded to Y.

[0250] In certain embodiments, the present invention provides a series of compounds having the structure of formula (2-5):

[0251]

[0252] or a pharmaceutically acceptable salt thereof, wherein:

[0253] R 1 is -NH2, -NH(CH3), -O-CH3 or -NH-CH2-C(O)-NH2;

[0254] R 2 is -H, -CH3, =CH2 or =CH(alkyl);

[0255] R 3 is -H, -CH3, =CH2 or =CH(alkyl);

[0256] R 5 is -C(O)-R1 or -CN;

[0257] Ring A is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0258] Ring B-NHR 4 Selected from

[0259] R 4 is hydrogen, a protecting group, -C(O)-CH3, -L' or -LY;

[0260] L', when present, is a reactive linker moiety comprising an alkynyl group;

[0261] L, when present, is a linker moiety;

[0262] Y, when present, is the mitochondrial targeting moiety;

[0263] Each are independently a single bond or a double bond; and

[0264] Any hydrogen atoms are optionally replaced with deuterium.

[0265] In certain embodiments, L is T is selected from alkyl and (-CH2CH2-O-) v -; v is an integer selected from 3-9; and T is bonded to Y.

[0266] In certain embodiments, the present invention provides a series of compounds having the structure of formula (2-6):

[0267]

[0268] or a pharmaceutically acceptable salt thereof, wherein:

[0269] R 1 is -NH2 or -O-CH3;

[0270] R 2 is =CH2 or =CH(alkyl);

[0271] R 3 is =CH2 or =CH(alkyl);

[0272] Ring A is thiazolyl substituted by alkyl or thienyl;

[0273] Ring B-NHR 4 yes

[0274] R 4 is a protecting group or -L';

[0275] L', when present, is a reactive linker moiety comprising an alkynyl group;

[0276] Any hydrogen atoms are optionally replaced with deuterium.

[0277] In certain embodiments, R 5 Yes-C(O)-R 1 ; and R 1 is -OCH3. In other embodiments, R 5 Yes-C(O)-R 1 ; and R 1 It is -NH2.

[0278] In certain embodiments, when Ring A is polycyclic, then Ring B is absent.

[0279] In certain embodiments, Ring A is a 5-membered ring or a 5-membered ring fused to a second ring.

[0280] In certain embodiments, Ring A is a 5-membered heteroaryl. In some embodiments, Ring A is a bicyclic heteroaryl.

[0281] In certain embodiments, Ring A is thiazolyl, thienyl, oxazolyl, or imidazolyl. In other embodiments, Ring A is thiazolyl, thienyl, or oxazolyl. In some embodiments, Ring A is pyrrolidinyl. In other embodiments, Ring A is thiazolyl.

[0282] In certain embodiments, Ring A is:

[0283] Wherein * represents the bond to ring B.

[0284] In certain embodiments, Ring A is: Wherein * represents the bond to ring B.

[0285] In certain embodiments, Ring A is: Wherein * represents the bond to ring B.

[0286] In certain embodiments, Ring A is: Wherein * represents the bond to ring B.

[0287] In certain embodiments, Ring A is a 5-membered cycloalkyl or heterocyclyl.

[0288] In certain embodiments, Ring A is cyclopentyl or tetrahydrofuranyl.

[0289] In certain embodiments, Ring A is Wherein * represents the bond to ring B.

[0290] In certain embodiments, Ring A is a bridged bicyclic cycloalkyl or heterocyclyl.

[0291] In certain embodiments, Ring A is bicyclo[2.1.1]hexyl or oxabicyclo[2.1.1]hexyl.

[0292] In certain embodiments, Ring A is Wherein * represents the bond to ring B.

[0293] In certain embodiments, wherein Ring A is phenyl.

[0294] In certain embodiments, Ring A is Wherein * represents the bond to ring B.

[0295] In certain embodiments, Ring A is a polycyclic aryl, heteroaryl, cycloalkyl, or heterocyclyl.

[0296] In certain embodiments, Ring A is a bicyclic heteroaryl.

[0297] In certain embodiments, Ring A is: Where *** and ** represent ring B or -NH-R 4 .

[0298] In certain embodiments, Ring B is a 6-membered ring.

[0299] In certain embodiments, B is phenyl.

[0300] In certain embodiments, Ring B is unsubstituted phenyl. In other embodiments, Ring B is phenyl substituted with halogen.

[0301] In certain embodiments, Ring B is: wherein Z is halo and ** represents a bond to ring A. In some embodiments, Z is selected from fluoro or chloro.

[0302] In certain embodiments, Ring B is: wherein Z is an alkoxy group, such as methoxy, or an alkyl group, such as methyl.

[0303] In other embodiments, Ring B is

[0304] In certain embodiments, Ring B is a 6-membered heteroaryl.

[0305] In certain embodiments, Ring B is pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl.

[0306] In certain embodiments, Ring B is pyridinyl, pyrimidinyl, or pyrazinyl.

[0307] In certain embodiments, Ring B is: Wherein * represents the bond to ring A.

[0308] In certain embodiments, Ring B is a bridged bicyclic cycloalkyl.

[0309] In certain embodiments, Ring A is bicyclo[2.2.2]octyl or bicyclo[1.1.1]pentyl.

[0310] In certain embodiments, Ring B is: wherein Z is a halogen group and ** represents a bond to ring A.

[0311] In certain embodiments, Ring B is selected from Where * indicates -NHR 4 key.

[0312] In certain embodiments, R 5 Yes-CN.

[0313] In certain embodiments, the compound of formula (IA) has the structure of formula (IA-1):

[0314] or a pharmaceutically acceptable salt thereof, wherein

[0315] R 2 is -H or -CH3; and

[0316] R 3 is =CH2 or =CH(alkyl); and

[0317] Ring A, Ring B, R 1 , R 4 and R 5 Each of which is any one of the embodiments described above for Formula (IA).

[0318] In certain embodiments of Formula (IA-1), R 2 Yes -H and R 3 =CH2, or R 2 is -CH3 and R 3 Yes =CH2.

[0319] In certain embodiments, the compound of formula (IA) has the structure of formula (IA-2):

[0320] or a pharmaceutically acceptable salt thereof,

[0321] in

[0322] R 2 is =CH2 or =CH(alkyl); and

[0323] R 3is -H or -CH3; and

[0324] Ring A, Ring B, R 1 , R 4 and R 5 Each of which is any one of the embodiments described above for Formula (IA).

[0325] In certain embodiments of Formula (IA-2), R 2 =CH2 and R 3 Yes -H, or R 2 =CH2 and R 3 It is -CH3.

[0326] In certain embodiments, the compound of formula (IA) has the structure of formula (IA-3):

[0327] or a pharmaceutically acceptable salt thereof,

[0328] in

[0329] R 2 is =CH2 or =CH(alkyl); and

[0330] R 3 is =CH2 or =CH(alkyl); and

[0331] Ring A, Ring B, R 1 , R 4 and R 5 Each of which is any one of the embodiments described above for Formula (IA).

[0332] In certain embodiments of Formula (IA-3), R 2 =CH2 and R 3 Y = CH2, R 2 =CH2 and R 3 =CH(CH3), or R 2 =CH(CH3) and R 3 Yes =CH2.

[0333] In certain embodiments, the compound of formula (IA) has the structure of formula (IB):

[0334] or a pharmaceutically acceptable salt thereof, wherein

[0335] Ring A, Ring B, R 1 , R 2 , R 3 and R 4 Each of which is any one of the embodiments described above for Formula (IA).

[0336] In certain embodiments, the compound of formula (IA) has the structure of formula (IB):

[0337]

[0338] or a pharmaceutically acceptable salt thereof, wherein:

[0339] R 1 is -NH2 or -O-CH3;

[0340] R 2 is CH2 or CH(alkyl);

[0341] R 3 is CH2 or CH(alkyl);

[0342] Ring A is heteroaryl, cycloalkyl or heterocyclyl;

[0343] Ring B is aryl, heteroaryl, cycloalkyl or heterocyclyl;

[0344] R 4 is hydrogen, a protecting group, -C(O)-CH3, -L' or -LY;

[0345] L', when present, is a reactive linker moiety;

[0346] L, when present, is a linker moiety;

[0347] Y, when present, is a mitochondrial targeting moiety; and

[0348] Any hydrogen atoms are optionally replaced with deuterium.

[0349] In certain embodiments of formula (IB), Ring A is a 5-membered ring or a 5-membered ring fused or bridged to a second ring. In certain embodiments, Ring A is thiazolyl, thienyl, oxazolyl, cyclopentyl, or bicyclo[2.1.1]hexyl.

[0350] In certain embodiments of Formula (IB), Ring A is thiazolyl.

[0351] In certain embodiments of Formula (IB), Ring A is:

[0352]

[0353] In certain embodiments of Formula (IB), Ring A is:

[0354] Wherein * represents the bond to ring B.

[0355] In certain embodiments of Formula (IB), Ring B is a six-membered ring. In certain embodiments, Ring B is phenyl or pyridinyl. In certain embodiments, Ring B is unsubstituted phenyl.

[0356] In certain embodiments of Formula (IB), Ring B is phenyl substituted with halogen.

[0357] In certain embodiments of Formula (IB), R 2 and R 3 are different.

[0358] In certain embodiments of Formula (IB), R 2 and R 3 In certain embodiments, R 2 It is CH(Me) or -CH2.

[0359] In certain embodiments of Formula (IB), R 3 It is CH(Me) or -CH2.

[0360] In certain embodiments of Formula (IB), R 2 and R 3 Each is CH2.

[0361] In certain embodiments of Formula (IB), R 4 is hydrogen, a protecting group or -C(O)-CH3.

[0362] In certain embodiments, the compound is selected from:

[0363]

[0364] Where R 4 is hydrogen, a protecting group or -C(O)-CH3; and

[0365] The drawing is The bond represents either possible stereochemistry of the attached olefin, E or Z.

[0366] In certain embodiments, the compound is selected from:

[0367]

[0368]

[0369] Where R 4 is hydrogen, a protecting group or -C(O)-CH3. In certain embodiments, the compound is selected from:

[0370]

[0371] In certain embodiments, the compound is selected from:

[0372]

[0373] Where R 4 is hydrogen, a protecting group or -C(O)-CH3; and wherein Z is selected from fluorine, chlorine, bromine and iodine. In certain embodiments, the compound is selected from:

[0374]

[0375]

[0376] Where R 4 is hydrogen, a protecting group or -C(O)-CH3.

[0377] In certain embodiments, the compound is selected from:

[0378]

[0379]

[0380] Where R 4 is hydrogen, a protecting group or -C(O)-CH3.

[0381] In certain embodiments, the protecting group is Boc.

[0382] In certain embodiments, R 4 In certain embodiments, L' is -C(O)-XC(O)OH or -C(O)-XC(O)NH2; X is -(CH2) n -, and n is 2, 3, 4 or 5.

[0383] In certain embodiments, the compound is selected from:

[0384]

[0385]

[0386] Where X is -(CH2) n -; and n is 2, 3, 4 or 5.

[0387] In certain embodiments, the compound is selected from:

[0388]

[0389]

[0390]

[0391] Where X is -(CH2)n -;and

[0392] n is 2, 3, 4 or 5. In certain embodiments, R 4 In some embodiments, L is a cleavable linker. In some embodiments, L is a non-cleavable linker. In some embodiments, L has a chain length of about 2 to about 30 atoms. In some embodiments, L has a chain length of about 5 to about 20 atoms.

[0393] In certain embodiments, L is -C(O)-XC(O)-; X is -(CH2) n -; and n is 2, 3, 4 or 5.

[0394] In certain embodiments, L is -C(O)-XC(O)-; X is -(CH2CH2-O-) m -(C H2CH2)-; and m is 2, 3, 4, 5 or 6.

[0395] In certain embodiments, R 4 is L', wherein the alkynyl group is

[0396] In certain embodiments, the compound is selected from:

[0397]

[0398]

[0399] Where X is -(CH2) n -; and n is 2, 3, 4 or 5. In certain embodiments, the compound is selected from:

[0400]

[0401]

[0402]

[0403] Where X is -(CH2) n -;

[0404] m is 2, 3, 4, 5, or 6; and

[0405] n is 2, 3, 4, or 5.

[0406] In certain embodiments, L' comprises an alkynyl group or an azide group.

[0407] In certain embodiments, wherein R 4is -C(O)-X'-C≡CH or -C(O)-X'-N3; X' is -(CH2) n -; and n is 2, 3, 4 or 5.

[0408] In certain embodiments, L comprises a heteroaryl group. In other embodiments, L comprises a triazolyl group.

[0409] In certain embodiments, R 4 yes

[0410] R 6 is -H or -C(O)CH3;

[0411] X' is -(CH2) n -;

[0412] X" is -(CH2) o -;

[0413] n is 2, 3, 4, or 5; and

[0414] o is 2, 3, 4, or 5.

[0415] In certain embodiments, R 4 yes

[0416] R 6 is -H or -C(O)CH3;

[0417] X' is -(CH2) n -;

[0418] X" is -(CH2) o -;

[0419] n is 2, 3, 4, or 5; and

[0420] o is 2, 3, 4, or 5.

[0421] In certain embodiments, R 4 yes

[0422] X' is -(CH2) n -;

[0423] X" is -(CH2) o -;

[0424] n is 2, 3, 4, or 5; and

[0425] o is 2, 3, 4, or 5.

[0426] In certain embodiments, Y is a berberine cation, a rhodamine cation, an indolium cation, a pyridinium cation, a tetraguanidinium cation, a cyanine derivative, a guanidinium cation, a biguanidinium cation, a triphenylphosphonium cation, a triethylammonium cation, a triphenylamine, a tetraphenylethylene moiety, an arylphosphonium cation, an SS peptide, a mitochondrial penetrating peptide (MPP), a mitochondrial targeting sequence (MTS) peptide, a hemigramicidin S-linked nitroxide, a dequalinium chloride (DQA) cation, a nonlocalized lipophilic cation, a F16 ( (E)-4-(1H-indol-3-ylvinyl)-N-methylpyridinium iodide), (L-cyclohexylalanine-D-arginine)3, mitochondrial targeting nanocarriers, DDDK peptide, glycyrrhetinic acid, α-tocopheryl succinate (α-TOS), graphene oxide nanocarriers, PEG-pro-apoptotic peptide (KLAKLAK)2, Dmt-D-Arg-Phe-Lys-NH2 peptide, methylglyoxal, N-nonylacridine orange, quinoline, styryl fluorophore or 15d-PGJ2.

[0427] In certain embodiments, Y is a mitochondrial penetrating peptide.

[0428] In certain embodiments, Y has structural formula (V):

[0429]

[0430] In certain embodiments, the compound is selected from:

[0431]

[0432] or a pharmaceutically acceptable salt thereof; wherein X is [-(CH2) n -] or [-(CH2CH2-O-) m -(C H2CH2)-]; n is 3, 4 or 5; and m is 2, 3, 4, 5 or 6.

[0433] In certain embodiments, the present invention provides a pharmaceutically acceptable composition comprising any one of the compounds described herein; and a pharmaceutically acceptable carrier.

[0434] In certain embodiments, the compositions are formulated for oral or parenteral delivery.

[0435] In certain embodiments, the compound is contained within a nanoparticle, a liposome, or a micelle, wherein the nanoparticle, the liposome, or the micelle is conjugated to a mitochondrial targeting moiety.

[0436] In certain embodiments, the present invention discloses a composition comprising a compound of formula (IA) or any subformula thereof, wherein R 4is hydrogen, a protecting group or -C(O)-CH3 or any of the more specific embodiments thereof described herein, the compound is contained in a nanoparticle, a liposome or a micelle, wherein the nanoparticle, the liposome or the micelle is conjugated to a mitochondrial targeting moiety.

[0437] In certain embodiments, the compositions are formulated for oral or parenteral delivery.

[0438] In certain embodiments, the present invention discloses a composition comprising a compound of formula (IB), wherein R 4 is hydrogen, a protecting group or -C(O)-CH3 or any of the more specific embodiments thereof described herein, the compound is contained in a nanoparticle, a liposome or a micelle, wherein the nanoparticle, the liposome or the micelle is conjugated to a mitochondrial targeting moiety.

[0439] In certain embodiments, the nanoparticle, liposome, or micelle is selected from poly(ethylene glycol), poly(ε-caprolactone), a polysaccharide, poly[(2-hydroxypropyl)-methacrylic acid], poly(lactic-co-glycolic acid), and any combination of the foregoing.

[0440] In certain embodiments, the present invention discloses a method for treating cancer (e.g., solid tumors or blood cancers), the method comprising administering to a subject in need thereof a therapeutically effective amount of any one of the compounds described herein or a composition of the compounds. In certain embodiments, the cancer (solid tumor or blood cancer) is selected from lung cancer, breast cancer, prostate cancer, melanoma, esophageal cancer, leukemia, cervical cancer, liver cancer, colon cancer, gastric cancer, colorectal cancer, glioblastoma, head and neck cancer, pancreatic cancer, mesothelioma and ovarian cancer. In certain embodiments, the cancer is selected from mesothelioma, lung cancer, ovarian cancer and breast cancer.

[0441] In certain embodiments, the present invention discloses a compound selected from the group consisting of:

[0442]

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449]

[0450]

[0451]

[0452]

[0453] Example

[0454] General Experimental Procedure 1

[0455] Suzuki coupling

[0456] The starting boronic acid or boronic ester (1 equivalent) is introduced into a flask under N2. 1,2-dimethoxyethane (0.1M), bromide (1.05 equivalents), 2M aqueous sodium carbonate solution (5 equivalents) are added. The mixture is then degassed with N2 for several minutes. Tetrapalladium (0.05 equivalents) is then added, and the mixture is stirred at 90°C (external) until complete conversion (overnight). The reaction is then stopped. Water is added, and the mixture is extracted with EtOAc. Brine is added to improve separation. The organic layer is dried over Na2SO4, filtered and concentrated. The crude material is purified by automated FCC. The desired product is obtained in a yield of 40-70%. Note: Hydrolysis products are also observed in some reactions. This is separated by acidifying the aqueous phase and extracted twice with EtOAc. The combined organics are washed with brine, dried over Na2SO4, filtered and concentrated to obtain hydrolysis products.

[0457] General Experimental Procedure 2

[0458] Ester hydrolysis

[0459] To a solution of the ester (1 eq.) in THF (0.05 M) was added a solution of lithium hydroxide monohydrate (4 eq.) in water (2 M), and the mixture was stirred at room temperature for 2-16 h. The mixture was diluted with EtOAc and washed with 1 M HCl and brine (30 mL each), dried over NaSO, filtered and concentrated to give the desired product.

[0460] General Experimental Procedure 3

[0461] EDCI-mediated amide coupling

[0462] To a solution of the acid (1 eq) and the amine (1 eq) in DCM (0.05 M) was added DiPEA (3 eq), cyano (E)-2-(hydroxyimino)butyric anhydride (1.3 eq) and EDCI (1.3 eq) and the resulting mixture was stirred at room temperature for 16 hours. The mixture was washed with 1 M HCl, water, NaHCO3 and brine, dried over Na2SO4, filtered and concentrated. The crude material was purified by automated FCC to give the desired product.

[0463] General Experimental Procedure 4

[0464] HATU-mediated amide coupling

[0465] To a solution of the acid (1 eq.) and the amine (1 eq.) in DCM (0.05 M) was 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 1 M HCl, water, NaHCO and brine, dried over NaSO, filtered and concentrated. The crude material was purified by automated FCC to give the desired product.

[0466] General Experimental Procedure 5

[0467] Acetylation of serine side chains

[0468] To the solution of free alcohol (1 equivalent) in MeCN (0.05M), triethylamine (2 equivalents / alcohol), DMAP (0.2 equivalent) and acetic anhydride (1.1 equivalents / alcohol) are added, and the resulting mixture is stirred for 2 hours. The reaction mixture is diluted with water and MeCN is removed in a vacuum. The mixture is extracted with EtOAc, and the organic phase is washed with salt water, through Na2SO4 is dried, filtered and concentrated. By automation FCC purification of crude material, desired product is obtained.

[0469] General Experimental Procedure 6

[0470] Stepwise elimination of mono-TBDPS-protected bis-serine

[0471] Triethylamine (10 equivalents) and methanesulfonyl chloride (1.2 equivalents) are added to a solution of a mono-TBDPS protected bisserine derivative (1 equivalent) in DCM (0.05M), and the resulting mixture is stirred at room temperature for 1 hour. The reaction is tracked by LCMS. Optionally triethylamine is added again. After completion, a solution of tetrabutylammonium fluoride in THF (1M, 2 equivalents) is added, and the mixture is stirred for 1 hour. The mixture is washed with 1M HCl and brine, dried over Na2SO4, filtered and concentrated. The residue is dissolved in CH2Cl2, and triethylamine (10 equivalents) and methanesulfonyl chloride (1.2 equivalents) are added. The resulting mixture is stirred at room temperature for 1 hour. The reaction is tracked by LCMS. Optionally triethylamine is added again. The mixture is washed with 1M HCl and brine, dried over Na2SO4, filtered and concentrated. The crude material is purified by automated reverse phase FCC.

[0472] General Experimental Procedure 7

[0473] Stepwise elimination of mono-TBDPS, mono-acetate

[0474] To the solution of mono TBDPS monoacetyl bisserine derivative (1 equivalent) in CH2Cl2 (0.05M), DBU (2 equivalents) is added, and the mixture is stirred at room temperature for 1-16 hours. Then a solution of TBAF in THF (1M, 2 equivalents) is added, and the mixture is stirred for 2 hours. The concentrated mixture is purified by automation FCC. To the intermediate of the single elimination in CH2Cl2 (0.05M), triethylamine (6 equivalents) and mesyl chloride (1.5 equivalents) are added. Optionally, triethylamine is added again. The mixture is washed with 1MHCl and salt water, dried over Na2SO4, filtered and concentrated. The crude material is purified by automation reverse phase FCC.

[0475] General Experimental Procedure 8

[0476] Elimination of diacetate

[0477] A solution of diacetate in THF / CH2Cl2 (1 / 1, v / v, 0.05M) was cooled to 0°C and DBU (4 equivalents) was added. The mixture was stirred at 0°C for 2 hours. The mixture was diluted with CH2Cl2, and water and HCl (1M aqueous solution) were added to acidify the mixture to pH=4. The mixture was extracted with CH2Cl2 (3×), and the combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The crude material was purified by automated reverse phase FCC.

[0478] General Experimental Procedure 9

[0479] Preparation of primary amides

[0480] To a solution of formic acid (1 equivalent) in DMF (0.1M) or CH2Cl2 (0.15M) add NH4Cl (3 equivalents), DiPEA (3.5 equivalents) and HATU (1.1 equivalents). Stir the mixture at room temperature overnight. Quench the reactant with water. In the case of a clear solution, then extract the mixture with EtOAc. Wash the organic layer with brine, dry over Na2SO4, filter and concentrate. Purify the crude material by automated FCC. In the case of a precipitate, then filter the mixture, and wash with water and the solvent used in the reaction. Recover the solid, concentrate to remove trace solvents, and obtain the required primary amide.

[0481] General Experimental Procedure 10

[0482] Thiohexiation of primary amides

[0483] To a solution of primary amide (1 eq.) in THF (0.2 M) was added Lawesson's reagent (0.7 eq.) and the mixture was stirred at 80 °C (external) until complete conversion. The reaction was then quenched with a saturated solution of NaHCO and the mixture was extracted with EtOAc. The organic layer was washed with brine, dried over NaSO, filtered and concentrated. The crude material was purified by automated FCC.

[0484] General Experimental Procedure 11

[0485] Formation of thiazole from sulfamide

[0486] To a solution of primary thioamide (1 eq.) in EtOH (0.2 M) was added ethyl bromopyruvate (1.1 eq.) and the resulting mixture was stirred at room temperature or 50 ° C (external) until complete conversion. The reactants were quenched with a saturated solution of NaHCO and the mixture was extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. The crude material was purified by automated FCC.

[0487] Analytical methods

[0488] LCMS method "21020335B TFA LCMS-5C3"

[0489]

[0490] Time (min) ％A ％B 0.0 95 5 0.5 95 5 2.5 10 90 3.5 10 90

[0491] LCMS method "21020335C TFA LCMS-5C4"

[0492] System: with UV detector and HP 6130MSD mass spectrometer

[0493]

[0494] Time (min) ％A ％B 0.0 80 20 1.5 10 90 3.0 10 90

[0495] LCMS method "22010199LCMS-5C3"

[0496]

[0497] Time (min) ％A ％B 0.0 80 20 1.5 0 100 3.0 0 100

[0498] LCMS method "22010199A TFA LCMS-5C1"

[0499]

[0500]

[0501] Time (min) ％A ％B 0.0 50 50 1.5 10 90 3.0 10 90

[0502] LCMS method "22010199C TFA LCMS-5C3"

[0503]

[0504] Time (min) ％A ％B 0.0 80 20 1.5 10 90 3.0 10 90

[0505] LCMS method "22010199D TFA LCMS-5C8"

[0506]

[0507]

[0508] Time (min) ％A ％B 0.0 80 20 1.5 10 90 3.0 10 90

[0509] LCMS method "30833LCMS-6"

[0510]

[0511] Time (min) ％A ％B 0.0 80 20 0.5 80 20 3.5 10 90 6.0 10 90

[0512] LCMS method "General 3 basic"

[0513]

[0514]

[0515] Time (min) ％A ％B 0.0 80 20 1.5 5 95 2.5 5 95 2.6 85 15

[0516] LCMS method "General 3acidic"

[0517]

[0518] Time (min) ％A ％B 0.0 80 20 1.5 5 95 2.5 5 95 2.6 85 15

[0519] Example 1A: Exemplary Synthesis of Thiostrepton-Based Structure A1

[0520]

[0521] Scheme 1A: Synthesis of Compound Compound A-1

[0522] Commercially available thiazole ethyl ester SM1a was hydrolyzed with LiOH to give formic acid Int2a. Subsequent amide coupling with serine methyl ester (SM3) gave amide Int4a. The free hydroxyl group was protected by treatment with TBDPS-Cl, and compound Int5a was obtained after column chromatography (79% in 3 steps). The methyl ester was hydrolyzed with LiOH to give the formic acid derivative Int6a, which was subsequently coupled with serine methyl ester SM7a to give dipeptide Int8a, which was purified by column chromatography (47% in 2 steps). The hydroxyl moiety was eliminated by treatment with methanesulfonyl chloride and NEt3, and the dehydroalanine derivative Int9a was obtained. A second elimination reaction was performed after TBAF-mediated TBDPS removal to give the didehydroalanine derivative Int11a.

[0523] Example 1B: Exemplary Synthesis of Thiostrepton-Based Structure A1((5)-HCl)

[0524]

[0525] Scheme 1B: Synthesis of Compound A-1

[0526] Suzuki coupling between boronate SM1B and bromide SM2B gave compound Int3B in moderate yield (65%). The Boc protecting group was exchanged to Alloc in 95% yield to give Alloc-protected compound Int4B. Methyl ester hydrolysis and subsequent EDCI-mediated coupling with serine methyl ester gave compound Int6B. The free alcohol was protected with TBDPS, and compound Int7B was isolated in 79% in 3 steps after purification. Methyl ester hydrolysis and subsequent EDCI-mediated coupling with serine methyl ester gave compound Int9B (45% yield). The hydroxyl group was eliminated by treatment with methanesulfonyl chloride and NEt3. A second elimination reaction was performed after TBAF-mediated TBDPS removal to give the didehydroalanine target. Finally, the Alloc group was removed by treatment with tetrapalladium in the presence of a scavenger to give compound A-1.

[0527] Alternative synthesis:

[0528] 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid intermediate 2

[0529]

[0530] Step 1. Ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylate (Intermediate 1) was prepared following General Experimental Procedure 1. 4-(tert-Butoxycarbonylamino)phenylboronic acid (2.5 g, 11 mmol) and ethyl 2-bromothiazole-4-carboxylate (2.5 g, 11 mmol) gave ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylate (Intermediate 1) (2.4 g, 6.9 mmol, 65%) as an off-white solid. 1 H NMR (400MHz, CDCl3) δ8.08 (s, 1H), 7.96-7.88 (m, 2H), 7.47-7.40 (m, 2H), 6.59 (s, 1H), 4.42 (q, J = 7.1Hz, 2H), 1.51 (s, 9H), 1.41 (t, J = 7.1Hz, 3H).

[0531] Step 2. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) was prepared following General Experimental Procedure 2. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylate (Intermediate 1) (0.93 g, 2.7 mmol) gave 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) (0.82 g, 2.6 mmol, 97%) as an off-white solid. 1 H NMR (400MHz, CDCl3) δ8.09(s,1H),7.89-7.82(m,2H),7.45-7.34(m,2H),1.47(s,9H),1.24-1.13(m,1H).

[0532] Methyl 2-(2-(2-(4-aminophenyl)thiazole-4-carboxamido)acrylamido)acrylate hydrochloride

[0533] (Compound A1)

[0534]

[0535] Step 1. Prepare ethyl 2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylate using the following procedure. To a solution of ethyl 2-(4-((tert-butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylate (Intermediate 1) (2.4 g, 1 eq., 6.9 mmol) in CH2Cl2 (10 mL) was added TFA (10 mL), and the resulting mixture was stirred at room temperature for 2 hours. The mixture was concentrated in vacuo, and the residue was dissolved in THF (50 mL). Pyridine (2.8 mL, 5 eq., 34 mmol) was added to the mixture, followed by allyl chloroformate (0.96 mL, 1.3 eq., 9.0 mmol), and the resulting mixture was stirred for 1 hour. The mixture was diluted with EtOAc, washed with 1M HCl and brine, dried over Na2SO4 and concentrated to give crude ethyl 2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylate as a yellow solid. The crude material was used as is in the next step.

[0536] Step 2. 2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylic acid was prepared following general experimental procedure 2. Crude ethyl 2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylate gave 2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylic acid (2.0 g, 6.6 mmol, 95%) as an orange solid. 1 H NMR (400MHz, CDCl3) δ8.20 (s, 1H), 7.95-7.87 (m, 2H), 7.50 (d, J = 8.5Hz, 2H), 6.83 (s, 1H), 5.96 (ddt, J =17.2,10.4,5.8Hz,1H),5.37(dq,J=17.2,1.5Hz,1H),5.32-5.24(m,1H),4.68(dt,J=5.8,1.4Hz,2H)

[0537] Step 3. N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 3. 2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylic acid 2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylic acid (1.1 g, 1 eq, 3.6 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (1.7 g, 1.2 eq, 4.3 mmol) gave N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.9 g, 3.0 mmol, 82%).

[0538] Step 4. N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine was prepared following General Experimental Procedure 2. N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.9 g, 1 eq, 3.0 mmol) afforded N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (1.8 g, 2.8 mmol, 93%) as a clear oil. The crude material was used as is in the next step.

[0539] Step 5. N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine methyl ester was prepared following general experimental procedure 3. N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (1.8 g, 2.8 mmol) gave N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine methyl ester (0.75 g, 1.0 mmol, 35%) as a yellow oil.

[0540] Step 6. Methyl 2-(2-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 6. Methyl N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine ester (0.75 g, 1 eq, 1.0 mmol) gave crude material of methyl 2-(2-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate (0.9 g) which was used in the next step without purification.

[0541] Step 7. Prepare methyl 2-(2-(2-(4-aminophenyl)thiazole-4-carboxamido)acrylamido)acrylate using the following procedure. To a solution of crude methyl 2-(2-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate (0.90 g) in DCM (20 mL) was added phenylsilane (1.1 g, 1.2 mL, 5 eq., 9.9 mmol), followed by tetrakis(triphenylphosphine)-palladium(0) (0.23 g, 0.1 eq., 0.20 mmol), and the resulting mixture was stirred for 1 hour. MeCN (4 mL) was added, and CH2Cl2 was removed in vacuo. The mixture was directly submitted to automated reverse phase FCC to give methyl 2-(2-(2-(4-aminophenyl)thiazole-4-carboxamido)acrylamido)acrylate (8.5 mg, 23 μmol, 1.2%) as an off-white solid. LCMS (21020335C TFA LCMS-5C4) RT: 1.274 min: Area % (215 nm): 79.1%; m / z = 373.2 [M+H] + . 1 H NMR(400MHz,MeOD)δ8.38(s,1H),8.33-8.17(m,2H),7.57-7.52(m,2H),6.60( d,J=1.8Hz,1H),6.29(s,1H),6.00(s,1H),5.76(d,J=1.9Hz,1H),3.86(s,3H).

[0542] Example 2A: Exemplary synthesis of structure A2 ((1)) based on thiostrepton

[0543]

[0544] Scheme 2A: Synthesis of Compound A-2 via Dipeptide Building Blocks

[0545] Compound A-2 is obtained by coupling between thiazole derivatives and dipeptides. The dipeptide is synthesized starting with Boc-Ser-OMe. The hydroxyl group is first protected by treatment with TBDPS-Cl to obtain a serine derivative. The methyl ester is hydrolyzed by treatment with LiOH to obtain formic acid, which is then coupled with serine amide to obtain a dipeptide (27% in 3 steps) after column chromatography. The Boc protecting group is removed by treatment with HCl and a dipeptide is obtained. EDCI-mediated coupling of a dipeptide with formic acid gives a thiazole-coupled dipeptide (51%) after column chromatography. Hydroxyl elimination mediated by methanesulfonyl chloride and NEt3, followed by removal of TBDPS by TBAF and a second elimination reaction, gives a Boc-protected compound after preparative HPLC purification. The Boc protecting group is removed by treatment with HCl and compound A-2 is obtained.

[0546] Example 2B: Exemplary Synthesis of Thiostrepton-Based Structure A2

[0547]

[0548] Scheme 2B: Synthesis of Compound A-2 via a Dipeptide Building Block

[0549] The hydroxyl portion of Boc-Ser-OMe was protected with TBDPS to obtain Boc-Ser(TBDPS)-OMe. The methyl ester was subsequently removed by treatment with LiOH to obtain the acid. EDCI-mediated peptide coupling with serinamide gave a dipeptide at 27% in 3 steps. The Boc group was removed by treatment with HCl to obtain the amine. The EDCI-mediated coupling between the amine and the acid (whose synthesis is shown above) gave the product at 75% in 2 steps. The hydroxyl group was eliminated by treatment with methanesulfonyl chloride and NEt3. A second elimination reaction was performed after TBAF-mediated TBDPS removal to obtain the didehydroalanine target. Finally, the Alloc group was removed by treatment with palladium in the presence of a scavenger to obtain compound A-2.

[0550] Alternative synthesis of tautomers of compound A-2 / tautomer ((1) / tautomer):

[0551] Methyl 2-(2-(2-(4-aminophenyl)thiazole-4-carboxamido)acrylamido)acrylate hydrochloride

[0552]

[0553] Step 1. General Experimental Procedure 3 was followed to prepare allyl (4-(4-(((S)-1-(((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-3-((tert-butyldiphenylsilyl)oxy)-1-oxopropan-2-yl)carbamoyl)thiazol-2-yl)phenyl)carbamate. 2-(4-(((Allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylic acid (1.2 g, 1 eq, 3.8 mmol) (prepared as in step 2) and (S)-2-amino-N-((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)propanamide hydrochloride (prepared as in 7) (2.2 g, 1.2 eq, 4.6 mmol) gave allyl (4-(4-(((S)-1-(((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-3-((tert-butyldiphenylsilyl)oxy)-1-oxopropan-2-yl)carbamoyl)thiazol-2-yl)phenyl)carbamate (1.5 g, 2.1 mmol, 55%) as a white foam. LCMS (General 3acidic) RT: 1.44 min; Area % (254 nm): 92%; m / z = 716.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.35 (d, J=6.6Hz, 1H), 8.06 (s, 1H), 7.89-7.81 (m, 2H), 7.65 (tt, J=6.2, 1.5Hz, 4H),7.38(dddd,J=22.2,20.7,8.4,6.7Hz,8H),6.85(s,1H),6.79(s,1H),6.03-5.89(m,1H),5.42-5 .32(m,2H),5.31-5.23(m,1H),4.71-4.59(m,3H),4.56(dt,J=7.7,4.1Hz,1H),4.22(ddd,J=17.4,10 .8,3.3Hz,2H),3.95(dd,J=10.2,4.9Hz,1H),3.63(dd,J=11.4,4.5Hz,1H),3.47(s,1H),1.09(s,9H).

[0554] Step 2. General Experimental Procedure 6 was followed to prepare allyl (4-(4-((3-((3-amino-3-oxoprop-1-en-2-yl)amino)-3-oxoprop-1-en-2-yl)carbamoyl)thiazol-2-yl)phenyl)carbamate. Allyl (4-(4-(((S)-1-(((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-3-((tert-butyldiphenylsilyl)oxy)-1-oxopropan-2-yl)carbamoyl)thiazol-2-yl)phenyl)carbamate (0.35 g, 0.49 mmol) gave crude material (4-(4-((3-((3-amino-3-oxoprop-1-en-2-yl)amino)-3-oxopropyl-1-en-2-yl)carbamoyl)thiazol-2-yl)phenyl)carbamate (0.35 g) which was used as is in the next step.

[0555] Step 3. Prepare N-(3-((3-amino-3-oxoprop-1-ene-2-yl)amino)-3-oxoprop-1-ene-2-yl)-2-(4-aminophenyl)thiazole-4-carboxamide hydrochloride using the following procedure. To a solution of crude material (4-(4-((3-((3-amino-3-oxoprop-1-ene-2-yl)amino)-3-oxoprop-1-ene-2-yl)carbamoyl)thiazole-2-yl)phenyl)carbamic acid allyl ester (0.35 g, 0.79 mmol) in DCM (10 mL), silylbenzene (0.49 mL, 5 eq., 4.0 mmol) and tetrapalladium (92 mg, 0.1 eq., 79 μmol) were added, and the resulting mixture was stirred for 1 hour. MeCN (4 mL) was added, and DCM was removed in vacuo. The mixture was submitted directly to automated reverse phase FCC to give methyl 2-(2-(2-(4-aminophenyl)thiazole-4-carboxamido)acrylamido)acrylate hydrochloride as a white solid. LCMS (21020335B TFA LCMS-5C3) RT: 1.138 min; Area: 63.7% (215 nm); m / z = 458.2 [M+H] + .

[0556] Example 3: Exemplary Synthesis of Thiostrepton-Based Structure B

[0557]

[0558] Scheme 3: Synthesis of Hexapeptide Compound B

[0559] The sequential construction SPPS of the hexapeptide compound B was started using 5 grams of Rink amide resin (0.7 mmol / g) and a standard SPPS protocol. After complete deprotection and simultaneous cleavage from the resin using TFA / TIPS / water (95 / 2.5 / 2.5), the crude peptide was obtained by precipitation from MTBE: heptane (1: 1) and lyophilization. The crude peptide was purified using preparative HPLC to obtain compound B in the form of a TFA salt with a purity of >95%. For conversion to the HCl salt, the purified peptide was basified with NaOH and re-acidified with HCl, followed by preparative HPLC using HCl buffer to obtain the pure peptide in the form of the HCl salt.

[0560] Alternative synthesis:

[0561] H-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2 HCl Salt

[0562]

[0563] H-Cha-D-Ar g-Cha-D-Arg-Cha-D-Arg-NH2 HCl salt was prepared on a 2.8 mmol scale following the general procedure for solid phase peptide synthesis. Purification was performed using HCl buffer to obtain the desired HCl salt form. H-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2 HCl salt (0.89 g, 0.82 mmol, 29%) was isolated as a white solid. LCMS (30833LCMS-6): RT: 1.819 min; Area: 92.5% (215 nm), 99.7% (ELSD); m / z = 945.9 [M+H] +

[0564] Example 4: Compounds 1 and 2

[0565] tert-Butyl (4-(4-((3-((3-amino-3-oxoprop-1-en-2-yl)amino)-3-oxoprop-1-en-2-yl)carbamoyl)thiazol-2-yl)phenyl)carbamate and tert-butyl (4-(4-(((E)-1-(((E)-1-amino-1-oxoprop-2-ylidene)amino)-1-oxoprop-2-ylidene)carbamoyl)thiazol-2-yl)phenyl)carbamate

[0566]

[0567] Step 1. Prepare N-(tert-butyloxycarbonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester using the following procedure. Add TBDPS-Cl (17 g, 15 mL, 1.1 equivalents, 60 mmol) to a solution of (tert-butyloxycarbonyl)-L-serine methyl ester (12 g, 1 equivalent, 55 mmol) in DCM (250 mL) and imidazole (8.2 g, 2.2 equivalents, 0.12 mol), and stir the resulting mixture at room temperature overnight. The mixture was washed with 1M HCl (200 mL) and brine (200 mL), dried over Na2SO4, filtered and concentrated to give N-(tert-butyloxycarbonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (24 g, 52 mmol, 96%) as a turbid oil, which was left to solidify. 1 H NMR (400MHz, CDCl3) δ7.59 (m, 4H), 7.46-7.32 (m, 6H), 5.39 (d, J = 8.8Hz, 1H), 5.28 (d, J = 1.0Hz, 1H), 4.38 (dt, J = 9. 2, 3.1Hz, 1H), 4.05 (dd, J = 10.0, 3.0Hz, 1H), 3.87 (dd, J = 10.1, 3.1Hz, 1H), 3.72 (s, 3H), 1.44 (s, 9H), 1.01 (s, 9H).

[0568] Step 2. N-(tert-Butyloxycarbonyl)-O-(tert-Butyldiphenylsilyl)-L-serine was prepared following General Experimental Procedure 2. N-(tert-Butyloxycarbonyl)-O-(tert-Butyldiphenylsilyl)-L-serine methyl ester (12 g, 26 mmol) gave N-(tert-Butyloxycarbonyl)-O-(tert-Butyldiphenylsilyl)-L-serine (11.9 g, 26.8 mmol, quantitative). The material was used in the next step without purification.

[0569] Step 3. Tert-butyl ((S)-1-(((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-3-((tert-butyldiphenylsilyl)oxy)-1-oxopropan-2-yl)carbamate was prepared following General Experimental Procedure 3. N-(tert-Butyloxycarbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (10 g, 1 eq, 23 mmol) and L-serinamide hydrochloride (6.5 g, 2 eq, 46 mmol) gave tert-butyl ((S)-1-(((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-3-((tert-butyldiphenylsilyl)oxy)-1-oxopropan-2-yl)carbamate (2.5 g, 4.7 mmol, 20%) as a slightly pink solid. 1HNMR(400MHz, CDCl3)δ7.59(m,4H),7.48-7.33(m,6H),7.21(m,1H),6.83(s,1H),5.33(s,1H),5.23(s,1H),4.50(s,1H),4.22(dd,J=11.3,2.7Hz ,1H),4.16(d,J=5.8Hz,1H),4.09(dd,J=10.3,4.1Hz,1H),3.83(dd,J=10.4,4.4Hz,1H),3.60(dd,J=11.4,4.3Hz,1H),1.43(s,9H),1.04(s,9H).

[0570] Step 4. (S)-2-amino-N-((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)propanamide hydrochloride was prepared using the following procedure. Tert-butyl ((S)-1-(((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-3-((tert-butyldiphenylsilyl)oxy)-1-oxopropan-2-yl)carbamate (2.5 g, 1 eq, 4.7 mmol) was dissolved in 15 mL of 4M HCl in dioxane, and the resulting mixture was stirred at room temperature for 90 minutes. The mixture was concentrated to give (S)-2-amino-N-((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)propanamide hydrochloride (2.2 g, 4.6 mmol, 98%) as a white solid. 1 H NMR(400MHz,MeOD)δ7.73-7.62(m,4H),7.52-7.39(m,6H),4.54(t,J=5.4Hz,1H ), 4.11 (t, J = 4.5Hz, 1H), 4.03 (d, J = 4.5Hz, 2H), 3.86-3.74 (m, 2H), 1.07 (s, 9H).

[0571] Step 5. General Experimental Procedure 3 was followed to prepare tert-butyl (4-(4-(((S)-1-(((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-3-((tert-butyldiphenylsilyl)oxy)-1-oxopropan-2-yl)carbamoyl)thiazol-2-yl)phenyl)carbamate. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) (0.70 g, 2.2 mmol) and (S)-2-amino-N-((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)propanamide hydrochloride (2.0 g, 2 equiv., 4.4 mmol) gave tert-butyl (4-(4-(((S)-1-(((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-3-((tert-butyldiphenylsilyl)oxy)-1-oxopropan-2-yl)carbamoyl)thiazol-2-yl)phenyl)carbamate (1.3 g, 1.8 mmol, 81%). LCMS (General 3acidic) RT: 1.59 min; Area % (214 nm): 75%; m / z = 732.2 [M+H] + .

[0572] Step 6. Tert-butyl (4-(4-((3-((3-amino-3-oxoprop-1-en-2-yl)amino)-3-oxoprop-1-en-2-yl)carbamoyl)thiazol-2-yl)phenyl)carbamate and tert-butyl (4-(4-(((E)-1-(((E)-1-amino-1-oxoprop-2-ylidene)amino)-1-oxoprop-2-ylidene)carbamoyl)thiazol-2-yl)phenyl)carbamate were prepared using the following procedure. To a solution of tert-butyl (4-(4-(((S)-1-(((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-3-((tert-butyldiphenylsilyl)oxy)-1-oxopropan-2-yl)carbamoyl)thiazol-2-yl)phenyl)carbamate (0.13 g, 0.18 mmol) in CHCl (2 mL) was added triethylamine (0.12 mL, 5 eq, 0.89 mmol) and methanesulfonyl chloride (21 μL, 1.5 eq, 0.27 mmol), and the resulting mixture was stirred at room temperature for 3 h. The mixture was diluted with CHCl (25 mL) and washed with 1 M HCl and brine, dried over NaSO, filtered and partially concentrated to a volume of about 10 mL. Then, DBU (54 μL, 2 eq, 0.36 mmol) was added, and the mixture was stirred for 1 h. Add the solution (0.36mL, 1.0 molar concentration, 2 equivalents, 0.36mmol) of TBAF in THF, and stir the mixture for 30 minutes.Use CH2Cl2 (25mL) dilute mixture and use 1MHCl and salt water washing, through Na2SO4 dry, filter and partially concentrate to reach the volume of about 10mL.In gained mixture, add triethylamine (0.12mL, 5 equivalents, 0.89mmol) and mesyl chloride (21 μ L, 1.5 equivalents, 0.27mmol), and at room temperature stir gained mixture 1 hour.Use CH2Cl2 (25mL) dilute mixture and use 1M HCl and salt water washing, through Na2SO4 dry, filter and partially concentrate to reach the volume of about 10mL.Add DBU (54 μ L, 2 equivalents, 0.36mmol), and stir gained mixture 1 hour.Use CH2Cl2 (25mL) dilute mixture and use 1M HCl and salt water washing, through Na2SO4 dry, filter and concentrate. The crude material was purified by automated reverse phase FCC to afford 1 (2.0 mg, 4.4 μmol, 2.4%) and 2 (8.0 mg, 17 μmol, 9.7%).

[0573] 1: LCMS (22010199A TFA LCMS-5C1) RT: 1.138 min; area: 63.7% (215 nm); m / z 458.2 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.99 (s, 1H), 8.92 (s, 1H), 8.07 (d, J = 5.2Hz, 1H), 7.96-7.86 (m, 2H), 7.45 (m, 2H), 6.76 (d, J = 2 .2Hz,1H),6.65(d,J=2.1Hz,1H),6.62(s,1H),5.53(t,J=1.9Hz,1H),5.43-5.36(m,1H),1.65(s,13H),1.52(s,9H).

[0574] 2: LCMS (22010199LCMS-5C3) RT: 1.138 min; area: 63.7% (215 nm); m / z 458.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ10.84(s,1H),8.22(s,1H),7.93-7.86(m,2H),7.53-7. 47(m,2H),7.36(s,1H),6.66(s,1H),2.33(s,3H),1.78(s,3H),1.53(s,9H).

[0575] Example 5: Compound 4

[0576] Methyl 2-(2-(2-(4-((tert-butyloxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate

[0577]

[0578] Step 1. N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 3. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) (2.0 g, 6.2 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (2.5 g, 1 eq, 6.2 mmol) gave N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (3.5 g, 5.3 mmol, 85%).

[0579] Step 2. N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (Intermediate 3) was prepared following General Experimental Procedure 2. N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (3.5 g, 5.3 mmol) afforded the crude material N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (Intermediate 3) as a clear oil which was used as is in the next step.

[0580] Step 3. N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine methyl ester was prepared following General Experimental Procedure 3. Crude material N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (Intermediate 3) (obtained from Step 2) and L-serine methyl ester hydrochloride (1.2 g, 1.5 equiv., 8.0 mmol) gave N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine methyl ester (1.0 g, 1.3 mmol, 25%) as a yellow oil. LCMS (General 3acidic) RT: 1.74 min; Area % (254 nm): 87%; m / z = 747.2 [M+H] + .

[0581] Step 4. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 6. Methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine ester (0.10 g, 1 eq, 0.13 mmol) afforded methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate (22 mg, 47 μmol, 35%) as a white solid. LCMS (22010199A TFA LCMS-5C1) RT: 1.6884 min; Area % (215 nm): 97.1%; m / z = 473.2 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.98(s,1H),8.52(s,1H),8.07(s,1H),7.96-7.88(m,2H),7.45(d,J=8.7Hz,2H),6.75(d, J=2.2Hz,1H),6.69(s,1H),6.61(s,1H),6.01(d,J=1.3Hz,1H),5.48(t,J=1.9Hz,1H),3.88(s,3H),1.52(s,9H).

[0582] Alternative synthesis:

[0583]

[0584] Step 1. Ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylate was prepared following General Experimental Procedure 1. (4-((tert-butoxycarbonyl)amino)phenyl)boronic acid (2.0 g, 8.4 mmol) and ethyl 2-bromothiazole-5-carboxylate (2.0 g, 8.4 mmol) gave ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylate (1.26 g, 3.62 mmol, 43%) as a yellow solid. 1 H NMR(299MHz, CDCl3)δ8.39(s,1H),7.94(d,J=8.8Hz,2H),7.49(d,J=8.6Hz,2H),6 .66 (s, 1H), 4.40 (q, J = 7.1Hz, 2H), 1.55 (d, J = 1.0Hz, 10H), 1.42 (t, J = 7.1Hz, 3H).

[0585] Step 2. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid was prepared following general experimental procedure 2. Ethyl 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carboxylate (1.2 g, 3.4 mmol) gave 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid (1.0 g, 3.1 mmol, 91%) as a white solid.

[0586] Step 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid (1.6 g, 5.0 mmol) and H-Ser-OMe.HCl (0.93 g, 1.2 eq., 6.0 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine methyl ester (1.4 g, 3.3 mmol, 67%) as an orange oil. LCMS (General 3acidic) RT: 0.99 min; Area % (254 nm): 88%; m / z = 422.3 [M+H] + .

[0587] Step 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine methyl ester (1.4 g, 3.3 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine (0.98 g, 2.4 mmol, 72%).

[0588] Step 5. N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following general experimental procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine (0.98 g, 2.4 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (1.1 g, 1.2 equiv, 2.9 mmol) gave crude material N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (2.25 g) which was used as is in the next step.

[0589] Step 6. N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 5. The crude material N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (2.25 g) and acetic anhydride (0.34 mL, 3.6 mmol) gave N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.40 g, 0.51 mmol, 21% over 2 steps). LCMS (General 3acidic) RT: 1.78 min; Area % (214 nm): 100%; m / z = 789.6 [M+H] + .

[0590] Step 7. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 7. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine (0.19 g, 0.24 mmol) afforded methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxamido)acrylamido)acrylate (7.8 mg, 16 μmol, 27%) as a white solid. LC-MS (22010199A TFA LCMS-5C1): RT: 1.353 min; area 97.7% (215 nm), 96.7% (304 nm); m / z = 473.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ (ppm) 8.66 (s, 1H), 8.57 (s, 1H), 8.24 (s, 1H), 7.92 (d, J = 8.8Hz, 2H), 7.48 (d, J = 8.8Hz, 2H), 6.72 (d, J =2.5Hz,1H),6.67(s,1H),6.63(s,1H),6.04(d,J=1.0Hz,1H),5.49(dd,J=2.3Hz,J=1.3Hz,1H),3.91(s,1H),1.54(s,9H).

[0591] Example 6: Compound 6

[0592] Methyl 2-(2-(2-(4-acetylaminophenyl)thiazole-4-carboxamido)acrylamido)acrylate

[0593]

[0594] Step 1. use the following procedure to prepare N-(2-(4-aminophenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine methyl ester.N-(2-(4-((tert-butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine methyl ester (prepared as in step 3) (0.80g, 1 equivalent, 1.1mmol) is dissolved in CH2Cl2 (5mL) and TFA (5mL) is added.The resulting mixture is stirred at room temperature for 1 hour.Use CH2Cl2 (50mL) dilute the mixture and pour into 1M NaOH (150mL aqueous solution).Separate each layer, and use CH2Cl2 (50mL) to extract aqueous phase. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated to give N-(2-(4-aminophenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine methyl ester (0.65 g, 1.0 mmol, 94%) as a yellow oil.

[0595] Step 2. (S)-methyl 2-(2-(2-(4-acetamidophenyl)thiazole-4-carboxamido)-3-((methylsulfonyl)oxy)propionamido)acrylate was prepared using the following procedure. Methyl N-(2-(4-aminophenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine (0.22 g, 1 eq, 0.34 mmol) was dissolved in CH2Cl2 (5 mL), and DIPEA (0.18 mL, 3 eq, 1.0 mmol) and acetic acid (41 mg, 39 μL, 2 eq, 0.68 mmol) were added. Then, COMU (0.17 g, 1.2 eq, 0.41 mmol) was added, and the resulting mixture was stirred at room temperature for 16 hours. LCMS monitoring showed partial conversion to diacetylated product. DiPEA (0.18 mL, 3 eq., 1.0 mmol), acetic acid (39 μL, 2 eq., 0.68 mmol) and COMU (0.17 g, 1.2 eq., 0.41 mmol) were added, and the mixture was stirred for 24 hours. The mixture was diluted with CH2Cl2 (20 mL) and washed with HCl (1 M, 20 mL), NaHCO3 (20 mL saturated aqueous solution) and brine, dried over Na2SO4, filtered and concentrated. The residue was dissolved in CH2Cl2 (5 mL) and DBU (0.15 mL, 3 eq., 1.0 mmol) was added. The resulting mixture was stirred for 1 hour. A solution of tetrabutylammonium fluoride in THF (0.51 mL, 1.0 M, 1.5 eq., 0.51 mmol) was added, and the resulting mixture was stirred for 1 hour. The mixture was diluted with CH2Cl2 (20 mL) and washed with HCl (1 M, 20 mL) and brine, dried over Na2SO4, filtered and concentrated. Residue is dissolved in CH2Cl2 (5mL) in, and add mesyl chloride (40 μ L, 1.5 equivalents, 0.51mmol) and triethylamine (0.24mL, 5 equivalents, 1.7mmol).At room temperature stir gained mixture 90 minutes, after this, observe complete mesylation and partial elimination by LCMS monitoring.Use CH2Cl2 (20mL) dilute mixture and use HCl (1M, 20mL) and salt water washing, through Na2SO4 dry, filter and concentrate.Residue is dissolved in CH2Cl2 (5mL) in and add DBU (0.16g, 0.15mL, 3 equivalents, 1.0mmol).At room temperature stir gained mixture 90 minutes.Use CH2Cl2 (20mL) dilute mixture and use HCl (1M, 20mL) and salt water washing, through Na2SO4 dry, filter and concentrate. The crude material was purified by automated reverse phase FCC to afford methyl 2-(2-(2-(4-acetamidophenyl)thiazole-4-carboxamido)acrylamido)acrylate (5.2 mg, 0.34 mmol, 3.7%) as a white solid.LCMS (22010199C TFA LCMS-5C3) RT: 1.062 min; Area % (215 nm): 91.0%; m / z = 415.0 [M+H]. + . 1 H NMR (400MHz, CDCl3) δ9.99(s,1H),8.53(s,1H),8.10(s,1H),8.00-7.92(m,2H),7.61(d,J=8.3Hz,2H),7.29(s, 1H), 6.76 (d, J = 2.2Hz, 1H), 6.69 (s, 1H), 6.01 (d, J = 1.4Hz, 1H), 5.48 (t, J = 1.9Hz, 1H), 3.88 (s, 3H), 2.21 (s, 3H).

[0596] Example 7: Compound 16

[0597] Methyl 2-(2-(2-phenylthiazole-4-carboxamido)acrylamido)acrylate

[0598]

[0599] Step 1. 2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carboxylic acid was prepared using the following procedure. To a solution of (4-((tert-butoxycarbonyl)amino)phenyl)boronic acid (1.0 g, 4.2 mmol) and ethyl 2-bromooxazole-4-carboxylate (0.93 g, 4.2 mmol) in 1,2-dimethoxyethane (25 mL) was added sodium carbonate (aqueous solution, 2M, 21 mL, 10 eq., 42 mmol) and tetrapalladium (0.24 g, 0.05 eq., 0.21 mmol). The resulting mixture was stirred at 80 °C for 16 hours. The mixture was cooled to room temperature and diluted with EtOAc. The mixture was washed with water. The aqueous layer was acidified by adding 1M HCl. The precipitate formed was isolated by filtration and dried under reduced pressure to give 2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carboxylic acid (0.66 g, 2.2 mmol, 51%) as a white solid. 1 H NMR (299MHz, CD3OD) δ8.52 (s, 1H), 8.00 (m, 2H), 7.60 (m, 2H), 1.55 (s, 9H).

[0600] Step 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)oxazole-4-carboxylic acid (0.66 g, 2.2 mmol) and L-serine methyl ester hydrochloride (0.41 g, 1.2 equiv, 2.6 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serine methyl ester (0.98 g, 2.4 mmol, quantitative) as an orange oil.

[0601] Step 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serine methyl ester (0.98 g, 2.4 mmol) afforded (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serine (0.64 g, 1.6 mmol, 68%) as an off-white solid. 1 H NMR (299MHz, cd3od) δ8.43(s,1H),8.05-7.96(m,2H),7.60(m,2H),4.71m,1H),4.14-3.94(m,2H),3.79-3.69(m,1H),1.55(s,9H).

[0602] Step 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl-L-serine methyl ester was prepared following General Experimental Procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serine (0.20 g, 0.51 mmol) and L-serine methyl ester hydrochloride (95 mg, 1.2 equiv, 0.61 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl-L-serine methyl ester (0.18 g, 0.37 mmol, 72%) as a colorless oil.

[0603] Step 5. O-Acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl)-L-serine methyl ester was prepared following general experimental procedure 5. (2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl-L-serine methyl ester (0.18 g, 0.37 mmol) and acetic anhydride (90 μL, 2.6 equiv, 0.95 mmol) gave O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl)-L-serine methyl ester (0.12 g, 0.20 mmol, 56%). 1 H NMR (299MHz, CDCl3) δ8.26(s,1H),7.98(d,J=8.6Hz,2H),7.79(d,J=8.0Hz,1H),7.51(d,J=8.6Hz,2H),7.32(d,J=7 .8Hz,1H),6.88(s,1H),5.04(m,1H),4.87(m,1H),4.58-4.36(m,4H),3.80(m,3H),2.15-1.98(m,6H),1.54(s,9H).

[0604] Step 6. (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carboxamido)acryloyl)-L-serine methyl ester was prepared following General Experimental Procedure 8. O-Acetyl-N-(O-Acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl)-L-serine methyl ester (0.12 g, 0.20 mmol) afforded (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carboxamido)acryloyl)-L-serine methyl ester (29 mg, 64 μmol, 32%) as a white solid. LC-MS (22010199ATFA LCMS-5C1): rt 1.560 min; area 99.7% (215 nm), 99.2% (304 nm); m / z = [M+H] + =457.2 1 H NMR (400MHz, CDCl3) δ (ppm) 9.57 (s, 1H), 8.54 (s, 1H), 8.25 (s, 1H), 8.02 (d, J = 8.7Hz, 2H), 7.49 (d, J = 8.7Hz, 2H), 6.75(d,J=2.2Hz),6.71(s,1H),6.64(s,1H),6.03(d,J=1.0Hz,1H),5.49(t,J=1.9Hz,1H),3.90(s,3H),(s,9H).

[0605] Example 8: Compound 49

[0606]

[0607] Step 1. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) (0.68 g, 2.1 mmol) and L-serine methyl ester hydrochloride (0.40 g, 1.2 equiv, 2.6 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine methyl ester (0.87 g, 2.1 mmol, 97%) as an off-white solid. LCMS (General 3 basic) RT: 1.44 min; Area % (254 nm): 98%; m / z = 420.3 [MH] - .

[0608] Step 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-serine (Intermediate 4) was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-serine methyl ester (0.88 g, 2.1 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-serine (Intermediate 4) (0.84 g, 2.1 mmol, quantitative) as a white solid.

[0609] Step 3. General experimental procedure 4 was followed to prepare N-((2-(4-((tert-butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-serine (Intermediate 4) (0.87 g, 2.1 mmol) and L-Ser(OTBDPS)-OMe.HCl (1.0 g, 1.2 eq., 2.6 mmol) gave N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.5 g, 1.9 mmol, 91%) LCMS (General 3acidic) RT: 1.75 min; Area % (254 nm): 98%; m / z = 747.6 [M+H] + .

[0610] Step 4. N-(O-Acetyl-N-(2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 5. N-((2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.5 g, 1.9 mmol) gave N-(O-acetyl-N-(2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.6 g, 2.0 mmol, quantitative) as a yellow foam. LCMS (General 3acidic) RT: 1.84 min; Area % (254 nm): 98%; m / z = 789.6 [M+H] + .

[0611] Step 5. prepare N-(O-acetyl-N-(2-(4-aminophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester using the following procedure. To N-(O-acetyl-N-(2-(4-((tert-butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.6g, 2.0mmol) in CH2Cl2(30mL) in a solution of TFA (4.6mL, 30 equivalents, 60mmol), and stir the resulting mixture for 1 hour. Use saturated NaHCO3aqueous solution to quench the reaction mixture until gas formation stops and pH test paper shows that the mixture pH is about 7-8. The mixture was extracted with CH2Cl2 (3×100 ml) and the combined organics were dried over Na2SO4, filtered and concentrated to give N-(O-acetyl-N-(2-(4-aminophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.3 g, 1.8 mmol, 91%). LCMS (General 3basic) RT: 2.01 min; Area % (254 nm): 72%; m / z=687.4 [MH] - .

[0612] Step 6. The DBCO coupled intermediate was prepared using the following procedure. Propyl chloroformate (0.15 mL, 1.1 eq., 1.3 mmol) was added to a solution of DBCO acid (0.37 g, 1.2 mmol) and triethylamine (0.19 mL, 1.1 eq., 1.3 mmol) in CH2Cl2 (6 mL) at 0°C, and the resulting mixture was stirred at 0°C for 1 hour. A solution of N-(O-acetyl-N-(2-(4-aminophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.61 g, 0.73 eq., 0.88 mmol) in CH2Cl2 (3 mL) was added, and the resulting mixture was stirred at room temperature for 2 hours. The mixture was washed with NaHCO3 (saturated, 3×10 mL) and HCl (1 M, 3×10 mL), dried over Na2SO4, filtered and concentrated. The crude material was purified by automated FCC to give the desired product (0.52 g, 0.54 mmol, 45%) as a slightly yellow foam. LCMS (General 3 acidic) RT: 1.86 min; Area % (254 nm): 86%; m / z = 976.5 [M+H] + .

[0613] Step 7. The DBCO-coupled bis-dehydroalanine compound was prepared following General Experimental Procedure 5. The DBCO-coupled intermediate (0.52 g, 0.54 mmol) gave the DBCO-coupled bis-dehydroalanine compound (24 mg, 37 μmol, 6.9%) as a white solid. LCMS (22010199D TFA LCMS-5C8) RT: 1.642 min; Area % (215 nm): 79.2%; m / z = 660.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ10.05(s,1H),8.56(s,1H),8.42(s,1H),8.09(s,1H),7.84(d,J=8 .6Hz,2H),7.71(d,J=7.4Hz,1H),7.50-7.28(m,8H),7.19(dd,J=7.4,1.5Hz,1H),6.78( d,J=2.2Hz,1H),6.73(s,1H),6.03(d,J=1.3Hz,1H),5.50(t,J=1.9Hz,1H),5.19(d,J=1 3.9Hz, 1H), 3.91 (s, 3H), 3.73 (d, J = 13.8Hz, 1H), 2.86 (m, 1H), 2.45 (m, 2H), 2.07 (m, 1H).

[0614] Example 9: Compound 50

[0615] Methyl 2-(2-(2-(4-(hex-5-ynylamino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate

[0616]

[0617] Step 1. Prepare methyl 2-(2-(2-(4-(hex-5-ynylamino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate using the following procedure. Methyl N-(2-(4-aminophenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serine (prepared as in step 1) (0.22 g, 0.334 mmol) was dissolved in CH2Cl2 (5 mL), and DiPEA (0.18 mL, 3 eq., 1.0 mmol) and hex-5-ynyl acid (56 μL, 1.5 eq., 0.51 mmol) were added. COMU (0.18 g, 1.2 eq., 0.41 mmol) was then added, and the resulting mixture was stirred at room temperature for 16 hours. LCMS monitoring showed partial conversion to the bis-acylated product. DiPEA (0.18 mL, 3 eq., 1.0 mmol), hex-5-ynoic acid (57 mg, 56 μL, 1.5 eq., 0.510 mmol) and COMU (0.18 mg, 1.2 eq., 0.41 mmol) were added, and the resulting mixture was stirred for 24 hours. The mixture was diluted with CH2Cl2 (20 mL) and washed with HCl (1 M, 20 mL), NaHCO3 (20 mL saturated aqueous solution) and brine, dried over Na2SO4, filtered and concentrated. The residue was dissolved in CH2Cl2 (5 mL) and DBU (0.15 mL, 3 eq., 1.0 mmol) was added. The resulting mixture was stirred for 1 hour. A solution of tetrabutylammonium fluoride in THF (0.51 mL, 1.0 M, 1.5 eq., 0.51 mmol) was added, and the resulting mixture was stirred for 1 hour. With CH2Cl2 (20mL) dilute mixture and with HCl (1M, 20mL) and salt water washing, through Na2SO4 dry, filter and concentrate.Residue is dissolved in CH2Cl2 (5mL) in, and add methylsulfonyl chloride (40 μ L, 1.5 equivalents, 0.51mmol) and triethylamine (0.17g, 0.24mL, 5 equivalents, 1.7mmol).At room temperature stir gained mixture 90 minutes, after this, observe complete mesylation and partial elimination by LCMS monitoring.With CH2Cl2 (20mL) dilute mixture and with HCl (1M, 20mL) and salt water washing, through Na2SO4 dry, filter and concentrate.Residue is dissolved in CH2Cl2 (5mL) in and add DBU (0.16g, 0.15mL, 3 equivalents, 1.0mmol).At room temperature stir gained mixture 90 minutes. The mixture was diluted with CH2Cl2 (20 mL) and washed with HCl (1 M, 20 mL) and brine, dried over Na2SO4, filtered and concentrated.The crude material was purified by automated reverse phase FCC to give methyl 2-(2-(2-(4-(hex-5-ynylamino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate (4.5 mg, 9.6 μmol, 2.8%) as a white solid. LCMS (22010199C TFA LCMS-5C3) RT: 1.287 min; Area % (215 nm): 93.0%; m / z = 467.2 [M+H]. + . 1 HNMR (400MHz, CDCl3) δ9.98 (s, 1H), 8.53 (s, 1H), 8.00-7.92 (m, 2H), 7.62 (d, J = 8.3Hz, 2H), 7.31 (s, 1H), 6.76 (d, J = 2.2Hz, 1H), 6.69 (s ,1H),6.01(d,J=1.2Hz,1H),5.48(t,J=1.9Hz,1H),3.88(s,3H),2.55(t,J=7.3Hz,2H),2.34(td,J=6.7,2.6Hz,2H),2.04-1.92(m,3H).

[0618] Example 10: Compound 51

[0619]

[0620] Compound 51 was prepared following the general procedure of the conjugation chemistry between azide and peptides. 49 (3.0 mg, 4.5 μmol) and 81 (5.4 mg, 4.5 μmol) gave 51 (3.5 mg, 1.9 μmol, 41%) as a white solid. LCMS (30833 LCMS-6): RT: 2.788 min and 2.822 min (regional isomers); Area: 95.3% (215 nm), 99.6% (ELSD); m / z = 1744.1 [M+H] +

[0621] Example 11: Compound 52

[0622] Methyl 2-(2-(2-(4-((tert-butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxamido)acrylamido)acrylate

[0623]

[0624] Step 1. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylic acid methyl ester was prepared following General Experimental Procedure 1. 2-Bromo-5-methyl-thiazole-4-carboxylic acid methyl ester (0.91 g, 3.8 mmol) and (4-((tert-Butyloxycarbonyl)amino)phenyl)boronic acid (0.86 g, 3.6 mmol) gave 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylic acid methyl ester (1.1 g, 3.1 mmol, 88%). LCMS (General 3 basic) RT: 1.80 min; Area % (214 nm): 87%; m / z = 347.2 [MH] - .

[0625] Step 2. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylic acid was prepared following General Experimental Procedure 2. Methyl 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylate (1.1 g, 3.1 mmol) gave methyl 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylate (1.0 g, 3.1 mmol, quantitative). LCMS (General 3 basic) RT: 1.17 min; Area % (214 nm): 81%; m / z = 335.4 [M+H] + .

[0626] Step 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serine methyl ester was prepared following General Experimental Procedure 3. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylic acid (1.1 g, 3.2 mmol) and L-Ser-OMe.HCl (0.60 g, 1.2 eq, 3.9 mmol) gave crude (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serine methyl ester (2.6 g) which was used in the next step without purification. LCMS (General 3 basic) RT: 1.61 min; Area % (214 nm): 69%; m / z = 436.2 [M+H] + .

[0627] Step 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serine was prepared following General Experimental Procedure 2. Crude (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serine methyl ester (2.6 g crude) gave crude (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serine (1.8 g) which was used in the next step without purification. LCMS (General 3 basic) RT: 1.06 min; Area % (214 nm): 69%; m / z = 422.4 [M+H] + .

[0628] Step 5. N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following general experimental procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serine (1.4 g, crude) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (1.6 g, 4.0 mmol) gave N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.2 g, 1.5 mmol) as a clear yellow oil. LCMS (General 3acidic) RT: 1.85 min; Area % (214 nm): 73%; m / z = 761.6 [M+H] + .

[0629] Step 6. General Experimental Procedure 5 was followed to prepare N-(O-acetyl-N-(2-(4-((tert-butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester. N-((2-(4-((tert-butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.2 g, 1.5 mmol) and acetic anhydride (0.16 mL, 1.1 eq., 1.7 mmol) gave N-(O-acetyl-N-(2-(4-((tert-butyloxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.1 g, 1.3 mmol, 87%) as a yellow viscous oil. LCMS (General 3 basic) RT: 2.31 min; Area % (214 nm): 86%; m / z = 801.5 [M+H] + .

[0630] Step 7. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 7. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serine ester (0.21 g, 0.26 mmol) afforded methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxamido)acrylamido)acrylate (11 mg, 23 μmol, 13%) as a white solid. LCMS (22010199C TFA LCMS-5C3) RT: 1.702 min; Area: 91.1% (215 nm); m / z 487.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ10.08(s,1H),8.51(s,1H),7.88-7.80(m,2H),7.42(d,J=8.6Hz,2H),6.72-6.66(m ,2H),6.57(s,1H),6.00(d,J=1.2Hz,1H),5.42(t,J=1.8Hz,1H),3.88(s,3H),2.85(s,3H),1.52(s,9H).

[0631] Example 12: Compound 53

[0632] (Z)-2-(2-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoic acid methyl ester

[0633]

[0634] Step 1. N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl-L-threonine methyl ester was prepared following General Experimental Procedure 3. N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)serine (Intermediate 3) (0.90 g, 1.4 mmol) and H-Thr-OMe.HCl (0.29 g, 1.2 equiv, 1.7 mmol) gave N-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl-L-threonine methyl ester (0.29 g, 0.38 mmol, 27%) as a light yellow foam. LCMS (General 3 basic) RT: 2.20 min; Area % (254 nm): 81%; m / z = 761.3 [M+H] + .

[0635] Step 2. O-Acetyl-N-(N-(2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl)-L-threonine methyl ester was prepared following General Experimental Procedure 5. N-(2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl-L-threonine methyl ester (0.29 g, 0.38 mmol) and acetic anhydride (37 μL, 1.05 equiv, 0.39 mmol) gave O-acetyl-N-(N-(2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl)-L-threonine methyl ester (0.30 g, 0.37 mmol, 98%) as a light yellow foam.

[0636] Step 3. (Z)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoic acid methyl ester was prepared following General Experimental Procedure 7. O-Acetyl-N-(N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl)-L-threonine methyl ester (0.10 g, 0.13 mmol) gave (Z)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoic acid methyl ester (22 mg, 45 μmol, 36%). LCMS (22010199C TFA LCMS-5C3) RT: 1.424 min; Area % (215 nm): 92.9%; m / z = 487.2 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.97(s,1H),8.07(s,1H),7.94-7.86(m,2H),7.48(s,1H),7.42(d,J=8.6Hz,2H),6.93(q,J=7.2Hz ,1H),6.76(d,J=1.8Hz,1H),6.59(s,1H),5.53(t,J=1.7Hz,1H),3.79(s,3H),1.82(dd,J=7.2,0.7Hz,3H),1.51(s,9H).

[0637] Example 13: Compound 55

[0638] (E)-2-(2-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoic acid methyl ester

[0639]

[0640] Step 1. N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester was prepared following General Experimental Procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) (785 mg, 1.93 mmol) and O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester (859 mg, 1.2 equiv, 2.31 mmol) gave N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester (1.54 g, 2.02 mmol, quantitative) as a viscous oil. LCMS (General 3acidic) RT: 1.81 min; Area % (254 nm): 96%; m / z = 761.3 [M+H] + .

[0641] Step 2. N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester was prepared following General Experimental Procedure 5. N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester (1.6 g, 2.1 mmol) and acetic anhydride (0.22 mL, 1.1 equiv, 2.3 mmol) afforded N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester (1.37 g, 1.71 mmol, 81%) as a yellow solid. LCMS (General 3acidic) RT: 1.88 min; Area % (254 nm): 75%; m / z = 803.5 [M+H] + .

[0642] Step 3. (E)-methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoate was prepared following General Experimental Procedure 7. N-(O-Acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester (306 mg, 0.38 mmol) gave (E)-methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoate (4.6 mg, 9.5 μmol, 2.5%). LCMS (22010199C TFA LCMS-5C3) RT: 1.470 min; Area: 93.8% (215 nm); m / z 487.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.98(s,1H),8.17(s,1H),8.06(s,1H),7.95-7.88(m,2H),7.44(d,J=8.6Hz,2H),7.33(q, J=7.7Hz,2H),6.70(d,J=2.0Hz,1H),6.59(s,1H),5.42(s,1H),3.88(s,3H),2.15(d,J=7.7Hz,3H),1.52(s,9H).

[0643] Example 14: Compound 56

[0644] Methyl 2-(2-(2-(4-((tert-butyloxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acetamido)acrylate

[0645]

[0646] Step 1. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycine methyl ester was prepared following General Experimental Procedure 3. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) (0.41 g, 1 eq, 1.3 mmol) and glycine methyl ester hydrochloride (0.17 g, 1 eq, 1.3 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycine methyl ester (0.37 g, 0.94 mmol, 74%) as a white foam. LCMS (General 3 basic) RT: 1.63 min; Area % (254 nm): 92%; m / z = 392.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.03 (s, 1H), 7.91-7.82 (m, 3H), 7.49-7.41 (m, 2H), 6.71 (s, 1H), 4.26 (d, J = 5.7Hz, 2H), 3.78 (s, 3H), 1.51 (s, 9H).

[0647] Step 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycine was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycine methyl ester (0.37 mg, 1 eq, 0.94 mmol) afforded (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycine (0.32 g, 0.85 mmol, 90%) as a white solid. LCMS (General 3 basic) RT: 1.12 min; Area % (254 nm): 96%; m / z = 378.2 [M+H] + . 1 H NMR (400MHz, DMSO) δ9.69 (s, 1H), 8.71 (t, J = 6.1 Hz, 1H), 8.24 (s, 1H), 7.98-7.90 (m, 2H), 7.65-7.57 (m, 2H), 3.96 (d, J = 6.1Hz, 2H), 1.49 (s, 9H).

[0648] Step 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycyl-L-serine methyl ester was prepared following General Experimental Procedure 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycine (0.31 g, 1 eq, 0.83 mol) and L-serine methyl ester hydrochloride (0.16 g, 1.2 eq, 1.0 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycylserine methyl ester (0.29 mg, 0.60 mmol, 72%). LCMS (General 3 basic) RT: 1.47 min; Area % (254 nm): 93%; m / z = 479.3 [M+H] + . 1 H NMR(400MHz,MeOD)δ8.07(s,1H),7.90-7.82(m,2H),7.52-7.44(m,2H),4.56(t,J =4.3Hz,1H),4.15(d,J=2.1Hz,2H),3.92-3.76(m,2H),3.71(s,3H),1.49(s,9H).

[0649] Step 4. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acetamido)acrylate was prepared using the following procedure. To a solution of methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycylserine (0.25 g, 1 eq, 0.51 mmol) in MeCN (30 mL) was added triethylamine (0.14 mL, 2 eq, 1.0 mmol) and the resulting mixture was stirred at room temperature for 6 days. The mixture was concentrated and purified by automated FCC and automated reverse phase FCC to give methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acetamido)acrylate (28 mg, 61 μmol, 12%) as a white solid. LCMS (22010199D TFA LCMS-5C8) RT: 1.382 min; Area % (215 nm): 98.0%; m / z = 461.2 [M+H] + . 1H NMR (400MHz, CDCl3) δ8.36 (s, 1H), 8.09 (d, J = 2.4Hz, 1H), 8.00 (d, J = 6.5Hz, 1H), 7.91-7.80 (m, 2H), 7.45 (d, J=8.3Hz,2H),6.61(d,J=2.5Hz,2H),5.92(d,J=1.5Hz,1H),4.24(d,J=5.9Hz,2H),3.82(s,3H),1.52(s,9H).

[0650] Example 15: Compound 57

[0651] (S)-methyl 2-(2-(2-(4-((tert-butyloxycarbonyl)amino)phenyl)thiazole-4-carboxamido)propionamido)acrylate

[0652]

[0653] Step 1. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-alanine methyl ester was prepared following General Experimental Procedure 3. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) (0.53 g, 1.7 mmol) and H-Ala-OMe.HCl (0.27 g, 1.2 eq, 1.9 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-alanine methyl ester (0.43 mg, 1.1 mmol, 63%) LCMS (General 3 basic) RT: 1.77 min; Area % (254 nm): 86%; m / z = 404.2 [M+H] + .

[0654] Step 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-D-alanine was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-D-alanine methyl ester (0.43 g, 1.1 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-D-alanine (0.41 g, 1.1 mmol, quantitative) as a yellow foam. LCMS (General 3 basic) RT: 1.16 min; Area % (254 nm): 95%; m / z = 392.2 [M+H] + .

[0655] Step 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-alanyl-L-serine methyl ester was prepared following General Experimental Procedure 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-D-alanine (0.41 g, 1.1 mmol) and H-Ser-OMe.HCl (0.25 mg, 1.2 equiv, 1.38 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-alanyl-L-serine methyl ester (0.24 mg, 0.50 mmol, 47%) as a yellow foam. LCMS (General 3 basic) RT: 1.54 min; Area % (254 nm): 85%; m / z = 493.2 [M+H] + .

[0656] Step 4. (S)-methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)propionamido)acrylate was prepared using the following procedure. To a solution of (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-alanylserine methyl ester (0.24 mg, 1 eq, 0.50 mmol) in MeCN (30 mL) was added triethylamine (0.14 mL, 2 eq, 1.0 mmol) and the resulting mixture was stirred at room temperature for 6 days. The mixture was concentrated and purified by automated FCC and automated reverse phase FCC to give (S)-methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)propionamido)acrylate as a white solid. LCMS (22010199D TFA LCMS-5C8) RT: 1.453 min; Area % (215 nm): 94.9%; m / z = 475.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.54(s,1H),8.08(s,1H),7.90-7.80(m,3H),7.49-7.42(m,2H),6.70(s,1H),6.5 9(s,1H),5.91(d,J=1.4Hz,1H),4.77(p,J=7.1Hz,1H),3.81(s,3H),1.56(d,J=7.0Hz,3H),1.52(s,9H).

[0657] Example 16: Compound 58

[0658] (2-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)glycine methyl ester

[0659]

[0660] Step 1. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine methyl ester was prepared following General Experimental Procedure 3. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) (1.0 g, 3.2 mmol) and L-Ser-OMe.HCl (0.59 g, 1.2 eq, 3.8 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine methyl ester (0.88 g, 2.1 mmol, 66%). LCMS (General 3 basic) RT: 1.54 min; Area % (254 nm): 64%; m / z = 420.2 [MH] - .

[0661] Step 2. (2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) was prepared following General Experimental Procedure 2. (2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine methyl ester (0.88 g, 2.1 mmol) afforded (2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) (0.84 mg, 2.1 mmol, 99%) as a white solid.

[0662] Step 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serylglycine methyl ester was prepared following General Experimental Procedure 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) (0.42 g, 1.0 mmol) and H-Gly-OMe.HCl (0.18 g, 1.4 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serylglycine methyl ester (0.35 g, 0.72 mmol, 70%) as a yellow solid. 1 H NMR (400MHz, MeOD) δ8.16 (s, 1H), 7.98-7.88 (m, 2H), 7.59-7.51 (m, 2H), 4.71 (t, J = 4. 9Hz, 1H), 4.04-3.96 (m, 3H), 3.92 (dd, J = 11.3, 4.8Hz, 1H), 3.73 (s, 3H), 1.54 (s, 9H).

[0663] Step 4. O-Acetyl-N-(2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serylglycine methyl ester was prepared following General Experimental Procedure 5. (2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serylglycine methyl ester (0.13 mg, 0.27 mmol) and acetic anhydride (26 μL, 1 eq, 0.27 mmol) gave O-Acetyl-N-(2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serylglycine methyl ester (153 mg, quantitative). LCMS (General 3acidic) RT: 1.14 min; Area % (254 nm): 92%; m / z = 521.5 [M+H] + .

[0664] Step 5. (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)glycine methyl ester was prepared using the following procedure. To a solution of (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)glycine methyl ester (51 mg, 99 μmol) in CHCl (1 mL) was added DBU (30 μL, 2 eq, 197 μmol) and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated in vacuo and the crude material was purified by automated reverse phase FCC to give (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)glycine methyl ester (12.5 mg, 27.1 μmol, 28%) as a white solid. LCMS (22010199D TFA LCMS-5C8) RT: 1.403 min; Area % (215 nm): 93.2%; m / z = 461.2 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.98(s,1H),8.06(s,1H),7.94-7.82(m,2H),7.48-7.39(m,2H),6.73(s, 1H), 6.71-6.66 (m, 2H), 5.43 (t, J = 1.7Hz, 1H), 4.18 (d, J = 5.0Hz, 2H), 3.79 (s, 3H), 1.52 (s, 9H).

[0665] Example 17: Compound 59

[0666] (2-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)-L-alanine methyl ester

[0667]

[0668] Step 1. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl-L-alanine methyl ester was prepared following General Experimental Procedure 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) (0.42 g, 1.0 mmol) and H-Ala-OMe.HCl (0.19 g, 1.3 eq, 1.3 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl-L-alanine methyl ester (0.24 g, 0.49 mmol, 48%) as a yellow foam. LCMS (General 3 basic) RT: 1.60 min; Area % (254 nm): 81%; m / z = 493.2 [M+H] + .

[0669] Step 2. O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl-L-alanine methyl ester was prepared following General Experimental Procedure 5. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl-L-alanine methyl ester (0.20 g, 0.41 mmol) and acetic anhydride (42 μL, 0.44 mmol) gave O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl-L-alanine methyl ester (0.23 g, 0.44 mmol, quantitative) as a light yellow foam. LCMS (General 3 basic) RT: 1.59 min; Area % (254 nm): 86%; m / z = 535.2 [M+H] + .

[0670] Step 3. (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)-L-alanine methyl ester was prepared using the following procedure. To a solution of O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl-L-alanine methyl ester (52 mg, 98 μmol) in CH2Cl2 (1 mL) was added DBU (30 μL, 0.20 mmol) and the resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated in vacuo and the crude material was purified by automated reverse phase FCC to give (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)-L-alanine methyl ester (10.8 mg, 22.8 μmol, 23%). LCMS (22010199D TFA LCMS-5C8) RT: 1.472 min; Area % (215 nm): 97.4%; m / z = 475.2 [M+H]+ . 1 H NMR (400MHz, CDCl3) δ9.99 (s, 1H), 8.05 (s, 1H), 7.90 (d, J = 8.4Hz, 2H), 7.43 (d, J = 8.3Hz, 2H), 6.80 (d, J = 7.2Hz ,1H),6.72(s,1H),6.67(d,J=1.8Hz,1H),5.44-5.39(m,1H),4.69(p,J=7.2Hz,1H),3.79(s,3H),1.50(m,12H).

[0671] Example 18: Compound 60

[0672] N3(CH2CH2O)3CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2 HCl salt

[0673]

[0674] N3(CH2CH2O)3CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2 HCl salt was prepared on a 0.25 mmol scale following the general procedure for solid phase peptide synthesis. Purification using HCl buffer afforded the desired HCl salt form. N3(CH2CH2O)3CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2 HCl salt (53 mg, 42 μmol, 17%) was isolated as a white solid. LCMS (30833LCMS-6): RT: 2.365 min; Area: 83.7% (215 nm), 99.8% (ELSD); m / z = 1160.8 [M+H] +

[0675] Example 19: Compound 61

[0676]

[0677] Compound 61 was prepared following the general procedure of the conjugation chemistry between azide and peptides. Compound 49 (3.0 mg, 4.5 μmol) and compound 60 (5.8 mg, 4.5 μmol) gave compound 61 (4.5 mg, 2.3 μmol, 51%) as a white solid. LCMS (30833 LCMS-6): RT: 2.781 min; Area: 91.1% (215 nm), 99.4% (ELSD); m / z = 1820.1 [M+H] +

[0678] Example 20: Compound 62

[0679] (E)-2-(2-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoic acid methyl ester

[0680]

[0681] Step 1. N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester was prepared following General Experimental Procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) (785 mg, 1.93 mmol) and O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester (859 mg, 1.2 equiv, 2.31 mmol) gave N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester (1.54 g, 2.02 mmol, quantitative) as a viscous oil. LCMS (General 3acidic) RT: 1.81 min; Area % (254 nm): 96%; m / z = 761.3 [M+H] + .

[0682] Step 2. N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester was prepared following General Experimental Procedure 5. N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester (1.6 g, 2.1 mmol) and acetic anhydride (0.22 mL, 1.1 equiv, 2.3 mmol) afforded N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester (1.37 g, 1.71 mmol, 81%) as a yellow solid. LCMS (General 3acidic) RT: 1.88 min; Area % (254 nm): 75%; m / z = 803.5 [M+H] + .

[0683] Step 3. (E)-methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoate was prepared following General Experimental Procedure 7. N-(O-Acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreonine methyl ester (306 mg, 0.38 mmol) gave (E)-methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoate (4.6 mg, 9.5 μmol, 2.5%). LCMS (22010199C TFA LCMS-5C3) RT: 1.470 min; Area: 93.8% (215 nm); m / z 487.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.98(s,1H),8.17(s,1H),8.06(s,1H),7.95-7.88(m,2H),7.44(d,J=8.6Hz,2H),7.33(q, J=7.7Hz,2H),6.70(d,J=2.0Hz,1H),6.59(s,1H),5.42(s,1H),3.88(s,3H),2.15(d,J=7.7Hz,3H),1.52(s,9H).

[0684] Example 21: Compound 63

[0685] (E)-2-(2-(2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoic acid methyl ester

[0686]

[0687] Step 1. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonine methyl ester was prepared following General Experimental Procedure 4. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) (0.98 g, 3.1 mmol) and L-allothreonine methyl ester hydrochloride (0.57 g, 1.1 equiv, 3.4 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonine methyl ester (1.6 g, 3.2 mmol, quantitative) as a white solid.

[0688] Step 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonine was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonine methyl ester (1.6 g, 3.2 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonine (1.4 g, 3.2 mmol, quantitative) as a white solid.

[0689] Step 3. N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonine (1.4 g, 3.2 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (1.4 g, 1.1 equiv., 3.5 mmol) gave N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (2.3 g, 3.0 mmol, 93%).

[0690] Step 4. N-(O-Acetyl-N-(2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following general experimental procedure 5. N-((2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (2.3 g, 3.0 mmol) gave N-(O-acetyl-N-(2-(4-((tert-Butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (2.2 g, 2.7 mmol, 90%).

[0691] Step 5. O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-L-serine methyl ester was prepared using the following procedure. To a solution of N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.30 g, 0.37 mmol) in THF (6 mL) was added a solution of TBAF in THF (0.56 mL, 1 M, 1.5 eq, 0.56 mmol) and the resulting mixture was stirred for 1 h. The reaction mixture was concentrated in vacuo and the residue was dissolved in MeCN (6 mL). Triethylamine (0.13mL, 2.5 equivalents, 0.93mmol) and acetic anhydride (48 μL, 1.35 equivalents, 0.50mmol) were added, and the resulting mixture was stirred at room temperature for 45 minutes. Water was added and the mixture was concentrated in vacuo to remove MeCN. The mixture was then extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. The crude material was purified by automated FCC to obtain O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allethroyl)-L-serine methyl ester (0.16g, 0.27mmol, 72%) as a white solid.

[0692] Step 6. (Z)-methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)but-2-enoylamino)acrylate (E / Z stereochemistry arbitrarily assigned) was prepared following General Experimental Procedure 8. O-Acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-L-serine methyl ester (0.15 g, 0.25 mmol) afforded methyl (Z)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)but-2-enoylamino)acrylate (30 mg, 62 μmol, 25%) as a white solid. LCMS (22010199C TFALCMS-5C3) RT: 1.387 min; Area: 99.6% (215 nm), 99.3% (306 nm); m / z 487.2 [M+H] + . 1H NMR (400MHz, CDCl3) δ8.73(s,1H),8.48(s,1H),8.13(s,1H),7.95-7.88(m,2H),7.48(d,J=8.5Hz,2H),6.73(q, J=7.1Hz,1H),6.69(s,1H),6.63(s,1H),5.94(d,J=1.4Hz,1H),3.81(s,3H),1.89(d,J=7.1Hz,3H),1.54(s,9H).

[0693] Example 22: Compound 67

[0694] Methyl 2-(2-(2-(4-((tert-butyloxycarbonyl)amino)phenyl)thiazole-5-carboxamido)acrylamido)acrylate

[0695]

[0696] Step 1. Ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylate was prepared following General Experimental Procedure 1. (4-((tert-butoxycarbonyl)amino)phenyl)boronic acid (2.0 g, 8.4 mmol) and ethyl 2-bromothiazole-5-carboxylate (2.0 g, 8.4 mmol) gave ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylate (1.26 g, 3.62 mmol, 43%) as a yellow solid. 1 H NMR(299MHz, CDCl3)δ8.39(s,1H),7.94(d,J=8.8Hz,2H),7.49(d,J=8.6Hz,2H),6 .66 (s, 1H), 4.40 (q, J = 7.1Hz, 2H), 1.55 (d, J = 1.0Hz, 10H), 1.42 (t, J = 7.1Hz, 3H).

[0697] Step 2. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid was prepared following general experimental procedure 2. Ethyl 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carboxylate (1.2 g, 3.4 mmol) gave 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid (1.0 g, 3.1 mmol, 91%) as a white solid.

[0698] Step 3. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. 2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid (1.6 g, 5.0 mmol) and H-Ser-OMe.HCl (0.93 g, 1.2 eq., 6.0 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine methyl ester (1.4 g, 3.3 mmol, 67%) as an orange oil. LCMS (General 3acidic) RT: 0.99 min; Area % (254 nm): 88%; m / z = 422.3 [M+H] + .

[0699] Step 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine methyl ester (1.4 g, 3.3 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine (0.98 g, 2.4 mmol, 72%).

[0700] Step 5. N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following general experimental procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine (0.98 g, 2.4 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (1.1 g, 1.2 equiv, 2.9 mmol) gave crude material N-((2-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (2.25 g) which was used as is in the next step.

[0701] Step 6. N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 5. The crude material N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (2.25 g) and acetic anhydride (0.34 mL, 3.6 mmol) gave N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.40 g, 0.51 mmol, 21% over 2 steps). LCMS (General 3acidic) RT: 1.78 min; Area % (214 nm): 100%; m / z = 789.6 [M+H] + .

[0702] Step 7. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 7. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine (0.19 g, 0.24 mmol) afforded methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxamido)acrylamido)acrylate (7.8 mg, 16 μmol, 27%) as a white solid. LC-MS (22010199A TFA LCMS-5C1): RT: 1.353 min; area 97.7% (215 nm), 96.7% (304 nm); m / z = 473.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ (ppm) 8.66 (s, 1H), 8.57 (s, 1H), 8.24 (s, 1H), 7.92 (d, J = 8.8Hz, 2H), 7.48 (d, J = 8.8Hz, 2H), 6.72 (d, J =2.5Hz,1H),6.67(s,1H),6.63(s,1H),6.04(d,J=1.0Hz,1H),5.49(dd,J=2.3Hz,J=1.3Hz,1H),3.91(s,1H),1.54(s,9H).

[0703] Example 23: Compound 68

[0704] Methyl 2-(2-(5-(4-((tert-butyloxycarbonyl)amino)phenyl)thiophene-3-carboxamido)acrylamido)acrylate

[0705]

[0706] Step 1. Ethyl 5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxylate was prepared following General Experimental Procedure 1. Ethyl 5-bromothiophene-3-carboxylate (1.0 g, 4.2 mmol) and (4-((tert-butoxycarbonyl)amino)phenyl)boronic acid (1.0 g, 4.2 mmol) gave ethyl 5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxylate (1.1 g, 3.2 mmol, 76%) as a white solid. 1 H NMR (299MHz, CDCl3) δ8.00(m,1H),7.65(m,1H),7.55(d,J=8.7Hz,2H),7.41(d,J=8.5Hz, 2H), 7.28 (s, 4H), 6.54 (s, 1H), 4.37 (q, J = 7.1Hz, 2H), 1.55 (s, 9H), 1.40 (t, J = 7.1Hz, 3H).

[0707] Step 2. 5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carboxylic acid was prepared following General Experimental Procedure 2. Ethyl 5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carboxylate (1.1 g, 3.2 mmol) and 5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carboxylic acid (0.90 g, 2.8 mmol, 89%) were prepared as white solids. 1 H NMR (299MHz, CD3OD) δ8.08 (m, 1H), 7.65 (m, 1H), 7.62-7.52 (m, 2H), 7.48 (d, J = 8.7Hz, 2H), 1.54 (s, 8H).

[0708] Step 3. (5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 3. 5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carboxylic acid (0.90 g, 2.8 mmol) and L-serine methyl ester hydrochloride (0.44 g, 2.8 mmol) gave (5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serine methyl ester (1.0 g, 2.4 mmol, 84%) as a yellow oil. 1H NMR (299MHz, CDCl3) δ7.84(m,1H),7.56-7.46(m,3H),7.40(d,J=8.5Hz,2H),7.03(d,J=7.2Hz,1H),6.64(s ,1H),4.87(dt,J=7.2,3.6Hz,1H),4.39(q,J=7.1Hz,1H),4.09(t,J=4.0Hz,2H),3.85(s,3H),1.55(s,9H).

[0709] Step 4. (5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serine methyl ester (1.0 g, 0.24 mmol) afforded (5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serine (0.90 g, 2.2 mmol, 93%) as a yellow solid. 1 H NMR (299MHz, CD3OD) δ9.05 (s, 1H), 8.25 (d, J = 8.0Hz, 0H), 8.04 (d, J = 1.4Hz, 1H), 7.74 (d, J = 1.4Hz, 1H),7.63-7.52(m,2H),7.48(d,J=8.6Hz,2H),4.76-4.67(m,1H),4.10-3.92(m,2H),1.55(s,9H).

[0710] Step 5. N-((5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. (5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serine (0.20 g, 0.49 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (0.23 g, 1.2 equiv, 0.59 mmol) gave crude material N-((5-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (233 mg) which was used in the next step without purification.

[0711] Step 6. N-(O-acetyl-N-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 5. The crude material N-((5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (233 mg) gave N-(O-acetyl-N-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.10 g, 0.13 mmol, 26% over 2 steps)

[0712] Step 7. Methyl 2-(2-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 7. Methyl N-(O-acetyl-N-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine ester (50 mg, 63 μmol) gave methyl 2-(2-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxamido)acrylamido)acrylate (3.9 mg, 8.3 μmol, 31%) as a white solid. LC-MS (22010199A TFA LCMS-5C1): RT 1.483 min; area 84.0% (215 nm), 88.7% (304 nm); m / z = 472.0 [M+H] +

[0713] Example 24: Compound 69

[0714] Methyl 2-(2-(4'-((tert-butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxamido)acrylamido)acrylate

[0715]

[0716] Step 1. Prepare 4'-((tert-butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-methyl formate using the following procedure. To a solution of 4'-amino-[1,1'-biphenyl]-4-methyl formate (0.50 g, 2.2 mmol) in CH2Cl2 (20 mL), DMAP (0.54 g, 2 equivalents, 4.4 mmol) and Boc2O (0.48 g, 1 equivalent, 2.2 mmol) were added, and the resulting mixture was stirred at room temperature for 16 hours. The mixture was filtered and HCl (1M, 50 mL aqueous solution) was added. The mixture was extracted with EtOAc, and the organic layer was dried over Na2SO4, filtered and concentrated. The crude material was purified by FCC to give 4'-((tert-butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-methyl formate (0.30 g, 0.92 mmol, 42%). 1 H NMR (299MHz, CDCl3) δ8.10(m,2H),7.62(m,5H),7.53-7.44(m,2H),6.70(s,1H),3.95(s,3H),1.55(s,9H).

[0717] Step 2. 4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxylic acid was prepared following General Experimental Procedure 2. Methyl 4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxylate (0.52 g, 1.6 mmol) gave 4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxylic acid (0.34 g, 1.1 mmol, 66%) 1 H NMR (299MHz, CD3OD) δ8.13-8.00(m,2H),7.77-7.58(m,4H),7.53(m,2H),1.55(s,9H).

[0718] Step 3. (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. 4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxylic acid (90 mg, 0.29 mmol) and L-serine methyl ester hydrochloride (90 mg, 2 eq, 0.57 mmol) gave (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serine methyl ester (34 mg, 82 μmol, 29%). LCMS (General 3acidic) RT: 1.07 min; Area % (254 nm): 100%; m / z = 415.1 [M+H] + .

[0719] Step 4. (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serine methyl ester (34 mg, 82 μmol) gave crude material (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serine (77 mg) which was used as is in the next step. LCMS (General 3acidic) RT: 0.95 min; Area % (254 nm): 82%; m / z = 401.3 [M+H] + .

[0720] Step 5. N-((4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serine (100 mg, 0.25 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (118 mg, 0.30 mmol) gave N-((4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (66 mg, 89 μmol, 36%). LCMS (General 3acidic) RT: 1.72 min; Area % (254 nm): 77%; m / z = 740.6 [M+H] + .

[0721] Step 6. N-(O-Acetyl-N-(4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 5. N-((4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (66 mg, 89 μmol) gave N-(O-acetyl-N-(4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (44 mg, 56 μmol, 63%). LCMS (General 3acidic) RT: 1.81 min; Area % (254 nm): 100%; m / z = 740.6 [M+H] + .

[0722] Step 7. Methyl 2-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 7. Methyl N-(O-acetyl-N-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine ester (42 mg, 54 μmol) gave methyl 2-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxamido)acrylamido)acrylate (5.0 mg, 11 μmol, 20%). LC-MS (22010199A TFA LCMS-5C1): rt 1.483 min; area 78.4% (215 nm), 83.7% (304 nm); m / z = [M+H] + =466.2. 1 H NMR(CDCl3)δ(ppm)8.92(s,1H),8.60(s,1H),7.92(d,J=8.3Hz,2H),7.67(d,J=8.1Hz,2H),7.57(d,J=8.4Hz,2H), 7.47(d,J=8.4Hz,2H),6.82(m,1H),6.67(s,1H),6.56(s,1H),6.03(s,1H),5.49(s,1H),3.91(s,3H),1.54(s,9H).

[0723] Example 25: Compound 70

[0724] Methyl 2-(2-(2-(4-((tert-butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxamido)acrylamido)acrylate

[0725]

[0726] Step 1. Ethyl 2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylate was prepared following General Experimental Procedure 1. Ethyl 2-bromothiazole-4-carboxylate (0.20 g, 0.85 mmol) and (4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)boronic acid (0.20 g, 0.78 mmol) gave ethyl 2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylate (0.15 g, 0.42 mmol, 53%) 1H NMR (400MHz, CDCl3) δ8.20(t,J=8.2Hz,1H),8.10(s,1H),7.79(dd,J=11.8,2.0Hz,1H),7.6 8(d,J=8.6Hz,1H),6.82(s,1H),4.43(q,J=7.1Hz,2H),1.52(s,9H),1.41(t,J=7.1Hz,3H).

[0727] Step 2. 2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylic acid was prepared following General Experimental Procedure 2. Ethyl 2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylate (0.71 g, 1.9 mmol) gave 2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylic acid (0.63 g, 1.9 mmol, 97%). 1 H NMR (400MHz, CDCl3) δ8.25 (d, J = 8.3 Hz, 1H), 8.21 (s, 1H), 7.74 (dd, J = 11.7, 2.0 Hz, 1H), 7.70-7.63 (m, 1H), 6.86 (d, J = 3.6 Hz, 1H), 1.53 (s, 9H).

[0728] Step 3. (2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serine methyl ester was prepared following General Experimental Procedure 4. 2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylic acid (0.63 g, 1.9 mmol) and L-serine methyl ester hydrochloride (0.32 g, 1.1 equiv., 2.1 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serine methyl ester (0.87 g, 90% purity, 1.8 mmol, 96%)

[0729] Step 4. (2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serine was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serine methyl ester (0.87 g, purity 90%, 1.8 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serine (0.85 g, purity: 90%, 1.8 mmol, quantitative) as an off-white solid.

[0730] Step 5. N-((2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serine (0.76 g, 1.8 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (0.78 g, 1.1 equiv., 2.0 mmol) gave N-((2-(4-((tert-Butyloxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.2 g, 1.5 mmol, 84%)

[0731] Step 6. N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following general experimental procedure 5. N-((2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.2 g, 1.5 mmol) gave N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.1 g, 1.4 mmol, 93%). 1 H NMR (400MHz, CDCl3) δ8.21(t,J=8.2Hz,1H),8.09(d,J=7.7Hz,1H),8.04(s,1H),7.72(dd,J=11.7,2.0Hz,1H) ,7.63(d,J=8.4Hz,1H),7.61-7.50(m,5H),7.44-7.26(m,7H),7.12(d,J=8.1Hz,1H),6.83(d,J=3.5Hz,1H),4 .95-4.85(m,1H),4.67(dt,J=8.2,2.9Hz,1H),4.52(dd,J=11.3,5.6Hz,1H),4.39(dd,J=11.3,6.2Hz,1H),4. 11(dd,J=10.3,2.7Hz,2H),3.88(dd,J=10.3,3.0Hz,1H),3.73(s,3H),2.08(s,3H),1.53(s,9H),0.96(s,9H).

[0732] Step 7. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 7. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine ester (0.30 g, 0.37 mmol) afforded methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxamido)acrylamido)acrylate (30 mg, 59 μmol, 16%) as a white solid. LCMS (22010199A TFA LCMS-5C1): RT: 1.815 min; Area: 98.1% (215 nm), 96.8% (306 nm); m / z = 491.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.97(s,1H),8.55(s,1H),8.23(t,J=8.3Hz,1H),8.12(s,1H),7.81(dd,J=11.7,2.0Hz,1H),7.68(dd,J=8.6,1.3H z,1H),6.86(m,1H),6.77(d,J=2.2Hz,1H),6.72(s,1H),6.04(d,J=1.4Hz,1H),5.51(t,J=1.9Hz,1H),f3.90(s,3H),1.57-1.49(m,9H).

[0733] Example 26: Compound 71

[0734] Methyl 2-(2-(2-(3-((tert-butyloxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate

[0735]

[0736] Step 1. Ethyl 2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylate was prepared following General Experimental Procedure 1. Ethyl 2-bromothiazole-4-carboxylate (2.3 g, 1.05 equiv, 9.7 mmol) and (3-((tert-butoxycarbonyl)amino)phenyl)boronic acid (2.2 g, 9.3 mmol) gave ethyl 2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylate (1.4 g, 4.0 mmol, 43%) as a white solid. 1H NMR (400MHz, CDCl3) δ8.13(s,1H),7.94(t,J=2.0Hz,1H),7.64(ddd,J=7.7,1.7,1.0Hz,1H),7.55(d,J=8 .2Hz,1H),7.35(t,J=8.0Hz,1H),6.56(s,1H),4.43(q,J=7.1Hz,2H),1.51(s,9H),1.41(t,J=7.1Hz,3H).

[0737] Step 2. 2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid was prepared following general experimental procedure 2. Ethyl 2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylate (1.4 g, 4.0 mmol) gave 2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (1.3 g, 3.8 mmol, 95%) as a white solid. 1 H NMR (400MHz, CDCl3) δ8.22 (s, 1H), 8.05 (s, 1H), 7.59 (d, J = 7.6Hz, 1H), 7.46 (d, J = 8.2Hz, 1H), 7.37 (t, J = 7.9Hz, 1H), 1.53 (s, 9H).

[0738] Step 3. (2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine methyl ester was prepared following General Experimental Procedure 4. 2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (1.3 g, 3.8 mmol) and L-serine methyl ester hydrochloride (0.65 g, 1.1 equiv., 4.2 mmol) gave (2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine methyl ester (1.5 g, 3.5 mmol, 93%). 1 H NMR (299MHz, CDCl3) δ8.27(d,J=7.6Hz,1H),8.14(d,J=1.5Hz,1H),7.98(d,J=1.9Hz,1H),7.61(m,2H),7. 41(t,J=8.0Hz,1H),7.28(s,5H),6.66(s,1H),5.00-4.84(m,1H),4.12(m,2H),3.87(s,3H),1.57(s,9H).

[0739] Step 4. (2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine was prepared following General Experimental Procedure 2. (2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine methyl ester (1.5 g, 3.5 mmol) gave the crude material (2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (1.5 g, 3.8 mmol, quantitative) which was used as is in the next step.

[0740] Step 5. N-((2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. (2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (0.80 g, 2.0 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (0.75 g, 1.9 mmol) gave N-((2-(3-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.2 g, 1.6 mmol, 79%).

[0741] Step 6. N-(O-acetyl-N-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following general experimental procedure 5. N-((2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.2 g, 1.6 mmol) gave N-(O-acetyl-N-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (1.1 g, 1.4 mmol, 89%). 1H NMR (400MHz, CDCl3) δ8.11(d,J=7.8Hz,1H),8.07(s,1H),7.90(t,J=2.0Hz,1H),7.62-7.50(m,6 H),7.44-7.26(m,8H),7.13(d,J=8.2Hz,1H),6.57(s,1H),4.92(dt,J=7.9,5.9Hz,1H),4.67(dt ,J=8.1,2.8Hz,1H),4.51(dd,J=11.3,5.7Hz,1H),4.39(dd,J=11.3,6.2Hz,1H),4.11(dd,J=10. 3, 2.7Hz, 1H), 3.87 (dd, J = 10.3, 3.0Hz, 1H), 3.73 (s, 3H), 2.08 (s, 3H), 1.52 (s, 9H), 0.96 (s, 9H).

[0742] Step 7. Methyl 2-(2-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 7. Methyl N-(O-acetyl-N-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine ester (0.30 g, 0.38 mmol) afforded methyl 2-(2-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate (51 mg, 0.11 mmol, 57%) as a white solid. LCMS (22010199ATFALCMS-5C1): RT: 1.704 min; Area: 98.9% (215 nm), 98.6% (306 nm); m / z 473.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ10.00(s,1H),8.55(s,1H),8.15(s,1H),7.93(t,J=2.0Hz,1H),7.67(d,J=7.8Hz,1H),7.62(d,J=8.3Hz,1H),7.40( t,J=8.0Hz,1H),6.78(d,J=2.2Hz,1H),6.71(s,1H),6.66(s,1H),6.03(d,J=1.3Hz,1H),5.51(t,J=2.0Hz,1H),3.91(s,3H),1.55(s,9H).

[0743] Example 27: Compound 72

[0744] 2-(2-(2-(2-((tert-butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxamido)acrylamido)methyl acrylate

[0745]

[0746] Step 1. Ethyl 2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylate was prepared following General Experimental Procedure 1. (2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)boronic acid (1.2 g, 5.2 mmol) and ethyl 2-bromothiazole-4-carboxylate (1.4 g, 1.1 equiv, 5.8 mmol) gave ethyl 2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylate (1.1 g, 3.1 mmol, 61%) 1 H NMR (400MHz, CDCl3) δ9.15 (s, 2H), 8.62 (s, 1H), 8.18 (s, 1H), 4.43 (q, J = 7.1Hz, 2H), 1.63 (s, 1H), 1.56 (s, 9H), 1.41 (t, J = 7.1Hz, 3H).

[0747] Step 2. 2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylic acid was prepared following General Experimental Procedure 2. Ethyl 2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylate (2.1 g, 5.9 mmol) gave crude 2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylic acid (2.0 g) which was used in the next step without purification.

[0748] Step 3. (2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serine methyl ester was prepared following General Experimental Procedure 4. 2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylic acid (0.64 g, 2.0 mmol) and L-serine methyl ester hydrochloride (0.34 g, 1.1 equiv., 2.2 mmol) gave (2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serine methyl ester (0.93 g, 2.0 mmol, quantitative).

[0749] Step 4. (2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serine was prepared following General Experimental Procedure 2. (2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serine methyl ester (0.84 g, 2.0 mmol) gave (2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serine (0.55 g, 1.3 mmol, 67%). 1H NMR (400MHz, DMSO) δ10.48(s,1H),9.19(s,1H),8.41(s,1H),8.37(d,J=8.1Hz,1H),4.49(dt,J=8.3,4.2Hz,1H),3.84(m,2H),1.48(s,9H).

[0750] Step 5. N-((2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. (2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serine (0.54 g, 1.3 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (0.57 g, 1.1 equiv, 1.4 mmol) afforded N-((2-(2-((tert-Butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.42 g, 0.56 mmol, 43%) as an off-white solid. LCMS (General 3acidic) RT: 1.53 min; Area % (254 nm): 76%; m / z = 749.7 [M+H] + .

[0751] Step 6. N-(O-acetyl-N-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following general experimental procedure 5. N-((2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.40 g, 0.53 mmol) gave N-(O-acetyl-N-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.24 g, 0.30 mmol, 57%). LCMS (General 3acidic) RT: 1.63 min; Area % (254 nm): 96%; m / z = 691.5 [M-Boc+H] + .

[0752] Step 7. O-acetyl-N-(O-acetyl-N-(2-(2-((tert-butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-L-serine methyl ester was prepared using the following procedure. To a solution of N-(O-acetyl-N-(2-(2-((tert-butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.24 g, 0.30 mmol) in THF (5 mL) was added a solution of TBAF in THF (1 M, 0.45 mL, 1.5 eq, 0.45 mmol), and the resulting mixture was stirred for 1 hour. The mixture was concentrated and the residue was dissolved in MeCN (5 mL). Triethylamine (0.10mL, 2.4 equivalents, 0.72mmol) and acetic anhydride (38 μL, 1.3 equivalents, 0.40mmol) were added, and the resulting mixture was stirred at room temperature for 45 minutes. Water was added and the mixture was extracted with EtOAc. The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. The crude material was purified by automated FCC to obtain O-acetyl-N-(O-acetyl-N-(2-(2-((tert-butyloxycarbonyl)amino)pyrimidine-5-yl)thiazole-4-carbonyl)-L-seryl)-L-serine methyl ester (87mg, 0.15mmol, 48%).

[0753] Step 8. Methyl 2-(2-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxamido)acrylamido)acrylate was prepared using the following procedure. To a solution of O-acetyl-N-(O-acetyl-N-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-L-serine methyl ester (86 mg, 0.14 mmol) in CHCl (1.5 mL) and THF (1.5 mL) was added DBU (87 μL, 4 eq, 0.58 mmol) at 0°C and the mixture was stirred at 0°C for 2 hours. Water was added and the aqueous phase was adjusted to pH 4 by adding 1 M HCl. The mixture was extracted with CHCl and the organic layer was washed with brine, dried over NaSO, filtered and concentrated. The crude material was purified by automated reverse phase FCC to give methyl 2-(2-(2-(2-((tert-butyloxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxamido)acrylamido)acrylate (27 mg, 57 μmol, 39%). LCMS (22010199A TFA LCMS-5C1): RT: 1.460 min; Area: 96.8% (215 nm), 96.2% (306 nm); m / z 473.0 [MH] - . 1H NMR (400MHz, CDCl3) δ10.01(s,1H),9.17(s,2H),8.54(s,1H),8.20(s,1H),7.96(s,1H),6.77(d,J =2.3Hz, 1H), 6.70 (s, 1H), 6.03 (d, J = 1.3Hz, 1H), 5.51 (t, J = 1.9Hz, 1H), 3.90 (s, 3H), 1.57 (s, 9H).

[0754] Example 28: Compound 75

[0755] Methyl 2-(2-(4-(4-((tert-butyloxycarbonyl)amino)phenyl)thiazole-2-carboxamido)acrylamido)acrylate

[0756]

[0757] Step 1. 4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carboxylic acid was prepared using the following procedure. To a solution of ethyl 4-bromothiazole-2-carboxylate (1.0 g, 4.2 mmol) and (4-((tert-butoxycarbonyl)amino)phenyl)boronic acid in 1,2-dimethoxyethane (25 mL) was added sodium carbonate (2M in water, 10 eq., 42 mmol) and tetrapalladium (0.24 g, 0.05 eq., 0.21 mmol), and the resulting mixture was stirred at 80 °C for 16 hours. The mixture was cooled to room temperature, diluted with ethyl acetate and washed with water. The aqueous phase was acidified with 1 M HCl solution and extracted with ethyl acetate. The organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to give 4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carboxylic acid (1.1 g, 43% purity, 1.4 mmol, 35%) as a mixture with the decarboxylated product. The mixture was used as is in the next step.

[0758] Step 2. (4-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. Crude 4-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-2-carboxylic acid (1.1 g, 43% purity, 1.4 mmol) and L-serine methyl ester hydrochloride (0.26 g, 1.2 equiv, 1.7 mmol) gave (4-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serine methyl ester (0.61 g, 1.45 mmol, quantitative).

[0759] Step 3. (4-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (4-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serine methyl ester (0.60 g, 1.4 mmol) gave (4-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serine (0.59 g, 1.4 mmol, quantitative).

[0760] Step 4. (4-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl-L-serine methyl ester was prepared following General Experimental Procedure 4. (4-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serine (0.58 g, 1.4 mmol) and L-serine methyl ester hydrochloride (0.27 g, 1.2 equiv, 1.7 mmol) gave (4-(4-((tert-Butyloxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl-L-serine methyl ester (0.52 g, 1.0 mmol, 72%). LCMS (General 3acidic) RT: 0.94 min; Area % (254 nm): 91%; m / z = 509.4 [M+H] + .

[0761] Step 5. O-Acetyl-N-(O-acetyl-N-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl)-L-serine methyl ester was prepared following General Experimental Procedure 5. (4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl-L-serine methyl ester (0.52 g, 1.0 mmol) and acetic anhydride (0.25 mL, 4 equiv, 2.7 mmol) gave O-acetyl-N-(O-acetyl-N-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl)-L-serine methyl ester (0.33 g, 0.56 mmol, 55%)

[0762] Step 6. Methyl 2-(2-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 8. Methyl O-acetyl-N-(O-acetyl-N-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl)-L-serine ester (0.35 g, 0.59 mmol) afforded methyl 2-(2-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carboxamido)acrylamido)acrylate (85 mg, 0.18 mmol, 30%) as a white solid. LC-MS (22010199A TFA LCMS-5C1): RT: 1.731 min; area 98.6% (215 nm), 97.8% (304 nm); m / z = 473.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ (ppm) 9.92 (s, 1H), 8.55 (s, 1H), 7.89 (d, J = 8.7Hz, 2H), 7.67 (s, 1H), 7.45 (d, J = 8.6Hz, 2H), 6.76 (d, J =2.4Hz, 1H), 6.71 (s, 1H), 6.59 (s, 1H), 6.04 (d, J = 1.0Hz, 1H), 5.54 (dd, J = 2.3Hz, J = 1.6Hz, 1H), 3.90 (s, 3H), 1.54 (s, 9H).

[0763] Example 29: Compound 76

[0764] Methyl 2-(2-(2-(4-((tert-butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxamido)acrylamido)acrylate

[0765]

[0766] Step 1. Ethyl 2-(4-((tert-butoxycarbonyl)amino)-2-fluorophenyl)thiazole-4-carboxylate was prepared following General Experimental Procedure 1. Ethyl 2-bromothiazole-4-carboxylate (0.47 g, 2.0 mmol) and tert-butyl (3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)carbamate (0.70 g, 2.0 mmol) gave ethyl 2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxylate (0.38 g, 1.0 mmol, 51%). LCMS (General 3acidic) RT: 1.58 min; Area % (254 nm): 96%; m / z = 383.2 [M+H] + .

[0767] Step 2. 2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxylic acid was prepared following General Experimental Procedure 2. Ethyl 2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxylate (0.38 g, 0.99 mmol) gave 2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxylic acid (0.30 g, 0.84 mmol, 85%) 1 H NMR (299MHz, CDCl3) δ8.36 (s, 1H), 8.25 (d, J = 8.9Hz, 1H), 7.82 (s, 1H), 7.31-7.22 (m, 2H), 6.78 (s, 1H), 1.56 (s, 9H).

[0768] Step 3. (2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. 2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxylic acid (0.44 g, 1.2 mmol) and L-serine methyl ester hydrochloride (0.23 g, 1.2 equiv, 1.5 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serine methyl ester (0.42 g, 0.92 mmol, 75%). LCMS (General 3acidic) RT: 1.23 min; Area % (254 nm): 93%; m / z = 456.3 [M+H] + .

[0769] Step 4. (2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serine methyl ester (0.42 g, 0.92 mmol) afforded (2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serine (0.33 g, 0.75 mmol, 81%) as an orange solid.

[0770] Step 5. N-((2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serine (0.33 g, 0.75 mmol) and O-(tert-butyldiphenylsilyl)-L-serine methyl ester hydrochloride (0.35 g, 1.2 equiv., 0.90 mmol) gave N-((2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.37 g, 0.48 mmol, 64%).

[0771] Step 6. General Experimental Procedure 5 was followed to prepare N-(O-acetyl-N-(2-(4-((tert-butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester. N-((2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.37 g, 0.48 mmol) and acetic anhydride (68 μL, 1.5 equiv., 0.72 mmol) gave N-(O-acetyl-N-(2-(4-((tert-Butyloxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine methyl ester (0.22 g, 0.27 mmol, 57%). 1 H NMR (299MHz, CDCl3) δ8.28-8.12(m,3H),7.85(s,1H),7.57(m,4H),7.46-7.30(m ,6H),7.19(m,2H),6.66(s,1H),4.95(q,J=6.3Hz,1H),4.70(d,J=8.1Hz,1H),4. 56(dd,J=11.3,5.7Hz,1H),4.43(dd,J=11.3,6.2Hz,1H),4.14(d,J=8.1Hz,1H), 3.96-3.86(m,1H),3.77(d,J=1.2Hz,3H),2.11(s,3H),1.56(s,9H),1.00(s,9H).

[0772] Step 7. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 7. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serine ester (0.22 g, 0.27 mmol) afforded methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxamido)acrylamido)acrylate (15 mg, 30 μmol, 29%) as a white solid. LCMS (22010199A TFA LCMS-5C1): rt 1.880 min; area 96.5% (215 nm), 95.6% (304 nm); m / z = 507.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ (ppm) 10.02 (s, 1H), 8.55 (s, 1H), 8.31 (d, J = 8.7Hz, 1H), 8.23 ​​(s, 1H), 7.84 (d, J = 1.8Hz, 1H), 7.21 (dd, J = 8.7Hz, J =2.3Hz, 1H), 6.78 (d, J = 2.2Hz, 1H), 6.70 (s, 1H), 6.66 (s, 1H), 6.02 (d, J = 1.1Hz, 1H), 5.48 (t, J = 1.8Hz, 1H), 3.90 (s, 3H), 1.54 (s, 9H).

[0773] Example 30: Compound 77

[0774] Methyl 2-(2-(4'-((tert-butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxamido)acrylamido)acrylate

[0775]

[0776] Step 1. 4'-((tert-butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxylic acid was prepared using the following procedure. DMAP (0.54 g, 2 equivalents, 4.4 mmol) was added to a solution of 4'-amino-[1,1'-biphenyl]-3-carboxylic acid methyl ester (0.50 g, 2.2 mmol) in CH2Cl2 (20 mL). Boc2O (0.72 g, 1.5 equivalents, 3.3 mmol) was then added portionwise, and the mixture was stirred at room temperature for 16 hours. The mixture was washed with water and brine, dried over Na2SO4 and concentrated in vacuo. The crude material was dissolved in THF (20 mL), and NaOH solution (1M aqueous solution, 7 equivalents, 15.4 mL, 15 mmol) was added, and the resulting mixture was stirred for 64 hours. The mixture was quenched with 1M HCl and extracted with EtOAc. The organic phase was dried over Na 2 SO 4 , filtered and concentrated in vacuo to give 4′-((tert-butoxycarbonyl)amino)-[1,1′-biphenyl]-3-carboxylic acid (0.47 g, 1.5 mmol, 68%) as a yellow solid.

[0777] Step 2. (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. 4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxylic acid (0.45 g, 1.4 mmol) and L-serine methyl ester hydrochloride (0.27 g, 1.2 equiv, 1.7 mmol) gave crude material (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serine methyl ester (0.60 g) which was used as is in the next step.

[0778] Step 3. (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serine methyl ester (0.58 g, 1.4 mmol) afforded (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serine (0.37 g, 0.92 mmol, 66% over 2 steps) as a yellow oil. LCMS (General 3acidic) RT: 0.97 min; Area % (254 nm): 90%; m / z = 401.3 [M+H] + .

[0779] Step 4. (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl-L-serine methyl ester was prepared following General Experimental Procedure 4. (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serine (0.20 g, 0.50 mmol) and L-serine methyl ester hydrochloride (0.93 mg, 1.2 eq, 0.60 mmol) gave the crude material (4'-((tert-Butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl-L-serine methyl ester (0.16 g) which was used as is in the next step. LCMS (General 3acidic) RT: 0.95 min; Area % (254 nm): 84%; m / z = 502.4 [M+H] + .

[0780] Step 5. General Experimental Procedure 5 was followed to prepare O-acetyl-N-(O-acetyl-N-(4'-((tert-butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl)-L-serine methyl ester. The crude material (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl-L-serine methyl ester (0.16 g) and acetic anhydride (0.12 mL, 2.6 eq., 1.3 mmol) gave O-acetyl-N-(O-acetyl-N-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl)-L-serine methyl ester (130 mg, 0.22 mmol, 44%) and the monoelimination product O-acetyl-N-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxamido)acryloyl)-L-serine methyl ester (0.10 g, 0.19 mmol, 38%)

[0781] Step 6. General Experimental Procedure 8 was followed to prepare methyl 2-(2-(4'-((tert-butyloxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxamido)acrylamido)acrylate. O-Acetyl-N-(O-acetyl-N-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl)-L-serine methyl ester (130 mg, 0.22 mmol) and O-acetyl-N-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxamido)acryloyl)-L-serine methyl ester (0.10 g, 0.19 mmol) (prepared as in step 5) gave methyl 2-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxamido)acryloylamino)acrylate (6.6 mg, 14 μmol) as a white solid. LCMS (22010199A TFA LCMS-5C1): RT: 1.468 min; area 98.0% (215 nm), 97.2% (304 nm); m / z = 466.2 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.95(s,1H),8.61(s,1H),8.08(t,J=1.9Hz,1H),7.78(ddt,J=15.4,7.9,1.3Hz,2H),7.62-7.45(m,6H ), 6.85 (d, J = 2.3Hz, 1H), 6.68 (s, 1H), 6.60 (s, 1H), 6.05 (d, J = 1.4Hz, 1H), 5.53 (t, J = 1.8Hz, 1H), 3.93 (s, 3H), 1.56 (s, 9H).

[0782] Example 31: Compound 78

[0783] 2-(2-(2-(3-((tert-butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxamido)acrylamido)methyl acrylate

[0784]

[0785] Step 1. tert-Butyl (3-carbamoylbicyclo[1.1.1]pentan-1-yl)carbamate was prepared following General Experimental Procedure 9. 3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentane-1-carboxylic acid (2.0 g, 8.8 mmol) gave tert-butyl (3-carbamoylbicyclo[1.1.1]pentan-1-yl)carbamate (1.8 g, 8.1 mmol, 92%) as a white solid.

[0786] Step 2. Prepare tert-butyl (3-thiocarbamoylbicyclo[1.1.1]pentan-1-yl)carbamate following General Experimental Procedure 10. tert-butyl (3-carbamoylbicyclo[1.1.1]pentan-1-yl)carbamate (1.8 g, 8.1 mmol) gave tert-butyl (3-thiocarbamoylbicyclo[1.1.1]pentan-1-yl)carbamate (1.5 g, 6.4 mmol, 78%) LCMS (General 3acidic) RT: 0.79 min; Area % (254 nm): 100%

[0787] Step 3. Ethyl 2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylate was prepared following General Experimental Procedure 11. Tert-butyl (3-thiocarbamoylbicyclo[1.1.1]pentan-1-yl)carbamate (1.5 g, 6.1 mmol) gave ethyl 2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylate (1.8 g, 5.4 mmol, 88%). 1 H NMR (400MHz, CDCl3) δ8.06 (s, 1H), 5.00 (s, 1H), 4.41 (q, J = 7.1Hz, 2H), 2.49 (s, 6H), 1.46 (s, 9H), 1.39 (t, J = 7.1Hz, 3H).

[0788] Step 4. 2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylic acid was prepared following General Experimental Procedure 2. Ethyl 2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylate (1.8 g, 5.4 mmol) gave 2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylic acid (1.7 g, 5.4 mmol, quantitative)

[0789] Step 5. (2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. 2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylic acid (1.7 g, 5.4 mmol) and L-serine methyl ester hydrochloride (0.92 g, 1.1 equiv., 5.9 mmol) afforded (2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serine methyl ester (1.6 g, 90% purity, 3.6 mmol, 67%) as a white solid. 1H NMR (400MHz, CDCl3) δ8.07 (d, J = 7.7Hz, 1H), 8.00 (s, 1H), 4.83 (m, 1H), 4.13-3.98 (m, 2H), 3.81 (s, 3H), 2.44 (s, 6H), 1.45 (s, 9H).

[0790] Step 6. (2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serine methyl ester (1.6 g, 90% purity, 3.6 mmol) gave (2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serine (1.5 g, 85% purity, 3.2 mmol, 89%)

[0791] Step 7. (2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-seryl-L-serine methyl ester was prepared following General Experimental Procedure 4. (2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serine (1.5 g, 3.2 mmol) and L-serine methyl ester hydrochloride (0.70 g, 1.4 equiv, 4.5 mmol) gave (2-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-seryl-L-serine methyl ester (1.6 g, 3.2 mmol, 98%)

[0792] Step 8. O-Acetyl-N-(O-acetyl-N-(2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-seryl)-L-serine methyl ester was prepared following general experimental procedure 5. (2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-seryl-L-serine methyl ester (1.6 g, 3.2 mmol) gave O-acetyl-N-(O-acetyl-N-(2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-seryl)-L-serine methyl ester (1.2 g, 2.1 mmol, 68%) as a white solid.

[0793] Step 9. Methyl 2-(2-(2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 8. Methyl O-acetyl-N-(O-acetyl-N-(2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-seryl)-L-serine ester (0.17 g, 0.32 mmol) afforded methyl 2-(2-(2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxamido)acrylamido)acrylate (85 mg, 0.18 mmol, 58%) as a white solid. LCMS (22010199ATFA LCMS-5C1): rt 1.319 min; area 99.0% (215 nm), 98.7% (304 nm); m / z = 463.2 [M+H] + . 1 HNMR (400MHz, CDCl3) δ9.81 (s, 1H), 8.51 (s, 1H), 8.05 (s, 1H), 6.73 (d, J = 2.1Hz, 1H), 6.69 (s, 1H), 6.01(d,J=1.3Hz,1H),5.47(t,J=1.9Hz,1H),5.02(s,1H),3.90(s,3H),2.48(s,6H),1.47(s,9H).

[0794] Example 32: Compound 79

[0795] 2-(2-(2-(4-((tert-butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carboxamido)acrylamido)methyl acrylate

[0796]

[0797] Step 1. tert-Butyl (trans-4-carbamoylcyclohexyl)carbamate was prepared following General Experimental Procedure 9. trans-4-((tert-Butyloxycarbonyl)amino)cyclohexane-1-carboxylic acid (2.0 g, 8.2 mmol) gave tert-butyl (trans-4-carbamoylcyclohexyl)carbamate (1.9 g, 7.7 mmol, 94%) as a white solid.

[0798] Step 2. tert-Butyl (4-thiocarbamoylcyclohexyl)carbamate was prepared following General Experimental Procedure 10. tert-Butyl (trans-4-carbamoylcyclohexyl)carbamate (1.9 g, 7.7 mmol) gave tert-butyl (4-thiocarbamoylcyclohexyl)carbamate (1.2 g, 4.7 mmol, 61%, trans / cis 97:3) as an off-white solid.

[0799] Step 3. Ethyl 2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylate was prepared following General Experimental Procedure 11. Tert-butyl (4-thiocarbamoylcyclohexyl)carbamate (1.2 g, 4.7 mmol) gave ethyl 2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylate (1.1 g, 3.0 mmol, 64%, trans / cis 9:1).

[0800] Step 4. 2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylic acid was prepared following General Experimental Procedure 2. Ethyl 2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylate (1.1 g, 3.0 mmol) gave 2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylic acid (0.97 g, 3.0 mmol, 99%, trans / cis 9:1)

[0801] Step 5. (2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. 2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylic acid (0.97 g, 3.0 mmol) and L-serine methyl ester hydrochloride (0.51 g, 1.1 equiv., 3.3 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serine methyl ester (1.1 g, 89% purity, 2.2 mmol, 75%)

[0802] Step 6. (2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serine methyl ester (1.1 g, 89% purity, 2.2 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serine (1.0 g, 88% purity, 2.1 mmol, 97%).

[0803] Step 7. (2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl-L-serine methyl ester was prepared following General Experimental Procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serine (1.0 g, 88% purity, 2.1 mmol) and L-serine methyl ester hydrochloride (0.36 g, 1.1 equiv., 2.3 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl-L-serine methyl ester (0.55 g, 90% purity, 0.96 mmol, 45%).

[0804] Step 8. O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl)-L-serine methyl ester was prepared following general experimental procedure 5. (2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl-L-serine methyl ester (0.55 g, 90% purity, 0.96 mmol) gave O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl)-L-serine methyl ester (0.46 g, 0.77 mmol, 80%, trans / cis 87:13)

[0805] Step 9. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 8. Methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl)-L-serine ester (0.20 g, 0.33 mmol) gave two batches of methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxamido)acrylamido)acrylate 79-1 and 79-2.

[0806] 79-1 (81 mg, 0.17 mmol, 51%, trans / cis 89:11) LCMS (22010199A TFA LCMS-5C1): RT: 1.492 min; area: 86.2% (215 nm), 88.4% (306 nm); m / z 479.2 (M-H+); RT: 1.520 min; area 10.80% (215 nm), 11.61% (306 nm); m / z = 479.2 [M+H] +1 H-NMR (main isomer) (400MHz, CDCl3) δ9.90 (s, 1H), 8.53 (s, 1H), 8.03 (s, 1H), 6.74 (d, J = 2.2 Hz, 1H), 6.70 (s, 1H), 6.01 (d, J = 1.3 Hz, 1H), 5.46 (t, J = 1.9 Hz, 1H), 4.43 (s, 1H), 3.90 (s,3H),3.51(s,1H),2.97(tt,J=12.1,3.6Hz,1H),2.25(d,J=13.4Hz,2H),2.17(d,J= 12.7Hz, 2H), 1.66 (qd, J=13.1, 3.2Hz, 2H), 1.46 (s, 9H), 1.27 (qd, J=12.7, 3.4Hz, 2H).

[0807] 79-2 14 mg, 29 μmol, 8.7%, trans / cis 63:37. LCMS-5 (22010199A TFA LCMS-5C1): RT: 1.493 min; Area: 55.3% (215 nm), 54.5% (306 nm); m / z 479.2 (M-H+); RT: 1.526 min; Area: 42.9% (215 nm), 43.6% (306 nm); m / z = 379.2 [M-Boc+H] + . 1 H NMR (400MHz, CDCl3) δ9.99 (s, 0H), 9.90 (s, 1H), 8.54 (m, 1H), 8.03 (m, 1H), 6.74 (d, J = 2.0 Hz,1H),6.70(d,J=3.7Hz,1H),6.01(s,1H),5.47(m,1H),4.79(s,0H),4.43(s,0H),3.90 (s,3H),3.78(s,0H),3.50(s,0H),3.16(s,0H),3.02-2.91(m,1H),2.25(m,1H),2.17(m, 1H),2.01(s,1H),1.97-1.74(m,2H),1.74-1.59(m,2H),1.45(m,9H),1.37-1.20(m,2H).

[0808] Example 33: Compound 80

[0809] 2-(2-(2-(4-((tert-butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxamido)acrylamido)methyl acrylate

[0810]

[0811] Step 1. tert-Butyl (4-carbamoylbicyclo[2.2.2]octan-1-yl)carbamate was prepared following General Experimental Procedure 9. 4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octane-1-carboxylic acid (1.0 g, 3.7 mmol) gave tert-butyl (4-carbamoylbicyclo[2.2.2]octan-1-yl)carbamate (1.2 g, 82% purity, 3.5 mmol, 95%)

[0812] Step 2. tert-Butyl (4-thiocarbamoylbicyclo[2.2.2]octan-1-yl)carbamate was prepared following General Experimental Procedure 10. tert-Butyl (4-carbamoylbicyclo[2.2.2]octan-1-yl)carbamate (1.2 g, 82% purity, 3.5 mmol) afforded tert-butyl (4-thiocarbamoylbicyclo[2.2.2]octan-1-yl)carbamate (0.48 g, 1.7 mmol, 47%) as a white solid.

[0813] Step 3. Ethyl 2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylate was prepared following General Experimental Procedure 11. Tert-butyl (4-thiocarbamoylbicyclo[2.2.2]octan-1-yl)carbamate (0.48 g, 1.7 mmol) gave ethyl 2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylate (0.31 g, 0.82 mmol, 49%) as a white solid. 1 H NMR (400MHz, CDCl3) δ8.00 (s, 1H), 4.37 (q, J = 7.1Hz, 2H), 2.12-1.99 (m, 6H), 1.99-1.91 (m, 6H), 1.41 (s, 9H), 1.36 (t, J = 7.1Hz, 3H).

[0814] Step 4. 2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylic acid was prepared following General Experimental Procedure 2. Ethyl 2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylate (0.31 g, 0.82 mmol) gave 2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylic acid (0.32 g, 90% purity, 0.82 mmol, quantitative)

[0815] Step 5. (2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serine methyl ester was prepared following General Experimental Procedure 4. 2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylic acid (0.32 g, 90% purity, 0.82 mmol) and L-serine methyl ester hydrochloride (0.14 g, 1.1 equiv., 0.90 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serine methyl ester (0.33, 88% purity, 0.64 mmol, 78%).

[0816] Step 6. (2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serine was prepared following General Experimental Procedure 2. (2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serine methyl ester (0.33 g, 0.64 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serine (0.30 g, 0.66 mmol, quantitative).

[0817] Step 7. (2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-seryl-D-serine methyl ester was prepared following General Experimental Procedure 4. (2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serine (0.30 g, 0.66 mmol) and L-serine methyl ester hydrochloride (0.11 g, 1.1 equiv, 0.73 mmol) gave (2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-seryl-D-serine methyl ester (0.30 g, 0.55 mmol, 83%). 1 H NMR (400MHz, CDCl3) δ8.25(d,J=7.4Hz,1H),7.96(s,1H),7.67(d,J=7.9Hz,1H),4.69(m,2H),4.36(s,1H),4.17-4.08(m, 1H),4.08-3.92(m,2H),3.88-3.81(m,1H),3.79(s,3H),3.77-3.70(m,1H),3.61(m,1H),2.07-1.92(m,12H),1.42(s,9H).

[0818] Step 8. General Experimental Procedure 5 was followed to prepare O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-seryl)-L-serine methyl ester. (2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-seryl-L-serine methyl ester (0.30 g, 0.55 mmol) and acetic anhydride (0.13 mL, 2.5 equiv, 1.4 mmol) gave O-acetyl-N-(O-acetyl-N-(2-(4-((tert-Butyloxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-seryl)-L-serine methyl ester (0.26 g, 0.41 mmol, 76%).

[0819] Step 9. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxamido)acrylamido)acrylate was prepared following General Experimental Procedure 8. Methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-seryl)-L-serine ester (0.15 g, 0.23 mmol) afforded methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxamido)acrylamido)acrylate (65 mg, 0.13 mmol, 55%) as a white solid. LCMS (22010199ATFA LCMS-5C1): RT: 1.783 min; Area: 99.9% (215 nm), 99.6% (306 nm); m / z = 505.2 [M+H] +

[0820] General procedure for solid phase peptide synthesis

[0821] Fmoc Rink amide AM resin (0.70 mmol / g) was swollen by washing with CH2Cl2 (2×1 min, 10 mL / g resin). The resin was washed with DMF (3×1 min, 10 mL / g resin) and treated with 20% piperidine in DMF (10 mL / g resin) for 30 minutes. The resin was washed with DMF (3×1 min, 10 mL / g resin). The resin was treated with a solution of Fmoc-AA(PG)-OH (3 equiv.), HATU (3 equiv.) and DiPEA (3 equiv.) for 2-3 hours or 16 hours. The resin was washed with DMF (3×1 min, 10 mL / g resin). The wash-deprotection-wash-coupling cycle was repeated for each amino acid coupling. After the coupling cycle was completed, the resin was treated with TFA:TIPS:water (95:2.5:2.5, 10 mL / g resin) for 2 hours. The resin was removed by filtration and the filtrate was precipitated with MTBE:heptane (1:1 v / v). After centrifugation (5 minutes, 3000rpm), the supernatant was discarded, and the residue was resuspended in MTBE: heptane (1: 1v / v) and centrifuged again (5 minutes, 3000rpm). The supernatant was discarded, and the pellet was dissolved in 1: 1 water / MeCN. The solution was partially concentrated and lyophilized. The crude peptide was purified by automated reverse phase FCC.

[0822] Example 34: Compound 81

[0823]

[0824] N3(CH2)5C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2 was prepared on a 2.5 mmol scale following the general procedure for solid phase peptide synthesis. Purification was performed using HCl buffer to give the desired HCl salt form. N3(CH2)5C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2 HCl salt (0.58 g, 0.50 mmol, 20%) was isolated as a white solid. LCMS (30833LCMS-6): RT: 2.403 min; Area: 78.0% (215 nm), 99.6% (ELSD); m / z = 1084.7 [M+H] +

[0825] Example 35: Compound 82

[0826] N3(CH2CH2O)6CH2CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2

[0827] HCl salt

[0828]

[0829] N3(CH2CH2O)6CH2CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2 HCl salt was prepared on a 0.25 mmol scale following the general procedure for solid phase peptide synthesis. Purification using HCl buffer afforded the desired HCl salt form. N3(CH2CH2O)6CH2CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl salt (39 mg, 28 μmol, 11%) was isolated as a white solid. LCMS (30833LCMS-6): RT: 2.365 min; Area: 97.8% (215 nm), 99.6% (ELSD); m / z = 1306.8 [M+H] +

[0830] Example 36: Compound 83

[0831] N3(CH2CH2O)9CH2CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2

[0832] HCl salt

[0833]

[0834] N3(CH2CH2O)9CH2CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2 HCl salt was prepared on a 0.25 mmol scale following the general procedure for solid phase peptide synthesis. Purification using HCl buffer afforded the desired HCl salt form. N3(CH2CH2O)9CH2CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl salt (34 mg, 22 μmol, 8.8%) was isolated as a white solid. LCMS (30833LCMS-6): RT: 2.415 min; Area: 98.4% (215 nm), 99.8% (ELSD); m / z = 1438.9 [M+H] +

[0835] General procedure for conjugation chemistry between azido-peptide and compound 49

[0836] To a solution of compound 49 in DMF / water (1:1, 2 mL) was added the azido-peptide and the resulting mixture was stirred for 3 h. The mixture was directly purified using automated reverse phase FCC.

[0837] Example 37: Compound 84

[0838] Coupling of compound 49 and compound 82

[0839]

[0840] Compound 84 was prepared following the general procedure of the conjugation chemistry between the azido-peptide and 49. Compound 49 (3.0 mg, 4.5 μmol) and compound 82 (6.4 mg, 4.5 μmol) gave compound 84 (4.3 mg, 2.1 μmol, 46%) as a white solid. LCMS (30833 LCMS-6): RT: 2.791 min; Area: 81.9% (215 nm), 98.1% (ELSD); m / z = 1966.2 [M+H] +

[0841] Example 38: Compound 85

[0842] Coupling of compound 49 and compound 83

[0843]

[0844] Compound 85 was prepared following the general procedure of the conjugation chemistry between azide and peptides. Compound 49 (3.0 mg, 4.5 μmol) and compound 83 (7.0 mg, 4.5 μmol) gave compound 85 (4.3 mg, 2.0 μmol, 43%) as a white solid. LCMS (30833 LCMS-6): RT: 2.802 min; Area: 59.7% (215 nm), 85.5% (ELSD); m / z = 1049.7 [M+2H] 2+

[0845] Example 39: Predictive Assay

[0846] To test the binding mechanism and explore the binding target, cell treatment and immunoblotting will be performed as described by Cunniff et al. 2015. Covalent cross-linking of the enzyme can be detected by western blotting using specific antibodies.

[0847] Briefly, human tumor cell lines (HMESO cell lines derived from patients with malignant mesothelioma) will be cultured in appropriate culture media and treated with different concentrations of test compounds (0.1 μM-100 μM) for 24 hours. After 24 hours of exposure to test compounds, cell lysates will be produced in standard lysis buffer (RIPA buffer). Protein abundance will be quantified, and equal protein concentrations will be separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Protein immunoblotting will be performed using antibodies specific for proteins, such as PRX1, PRX2, PRX3 and PRX4. Covalent cross-linking modifications can be detected by the presence of approximately 46 kD antibody reactive species on protein immunoblots.

[0848] Cell death assays will be performed as in Nelson et al. 2021. Briefly, human tumor cell lines (HMESO cell line derived from a patient with malignant mesothelioma) will be cultured in 96-well plates and incubated with test compounds for 48 hours. Residual amounts of cellular material will be stained with crystal violet and total cell technology will be performed to determine % cell viability.

[0849] Example 40: Cell viability study

[0850] The cell lines were seeded in 96-well plates (Corning, Kennebunk, ME, USA) at a density of 2500 cells per well. The next day, the cells were treated with test compounds diluted in complete medium and then incubated for 48 hours. After incubation, the cells were washed with PBS (Corning Cellgro, Manassas, VA, USA), fixed with PBS containing 3.0% formaldehyde (Fisher BioReagents, Fair Lawn, NJ, USA), and stained with water containing 0.1% crystal violet (Acros Organics, Fair Lawn, NJ, USA) for 30 minutes. The crystal violet stain was removed, and the plate was washed with H2O and dried. To quantify cell viability, plates were imaged using a Lionheart plate reader (BioTek Instruments, Winooski, VT, USA) and / or analyzed by absorbance at 540 nm (crystal violet dye dissolved in 100% methanol) using a Synergy HTX plate reader (BioTek Instruments, Winooski, VT, USA). To determine the effective cytotoxic concentration (IC) of the test compound, the plate was analyzed by measuring the absorbance at 540 nm using a Synergy HTX plate reader (BioTek Instruments, Winooski, VT, USA). 50 ), and the data were plotted using a 4-parameter nonlinear regression model using GraphPad Prism7 software (GraphPad Software, San Diego, CA, USA).

[0851] The cell viability assay data in HMESO mesothelioma cells treated with compound (1)-HCl or (5)-HCl are summarized in Table 2 below. 50 (Drug concentration required to kill 50% of cells) is shown in μM concentration

[0852] Table 2.

[0853] TS (1)-HCl (5)-HCl DMSO <![CDATA[IC 50 ]]> 1.013 747.8 1.930* Unstable

[0854] *95% confidence interval (CI) 1.699-2.187

[0855] Example 41: Cross-linking studies

[0856] Malignant mesothelioma (MM) cells (H-MESO cell line) were seeded into 6-well plates in complete tissue culture medium. Cells were allowed to adhere for 24 hours and then treated with the specified concentrations of thiostrepton (TS), (1) or (5) (DMSO stock solution) for 24 hours. Cell lysates were generated using standard RIPA buffer, protein concentrations were determined using the Bradford assay, and 20 μg of total protein per sample was separated by reduced SDS-polyacrylamide gel electrophoresis. Proteins were transferred to PVDF membranes, blocked for 1 hour with 5% bovine serum albumin (BSA), and incubated overnight with PRX3 primary antibodies in 1X Tris buffered saline with Tween (TBST) at 4°C. The membranes were washed 3 times with 1X TBST and incubated with horseradish peroxidase-coupled (HRP) secondary antibodies for 1 hour at room temperature. The membranes were washed 3 times in 1X TBST and the HRP signal was developed using enhanced chemiluminescence and visualized on a GE digital imager.

[0857] Treatment of cells with 2.5 or 5 μM TS resulted in covalent PRX3 cross-linking (PRX3-X-PRX3). Similar but less robust results were observed for cells treated with (5) Figure 3 No PRX3-X-PRX3 crosslinking was observed in cells treated with (1) (see also Figure 3 ).

[0858] Example 42: Testing the Cytotoxic Activity of Compounds in Malignant Mesothelioma Cell Lines

[0859] Malignant mesothelioma (HMESO cell line) cells were seeded in 96-well plates (Corning, Kennebunk, ME, USA) at a density of 2500 cells per well. The next day, cells were treated with test compounds diluted in complete medium and then incubated for 48 hours (with technical replicates). After incubation, cells were washed with PBS (Corning Cellgro, Manassas, VA, USA), fixed with PBS containing 3.0% formaldehyde (Fisher BioReagents, Fair Lawn, NJ, USA), and stained with water containing 0.1% crystal violet (Acros Organics, Fair Lawn, NJ, USA) for 30 minutes. The crystal violet stain was removed, and the plate was washed with H2O and dried. To quantify cell viability, plates were imaged using a Lionheart plate reader (BioTek Instruments, Winooski, VT, USA) and / or analyzed by absorbance at 540 nm (crystal violet dye dissolved in 100% methanol) using a Synergy HTX plate reader (BioTek Instruments, Winooski, VT, USA). To determine the effective cytotoxic concentration (EC 50 ), the data were plotted using GraphPad Prism 7 software (GraphPad Software, San Diego, CA, USA) using a 4-parameter nonlinear regression model. The results are listed in Figure 5 middle.

[0860] Example 43: Covalent cross-linking of test compounds to recombinant peroxiredoxin 3 (rPRX3)

[0861] The master mix reagents in Table 3 were combined for 1X reactions in Eppendorf tubes on ice. The reactions were scaled to the number of test compounds tested. 16 μL of the master mix was added to a new Eppendorf tube containing 1 μL of the test compound (10 mM stock diluted in DMSO) and mixed by flicking and rapid centrifugation at 1,000 RPM. The reactions were incubated at 37°C for 18 hours. The reactions were removed from the incubation and quenched by adding 2 μL of Laemmli buffer containing 0.2 M dithiothreitol (DTT) and 10% sodium dodecyl sulfate. The samples were boiled at 98°C for 5 minutes. The samples were separated by polyacrylamide gel electrophoresis, transferred to PVDF membranes, and protein immunoblotted using anti-PRX3 antibodies (AbFrontier, LF-PA0255). The membrane was incubated with ECL reagent (ThermoScientific, Rockford, IL, USA) and visualized using the GE Amersham Imager chemiluminescent detection system. Unmodified rPRX3 was visualized as a single band at approximately 23 kDa, and rPRX3 covalently modified by the test compound was shown as a band of approximately 45 kDa. This was a qualitative assay to assess the presence or absence of a band of approximately 45 kDA. The qualitative results are listed in Figure 5 middle.

[0862] Table 3: Master mix

[0863] Reagents u 20mM Tris pH 8 11 rPRX3 (1 mg / ml) 2 0.5M TCEP 3

[0864] Example 44: Covalent cross-linking of test compounds to peroxiredoxin 3 (PRX3) in malignant mesothelioma cells

[0865] Human malignant mesothelioma (HMESO cell line) cells were seeded in 6-well plates at a density of 200,000 cells per well. After 24 hours, cells were treated with DMSO and test compounds diluted in cell culture medium. Cell lysates were collected 24 hours after treatment with RIPA buffer (50mMTris-HCl, 150mM NaCl, 1mM EDTA, 1% NP-40, 0.25% sodium deoxycholate, 0.1% sodium dodecyl sulfate in deionized (DI) water) for reduction of samples to be analyzed by reducing SDS-PAGE. Protein concentration was determined by Bradford assay (ThermoScientific, Rockford, IL, USA). Lysates (15 μg protein / well) were resolved by SDS-PAGE on a 4-12% gradient Bis-Tris Midi gel (Invitrogen, Carlsbad, CA, USA) under reducing conditions at a constant 200V for 50 minutes. The gel was transferred to a PVDF membrane at constant 1A for 50 minutes, blocked with 5% BSA diluted in 1× Tris-buffered saline with 1% Tween-20 (TBS-T) for a minimum of 1 hour, and incubated with anti-PRX3 antibody in 5% BSA TBS-T at 4°C overnight. The membrane was washed with 1× TBS-T for 1 hour, incubated with appropriate secondary antibodies for 1 hour, and washed again with 1× TBS-T for 1 hour. The membrane was incubated with ECL reagent (ThermoScientific, Rockford, IL, USA) and visualized using a GE Amersham Imager chemiluminescent detection system. Qualitative results are listed in Figure 5 middle.

[0866] Incorporation by reference

[0867] All publications and patents mentioned herein are hereby incorporated by reference in their entirety to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In the event of a conflict, the present application, including any definitions herein, will control.

[0868] Equivalent

[0869] In addition, the present invention encompasses all variations, combinations and arrangements of one or more of the limitations, elements, clauses and descriptive terms of one or more of the listed claims introduced into another claim. For example, any claim that depends on another claim can be modified to include one or more limitations found in any other claim that depends on the same basic claim. In the case where the element is presented in a list form, for example, in the format of a Markush group, each subgroup of the element is also disclosed, and any one or more elements can be deleted from the group. It should be understood that, in general, in the case where the present invention or aspects of the present invention are referred to as including specific elements and / or features, certain embodiments of the present invention or aspects of the present invention are composed of such elements and / or features or are substantially composed of such elements and / or features. For simplicity, those embodiments are not specifically listed in the verbatim statements herein. It should also be noted that the terms "comprising" and "containing" are intended to be open-ended, and allow the inclusion of additional elements or steps. In the case of giving a range, endpoints are included. In addition, unless otherwise indicated or apparent from the context and understanding of one of ordinary skill in the art, values ​​expressed as ranges may assume, in different embodiments of the invention, any specific value or sub-range within the recited range, up to the tenth of the unit on the lower limit of the range, unless the context clearly dictates otherwise.

[0870] The foregoing written description should be considered sufficient to enable one skilled in the art to practice the invention. The scope of the invention is not limited to the examples provided, because 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, other than those shown and described herein, will be apparent to one skilled in the art from the foregoing description and are within the scope of the appended claims. The advantages and purposes of the invention are not necessarily covered by every embodiment of the invention.

Claims

1. A compound having the structure of formula (IA): or a pharmaceutically acceptable salt thereof, wherein: R 1 is -NH2, -NH(CH3), -O-CH3 or -NH-CH2-C(O)-NH2; R 2 is -H, -CH3, =CH2 or =CH(alkyl); R 3 is -H, -CH3, =CH2 or =CH(alkyl); R 5 Yes-C(O)-R 1 or -CN; Ring A is aryl, heteroaryl, cycloalkyl or heterocyclyl; Ring B is absent or present, and when present is aryl, heteroaryl, cycloalkyl or heterocyclyl; R 4 is hydrogen, a protecting group, -C(O)-CH3, -L' or -LY; L', when present, is a reactive linker moiety; L, when present, is a linker moiety; Y, when present, is the mitochondrial targeting moiety; Each are independently a single bond or a double bond; and Any hydrogen atoms are optionally replaced with deuterium.

2. The compound of claim 1, wherein R 5 Yes-C(O)-R 1 ; and R 1 It is -OCH3.

3. The compound as claimed in claim 1, wherein R 5 Yes-C(O)-R 1 ; and R 1 It is -NH2.

4. The compound of any one of claims 1 to 3, wherein when Ring A is polycyclic, Ring B is absent.

5. The compound according to any one of claims 1 to 4, wherein the compound has the structure of formula (IA-1): in R 2 is -H or -CH3; and R 3 It is =CH2 or =CH(alkyl).

6. The compound as claimed in claim 5, wherein R 2 Yes -H and R 3 =CH2, or R 2 is -CH3 and R 3 Yes =CH2.

7. The compound according to any one of claims 1 to 4, wherein the compound has the structure of formula (IA-2): in R 2 is =CH2 or =CH(alkyl); and R 3 It is -H or -CH3.

8. The compound of claim 7, wherein R 2 =CH2 and R 3 Yes -H, or R 2 =CH2 and R 3 It is -CH3.

9. The compound according to any one of claims 1 to 4, wherein the compound has the structure of formula (IA-3): in R 2 is =CH2 or =CH(alkyl); and R 3 It is =CH2 or =CH(alkyl).

10. The compound of claim 9, wherein R 2 =CH2 and R 3 Y = CH2, R 2 =CH2 and R 3 =CH(CH3), or R 2 =CH(CH3) and R 3 Yes =CH2.

11. The compound of claim 1, wherein the compound has the structure of formula (IB): or a pharmaceutically acceptable salt thereof, wherein: R 1 is -NH2 or -O-CH3; R 2 is CH2 or CH(alkyl); R 3 is CH2 or CH(alkyl); Ring A is heteroaryl, cycloalkyl or heterocyclyl; Ring B is aryl, heteroaryl, cycloalkyl or heterocyclyl; R 4 is hydrogen, a protecting group, -C(O)-CH3, -L' or -LY; L', when present, is a reactive linker moiety; L, when present, is a linker moiety; Y, when present, is a mitochondrial targeting moiety; and Any hydrogen atoms are optionally replaced with deuterium.

12. The compound of claim 11, wherein R 2 and R 3 are different.

13. The compound of claim 11, wherein R 2 and R 3 are the same.

14. A compound as described in any one of claims 11 to 13, wherein R 2 It is CH(Me) or -CH2.

15. A compound as described in any one of claims 11 to 13, wherein R 3 It is CH(Me) or -CH2.

16. The compound of claim 13, wherein R 2 and R 3 Each is CH2.

17. The compound of any one of claims 1 to 16, wherein Ring A is a 5-membered ring or a 5-membered ring fused to a second ring.

18. The compound of any one of claims 1-16, wherein Ring A is a 5-membered heteroaryl.

19. The compound of claim 18, wherein ring A is thiazolyl, thienyl, oxazolyl or imidazolyl.

20. The compound of claim 19, wherein Ring A is thiazolyl, thienyl or oxazolyl.

21. The compound of claim 20, wherein Ring A is thiazolyl.

22. The compound of claim 18, wherein Ring A is: Wherein * represents the bond to ring B.

23. The compound of claim 18, wherein Ring A is: Wherein * represents the bond to ring B.

24. The compound of any one of claims 1-16, wherein Ring A is a 5-membered cycloalkyl or heterocyclyl.

25. The compound of claim 24, wherein Ring A is cyclopentyl or tetrahydrofuranyl.

26. The compound of claim 25, wherein ring A is Wherein * represents the bond to ring B.

27. The compound of any one of claims 1-16, wherein Ring A is a bridged bicyclic cycloalkyl or heterocyclyl.

28. The compound of claim 27, wherein Ring A is bicyclo[2.1.1]hexyl or oxabicyclo[2.1.1]hexyl.

29. The compound of claim 28, wherein ring A is Wherein * represents the bond to ring B.

30. The compound of any one of claims 1-16, wherein Ring A is phenyl.

31. The compound of claim 30, wherein Ring A is Wherein * represents the bond to ring B.

32. The compound of any one of claims 1-16, wherein Ring A is a polycyclic aryl, heteroaryl, cycloalkyl or heterocyclyl.

33. The compound of claim 32, wherein Ring A is a bicyclic heteroaryl.

34. The compound of claim 33, wherein Ring A is: Where *** represents -NH-R 4 .

35. The compound of any one of claims 1-31, wherein Ring B is a 6-membered ring.

36. The compound of claim 35, wherein Ring B is phenyl.

37. The compound of claim 36, wherein Ring B is unsubstituted phenyl.

38. The compound of claim 36, wherein Ring B is phenyl substituted with halogen.

39. The compound of claim 37 or 38, wherein Ring B is: wherein Z is a halogen group and ** represents a bond to ring A.

40. The compound of claim 35, wherein Ring B is a 6-membered heteroaryl.

41. The compound of claim 40, wherein Ring B is pyridyl or pyrazinyl.

42. The compound of claim 41, wherein Ring B is: Wherein ** represents the bond to ring A.

43. The compound of any one of claims 1-31, wherein Ring B is a bridged bicyclic cycloalkyl.

44. The compound of claim 43, wherein Ring A is bicyclo[2.2.2]octyl or bicyclo[1.1.1]pentyl.

45. The compound of claim 44, wherein Ring B is: wherein Z is a halogen group and ** represents a bond to ring A.

46. ​​A compound as described in any one of claims 1-45, wherein R 4 is hydrogen, a protecting group or -C(O)-CH3.

47. The compound of claim 1, selected from the group consisting of: Where R 4 is hydrogen, a protecting group or -C(O)-CH3; and The drawing is The bond represents either possible stereochemistry of the attached olefin, E or Z.

48. The compound of claim 1, selected from the group consisting of: Where R 4 is hydrogen, a protecting group or -C(O)-CH3.

49. The compound of claim 1, selected from the group consisting of:

50. The compound of claim 1, selected from the group consisting of: Where R 4 is hydrogen, a protecting group or -C(O)-CH3; and wherein Z is selected from fluorine, chlorine, bromine and iodine.

51. The compound of claim 1, selected from the group consisting of: Where R 4 is hydrogen, a protecting group or -C(O)-CH3.

52. The compound of claim 51, wherein the protecting group is Boc.

53. A compound as described in any one of claims 1-45, wherein R 4 Yes -L'.

54. The compound of claim 53, wherein: L' is -C(O)-XC(O)OH or -C(O)-XC(O)NH2; X is -(CH2) n -;and n is 2, 3, 4, or 5.

55. The compound of claim 54, selected from the group consisting of: Where X is -(CH2) n -;and n is 2, 3, 4, or 5.

56. The compound of claim 54, selected from the group consisting of: Where X is -(CH2) n -;and n is 2, 3, 4, or 5.

57. A compound as described in any one of claims 1-45, wherein R 4 Yes-LY.

58. The compound of claim 57, wherein L is a cleavable linker.

59. The compound of claim 57, wherein L is a non-cleavable linker.

60. The compound of any one of claims 57-59, wherein L has a chain length of about 2 to about 30 atoms.

61. The compound of claim 60, wherein L has a chain length of about 5 to about 20 atoms.

62. The compound of claim 57, wherein: L is -C(O)-XC(O)-; X is -(CH2) n -;and n is 2, 3, 4, or 5.

63. The compound of claim 57, wherein: L is -C(O)-XC(O)-; X is -(CH2CH2-O-) m -(CH2CH2)-; and m is 2, 3, 4, 5 or 6.

64. The compound of claim 1, selected from the group consisting of: Where X is -(CH2) n -;and n is 2, 3, 4, or 5.

65. The compound of claim 1, selected from the group consisting of: Where X is -(CH2) n -; m is 2, 3, 4, 5, or 6; and n is 2, 3, 4, or 5.

66. The compound of claim 53, wherein L' comprises an alkynyl group or an azido group.

67. The compound of claim 66, wherein R 4 is -C(O)-X'-C≡CH or -C(O)-X'-N3; X' is -(CH2) n -;and n is 2, 3, 4, or 5.

68. The compound of claim 57, wherein L comprises a heteroaryl group.

69. The compound of claim 68, wherein L comprises a triazole group.

70. The compound of claim 69, wherein R 4 yes R 6 is -H or -C(O)CH3; X' is -(CH2) n -; X" is -(CH2) o -; n is 2, 3, 4, or 5; and o is 2, 3, 4, or 5.

71. The compound of claim 70, wherein R 4 yes R 6 is -H or -C(O)CH3; X' is -(CH2) n -; X" is -(CH2) o -; n is 2, 3, 4, or 5; and o is 2, 3, 4, or 5.

72. The compound of claim 69, wherein R 4 yes X' is -(CH2) n -; X" is -(CH2) o -; n is 2, 3, 4, or 5; and o is 2, 3, 4, or 5.

73. The compound of any one of claims 57-72, wherein: Y is a berberine cation, a rhodamine cation, an indolium cation, a pyridinium cation, a tetraguanidinium cation, a cyanine derivative, a guanidinium cation, a biguanidinium cation, a triphenylphosphonium cation, a triethylammonium cation, a triphenylamine, a tetraphenylethylene moiety, an arylphosphonium cation, an SS peptide, a mitochondrial penetrating peptide (MPP), a mitochondrial targeting sequence (MTS) peptide, a hemigramicidin S-linked nitroxide, a dequalinium chloride (DQA) cation, a nonlocalized lipophilic cation, F16((E)-4 -(1H-indol-3-ylvinyl)-N-methylpyridinium iodide), (L-cyclohexylalanine-D-arginine)3, mitochondrial targeting nanocarriers, DDDK peptide, glycyrrhetinic acid, α-tocopheryl succinate (α-TOS), graphene oxide nanocarriers, PEG-pro-apoptotic peptide (KLAKLAK)2, Dmt-D-Arg-Phe-Lys-NH2 peptide, methylglyoxal, N-nonylacridine orange, quinoline, styryl fluorophore or 15d-PGJ2.

74. The compound of claim 73, wherein Y is a mitochondrial penetrating peptide (MPP).

75. The compound of claim 73, wherein Y has the structural formula (V):

76. The compound of claim 1, selected from the group consisting of: or a pharmaceutically acceptable salt thereof; Where X is [-(CH2) n -] or [-(CH2CH2-O-) m -(CH2CH2)-]; n is 3, 4, or 5; and m is 2, 3, 4, 5 or 6.

77. A pharmaceutically acceptable composition comprising a compound according to any one of claims 1-76; and a pharmaceutically acceptable carrier.

78. The composition of claim 77, formulated for oral or parenteral delivery.

79. A composition comprising a compound as described in any one of claims 1-76, wherein the compound is contained within a nanoparticle, a liposome or a micelle, wherein the nanoparticle, the liposome or the micelle is conjugated to a mitochondrial targeting moiety.

80. A composition comprising a compound of formula (IA), wherein R 4 is hydrogen, a protecting group or -C(O)-CH3, wherein the compound is contained within a nanoparticle, a liposome or a micelle, wherein the nanoparticle, the liposome or the micelle is conjugated to a mitochondrial targeting moiety.

81. A composition comprising a compound of formula (IB), wherein R 4 is hydrogen, a protecting group or -C(O)-CH3, wherein the compound is contained within a nanoparticle, a liposome or a micelle, wherein the nanoparticle, the liposome or the micelle is conjugated to a mitochondrial targeting moiety.

82. The composition of any one of claims 79-81, wherein the nanoparticle, the liposome or the micelle is selected from poly(ethylene glycol), poly(ε-caprolactone), a polysaccharide, poly[(2-hydroxypropyl)-methacrylic acid], poly(lactic acid-co-glycolic acid), and any combination of the foregoing.

83. A method of treating cancer (e.g., a solid tumor or a hematological cancer), the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-76 or a composition of any one of claims 77-82.

84. The method of claim 83, wherein the cancer (solid tumor or hematological cancer) is selected from lung cancer, breast cancer, prostate cancer, melanoma, esophageal cancer, leukemia, cervical cancer, liver cancer, colon cancer, gastric cancer, colorectal cancer, glioblastoma, head and neck cancer, pancreatic cancer, mesothelioma and ovarian cancer.

85. The method of claim 84, wherein the cancer is selected from the group consisting of mesothelioma, lung cancer, ovarian cancer, and breast cancer.

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