Thiostrepton-inspired compounds for cancer treatment and their preparation

Abstract

Disclosed herein are structures, compositions, and syntheses of pharma- ceutically relevant compounds inspired by thiostrepton, and methods of using such compounds to treat cancer.

Description

[Background technology]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 310400, filed February 15, 2022, and U.S. Provisional Patent Application No. 63 / 341238, filed May 12, 2022, each of which is incorporated by reference in its entirety herein.

[0002] Thiostrepton is a cyclic oligopeptide antibiotic also known as briamycin, tiactin, alaninamide, and HR4S203Y18. Recent studies have shown that thiostrepton also has promising anticancer activity. There remains a need for thiostrepton derivatives with beneficial pharmacological properties. Summary of the Invention

[0003] In certain embodiments, the present invention provides a series of compounds having the structure of formula (IA): [ka] or a pharma- ceutically 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 is -C(O)-R 1 or -CN, Ring A is aryl, heteroaryl, cycloalkyl, or heterocyclyl; Ring B is absent or present and, if present, is aryl, heteroaryl, cycloalkyl, or heterocyclyl; R 4is hydrogen, a protecting group, -C(O)-CH3, -L', or -LY; L', if present, is a reactive linker moiety; L, when present, is a linker moiety; Y, if present, is a mitochondrial targeting moiety, [ka] are each independently a single bond or a double bond; and Any hydrogen atoms are optionally replaced with deuterium.

[0004] In certain embodiments, the present invention also provides a series of compounds having the structure of formula (IB): [ka] or a pharma- ceutically 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', if present, is a reactive linker moiety; L, when present, is a linker moiety; Y, if present, is a mitochondrial targeting moiety, and Any hydrogen atoms are optionally replaced with deuterium.

[0005] Also provided herein is a method of treating cancer, comprising administering to a subject in need of cancer treatment any of the pharmaceutical compositions described herein. [Brief description of the drawings]

[0006] [Figure 1] Cell viability assay data in malignant mesothelioma (MM) cells (HMESO cell line) treated with compounds (1)-HCl or (5)-HCl. N=2 technical replicates and 2 biological replicates performed by 2 different lab technicians. All compounds were dissolved in 100% DMSO. When (5)-HCl was added to complete tissue culture medium, it precipitated at concentrations of approximately 20-80 μM. [Figure 2A] (5)-Image of HMESO mesothelioma cells treated with HCl (magnification 40x). [Figure 2B] (5)-Image of HMESO mesothelioma cells treated with HCl (magnification 20x). [Diagram 3] Western blot of PRX3 from HMESO mesothelioma cells treated with (1) or (5). [Figure 4] Bar graph of PRX3 (PRX3-X-PRX3):monomer ratio in HMESO mesothelioma cells treated with (1) or (5). [Figure 5-1] 1 is a table summarizing the results of Examples 42-44 for various compounds described herein. NT indicates "not tested." For EC50 values, "A" represents an EC50 of 5.0 μM or less, "B" represents an EC50 of 5.1 μM to 10.0 μM, "C" represents an EC50 of 10.1 μM to 50.0 μM, and "D" represents an EC50 of greater than 50.0 μM. [Figure 5-2] 1 is a table summarizing the results of Examples 42-44 for various compounds described herein. NT indicates "not tested." For EC50 values, "A" represents an EC50 of 5.0 μM or less, "B" represents an EC50 of 5.1 μM to 10.0 μM, "C" represents an EC50 of 10.1 μM to 50.0 μM, and "D" represents an EC50 of greater than 50.0 μM. [Figure 5-3]1 is a table summarizing the results of Examples 42-44 for various compounds described herein. NT indicates "not tested." For EC50 values, "A" represents an EC50 of 5.0 μM or less, "B" represents an EC50 of 5.1 μM to 10.0 μM, "C" represents an EC50 of 10.1 μM to 50.0 μM, and "D" represents an EC50 of greater than 50.0 μM. [Figure 5-4] 1 is a table summarizing the results of Examples 42-44 for various compounds described herein. NT indicates "not tested." For EC50 values, "A" represents an EC50 of 5.0 μM or less, "B" represents an EC50 of 5.1 μM to 10.0 μM, "C" represents an EC50 of 10.1 μM to 50.0 μM, and "D" represents an EC50 of greater than 50.0 μM. [Figure 5-5] 1 is a table summarizing the results of Examples 42-44 for various compounds described herein. NT indicates "not tested." For EC50 values, "A" represents an EC50 of 5.0 μM or less, "B" represents an EC50 of 5.1 μM to 10.0 μM, "C" represents an EC50 of 10.1 μM to 50.0 μM, and "D" represents an EC50 of greater than 50.0 μM. [Figure 5-6] 1 is a table summarizing the results of Examples 42-44 for various compounds described herein. NT indicates "not tested." For EC50 values, "A" represents an EC50 of 5.0 μM or less, "B" represents an EC50 of 5.1 μM to 10.0 μM, "C" represents an EC50 of 10.1 μM to 50.0 μM, and "D" represents an EC50 of greater than 50.0 μM. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] Pharmaceutical Compositions The compositions and methods described herein may be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal, such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or compound is preferably administered as a pharmaceutical composition, for example, comprising a compound described herein and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, by way of non-limiting example, aqueous solutions, such as water or physiologically buffered saline, or other solvents or vehicles, such as glycols, glycerol, oils, such as olive oil, or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are for human administration, particularly invasive routes of administration (i.e., routes such as injection or implantation that avoid transport or diffusion through epithelial barriers), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients may be selected, for example, to provide delayed release of the agent or to selectively target one or more cells, tissues, or organs. The pharmaceutical composition may be in dosage unit form, such as tablets, capsules (including sprinkle capsules and gelatin capsules), granules, lyophilisates for reconstitution, powders, solutions, syrups, suppositories, injections, etc. The composition may also be present in a transdermal delivery system, such as a skin patch. The composition may also be present in a solution suitable for topical administration, such as a lotion, cream, or ointment.

[0008] A pharma- ceutically acceptable carrier can contain a physiologically acceptable agent that acts to stabilize, increase the solubility, or increase the absorption of a compound, such as, for example, a compound described herein. 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 pharma- ceutical carrier that includes a physiologically acceptable agent depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition can be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) can be a liposome or other polymer matrix, which can incorporate, for example, a compound described herein. For example, liposomes containing phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively easy to make and administer.

[0009] As used herein, the phrase "pharmacologically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are suitable, within the scope of sound medical judgment, 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.

[0010] The phrase "pharmacologically acceptable carrier" as used herein means a pharma- ceutically 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 harmful to the patient. Some examples of substances that can function as pharma-ceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and derivatives thereof, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil. (10) glycols such as propylene glycol; (11) polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffers; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0011] The pharmaceutical compositions (preparations) can be administered to a subject by any of a number of routes of administration, including, for example, orally (e.g., as drenches such as aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue), absorption through the oral mucosa (e.g., sublingually), subcutaneously, transdermally (e.g., as a patch applied to the skin), and topically (e.g., as a cream, ointment, or spray applied to the skin). The compounds may also be formulated for inhalation. In certain embodiments, the compounds may simply be dissolved or suspended in sterile water. Details of suitable routes of administration and compositions suitable therefor can be found, for example, in U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970, and 4,172,896, and patents cited therein.

[0012] The formulations may be conveniently provided in unit dosage form and may be prepared by any method well known in the art of pharmacy. The amount of active ingredient that can be combined with carrier materials to produce a single dosage form varies depending on the host treated, the particular 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 compound that produces a therapeutic effect. Generally, out of 100 percent, this amount will range from about 1 percent to about 99 percent of active ingredient, preferably from about 5 percent to about 70 percent, and most preferably from about 10 percent to about 30 percent.

[0013] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound described herein, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound described herein with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0014] Formulations described herein suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), lyophilisates, 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 troche (using an inert base such as gelatin and glycerin, or sucrose and acacia), and / or mouthwash, etc., each containing a predetermined amount of a compound described herein as an active ingredient. The composition or compound may also be administered as a bolus, electuary, or paste.

[0015] To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), the active ingredient can be combined with one or more pharma- ceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, such as glycerol; (4) disintegrants, such as agar, calcium carbonate, etc. , potato starch or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) dissolution retarders, such as paraffin; (6) absorption enhancers, such as quaternary ammonium compounds; (7) wetting agents, such as cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets, and pills, the pharmaceutical composition may also contain buffering agents. Solid compositions of a similar type may also be used as fillers for soft and hard-filled gelatin capsules, using excipients such as lactose or milk sugar, and high molecular weight polyethylene glycols, and the like.

[0016] Tablets can be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surface active agents or dispersants. Molded tablets can be made by molding a mixture of the powdered compound moistened with an inert liquid diluent in a suitable machine.

[0017] Tablets and other solid dosage forms of pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may be optionally scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may also be formulated to provide sustained or controlled release of the active ingredient therein, for example, with hydroxypropylmethylcellulose in various proportions to provide the desired release profile, with other polymer matrices, with liposomes, and / or with microspheres. They may be sterilized, for example, by filtration through a bacteria-retaining filter, or by 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 immediately before use. These compositions may also optionally contain opacifying agents, and may be of a composition that releases the active ingredient(s) 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. The active ingredient can also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0018] Liquid dosage forms useful for oral administration include pharma- ceutically acceptable emulsions, lyophilisates for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs.In addition to the active ingredient, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, cyclodextrin and its derivatives, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, sesame oil), glycerol, tetrahydrofuran alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof.

[0019] 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.

[0020] Suspensions may contain, in addition to the active compounds, suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.

[0021] 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 pharma- ceutical acceptable carrier, and any preservatives, buffers, or propellants which may be required.

[0022] The ointments, pastes, creams and gels may contain, in addition to the active compound, excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc and zinc oxide, or mixtures thereof.

[0023] Powders and sprays can contain, in addition to the active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0024] Transdermal patch has the additional advantage of providing controlled delivery of the compound described herein to the body.Such dosage forms can be made by dissolving or dispersing active compound in suitable medium.Absorption enhancers can also be used to increase the flux of compound through the skin.The rate of such flux can be controlled by either providing a rate-controlling membrane or dispersing compound in a polymer matrix or gel.

[0025] The phrases "parenteral administration" and "administered parenterally" as used herein refer to modes of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration include one or more active compounds in combination with one or more pharma- ceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted into a sterile injectable solution or dispersion immediately prior to use, which may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0026] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions described herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate, etc. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0027] These compositions may contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of microbial activity may be ensured by the inclusion of various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents in the composition, such as sugars, sodium chloride, and the like. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption, such as aluminum monostearate and gelatin.

[0028] In some cases, it is desirable to delay the absorption of a drug from subcutaneous or intramuscular injection in order to prolong the effect of the drug. This can be accomplished by using a liquid suspension of crystalline or amorphous material that is poorly water-soluble. In this case, the rate of absorption of the drug depends on the rate of dissolution, which in turn may depend on the crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form can be accomplished by dissolving or suspending the drug in an oil vehicle.

[0029] Injectable depot forms are made by forming microencapsulated matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. The rate of drug release can be controlled depending on the ratio of drug to polymer and the nature of the particular polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0030] For use in the methods described herein, the active compound may be provided per se or as a pharmaceutical composition containing, for example, 0.1-99.5% (more preferably 0.5-90%) of the active ingredient in combination with a pharma- ceutically acceptable carrier.

[0031] The introduction method may be provided by a rechargeable or biodegradable device. For the controlled delivery of drugs, including proteinaceous biopharmaceuticals, various sustained release polymeric devices have been developed and tested in vivo in recent years. Various biocompatible polymers, including both biodegradable and non-degradable polymers (including hydrogels), can be used to form implants for sustained release of compounds at specific target sites.

[0032] Actual dosage levels of the active ingredients in pharmaceutical compositions may be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without being toxic to the patient.

[0033] The selected dosage level will depend on a variety of factors, including the activity of the particular compound or combination of compounds, or esters, salts, or amides thereof, used, the route of administration, the time of administration, the excretion rate of the particular compound(s) used, the duration of treatment, other drugs, compounds, and / or materials used in combination with the particular compound(s) used, the age, sex, weight, condition, general health, and previous medical history of the patient being treated, and similar factors well known in the medical arts.

[0034] A physician or veterinarian of ordinary skill in the art can easily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, a physician or veterinarian can begin administration of a pharmaceutical composition or compound at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dose until the desired effect is achieved. By "therapeutically effective amount" is meant the concentration of the compound sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of a compound varies according to the subject's weight, sex, age, and medical history. Other factors that affect the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent administered together with the compound described herein. Multiple administrations of the drug can deliver a larger total dose. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13 ed., 1814-1882, incorporated herein by reference).

[0035] In general, a suitable daily dose of an active compound used in the compositions and methods described herein will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

[0036] If desired, the effective daily dose of active compound can be administered as 1, 2, 3, 4, 5, 6 or more separate partial doses at appropriate intervals throughout the day, optionally in unit dosage form.In certain embodiments, active compound can be administered 2 or 3 times a day.In preferred embodiments, active compound is administered once a day.

[0037] The patients receiving this treatment are animals in need of treatment, including primates, particularly humans, and other mammals such as horses, cows, pigs, sheep, cats, dogs, poultry, and pets in general.

[0038] In certain embodiments, the compounds described herein can be administered alone or in combination with another type of therapeutic agent.

[0039] The present disclosure includes the use of pharma- ceutically acceptable salts of the compounds described herein in the compositions and methods described herein.In certain embodiments, contemplated salts include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkyl ammonium salts.In certain embodiments, contemplated salts include, but are not limited to, L-arginine, benentamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, 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, contemplated salts include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.In certain embodiments, contemplated salts include 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, (+)-camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptone, Acids, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hipprinic 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 undecylenic acid salts.

[0040] Pharmaceutically acceptable acid addition salts may also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, etc. Mixtures of such solvates may also be prepared. The source of such solvates may be inherent in the solvent of crystallization, the solvent of preparation or crystallization, or adventitious to such solvent.

[0041] Wetting agents, emulsifying agents, and lubricating agents such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring, and perfuming agents, preservatives, and antioxidants can also be present in the composition.

[0042] Examples of pharma- ceutically acceptable antioxidants include: (1) water-soluble antioxidants such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium disulfite, and sodium sulfite; (2) lipid-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, and alpha-tocopherol; and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, and phosphoric acid.

[0043] definition Unless otherwise defined herein, 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 used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry described herein are those well known and commonly used in the art.

[0044] The methods and techniques of the present disclosure are generally carried out according to conventional methods well known in the art and as described in various general and more specific references cited and discussed throughout the specification, unless otherwise indicated.

[0045] Chemical terms used herein, unless otherwise defined herein, are used according to conventional usage in the art, as exemplified in "The McGraw-Hill Dictionary of Chemical Terms", Parker S., Ed., McGraw-Hill, San Francisco, CA (1985).

[0046] 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 case of conflict, the present specification, including its specific definitions, will control.

[0047] The term "agent" is used herein to denote a chemical compound (e.g., an organic or inorganic compound, a mixture of compounds), a biological macromolecule (e.g., nucleic acids, antibodies including portions thereof, as well as humanized, chimeric and human antibodies, and monoclonal antibodies, proteins or portions thereof, e.g., peptides, lipids, carbohydrates), or an extract made from biological material such as bacteria, plants, fungi, or animal (especially mammalian) cells or tissues. Agents include, for example, agents of known structure and agents of unknown structure.

[0048] "Patient," "subject," or "individual" are used interchangeably and refer to either a human or non-human animal. These terms include mammals, such as humans, primates, livestock animals (including cows, pigs, etc.), companion animals (e.g., dogs, cats, etc.), and rodents (e.g., mice and rats).

[0049] "Treating" a condition or patient refers to taking measures to obtain beneficial or desired results (including clinical results). As used herein, and as is well understood in the art, "treatment" is an approach to obtain beneficial or desired results (including clinical results). Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, whether detectable or undetectable, reduction in the extent of disease, stabilization of the disease state (i.e., not worsening), prevention of disease spread, delay or slowing of disease progression, improvement or palliation of the disease state, and remission (whether partial or total). "Treatment" can also mean prolonging survival compared to the expected survival if not receiving treatment.

[0050] The term "prevention" is art-recognized and, when used in reference to a condition, such as local recurrence (e.g., pain), a disease such as cancer, a complex syndrome such as heart failure, or any other medical condition, is well understood in the art and includes administration of a composition that reduces the frequency of or delays the onset of symptoms of a medical condition in a subject compared to subjects not receiving the composition. Thus, prevention of cancer includes, by way of example, reducing the number of detectable cancerous growths in a population of patients receiving a prophylactic treatment compared to an untreated control population, e.g., by a statistically and / or clinically significant amount, and / or delaying the appearance of detectable cancerous growths in a treated population versus an untreated control population.

[0051] "Administering" 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, ophthalmically, sublingually, orally (by ingestion), intranasally (by inhalation), intrathecally, intracerebrally, and transdermally (e.g., by absorption through the skin's channels). The compound or agent can also be suitably introduced by rechargeable or biodegradable polymeric or other devices, such as patches and pumps, or formulations that provide an extended, gradual or controlled release of the compound or agent. Administration can also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0052] The appropriate method of administering a substance, compound, or agent to a subject also depends, for example, on the age and / or physical condition of the subject, as well as the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability, and toxicity). In some embodiments, the compound or agent is administered orally, for example, by ingestion, to the subject. In some embodiments, the orally administered compound or agent is in a sustained or extended release formulation or is administered using a device for such sustained or extended release.

[0053] As used herein, the phrase "co-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 formulation is detected systemically, as per industry accepted methods, or when the two agents are effective simultaneously in the patient, which may include synergistic effects of the two agents). For example, different therapeutic compounds can be administered either simultaneously or sequentially, either in the same formulation or in separate formulations. In certain embodiments, different therapeutic compounds can be administered within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, or 1 week of each other. Thus, individuals receiving such treatment can benefit from the combined effects of different therapeutic agents.

[0054] A "therapeutically effective amount" or "therapeutically effective dose" of a compound or other agent described herein is an amount that, when administered to a subject, causes the drug or agent to have the intended therapeutic effect. The full therapeutic effect does not necessarily occur by administration of a single dose of such drug or agent, but may occur only after administration of a series of doses (multiple consecutive administrations). Thus, a therapeutically effective amount may be administered in one or more administrations. The exact 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 condition, such as cancer, being treated.

[0055] As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes examples when the event or circumstance occurs as well as examples when it does not occur. For example, "optionally substituted alkyl" refers to the fact that the alkyl may be substituted as well as the fact that the alkyl is not substituted.

[0056] 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.

[0057] The phrase "pharmacologically acceptable" is art-recognized. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers, and other materials and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0058] The terms "pharmacologically acceptable salt" or "salt" are used herein to refer to acid addition salts or base addition salts which are suitable or compatible with the treatment of a patient.

[0059] The term "pharmaceutically acceptable acid addition salt" as used herein means any non-toxic organic or inorganic salt of any base. Examples of inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Exemplary organic acids that form suitable salts include mono-, di-, 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, as well as sulfonic acids, such as p-toluenesulfonic acid and methanesulfonic acid. Mono- and di-acid salts may be formed, and such salts may exist in either hydrated, solvated, or substantially anhydrous form. In general, acid addition salts of compounds are more soluble in water and various hydrophilic organic solvents and generally exhibit higher melting points than their free base forms. The selection of suitable salts will be known to those skilled in the art. Other pharma-ceutically unacceptable salts, such as oxalates, may be used, for example, in the isolation of compounds for laboratory use or for subsequent conversion to a pharma-ceutically acceptable acid addition salt.

[0060] As used herein, the term "pharmaceutically acceptable base addition salt" refers to any non-toxic organic or inorganic base addition salt of any acid compound or any of its intermediates. Exemplary inorganic bases that form suitable salts include lithium, sodium, potassium, calcium, magnesium, or barium hydroxide. Exemplary organic bases that form suitable salts include aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine, and picoline or ammonia. The selection of appropriate salts is within the skill of the art.

[0061] Many of the compounds useful in the methods and compositions of the present disclosure have at least one stereocenter in their structure. This stereocenter may exist in either the R or S configuration, and the R and S designations are used according to the rules set forth in Pure Appl.Chem.(1976),45,11-30. The present disclosure contemplates all stereoisomeric forms, such as enantiomers and diastereomers, of the compounds, salts, prodrugs, or mixtures thereof, including all possible mixtures of stereoisomers. See, for example, WO01 / 062726.

[0062] Some compounds may exist in tautomeric forms and such forms, although not explicitly shown in the formulae set forth herein, are intended to be included within the scope of the present disclosure.

[0063] "Prodrug" or "Pharmaceutically acceptable prodrug" refers to a compound that is metabolized in the host after administration, e.g., hydrolyzed or oxidized, to form a compound of the present disclosure. Typical examples of prodrugs include compounds that have a biologically labile or cleavable (protecting) group on a functional moiety of the active compound. Prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated to yield an active compound. Examples of prodrugs that use esters or phosphoramidates as biologically labile or cleavable (protecting) groups are disclosed in U.S. Patent Nos. 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.

[0064] As used herein, the phrase "pharmaceutical acceptable carrier" means a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filter, diluent, excipient, solvent, or encapsulating material useful in formulating a drug for pharmaceutical or therapeutic use.

[0065] The term "linker," as used herein, means any chemical functional group that "connects" or chemically bonds any two or more other chemical functional groups in a pharma- ceutical related molecule. As a non-limiting example of the use of linkers in medicine, antibody-drug conjugations (ADCs) involve a pharma- ceutical active small molecule, drug, or toxin connected by a linker to a larger molecule, an antibody.

[0066] Examples of common linker types include both cleavable and non-cleavable linkers. Cleavable linkers contain chemical functional groups that can be cleaved in response to physiological stimuli such as chemical gradients, pH changes, or enzyme activity. Non-limiting examples include acid or base labile functional groups, pyrophosphate diesters, disulfide bonds, peptides, β-glucuronides, and the like. Non-cleavable linkers generally contain chemical functional groups that are less labile to the aforementioned physiological stimuli, non-limiting examples include certain alkyl groups and organic polymer functional groups.

[0067] As used herein, the term "reactive linker moiety" refers to a chemical structure having a terminal moiety that can react with another moiety (e.g., a mitochondrial targeting moiety) to form a covalent bond.

[0068] As used herein, the terms "mitochondrial targeting peptide", "mitochondrial targeting sequence", and "mitochondrial targeting moiety" are art-recognized terms that refer to chemical functional groups (peptides, sequences, or moieties) that are "targeted" to the mitochondrial membrane, i.e., are readily transported to and absorbed into the mitochondrial membrane (J. Zielonka, B. Kalyanaraman, et al., 2017). As used herein, mitochondrial targeting moieties may include, but are not limited to, the following species: berberine cation, rhodamine cation, indolium cation, pyridinium cation, tetraguanidinium cation, cyanine derivatives, guanidinium cation, biguanidinium cation, triphenylphosphonium cation, triethylammonium cation, triphenylamine, tetraphenylethene moiety, arylphosphonium cation, SS peptide, mitochondrial penetrating peptide (MPP), mitochondrial targeting sequence (MTS) peptide, hemigramicidin S-linked nitroxide, dequalinium (DQA) cation, delocalized lipophilic cation, F16 ((E)-4-(1H-indol-3-ylvinyl)-N-methylpyridinium iodide), (L-cyclohexylalanine-D-arginine)3, mitochondria-targeted nanocarriers, DDDK peptide, glycyrrhetinic acid, α-tocopheryl succinate (α-TOS), graphene oxide nanocarriers, PEG-proapoptotic peptide (KLAKLAK)2, Dmt-D-Arg-Phe-Lys-NH2 peptide, pyruvaldehyde, N-nonyl acridine orange, quinoline, styryl-azinium fluorophore, or 15d-PGJ2. Exemplary mitochondrial targeting moieties are listed in See. J Zielonka et al, Chem Rev 2017, 117, p10043-10120; KL Horton et al, Chemistry & Biology 2008, 15, pp375-382; 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.

[0069] It is understood that the substituents and substitution patterns on the compounds described herein can be selected by those skilled in the art to provide chemically stable compounds that can be readily synthesized from readily available starting materials using techniques known in the art as well as the methods described below. When a substituent is itself substituted with more than one group, it is understood that these multiple groups can be on the same carbon or on different carbons, so long as a stable structure results.

[0070] As used herein, the term "optionally substituted" refers to the replacement of one to six hydrogen radicals in a given structure with the radical of a specified substituent, including, but not limited to, hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH-O-alkyl, -OP(O)(O-alkyl), or -CH-OP(O)(O-alkyl).

[0071] Preferably, "optionally substituted" refers to the replacement of 1-4 hydrogen radicals in a given structure with a substituent as described above. More preferably, 1-3 hydrogen radicals are replaced with a substituent as described above. It is understood that the substituents may be further substituted.

[0072] The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0073] The term "acylamino" is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbylC(O)NH-.

[0074] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0075] 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.

[0076] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

[0077] As used herein, the term "alkyl" includes, but is not limited to, C1 to C 10 Straight chain alkyl group or C1-C 10 It refers to saturated aliphatic groups, including branched alkyl groups. Preferably, the "alkyl" group refers to a C1-C6 straight chain alkyl group or a C1-C6 branched chain alkyl group. Most preferably, the "alkyl" group refers to a C1-C4 straight chain alkyl group or a C1-C4 branched chain alkyl group. Examples of "alkyl" include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, 1-pentyl, 2-pentyl, 3-pentyl, neo-pentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl, or 4-octyl.

[0078] Furthermore, the term "alkyl," as used throughout the specification, examples, and claims, is intended to include both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl.

[0079] "C x~y " or "C x ~C yThe term "alkyl" when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups containing x to y carbons in the chain. CO alkyl indicates a hydrogen in the group's terminal position, and a bond if the group is internal. For example, C 1~6 An alkyl group contains 1 to 6 carbon atoms in the chain.

[0080] The term "alkylamino," as used herein, refers to an amino group substituted with at least one alkyl group.

[0081] The term "alkylthio," as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

[0082] As used herein, the term "amide" refers to the group: [ka] In the formula, R 9 and R 10 each independently represents hydrogen or a hydrocarbyl group, or R 9 and R 10 together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure.

[0083] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines and their salts, e.g., [ka] wherein R 9 , R 10 , and R 10 ' each independently represent a hydrogen or a hydrocarbyl group, or R 9 and R 10 together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure.

[0084] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0085] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0086] The term "aryl", used alone or as part of a larger moiety as in "aralkyl", "aralkoxy", or "aryloxyalkyl", refers to monocyclic and bicyclic ring systems having a total of 5 to 14 ring members, where each ring atom is carbon, at least one ring of the system is aromatic, and each ring of the system contains 3 to 7 ring members. The term "aryl" may be used synonymously 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, anthracyl, and the like, which may bear one or more substituents. The term "aryl" as used herein also includes within its scope groups in which an aromatic ring is fused to one or more non-aromatic carbon rings.

[0087] The term "carbamate" is art-recognized and refers to the group: [ka] In the formula, R 9 and R 10 independently represent hydrogen or a hydrocarbyl group.

[0088] The term "carbocyclylalkyl," as used herein, refers to an alkyl group substituted with a carbocycle group.

[0089] The term "carbocycle" includes 5-7 membered monocyclic and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings. Carbocycles include bicyclic molecules in which one, two, or more than two atoms are shared between the two rings. The term "fused carbocycle" refers to a bicyclic carbocycle in which each ring shares two adjacent atoms with the other ring. Each ring of a fused carbocycle may be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated, and aromatic bicyclic rings is included in the definition of carbocycle, as long as valences permit. Exemplary "carbocycles" include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. A "carbocycle" may be substituted at any one or more positions capable of carrying a hydrogen atom.

[0090] The term "carbocyclylalkyl," as used herein, refers to an alkyl group substituted with a carbocycle group.

[0091] The term "carbonate" is art-recognized and refers to a -OCO2- group.

[0092] The term "carboxy," as used herein, refers to a group represented by the formula -CO2H.

[0093] The term "ester" as used herein means an ester group consisting of -C(O)OR 9 R 9 represents a hydrocarbyl group.

[0094] The term "ether" as used herein refers to a hydrocarbyl group linked to another hydrocarbyl group through an oxygen. Thus, an ether substituent of a hydrocarbyl group can be hydrocarbyl-O-. Ethers can 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 can be represented by the general formula: alkyl-O-alkyl.

[0095] The terms "halo" and "halogen" as used herein mean halogen and include chloro, fluoro, bromo, and iodo.

[0096] The terms "hetarylalkyl" and "heteroarylalkyl," as used herein, refer to an alkyl group substituted with a hetaryl group.

[0097] 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, with 6, 10, or 14 pi electrons shared in a cyclic array, and having 1 to 5 heteroatoms in addition to the carbon atoms. The term "heteroatom" means nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of basic nitrogen. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl. The terms "heteroaryl" and "heteroar-" as used herein also include groups in which a heteroaromatic ring is fused to one or more aryl or heteroaryl rings, such that the resulting bicyclic or polycyclic ring system is fully aromatic. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, and phenoxazinyl. Heteroaryl groups may be monocyclic or bicyclic. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring", "heteroaryl group", or "heteroaromatic", all of which include optionally substituted rings. The term "heteroaralkyl" means an alkyl group substituted by a heteroaryl, where the alkyl and heteroaryl portions independently are optionally substituted.

[0098] The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0099] The term "heterocyclylalkyl," as used herein, refers to an alkyl group substituted with a heterocycle group.

[0100] As used herein, the terms "heterocycle", "heterocyclyl", "heterocyclic radical", and "heterocyclic ring" are used interchangeably and refer to a stable 5-7 membered monocyclic or 7-10 membered bicyclic heterocyclic moiety, saturated or partially unsaturated, having one or more, preferably 1-4 heteroatoms as defined above in addition to carbon atoms. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. By way of example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, or nitrogen, the nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or +NR (as in N-substituted pyrrolidinyl).

[0101] A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenylpyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle", "heterocyclyl", "heterocyclyl ring", "heterocyclic group", "heterocyclic moiety", and "heterocyclic radical" are used interchangeably herein and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or alicyclic rings, e.g., indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. Heterocyclyl groups can be monocyclic or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, where the alkyl and heterocyclyl moieties independently are optionally substituted.

[0102] The term "hydrocarbyl" as used herein refers to a group that is bonded through a carbon atom that does not have =O or =S substituents, and that usually has at least one carbon-hydrogen bond and a primarily carbon backbone, but may optionally contain heteroatoms. Thus, groups such as methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered hydrocarbyl for purposes of this application, while substituents such as acetyl (which has a =O substituent on the bonded carbon) and ethoxy (which is bonded through an oxygen rather than a carbon) are not. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

[0103] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group.

[0104] The term "lower" when used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups having 10 or fewer atoms in the substituent, preferably 6 or fewer atoms. For example, "lower alkyl" refers to an alkyl group containing 10 or fewer carbon atoms, preferably 6 or fewer carbon atoms. In certain embodiments, an acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituent as defined herein is a lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, respectively, whether appearing alone or in combination with other substituents (e.g., atoms in an aryl group are not counted when counting the carbon atoms of an alkyl substituent), such as in the descriptions hydroxyalkyl and aralkyl.

[0105] The terms "polycyclyl," "polycycle," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) in which two or more atoms are common to two adjacent rings, e.g., the rings are "fused rings." Each ring of a polycycle can be substituted or unsubstituted. In certain embodiments, each ring of a polycycle contains 3 to 10 atoms, preferably 5 to 7 atoms, within the ring.

[0106] The term "sulfate" is art-recognized and refers to the group -OSO3H, or a pharma- ceutically acceptable salt thereof.

[0107] The term “sulfonamide” is art recognized and refers to a group that can be represented by the general formula: [ka] In the formula, R 9 and R 10 independently represent hydrogen or hydrocarbyl.

[0108] The term "sulfoxide" is art-recognized and refers to the group --S(O)--.

[0109] The term "sulfonate" is art-recognized and refers to the group SO3H, or a pharma- ceutically acceptable salt thereof.

[0110] The term "sulfone" is art-recognized and refers to a -S(O)2- group.

[0111] The term "protecting group" is an art-recognized term that refers to a chemical functional group that can modify (usually covalently attach) an existing labile functional group on a target molecule. This modification "protects" the labile functional group during a subsequent reaction step, and the protecting group can be removed as needed (called "deprotection"). As a non-limiting example, the t-butyloxycarbonyl (Boc or boc) group is commonly used in synthetic chemistry to covalently modify and "protect" terminal amine groups.

[0112] The term "substituted" refers to a moiety having a substituent replacing a hydrogen on one or more carbons of the backbone. It is understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is based on the substituted atom and the possible valence of the substituent, and that the substitution results in a stable compound (e.g., not spontaneously altered by rearrangement, cyclization, elimination, etc.). As used herein, the term "substituted" is intended to include all possible substituents of organic compounds. In one broad aspect, the possible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, heteroatoms such as nitrogen can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. The substituents may include any of the substituents described herein, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxyl, 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. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain may themselves be substituted, where appropriate.

[0113] Suitable monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are independently halogen, -(CH) 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) 0~4 SR°, -(CH2) 0~4Ph (optionally substituted with R°), -(CH2) 0~4 O(CH2) 0~1 Ph (optionally substituted with R°), -CH=CHPh (optionally substituted with R°), -(CH2) 0~4 O(CH2) 0~1 -pyridyl (optionally substituted with R°), -NO2, -CN, -N3, -(CH2) 0~4 N(R°)2, -(CH2) 0~4 N(R°)C(O)R°, -N(R°)C(S)R°, -(CH2) 0~4 N(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 Linear or branched alkylene)ON(R°)2, or -(C1~4 linear or branched alkylene)C(O)ON(R°), where each R° may be substituted as defined below and is independently hydrogen, C 1~6 Aliphatic, -CH2Ph, -O(CH2) 0~1 Ph, -CH2- (a 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 definitions, two independent R° together with their intervening atom(s) form a 3-12 membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be optionally substituted as defined below.

[0114] Suitable monovalent substituents on R° (or the ring formed by two independent R° together with their intervening atoms) are independently halogen, -(CH2), 0~2 R ● , -(Halo R ● ), -(CH2) 0~2 OH, -(CH2) 0~2 OR ● , -(CH2) 0~2 CH(OR ● )2, -O(HaloR ● ), -CN, -N3, -(CH2) 0~2 C(O)R ● , -(CH2) 0~2 C(O)OH, -(CH2) 0~2 C(O)OR ● , -(CH 2)0~2 S.R. ● , -(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 1~4Linear or branched alkylene)C(O)OR ● , or -SSR ● where each R ● is unsubstituted or, if preceded by "halo", is substituted with one or more halogens only, and independently represents 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 =O and =S.

[0115] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include: ═O, ═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-, in the formula, R * each independently represents hydrogen, optionally substituted C 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. Preferred divalent substituents attached to adjacent substitutable carbons of the "optionally substituted" group include -O(CR * 2) 2~3 O-, wherein R * each independently represents hydrogen, optionally substituted C as defined below 1~6 It is selected from aliphatic or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0116] R *Suitable substituents on the aliphatic group include halogen, -R ● , -(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2, where each R ● is unsubstituted or, if preceded by "halo", is substituted with one or more halogens only, and independently represents 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.

[0117] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include -R † , -NR † 2. -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CH2C(O)R † , -S(O)2R † , -S(O)NR † 2. -C(S)NR † 2. -C(NH)NR † 2, or -N(R † )S(O)2R † wherein each R † are independently hydrogen, optionally substituted as defined below, 1~6 an aliphatic, unsubstituted -OPh, or substituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or, notwithstanding the above definitions, two independent R † together with their intervening atom(s) form an unsubstituted 3-12 membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0118] R † and substituted 5-6 membered saturated, partially unsaturated, or aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur are independently selected from halogen, -R ● , -(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2, where each R ● is unsubstituted or, if preceded by "halo", is substituted with one or more halogens only, and independently represents 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.

[0119] The term "thioalkyl," as used herein, refers to an alkyl group substituted with a thiol group.

[0120] The term "thioester" as used herein means -C(O)SR 9 Group or -SC(O)R 9 R 9 represents hydrocarbyl.

[0121] As used herein, the term "thioether" is the equivalent of an ether where the oxygen is replaced with a sulfur.

[0122] The term "urea" is art-recognized and can be represented by the general formula: [ka] In the formula, R 9 and R 10independently represent hydrogen or hydrocarbyl.

[0123] The term "tautomer" refers to each of two or more isomers of a compound that exist together in equilibrium and are interchanged by the migration of an atom or group within a molecule, such as a hydrogen atom. Exemplary tautomers of the present disclosure include: [ka] The depiction of one tautomer within a genus or compound species in this disclosure is intended to encompass the depicted compound and all of its tautomers. Specifically, in the above example, any of the structures may be represented as a set of 2n distinct species. [ka] also discloses, where n is the number of tautomeric sites on the molecule.

[0124] For purposes of this disclosure, any embodiment described herein also applies, where appropriate subject matter, to any of the general structural formulas described herein.

[0125] In certain embodiments, the present invention provides a series of compounds having the structure of formula (IA): [ka] or a pharma- ceutically 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 is -C(O)-R 1 or -CN, Ring A is aryl, heteroaryl, cycloalkyl, or heterocyclyl; Ring B is absent or present and, if present, is aryl, heteroaryl, cycloalkyl, or heterocyclyl; R 4 is hydrogen, a protecting group, -C(O)-CH3, -L', or -LY; L', if present, is a reactive linker moiety; L, when present, is a linker moiety; Y, if present, is a mitochondrial targeting moiety, [ka] are each independently a single bond or a double bond; and Any hydrogen atoms are optionally replaced with deuterium.

[0126] In certain embodiments, the present invention provides a series of compounds having the structure of formula (2-1): [ka] or a pharma- ceutically 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 is -C(O)-R 1 or -CN, Ring A is aryl, heteroaryl, cycloalkyl, or heterocyclyl; Ring B is absent or present and, if present, is aryl, heteroaryl, cycloalkyl, or heterocyclyl; R 4is hydrogen, a protecting group, -C(O)-CH3, -L', or -LY; L', if present, is a reactive linker moiety that contains an alkyne group; L, when present, is a linker moiety; Y, if present, is a mitochondrial targeting moiety, [ka] are each independently a single bond or a double bond; any hydrogen atom is optionally replaced with deuterium; however, Ring A is selected from unsubstituted thiazolyl, oxazolyl, cyclopentyl, and bicyclo[2.1.1]hexane; and ring B-NHR 4 but [ka] If a)R 4 is -L'-Y or L; or b)R 2 and R 3 are not both =CH2 or both =CH(alkyl).

[0127] In certain embodiments, L is [ka] T is alkyl and (-CH2CH2-O-) v -; v is an integer selected from 3 to 9, and T is attached to Y.

[0128] In certain embodiments, the present invention also provides a series of compounds having the structure of formula (2-2): [ka] or a pharma- ceutically 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', if present, is a reactive linker moiety that contains an alkyne group; L, when present, is a linker moiety; Y, if present, is a mitochondrial targeting moiety, and any hydrogen atom is optionally replaced with deuterium; however, Ring A is selected from unsubstituted thiazolyl, oxazolyl, cyclopentyl, and bicyclo[2.1.1]hexane; and ring B-NHR 4 but [ka] If a)R 4 is -L'-Y or L; or b)R 2 and R 3 are not both =CH2 or both =CH(alkyl).

[0129] In certain embodiments, L is [ka] T is alkyl and (-CH2CH2-O-) v -; v is an integer selected from 3 to 9, and T is attached to Y.

[0130] In certain embodiments, the present invention also provides a series of compounds having the structure of formula (2-3): [ka] or a pharma- ceutically 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, or -C(O)-CH3, and any hydrogen atom is optionally replaced with deuterium; however, Ring A is selected from unsubstituted thiazolyl, oxazolyl, cyclopentyl, and bicyclo[2.1.1]hexane; and ring B-NHR 4 but [ka] If R 2 and R 3 are not both =CH2 or both =CH(alkyl).

[0131] In certain embodiments, the present invention provides a series of compounds having the structure of formula (2-4): [ka] or a pharma- ceutically 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 3is -H, -CH3, =CH2, or =CH(alkyl); R 5 is -C(O)-R 1 or -CN, Ring A is [ka] where * indicates a bond to ring B, ** and *** are -NH-R 4 indicates, Ring B is absent or present and, if present, is aryl, heteroaryl, cycloalkyl, or heterocyclyl; R 4 is hydrogen, a protecting group, -C(O)-CH3, -L', or -LY; L', if present, is a reactive linker moiety that contains an alkyne group; L, when present, is a linker moiety; Y, if present, is a mitochondrial targeting moiety, [ka] are each independently a single bond or a double bond; and Any hydrogen atoms are optionally replaced with deuterium.

[0132] In certain embodiments, L is [ka] T is alkyl and (-CH2CH2-O-) v -; v is an integer selected from 3 to 9, and T is attached to Y.

[0133] In certain embodiments, the present invention provides a series of compounds having the structure of formula (2-5): [ka] or a pharma- ceutically acceptable salt thereof, wherein: R 1is -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 is -C(O)-R 1 or -CN, Ring A is aryl, heteroaryl, cycloalkyl, or heterocyclyl; Ring B-NHR 4 teeth, [ka] is selected from R 4 is hydrogen, a protecting group, -C(O)-CH3, -L', or -LY; L', if present, is a reactive linker moiety that contains an alkyne group; L, when present, is a linker moiety; Y, if present, is a mitochondrial targeting moiety, [ka] are each independently a single bond or a double bond; and Any hydrogen atoms are optionally replaced with deuterium.

[0134] In certain embodiments, L is [ka] T is alkyl and (-CH2CH2-O-) v -; v is an integer selected from 3 to 9, and T is attached to Y.

[0135] In certain embodiments, the present invention provides a series of compounds having the structure of formula (2-6): [ka] or a pharma- ceutically 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 thiazolyl substituted with alkyl or thiophenyl; Ring B-NHR 4 teeth, [ka] and R 4 is a protecting group or -L', L', if present, is a reactive linker moiety that contains an alkyne group; Any hydrogen atoms are optionally replaced with deuterium.

[0136] In certain embodiments, R 5 is -C(O)-R 1 and R 1 is -OCH3. In other embodiments, R 5 is -C(O)-R 1 and R 1 is -NH2.

[0137] In certain embodiments, when ring A is polycyclic, ring B is absent.

[0138] In certain embodiments, ring A is a 5-membered ring or a 5-membered ring fused to a second ring.

[0139] In certain embodiments, ring A is a 5-membered heteroaryl. In some embodiments, ring A is a bicyclic heteroaryl.

[0140] In certain embodiments, ring A is thiazolyl, thiophenyl, oxazolyl, or imidazolyl. In other embodiments, ring A is thiazolyl, thiophenyl, or oxazolyl. In some embodiments, ring A is pyrrolidinyl. In other embodiments, ring A is thiazolyl.

[0141] In certain embodiments, ring A is [ka] where * indicates the bond to ring B.

[0142] In certain embodiments, ring A is [ka] where * indicates the bond to ring B.

[0143] In certain embodiments, ring A is [ka] where * indicates the bond to ring B.

[0144] In certain embodiments, ring A is [ka] where * indicates the bond to ring B.

[0145] In certain embodiments, Ring A is a 5-membered cycloalkyl or heterocyclyl.

[0146] In certain embodiments, Ring A is cyclopentyl or tetrahydrofuranyl.

[0147] In certain embodiments, ring A is [ka] where * indicates the bond to ring B.

[0148] In certain embodiments, ring A is a bridged bicyclic cycloalkyl or heterocyclyl.

[0149] In certain embodiments, ring A is bicyclo[2.1.1]hexyl or oxabicyclo[2.1.1]hexyl.

[0150] In certain embodiments, ring A is [ka] where * indicates the bond to ring B.

[0151] In certain embodiments, ring A is phenyl.

[0152] In certain embodiments, ring A is [ka] where * indicates the bond to ring B.

[0153] In certain embodiments, Ring A is a polycyclic aryl, heteroaryl, cycloalkyl, or heterocyclyl.

[0154] In certain embodiments, ring A is a bicyclic heteroaryl.

[0155] In certain embodiments, ring A is [ka] *** and ** represent ring B or -NH-R 4 Shows.

[0156] In certain embodiments, ring B is a 6-membered ring.

[0157] In certain embodiments, B is phenyl.

[0158] In certain embodiments, Ring B is unsubstituted phenyl. In other embodiments, Ring B is halogen substituted phenyl.

[0159] In certain embodiments, ring B is [ka] and Z is halo and ** indicates the bond to ring A. In some embodiments, Z is selected from fluoro or chloro.

[0160] In certain embodiments, ring B is [ka] and Z is alkoxy, such as methoxy, or alkyl, such as methyl.

[0161] In other embodiments, Ring B is [ka] It is.

[0162] In certain embodiments, Ring B is a 6-membered heteroaryl.

[0163] In certain embodiments, Ring B is pyridinyl, pyrimidinylpyrazinyl, or pyridazinyl.

[0164] In certain embodiments, Ring B is pyridinyl, pyrimidinyl, or pyrazinyl.

[0165] In certain embodiments, ring B is [ka] where * indicates the bond to ring A.

[0166] In certain embodiments, Ring B is a bridged bicyclic cycloalkyl.

[0167] In certain embodiments, ring A is bicyclo[2.2.2]octanyl or bicyclo[1.1.1]pentanyl.

[0168] In certain embodiments, ring B is [ka] where Z is halo and ** indicates the bond to ring A.

[0169] In certain embodiments, ring B is [ka] * is selected from -NHR 4 The bond to is shown.

[0170] In certain embodiments, R 5 is -CN.

[0171] In certain embodiments, the compound of formula (IA) has the formula (IA-1): [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R 2 is -H or -CH3, R 3 is =CH2 or =CH(alkyl), Ring A, Ring B, R 1 , R 4 , and R 5 each of which is any of the embodiments described above for formula (IA).

[0172] In certain embodiments of formula (IA-1), R 2 is -H, R 3 is =CH2 or R 2 is -CH3, R 3 is =CH2.

[0173] In certain embodiments, the compound of formula (IA) has the formula (IA-2): [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R 2 is =CH2 or =CH(alkyl), R 3 is -H or -CH3, Ring A, Ring B, R 1 , R 4 , and R 5 each of which is any of the embodiments described above for formula (IA).

[0174] In certain embodiments of formula (IA-2), R 2 is =CH2 and R 3 is -H or R 2 is =CH2 and R 3 is -CH3.

[0175] In certain embodiments, the compound of formula (IA) has the formula (IA-3): [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R 2 is =CH2 or =CH(alkyl), R 3 is =CH2 or =CH(alkyl), Ring A, Ring B, R 1 , R 4 , and R 5 each of which is any of the embodiments described above for formula (IA).

[0176] In certain embodiments of formula (IA-3), R 2 is =CH2, R 3 =CH2 or R 2 is =CH2, R 3is =CH(CH3) or R 2 is =CH(CH3), and R 3 is =CH2.

[0177] In certain embodiments, the compound of formula (IA) has the formula (IB): [ka] or a pharma- ceutically acceptable salt thereof, wherein: Ring A, Ring B, R 1 , R 2 , R 3 and R 4 each of which is any of the embodiments described above for formula (IA).

[0178] In certain embodiments, the compound of formula (IA) has the formula (IB): [ka] or a pharma- ceutically 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', if present, is a reactive linker moiety; L, when present, is a linker moiety; Y, if present, is a mitochondrial targeting moiety, and Any hydrogen atoms are optionally replaced with deuterium.

[0179] 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, thiophenyl, oxazolyl, cyclopentyl, or bicyclo[2.1.1]hexyl.

[0180] In certain embodiments of formula (IB), ring A is thiazolyl.

[0181] In certain embodiments of formula (IB), ring A is [ka] It is.

[0182] In certain embodiments of formula (IB), ring A is [ka] where * indicates the bond to ring B.

[0183] In certain embodiments of formula (IB), ring B is a 6-membered ring. In certain embodiments, ring B is phenyl or pyridyl. In certain embodiments, ring B is unsubstituted phenyl.

[0184] In certain embodiments of formula (IB), ring B is halogen substituted phenyl.

[0185] In certain embodiments of formula (IB), R 2 and R 3 is different.

[0186] In certain embodiments of formula (IB), R 2 and R 3 are the same. In certain embodiments, R 2 is CH(Me) or -CH2.

[0187] In certain embodiments of formula (IB), R 3 is CH(Me) or -CH2.

[0188] In certain embodiments of formula (IB), R 2 and R 3 are CH2, respectively.

[0189] In certain embodiments of formula (IB), R 4 is hydrogen, a protecting group, or -C(O)-CH3.

[0190] In certain embodiments, the compound is [ka] is selected from R 4 is hydrogen, a protecting group, or -C(O)-CH3, [ka] The bond shown as indicates either possible stereochemistry of the attached alkene, E or Z.

[0191] In certain embodiments, the compound is [ka] is selected from R 4 is hydrogen, a protecting group, or -C(O)-CH3.

[0192] In certain embodiments, the compound is [ka] is selected from.

[0193] In certain embodiments, the compound is [ka] is selected from R 4 is hydrogen, a protecting group, or -C(O)-CH3, Z is selected from fluorine, chlorine, bromine, and iodine.

[0194] In certain embodiments, the compound is [ka] is selected from R 4 is hydrogen, a protecting group, or -C(O)-CH3.

[0195] In certain embodiments, the compound is [ka] is selected from R 4 is hydrogen, a protecting group, or -C(O)-CH3.

[0196] In certain embodiments, the protecting group is Boc.

[0197] In certain embodiments, R 4 In certain embodiments, L' is -C(O)-XC(O)OH or -C(O)-XC(O)NH2, where X is -(CH2). n and n is 2, 3, 4 or 5.

[0198] In certain embodiments, the compound is [ka] is selected from X is -(CH2) n and n is 2, 3, 4 or 5.

[0199] In certain embodiments, the compound is [ka] [ka] [ka] is selected from X is -(CH2) n - and n is 2, 3, 4, or 5. In certain embodiments, R 4 is -LY. In certain embodiments, L is a cleavable linker. In certain embodiments, L is a non-cleavable linker. In certain embodiments, L has a chain length of about 2 to about 30 atoms. In certain embodiments, L has a chain length of about 5 to about 20 atoms.

[0200] In certain embodiments, L is -C(O)-XC(O)-, where X is -(CH) n and n is 2, 3, 4 or 5.

[0201] In certain embodiments, L is -C(O)-XC(O)-, where X is -(CHCH-O-). m —(CH2CH2)—, and m is 2, 3, 4, 5 or 6.

[0202] In certain embodiments, R 4 is L', and the alkyne group is [ka] It is.

[0203] In certain embodiments, the compound is [ka] is selected from X is -(CH2) n and n is 2, 3, 4 or 5.

[0204] In certain embodiments, the compound is [ka] [ka] [ka] is selected from X is -(CH2) n - and m is 2, 3, 4, 5, or 6; and n is 2, 3, 4, or 5.

[0205] In certain embodiments, L' comprises an alkynyl or azido.

[0206] In certain embodiments, R 4 teeth, [ka] or -C(O)-X'-N3, where X' is -(CH2) n and n is 2, 3, 4 or 5.

[0207] In certain embodiments, L comprises heteroaryl. In other embodiments, L comprises triazolyl.

[0208] In certain embodiments, R 4 teeth, [ka] and R 6 is -H or -C(O)CH3, X' is -(CH2) n - and X” is -(CH2) o - and n is 2, 3, 4 or 5; o is 2, 3, 4 or 5.

[0209] In certain embodiments, R 4 teeth, [ka] and R 6 is -H or -C(O)CH3, X' is -(CH2)n - and X” is -(CH2) o - and n is 2, 3, 4 or 5; o is 2, 3, 4 or 5.

[0210] In certain embodiments, R 4 teeth, [ka] and X' is -(CH2) n - and X” is -(CH2) o - and n is 2, 3, 4 or 5; o is 2, 3, 4 or 5.

[0211] In certain embodiments, Y is selected from the group consisting of berberine cation, rhodamine cation, indolium cation, pyridinium cation, tetraguanidinium cation, cyanine derivatives, guanidinium cation, biguanidinium cation, triphenylphosphonium cation, triethylammonium cation, triphenylamine, tetraphenylethene moiety, arylphosphonium cation, SS peptide, mitochondrial penetrating peptide (MPP), mitochondrial targeting sequence (MTS) peptide, hemigramicidin S-linked nitroxide, dequalinium (DQA) cation, Delocalized lipophilic cations, F16 ((E)-4-(1H-indol-3-ylvinyl)-N-methylpyridinium iodide), (L-cyclohexylalanine-D-arginine)3, mitochondria-targeted nanocarriers, DDDK peptide, glycyrrhetinic acid, α-tocopheryl succinate (α-TOS), graphene oxide nanocarriers, PEG-proapoptotic peptide (KLAKLAK)2, Dmt-D-Arg-Phe-Lys-NH2 peptide, pyruvaldehyde, N-nonyl acridine orange, quinoline, styryl fluorophores, or 15d-PGJ2.

[0212] In certain embodiments, Y is a mitochondrial penetrating peptide.

[0213] In certain embodiments, Y has the structural formula (V). [ka]

[0214] In certain embodiments, the compound is [ka] or a pharma- ceutically acceptable salt thereof, wherein X is selected from [—(CH2) n -] or [-(CH2CH2-O-) m -(CH2CH2)-], n is 3, 4, or 5, and m is 2, 3, 4, 5, or 6.

[0215] In certain embodiments, the present invention provides a pharma- ceutically acceptable composition comprising any of the compounds described herein and a pharma- ceutically acceptable carrier.

[0216] In certain embodiments, the compositions are formulated for oral or parenteral delivery.

[0217] In certain embodiments, the compound is incorporated within a nanoparticle, liposome, or micelle, and the nanoparticle, liposome, or micelle is conjugated to a mitochondrial targeting moiety.

[0218] In certain embodiments, the present invention discloses a composition comprising a compound of formula (IA) or any subformula thereof, wherein R 4 is hydrogen, a protecting group, or -C(O)-CH3, or any of its more specific embodiments described herein is contained within a nanoparticle, liposome, or micelle, and the nanoparticle, liposome, or micelle is conjugated to a mitochondrial targeting moiety.

[0219] In certain embodiments, the compositions are formulated for oral or parenteral delivery.

[0220] 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 its more specific embodiments described herein is contained within a nanoparticle, liposome, or micelle, and the nanoparticle, liposome, or micelle is conjugated to a mitochondrial targeting moiety.

[0221] In certain embodiments, the nanoparticles, liposomes, or micelles are selected from poly(ethylene glycol), poly(ε-caprolactone), polysaccharides, poly[(2-hydroxypropyl)-methacrylic acid], poly(lactic-co-glycolic acid), and any combination of the foregoing.

[0222] In certain embodiments, the present invention discloses a method of treating cancer (e.g., a solid tumor or a hematological cancer), comprising administering to a subject in need of cancer treatment a therapeutically effective amount of any of the compounds described herein, or a composition of the compounds. In certain embodiments, the cancer (solid tumor or hematological cancer) is selected from lung, breast, prostate, melanoma, esophageal, leukemia, cervical, liver, colon, stomach, colon, glioblastoma, head and neck, pancreas, mesothelioma, and ovarian. In certain embodiments, the cancer is selected from mesothelioma, lung, ovarian, and breast.

[0223] In certain embodiments, the present invention discloses a compound selected from: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] EXAMPLES

[0224] General Experimental Procedure 1 Suzuki Coupling The starting boronic acid or boronic ester (1 equiv.) was introduced into the flask under N2. 1,2-Dimethoxyethane (0.1M), bromide (1.05 equiv.), and 2M aqueous sodium carbonate (5 equiv.) were added. The mixture was then degassed with N2 for a few minutes. Palladium tetrakis (0.05 equiv.) was then added and the mixture was stirred at 90 °C (external temperature) until complete conversion (overnight). The reaction was then quenched. Water was added and the mixture was extracted with EtOAc. Brine was added to improve separation. The organic layer was dried over Na2SO4, filtered and concentrated. The crude material was purified by automated FCC. The desired product was obtained in 40-70% yield. Note: In some reactions, hydrolysis products were also observed. This was isolated by acidifying the aqueous phase and extracting twice with EtOAc. The combined organics were washed with brine, dried over Na2SO4, filtered and concentrated to obtain the hydrolysis products.

[0225] General Experimental Procedure 2 Ester hydrolysis To a solution of the ester (1 equiv.) in THF (0.05 M) was added lithium hydroxide monohydrate (4 equiv.) as a solution in water (2 M) and the mixture was stirred at room temperature for 2-16 h. The mixture was diluted with EtOAc, washed with 1 M HCl and brine (30 mL each), dried over Na2SO4, filtered and concentrated to give the desired product.

[0226] General Experimental Procedure 3 EDCI-Mediated Amide Coupling To a solution of acid (1 eq.) and amine (1 eq.) in DCM (0.05M) was added DiPEA (3 eq.), cyano(E)-2-(hydroxyimino)butanoic anhydride (1.3 eq.) and EDCI (1.3 eq.), and the resulting mixture was stirred at room temperature for 16 h. The mixture was washed with 1M HCl, water, NaHCO3, and brine, dried over Na2SO4, filtered, and concentrated. The crude material was purified by automated FCC to give the desired product.

[0227] General Experimental Procedure 4 HATU-mediated amide coupling To a solution of acid (1 equiv.) and amine (1 equiv.) in DCM (0.05M) was added DiPEA (3 equiv.) and HATU (1.2 equiv.) and the resulting mixture was stirred at room temperature for 2-16 h. The mixture was washed with 1M HCl, water, NaHCO3, and brine, dried over Na2SO4, filtered, and concentrated. The crude material was purified by automated FCC to give the desired product.

[0228] General Experimental Procedure 5 Serine side chain acetylation To a solution of the free alcohol (1 eq.) in MeCN (0.05M) was added triethylamine (2 eq. / alcohol), DMAP (0.2 eq.) and acetic anhydride (1.1 eq. / alcohol) and the resulting mixture was stirred for 2 h. The reaction mixture was diluted with water and MeCN was removed in vacuo. The mixture was extracted with EtOAc and the organic phase was washed with brine, dried over Na2SO4, filtered and concentrated. The crude material was purified by automated FCC to give the desired product.

[0229] General Experimental Procedure 6 Stepwise removal of mono-TBDPS-protected bis-serine To a solution of the mono-TBDPS protected bis-serine derivative (1 eq.) in DCM (0.05M) was added triethylamine (10 eq.) and methanesulfonyl chloride (1.2 eq.) and the resulting mixture was stirred at room temperature for 1 h. The reaction was followed by LCMS. Optionally, additional triethylamine was added. After completion, a solution of tetrabutylammonium fluoride in THF (1M, 2 eq.) was added and the mixture was stirred for 1 h. The mixture was washed with 1M HCl and brine, dried over Na2SO4, filtered and concentrated. The residue was taken up in CH2Cl2 and triethylamine (10 eq.) and methanesulfonyl chloride (1.2 eq.) were added. The resulting mixture was stirred at room temperature for 1 h. The reaction was followed by LCMS. Optionally, additional triethylamine was added. The mixture was washed with 1M HCl and brine, dried over Na2SO4, filtered and concentrated. The crude material was purified by automated reverse phase FCC.

[0230] General Experimental Procedure 7 Stepwise removal of mono-TBDPS and mono-acetate To a solution of the mono-TBDPS monoacetyl bis-serine derivative (1 eq.) in CHCl (0.05M) was added DBU (2 eq.) and the mixture was stirred at room temperature for 1-16 h. Then a solution of TBAF in THF (1M, 2 eq.) was added and the mixture was stirred for 2 h. The mixture was concentrated and purified by automated FCC. To a solution of the mono-elimination intermediate in CHCl (0.05M) was added triethylamine (6 eq.) and methanesulfonyl chloride (1.5 eq.). Optionally, additional triethylamine was added. The mixture was washed with 1M HCl and brine, dried over NaSO, filtered and concentrated. The crude material was purified by automated reverse-phase FCC.

[0231] General Experimental Procedure 8 Bisacetate elimination A solution of the bisacetate in THF / CH2Cl2 (1 / 1, v / v, 0.05M) was cooled to 0°C and DBU (4 equiv.) was added. The mixture was stirred at 0°C for 2 h. The mixture was diluted with CH2Cl2 and water and HCl (1M aq.) were added to acidify the mixture to pH=4. The mixture was extracted with CH2Cl2 (3x) 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.

[0232] General Experimental Procedure 9 Preparation of primary amides To a solution of carboxylic acid (1 eq.) in DMF (0.1 M) or CH2Cl2 (0.15 M) was added NH4Cl (3 eq.), DiPEA (3.5 eq.) and HATU (1.1 eq.). The mixture was stirred at room temperature overnight. The reaction was quenched with water. If the solution was clear, 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. In case of a precipitate, the mixture was filtered and washed with water and the solvent used in the reaction. The solid was collected and concentrated to remove residual solvent to give the desired primary amide.

[0233] General Experimental Procedure 10 Thio-substituted primary amides To a solution of the primary amide (1 eq.) in THF (0.2M) was added Lawesson's reagent (0.7 eq.) and the mixture was stirred at 80° C. (external temperature) until complete conversion. The reaction was then quenched with saturated aqueous NaHCO3 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.

[0234] General Experimental Procedure 11 Formation of thiazoles from thioamides To a solution of the primary thioamide (1 equiv.) in EtOH (0.2 M) was added ethyl bromopyruvate (1.1 equiv.) and the resulting mixture was stirred at room temperature or 50° C. (external temperature) until complete conversion. The reaction was quenched with saturated aqueous NaHCO3 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.

[0235] Analysis method LCMS method "21020335B TFA LCMS-5 C3" System: Agilent 1290 series with UV detector and HP 6130 MSD mass detector Column: Waters XBridge BEH C18 XP (2.1 x 50 mm; 2.5 μm) Mobile phase A: Milli-Q containing 0.05% TFA Mobile phase B: Acetonitrile Pump flow rate: 0.6ml / min UV detection: 215.8 nm Injection volume: 0.1 μL Run Time: 3.5 minutes Column temperature: 35℃ Pump Program: Gradient [Table 1] LCMS method "21020335C TFA LCMS-5 C4" System: Agilent 1290 series with UV detector and HP 6130 MSD mass detector Column: Waters XBridge BEH C18 XP (2.1 x 50 mm; 2.5 μm) Mobile phase A: Milli-Q containing 0.05% TFA Mobile phase B: Acetonitrile Pump flow rate: 0.6ml / min UV detection: 215.8 nm Injection volume: 0.1 μL Run Time: 3.0 minutes Column temperature: 35℃ Pump Program: Gradient [Table 2] LCMS method "22010199 LCMS-5 C3" System: Agilent 1290 series with UV detector and HP 6130 MSD mass detector Column: Waters XBridge BEH C18 XP (2.1 x 50 mm; 2.5 μm) Mobile phase A: 10 mM NH4OAc (water / methanol / acetonitrile 900 / 60 / 40) Mobile phase B: 10 mM NH4OAc (water / methanol / acetonitrile 100 / 540 / 360) Pump flow rate: 0.6ml / min UV detection: 215.8 nm Injection volume: 0.3 μL Run Time: 3.5 minutes Column temperature: 35℃ Pump Program: Gradient [Table 3] LCMS method "22010199A TFA LCMS-5 C1" System: Agilent 1290 series with UV detector and HP 6130 MSD mass detector Column: Waters Acquity HSS T3 (2.1 x 75 mm; 1.8 μm) Mobile phase A: Milli-Q containing 0.05% TFA Mobile phase B: Acetonitrile Pump flow rate: 0.6ml / min UV detection: 215.8 nm Injection volume: 0.8 μL Run Time: 3.0 minutes Column temperature: 35℃ Pump Program: Gradient [Table 4] LCMS method "22010199C TFA LCMS-5 C3" System: Agilent 1290 series with UV detector and HP 6130 MSD mass detector Column: Waters XBridge BEH C18 XP (2.1 x 50 mm; 2.5 μm) Mobile phase A: Milli-Q containing 0.05% TFA Mobile phase B: Acetonitrile Pump flow rate: 0.6ml / min UV detection: 215.8 nm Injection volume: 0.8 μL Run Time: 3.0 minutes Column temperature: 35℃ Pump Program: Gradient [Table 5] LCMS method "22010199D TFA LCMS-5 C8" System: Agilent 1290 series with UV detector and HP 6130 MSD mass detector Column: Zorbax SB-C8 (2.1 x 50 mm; 1.8 μm) Mobile phase A: Milli-Q containing 0.05% TFA Mobile phase B: Acetonitrile Pump flow rate: 0.6ml / min UV detection: 215.8 nm Injection volume: 0.1 μL Run Time: 3.0 minutes Column temperature: 35℃ Pump Program: Gradient [Table 6] LCMS method "30833 LCMS-6" System: Agilent 1260 series with UV detector, ELSD 1260 detector, and Agilent 6120 mass detector Column: Waters XBridge BEH C18 XP (2.1 x 50 mm; 2.5 um) Mobile phase A: Milli-Q containing 0.1% TFA Mobile phase B: 0.1% TFA in acetonitrile Pump flow rate: 0.6ml / min UV detection: 215.8 nm Injection volume: 0.6 μL Run Time: 6.0 minutes Column temperature: 35℃ Pump Program: Gradient [Table 7] LCMS Method "General Tribasic" System: Waters Acquity UPLC with PDA detector and QDA mass detector Column: Waters XBridge BEH C18 (2.1 x 50 mm; 2.5 μm) Mobile phase A: 10 mM ammonium bicarbonate aqueous solution (pH 9.5) Mobile phase B: Acetonitrile Pump flow rate: 0.6ml / min UV detection: λmax Injection volume: 0.4 μL Run Time: 2.5 minutes Column temperature: 30℃ Pump Program: Gradient [Table 8] LCMS Method "General 3 Acids" System: Waters Acquity UPLC with PDA detector and QDA mass detector Column: Waters XBridge BEH C18 (2.1 x 50 mm; 2.5 μm) Mobile phase A: 0.1% (v / v) trifluoroacetic acid aqueous solution Mobile phase B: Acetonitrile Pump flow rate: 0.6ml / min UV detection: λmax Injection volume: 0.4 μL Run Time: 2.5 minutes Column temperature: 30℃ Pump Program: Gradient [Table 9]

[0236] Example 1A: Exemplary synthesis of thiostrepton-inspired structure A1 [ka] Scheme 1A: Synthesis towards compound A-1

[0237] Commercially available thiazole ethyl ester SM1a was hydrolyzed using LiOH to give the carboxylic acid Int 2a. Subsequent amide coupling with serine methyl ester (SM3) gave the amide Int 4a. The free hydroxyl group was protected by treatment with TBDPS-Cl to give compound Int 5a after column chromatography (79% over three steps). Hydrolysis of the methyl ester using LiOH gave the carboxylic acid derivative Int 6a, which was then coupled with serine methyl ester SM7a to give dipeptide Int 8a, which was purified by column chromatography (47% over two steps). Removal of the hydroxyl moiety was achieved by treatment with mesyl chloride and NEt3 to give the dehydroalanine derivative Int 9a. TBAF-mediated TBDPS removal followed by a second elimination reaction gave the bisdehydroalanine derivative Int 11a.

[0238] Example 1B: Exemplary synthesis of thiostrepton-inspired structure A1 ((5)-HCl) [ka] Scheme 1B: Synthesis towards compound A-1

[0239] Suzuki coupling of boronate ester SM1B with bromide SM2B gave compound Int 3B in moderate yield (65%). The Boc protecting group was converted to Alloc in 95% yield to give Alloc-protected compound Int 4B. Hydrolysis of the methyl ester followed by EDCI-mediated coupling of serine methyl ester gave compound Int6B. After protection of the free alcohol with TBDPS and purification, compound Int7B was isolated in 79% in three steps. Hydrolysis of the methyl ester followed by EDCI-mediated coupling of serine methyl ester gave compound Int9B (45% yield). Hydroxyl elimination was achieved by treatment with mesyl chloride and NEt3. TBAF-mediated TBDPS removal followed by a second elimination reaction gave the bisdehydroalanine motive. Finally, the Alloc group was removed by treatment with tetrakis in the presence of a scavenger to give compound A-1.

[0240] Alternative synthesis: 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid intermediate 2 [ka]

[0241] Step 1. Ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylate (Intermediate 1) was prepared according to 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 (400 MHz,CDCl3) δ 8.08 (s,1H),7.96 - 7.88 (m,2H),7.47 - 7.40 (m,2H),6.59 (s,1H),4.42 (q,J=7.1 Hz, 2H), 1.51 (s, 9H), 1.41 (t, J = 7.1Hz, 3H).

[0242] Step 2. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) was prepared according to General Experimental Procedure 2. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylate (Intermediate 1) (0.93 g, 2.7 mmol) gave 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) (0.82 g, 2.6 mmol, 97%) as an off-white solid. 1 H NMR (400 MHz, CDCl3) δ 8.09 (s,1H),7.89 - 7.82 (m,2H),7.45 - 7.34 (m,2H),1.47 (s,9H),1.24 - 1.13 (m,1H).

[0243] Methyl 2-(2-(2-(4-aminophenyl)thiazole-4-carboxamido)acrylamido)acrylate hydrochloride (Compound A1) [ka]

[0244] Step 1. Ethyl 2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylate was prepared as follows: To a solution of ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylate (Intermediate 1) (2.4 g, 1 equiv, 6.9 mmol) in CHCl (10 mL) was added TFA (10 mL) and the resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure and the residue was taken up in THF (50 mL). To the mixture was added pyridine (2.8 mL, 5 equiv, 34 mmol) followed by allyl chloroformate (0.96 mL, 1.3 equiv, 9.0 mmol) and the resulting mixture was stirred for 1 h. 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 directly in the next step.

[0245] Step 2. 2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylic acid was prepared according to 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 (400 MHz,CDCl3) δ 8.20 (s,1H),7.95 - 7.87 (m,2H),7.50 (d, J =8.5 Hz,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)

[0246] Step 3. Methyl N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to 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 methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (1.7 g, 1.2 eq, 4.3 mmol) gave methyl N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.9 g, 3.0 mmol, 82%).

[0247] Step 4. N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine was prepared according to general experimental procedure 2. Methyl N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.9 g, 1 eq, 3.0 mmol) gave 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 directly in the next step.

[0248] Step 5. Methyl N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate was prepared according to 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 methyl N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate (0.75 g, 1.0 mmol, 35%) as a yellow oil.

[0249] Step 6. Methyl 2-(2-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)acrylate was prepared according to General Experimental Procedure 6. Methyl N-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate (0.75 g, 1 eq, 1.0 mmol) gave crude methyl 2-(2-(2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)acrylate (0.9 g), which was used in the next step without purification.

[0250] Step 7. Methyl 2-(2-(2-(4-aminophenyl)thiazole-4-carboxamido)acrylamide)acrylate was prepared as follows: To a solution of crude methyl 2-(2-(2-(4-((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)acrylate (0.90 g) in DCM (20 mL) was added phenylsilane (1.1 g, 1.2 mL, 5 equiv, 9.9 mmol) followed by tetrakis(triphenylphosphine)-palladium(0) (0.23 g, 0.1 equiv, 0.20 mmol) and the resulting mixture was stirred for 1 h. MeCN (4 mL) was added and CHCl was removed in vacuo. The mixture was directly subjected 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-5 C4) RT: 1.274 min; Area % (215 nm): 79.1%; m / z = 373.2 [M+H] + . 1 H NMR (400 MHz,MeOD) δ 8.38 (s,1H),8.33 - 8.17 (m,2H),7.57 - 7.52 (m,2H),6.60 (d, J =1.8 Hz,1H),6.29 (s ,1H), 6.00 (s, 1H), 5.76 (d, J = 1.9 Hz, 1H), 3.86 (s, 3H).

[0251] Example 2A: Exemplary synthesis of thiostrepton-inspired structure A2 ((1)) [ka] Scheme 2A: Synthesis to Compound A-2 via Dipeptide Building Blocks

[0252] Compound A-2 was obtained by coupling of a thiazole derivative with a dipeptide. The dipeptide was synthesized starting from Boc-Ser-OMe. First, the hydroxyl was protected by treatment with TBDPS-Cl to give the serine derivative. The methyl ester was hydrolyzed by treatment with LiOH to give the carboxylic acid, which was then coupled with serine amide to give the dipeptide after column chromatography (27% for three steps). The Boc protecting group was removed by treatment with HCl to give the dipeptide. EDCI-mediated coupling of the dipeptide with the carboxylic acid afforded the thiazole-coupled dipeptide after column chromatography (51%). Removal of the hydroxyl mediated by mesyl chloride and NEt3, followed by TBDPS removal with TBAF, and a second elimination reaction gave the Boc-protected compound after preparative HPLC purification. Treatment with HCl removed the Boc protecting group to give compound A-2.

[0253] Example 2B: Exemplary synthesis of thiostrepton-inspired structure A2 [ka] Scheme 2B: Synthesis to Compound A-2 via Dipeptide Building Blocks

[0254] The hydroxyl moiety of Boc-Ser-OMe was protected with TBDPS to give Boc-Ser(TBDPS)-OMe. The methyl ester was then removed by treatment with LiOH to give the acid. EDCI-mediated peptide coupling with serinamide gave 27% of the dipeptide in three steps. Treatment with HCl removed the Boc group to give the amine. EDCI-mediated coupling of the amine with the acid (the synthesis of which is shown above) gave 75% of the product in two steps. Hydroxyl elimination was achieved by treatment with mesyl chloride and NEt3. TBAF-mediated TBDPS removal followed by a second elimination reaction gave the bisdehydroalanine motive. Finally, the Alloc group was removed by treatment with palladium in the presence of a scavenger to give compound A-2.

[0255] Alternative synthesis of tautomer / tautomer of compound A-2 ((1) / tautomer): Methyl 2-(2-(2-(4-aminophenyl)thiazole-4-carboxamido)acrylamido)acrylate hydrochloride [ka] Step 1. 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 was prepared according to General Experimental Procedure 3. 2-(4-(((allyloxy)carbonyl)amino)phenyl)thiazole-4-carboxylic acid (1.2 g, 1 equiv., 3.8 mmol) (as prepared in Step 2) and (S)-2-amino-N-((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)propanamide hydrochloride (as prepared in 7) (2.2 g, 1.2 equiv., 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 3 acid) RT: 1.44 min; Area % (254 nm): 92%; m / z=716.2 [M+H] + . 1H NMR (400 MHz,CDCl3) δ 8.35 (d, J =6.6 Hz,1H),8.06 (s,1H),7.89 - 7.81 (m,2H),7.65 (tt, J =6.2,1.5 Hz, 4H), 7.38 (dddd, J = 22.2, 20.7, 8.4, 6.7 Hz, 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, 4.1 Hz, 1H), 4.22 (ddd, J = 17.4, 10.8, 3.3 Hz, 2H), 3.95 (dd, J = 10.2, 4.9 Hz, 1H), 3.63 (dd, J = 11.4, 4.5 Hz, 1H), 3.47 (s, 1H), 1.09 (s, 9H).

[0256] Step 2. 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 was prepared according to general experimental procedure 6. 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 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 (0.35 g), which was used directly in the next step.

[0257] Step 3. N-(3-((3-amino-3-oxoprop-1-en-2-yl)amino)-3-oxoprop-1-en-2-yl)-2-(4-aminophenyl)thiazole-4-carboxamide hydrochloride was prepared using the following procedure. To a solution of crude 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 (0.35 g, 0.79 mmol) in DCM (10 mL) was added silylbenzene (0.49 mL, 5 eq, 4.0 mmol) and tetrakis (92 mg, 0.1 eq, 79 μmol) and the resulting mixture was stirred for 1 h. MeCN (4 mL) was added and DCM was removed in vacuo. The mixture was directly subjected 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-5 C3) RT: 1.138 min; Area: 63.7% (215 nm); m / z = 458.2 [M+H] + .

[0258] Example 3: Exemplary synthesis of thiostrepton-inspired structure B [ka] Scheme 3: Synthesis of compound hexapeptide compound B

[0259] Sequential build-up SPPS of hexapeptide compound B was initiated using 5 grams of Rink Amide resin (0.7 mmol / g) using standard SPPS protocols. 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 give compound B with a purity of >95% as the TFA salt. To convert to the HCl salt, the purified peptide was basified with NaOH and re-acidified with HCl, followed by preparative HPLC using HCl buffer to give the pure peptide as the HCl salt.

[0260] Alternative synthesis: H-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl salt [ka] H-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl salt was prepared on a 2.8 mmol scale following the general procedure for solid phase peptide synthesis. Purification using HCl buffer afforded the desired HCl salt form. H-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl salt (0.89 g, 0.82 mmol, 29%) was isolated as a white solid. LCMS (30833 LCMS-6): RT: 1.819 min; Area: 92.5% (215 nm), 99.7% (ELSD); m / z = 945.9 [M+H] +

[0261] Example 4: Compounds 1 and 2 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-oxopropan-2-ylidene)amino)-1-oxopropan-2-ylidene)carbamoyl)thiazol-2-yl)phenyl)carbamate [ka]

[0262] Step 1. Methyl N-(tert-butoxycarbonyl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared as follows: To a solution of methyl(tert-butoxycarbonyl)-L-serinate (12 g, 1 eq, 55 mmol) in DCM (250 mL) and imidazole (8.2 g, 2.2 eq, 0.12 mol) was added TBDPS-Cl (17 g, 15 mL, 1.1 eq, 60 mmol) and the resulting mixture was stirred at room temperature overnight. The mixture was washed with 1 M HCl (200 mL) and brine (200 mL), dried over Na2SO4, filtered and concentrated to give methyl N-(tert-butoxycarbonyl)-O-(tert-butyldiphenylsilyl)-L-serinate (24 g, 52 mmol, 96%) as a cloudy oil that solidified on standing. 1 H NMR (400 MHz,CDCl3) δ7.59(m,4H),7.46 - 7.32 (m,6H),5.39 (d, J =8.8 Hz,1H),5.28 (d, J = 1.0 Hz,1H) ,4.38 (dt, J = 9.2,3.1 Hz,1H),4.05 (dd, J = 10.0,3.0 Hz,1H),3.87 (dd, J = 10.1,3.1 Hz,1H),3.72 (s,3H), 1.44(s,9H),1.01(s,9H).

[0263] Step 2. N-(tert-butoxycarbonyl)-O-(tert-butyldiphenylsilyl)-L-serine was prepared according to General Experimental Procedure 2. Methyl N-(tert-butoxycarbonyl)-O-(tert-butyldiphenylsilyl)-L-serinate (12 g, 26 mmol) gave N-(tert-butoxycarbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (11.9 g, 26.8 mmol, quantitative). This material was used in the next step without purification.

[0264] 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 according to general experimental procedure 3. N-(tert-butoxycarbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (10 g, 1 equiv., 23 mmol) and L-serinamide hydrochloride (6.5 g, 2 equiv., 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. 1 H NMR (400 MHz,CDCl3) δ 7.59 (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, 4.4 Hz, 1H), 3.60 (dd, J = 11.4, 4.3 Hz, 1H), 1.43 (s, 9H), 1.04 (s, 9H).

[0265] Step 4. (S)-2-amino-N-((S)-1-amino-3-hydroxy-1-oxopropan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)propanamide hydrochloride was prepared according to 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 4 M HCl in dioxane (15 mL) and the resulting mixture was stirred at room temperature for 90 min. 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 HNMR (400 MHz,MeOD) δ 7.73 - 7.62 (m,4H),7.52 - 7.39 (m,6H),4.54 (t, J =5.4 Hz,1H),4.11 (t, J = 4.5 Hz,1H) ), 4.03 (d, J = 4.5 Hz, 2H), 3.86 - 3.74 (m, 2H), 1.07 (s, 9H).

[0266] Step 5. 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 was prepared according to general experimental procedure 3. 2-(4-((tert-butoxycarbonyl)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 eq, 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 3 acids) RT: 1.59 min; Area% (214nm): 75%; m / z=732.2[M+H] + .

[0267] 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-oxopropan-2-ylidene)amino)-1-oxopropan-2-ylidene)carbamoyl)thiazol-2-yl)phenyl)carbamate were prepared according to 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 equiv, 0.89 mmol) and methanesulfonyl chloride (21 μL, 1.5 equiv, 0.27 mmol) and the resulting mixture was stirred at room temperature for 3 h. The mixture was diluted with CHCl (25 mL), washed with 1 M HCl and brine, dried over NaSO, filtered, and partially concentrated to a volume of approximately 10 mL. DBU (54 μL, 2 equiv, 0.36 mmol) was then added and the mixture was stirred for 1 h. A solution of TBAF in THF (0.36 mL, 1.0 mol, 2 equiv, 0.36 mmol) was added and the mixture was stirred for 30 min. The mixture was diluted with CHCl (25 mL), washed with 1 M HCl and brine, dried over NaSO, filtered and partially concentrated to a volume of approximately 10 mL. To the resulting mixture was added triethylamine (0.12 mL, 5 equiv, 0.89 mmol) and methanesulfonyl chloride (21 μL, 1.5 equiv, 0.27 mmol) and the resulting mixture was stirred at room temperature for 1 h. The mixture was diluted with CHCl (25 mL), washed with 1 M HCl and brine, dried over NaSO, filtered and partially concentrated to a volume of approximately 10 mL. DBU (54 μL, 2 equiv, 0.36 mmol) was added and the resulting mixture was stirred for 1 h.The mixture was diluted with CH2Cl2 (25 mL), washed with 1M HCl and brine, dried over Na2SO4, filtered, and concentrated. The crude material was purified by automated reverse-phase FCC to give 1 (2.0 mg, 4.4 μmol, 2.4%) and 2 (8.0 mg, 17 μmol, 9.7%).

[0268] 1:LCMS(22010199A TFA LCMS-5 C1)RT:1.138min;Area:63.7%(215nm);m / z458.2[M+H] + . 1 H NMR (400 MHz,CDCl3) δ 9.99 (s,1H),8.92 (s,1H),8.07 (d, J =5.2 Hz,1H),7.96 - 7.86 (m,2H),7.45 (m, 2H), 6.76 (d, J = 2.2 Hz, 1H), 6.65 (d, J = 2.1 Hz, 1H), 6.62 (s, 1H), 5.53 (t, J = 1.9 Hz, 1H), 5.43 - 5.36 (m, 1H), 1.65 (s, 13H), 1.52 (s, 9H).

[0269] 2:LCMS(22010199 LCMS-5 C3)RT:1.138min;Area:63.7%(215nm);m / z458.2[M+H] + . 1 H NMR (400 MHz,CDCl3) δ 10.84 (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).

[0270] Example 5: Compound 4 Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate [ka]

[0271] Step 1. Methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 3. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (Intermediate 2) (2.0 g, 6.2 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (2.5 g, 1 eq, 6.2 mmol) gave methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serinate (3.5 g, 5.3 mmol, 85%).

[0272] Step 2. N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (Intermediate 3) was prepared according to general experimental procedure 2. Methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serinate (3.5 g, 5.3 mmol) gave crude N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (Intermediate 3) as a clear oil, which was used as such in the next step.

[0273] Step 3. Methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate was prepared according to General Experimental Procedure 3. Crude N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-serine (Intermediate 3) (obtained from Step 2) and methyl L-serinate hydrochloride (1.2 g, 1.5 eq, 8.0 mmol) gave methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate (1.0 g, 1.3 mmol, 25%) as a yellow oil. LCMS (general 3 acid) RT: 1.74 min; area % (254 nm): 87%; m / z=747.2 [M+H] + .

[0274] Step 4. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)acrylate was prepared according to General experimental procedure 6. Methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate (0.10 g, 1 eq, 0.13 mmol) gave methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)acrylate (22 mg, 47 μmol, 35%) as a white solid. LCMS(22010199A TFA LCMS-5 C1)RT:1.6884min;Area%(215nm):97.1%;m / z=473.2[M+H] + . 1H NMR (400 MHz,CDCl3) δ 9.98 (s,1H),8.52 (s,1H),8.07 (s,1H),7.96 - 7.88 (m,2H),7.45 (d, J =8.7 Hz, 2H), 6.75 (d, J = 2.2 Hz, 1H), 6.69 (s, 1H), 6.61 (s, 1H), 6.01 (d, J = 1.3 Hz, 1H), 5.48 (t, J = 1.9 Hz, 1H), 3.88 (s, 3H), 1.52 (s, 9H).

[0275] Alternative synthesis: [ka]

[0276] Step 1. Ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylate was prepared according to 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 (299 MHz,CDCl3) δ 8.39 (s,1H),7.94 (d, J =8.8 Hz,2H),7.49 (d, J = 8.6 Hz,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).

[0277] Step 2. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid was prepared according to general experimental procedure 2. Ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylate (1.2 g, 3.4 mmol) gave 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid (1.0 g, 3.1 mmol, 91%) as a white solid.

[0278] Step 3. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serinate was prepared according to general experimental procedure 4. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid (1.6 g, 5.0 mmol) and H-Ser-OMe.HCl (0.93 g, 1.2 equiv, 6.0 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serinate (1.4 g, 3.3 mmol, 67%) as an orange oil. LCMS (general 3 acid) RT: 0.99 min; Area % (254 nm): 88%; m / z=422.3 [M+H] + .

[0279] Step 4. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine was prepared according to general experimental procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serinate (1.4 g, 3.3 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine (0.98 g, 2.4 mmol, 72%).

[0280] Step 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 4. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine (0.98 g, 2.4 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (1.1 g, 1.2 equiv., 2.9 mmol) gave crude methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (2.25 g), which was used directly in the next step.

[0281] Step 6. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 5. Crude methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (2.25 g) and acetic anhydride (0.34 mL, 3.6 mmol) gave methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.40 g, 0.51 mmol, 21% in two steps). LCMS (general 3 acid) RT: 1.78 min; area % (214 nm): 100%; m / z=789.6 [M+H] + .

[0282] Step 7. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxamido)acrylamide)acrylate was prepared according to General experimental procedure 7. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.19 g, 0.24 mmol) gave methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxamido)acrylamide)acrylate (7.8 mg, 16 μmol, 27%) as a white solid. LC-MS (22010199A TFA LCMS-5 C1): RT: 1.353 min; area 97.7% (215nm), 96.7% (304nm); m / z=473.2[M+H] + . 1 H NMR (400 MHz,CDCl3) δ (ppm) 8.66 (s,1H),8.57 (s,1H),8.24 (s,1H),7.92 (d,J=8.8 Hz,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).

[0283] Example 6: Compound 6 Methyl 2-(2-(2-(4-acetamidophenyl)thiazole-4-carboxamido)acrylamido)acrylate [ka]

[0284] Step 1. Methyl N-(2-(4-aminophenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate was prepared using the following procedure. Methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate (as prepared in step 3) (0.80 g, 1 equiv., 1.1 mmol) was dissolved in CHCl (5 mL) and TFA (5 mL) was added. The resulting mixture was stirred at room temperature for 1 h. The mixture was diluted with CHCl (50 mL) and poured into 1 M NaOH (aq. 150 mL). The layers were separated and the aqueous phase was extracted with CHCl (50 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated to give methyl N-(2-(4-aminophenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate (0.65 g, 1.0 mmol, 94%) as a yellow oil.

[0285] Step 2. Methyl (S)-2-(2-(2-(4-acetamidophenyl)thiazole-4-carboxamido)-3-((methylsulfonyl)oxy)propanamido)acrylate was prepared as follows: Methyl N-(2-(4-aminophenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate (0.22 g, 1 eq, 0.34 mmol) dissolved in CHCl (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 h. LCMS monitoring showed partial conversion to the bis-acetylated product. Additional DiPEA (0.18 mL, 3 equiv, 1.0 mmol), acetic acid (39 μL, 2 equiv, 0.68 mmol) and COMU (0.17 g, 1.2 equiv, 0.41 mmol) were added and the mixture was stirred for 24 h. The mixture was diluted with CHCl (20 mL), washed with HCl (1 M, 20 mL), NaHCO (sat. aq. 20 mL) and brine, dried over NaSO, filtered and concentrated. The residue was dissolved in CHCl (5 mL) and DBU (0.15 mL, 3 equiv, 1.0 mmol) was added. The resulting mixture was stirred for 1 h. A solution of tetrabutylammonium fluoride in THF (0.51 mL, 1.0 M, 1.5 equiv, 0.51 mmol) was added and the resulting mixture was stirred for 1 h. The mixture was diluted with CH2Cl2 (20 mL), washed with HCl (1M, 20 mL) and brine, dried over Na2SO4, filtered and concentrated. The residue was dissolved in CH2Cl2 (5 mL) and methanesulfonyl chloride (40 μL, 1.5 equiv, 0.51 mmol) and triethylamine (0.24 mL, 5 equiv, 1.7 mmol) were added. The resulting mixture was stirred at room temperature for 90 min, after which complete mesylation and partial elimination was observed by LCMS monitoring. The mixture was diluted with CH2Cl2 (20 mL), washed with HCl (1M, 20 mL) and brine, dried over Na2SO4, filtered and concentrated.The residue was dissolved in CHCl (5 mL) and DBU (0.16 g, 0.15 mL, 3 eq, 1.0 mmol) was added. The resulting mixture was stirred at room temperature for 90 min. The mixture was diluted with CHCl (20 mL), washed with HCl (1M, 20 mL) and brine, dried over NaSO, filtered and concentrated. The crude material was purified by automated reverse phase FCC to give methyl 2-(2-(2-(4-acetamidophenyl)thiazole-4-carboxamido)acrylamide)acrylate (5.2 mg, 0.34 mmol, 3.7%) as a white solid. LCMS (22010199C TFA LCMS-5 C3) RT: 1.062 min; area % (215 nm): 91.0%; m / z = 415.0 [M+H]. + . 1 H NMR (400 MHz,CDCl3) δ 9.99 (s,1H),8.53 (s,1H),8.10 (s,1H),8.00 - 7.92 (m,2H),7.61 (d, J =8.3 Hz, 2H), 7.29 (s, 1H), 6.76 (d, J =2.2 Hz, 1H), 6.69 (s, 1H), 6.01 (d, J = 1.4 Hz, 1H), 5.48 (t, J = 1.9 Hz, 1H), 3.88 (s, 3H), 2.21 (s, 3H).

[0286] Example 7: Compound 16 Methyl 2-(2-(2-phenylthiazole-4-carboxamido)acrylamido)acrylate [ka]

[0287] 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 (aq, 2 M, 21 mL, 10 equiv, 42 mmol) and palladium tetrakis (0.24 g, 0.05 equiv, 0.21 mmol). The resulting mixture was stirred at 80° C. for 16 h. The mixture was cooled to room temperature and diluted with EtOAc. The mixture was washed with water. The aqueous layer was acidified by adding 1 M HCl. The precipitate that 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 (299 MHz,CD3OD) δ 8.52 (s,1H),8.00 (m,2H),7.60 (m,2H),1.55 (s,9H).

[0288] Step 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serinate was prepared according to general experimental procedure 4. 2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carboxylic acid (0.66 g, 2.2 mmol) and methyl L-serinate hydrochloride (0.41 g, 1.2 equiv, 2.6 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serinate (0.98 g, 2.4 mmol, quantitative) as an orange oil.

[0289] Step 3. (2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serine was prepared according to general experimental procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serinate (0.98 g, 2.4 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serine (0.64 g, 1.6 mmol, 68%) as an off-white solid. 1 H NMR (299 MHz,cd3od) δ 8.43 (s,1H),8.05 - 7.96 (m,2H),7.60 (m,2H),4.71 m,1H),4.14 - 3.94 (m,2H), 3.79 - 3.69 (m, 1H), 1.55 (s, 9H).

[0290] Step 4. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl-L-serinate was prepared according to general experimental procedure 4. (2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-serine (0.20 g, 0.51 mmol) and methyl L-serinate hydrochloride (95 mg, 1.2 equiv, 0.61 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl-L-serinate (0.18 g, 0.37 mmol, 72%) as a colorless oil.

[0291] Step 5. Methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl)-L-serinate was prepared according to the following general experimental procedure 5. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl-L-serinate (0.18 g, 0.37 mmol) and acetic anhydride (90 μL, 2.6 equiv, 0.95 mmol) gave methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl)-L-serinate (0.12 g, 0.20 mmol, 56%). 1 H NMR (299 MHz,CDCl3) δ 8.26 (s,1H),7.98 (d, J=8.6 Hz,2H ),7.79 (d,J = 8.0 Hz,1H),7.51 (d, J = 8.6 Hz) ,2H),7.32 (d,J =7.8 Hz,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).

[0292] Step 6. Methyl (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carboxamido)acryloyl)-L-serinate was prepared according to General experimental procedure 8. Methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carbonyl)-L-seryl)-L-serinate (0.12 g, 0.20 mmol) gave methyl (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)oxazole-4-carboxamido)acryloyl)-L-serinate (29 mg, 64 μmol, 32%) as a white solid. LC-MS (22010199A TFA LCMS-5 C1):rt 1.560 min;Area 99.7%(215nm),99.2%(304nm);m / z=[M+H]+ =457.2 1 H NMR (400 MHz, CDCl3) δ (ppm) 9.57 (s, 1H), 8.54 (s, 1H), 8.25 (s, 1H), 8.02 (d, J = 8.7 Hz, 2H), 7.49 (d, J = 8.7 Hz, 2H), 6.75 (d, J = 2.2 Hz), 6.71 (s, 1H), 6.64 (s, 1H), 6.03 (d, J = 1.0Hz, 1H), 5.49 (t, J = 1.9 Hz, 1H), 3.90 (s, 3H), (s, 9H).

[0293] Example 8: Compound 49 [ka]

[0294] Step 1. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-serinate was prepared according to general experimental procedure 4. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (intermediate 2) (0.68 g, 2.1 mmol) and methyl L-serinate hydrochloride (0.40 g, 1.2 equiv, 2.6 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serinate (0.87 g, 2.1 mmol, 97%) as an off-white solid. LCMS (general tribasic) RT: 1.44 min; area % (254 nm): 98%; m / z = 420.3 [MH] - .

[0295] Step 2. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-serine (Intermediate 4) was prepared according to General Experimental Procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-serinate (0.88 g, 2.1 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-serine (Intermediate 4) (0.84 g, 2.1 mmol, quantitative) as a white solid.

[0296] Step 3. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to general experimental procedure 4. (2-(4-((tert-butoxycarbonyl)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 equiv, 2.6 mmol) gave methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.5 g, 1.9 mmol, 91%). LCMS (general 3 acids) RT: 1.75 min; Area% (254nm): 98%; m / z=747.6[M+H] + .

[0297] Step 4. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.5 g, 1.9 mmol) gave methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.6 g, 2.0 mmol, quantitative) as a yellow foam. LCMS (general 3 acids) RT: 1.84 min; Area% (254nm): 98%; m / z=789.6[M+H] + .

[0298] Step 5. Methyl N-(O-acetyl-N-(2-(4-aminophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared using the following procedure. To a solution of methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.6 g, 2.0 mmol) in CHCl (30 mL), TFA (4.6 mL, 30 equiv, 60 mmol) was added and the resulting mixture was stirred for 1 h. The reaction mixture was quenched with saturated aqueous NaHCO until gas formation ceased and pH paper indicated the mixture had a pH of approximately 7-8. The mixture was extracted with CH2Cl2 (3x100ml) and the combined organics were dried over Na2SO4, filtered and concentrated to give methyl N-(O-acetyl-N-(2-(4-aminophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.3g, 1.8mmol, 91%). LCMS (general tribasic), RT: 2.01min, area% (254nm): 72%; m / z=687.4[MH]- .

[0299] Step 6. The DBCO coupling intermediate was prepared using the following procedure. To a solution of DBCO acid (0.37 g, 1.2 mmol) and triethylamine (0.19 mL, 1.1 equiv, 1.3 mmol) in CHCl (6 mL) was added propyl chloroformate (0.15 mL, 1.1 equiv, 1.3 mmol) at 0 °C and the resulting mixture was stirred at 0 °C for 1 h. A solution of methyl N-(O-acetyl-N-(2-(4-aminophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.61 g, 0.73 equiv, 0.88 mmol) in CHCl (3 mL) was added and the resulting mixture was stirred at room temperature for 2 h. The mixture was washed with NaHCO (sat, 3x10 mL) and HCl (1 M, 3x10 mL), dried over NaSO, 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 acid) RT: 1.86 min; Area % (254 nm): 86%; m / z=976.5 [M+H] + .

[0300] Step 7. DBCO-linked bisdehydroalanine compound was prepared according to general experimental procedure 5. DBCO-linked intermediate (0.52 g, 0.54 mmol) gave DBCO-linked bisdehydroalanine compound (24 mg, 37 μmol, 6.9%) as a white solid. LCMS (22010199D TFA LCMS-5 C8) RT: 1.642 min; Area % (215 nm): 79.2%; m / z = 660.2 [M+H] + . 1H NMR (400 MHz, CDCl3) δ 10.05 (s, 1H), 8.56 (s, 1H), 8.42 (s, 1H), 8.09 (s, 1H), 7.84 (d, J = 8.6 Hz, 2H), 7.71 (d, J = 7.4 Hz, 1H), 7.50 - 7.28 (m, 8H), 7.19 (dd, J = 7.4, 1.5 Hz, 1H), 6.78 (d, J = 2.2 Hz, 1H), 6.73 (s, 1H), 6.03 (d, J = 1.3 Hz, 1H), 5.50 (t, J = 1.9 Hz, 1H), 5.19 (d, J = 13.9 Hz, 1H), 3.91 (s, 3H), 3.73 (d, J = 13.8 Hz, 1H), 2.86 (m, 1H), 2.45 (m, 2H), 2.07 (m, 1H).

[0301] Example 9: Compound 50 Methyl 2-(2-(2-(4-(hex-5-ynamido)phenyl)thiazole-4-carboxamido)acrylamido)acrylate [ka]

[0302] Step 1. Methyl 2-(2-(2-(4-(hex-5-ynamido)phenyl)thiazole-4-carboxamido)acrylamide)acrylate was prepared as follows. Methyl N-(2-(4-aminophenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)-L-seryl-L-serinate (as prepared in step 1) (0.22 g, 0.334 mmol) dissolved in CHCl (5 mL) and DiPEA (0.18 mL, 3 equiv, 1.0 mmol) and hex-5-ynic acid (56 μL, 1.5 equiv, 0.51 mmol) were added. COMU (0.18 g, 1.2 equiv, 0.41 mmol) was then added and the resulting mixture was stirred at room temperature for 16 h. LCMS monitoring showed partial conversion to the bisacylated product. Additional DiPEA (0.18 mL, 3 equiv, 1.0 mmol), hex-5-ynoic acid (57 mg, 56 μL, 1.5 equiv, 0.510 mmol) and COMU (0.18 mg, 1.2 equiv, 0.41 mmol) were added and the resulting mixture was stirred for 24 h. The mixture was diluted with CHCl (20 mL), washed with HCl (1 M, 20 mL), NaHCO (sat. aq. 20 mL) and brine, dried over NaSO, filtered and concentrated. The residue was dissolved in CHCl (5 mL) and additional DBU (0.15 mL, 3 equiv, 1.0 mmol) was added. The resulting mixture was stirred for 1 h. A solution of tetrabutylammonium fluoride in THF (0.51 mL, 1.0 M, 1.5 equiv, 0.51 mmol) was added and the resulting mixture was stirred for 1 h. The mixture was diluted with CH2Cl2 (20 mL), washed with HCl (1M, 20 mL) and brine, dried over Na2SO4, filtered and concentrated. The residue was dissolved in CH2Cl2 (5 mL) and methanesulfonyl chloride (40 μL, 1.5 equiv, 0.51 mmol) and triethylamine (0.17 g, 0.24 mL, 5 equiv, 1.7 mmol) were added. The resulting mixture was stirred at room temperature for 90 min, after which complete mesylation and partial elimination was observed by LCMS monitoring. The mixture was diluted with CH2Cl2 (20 mL), washed with HCl (1M, 20 mL) and brine, dried over Na2SO4, filtered and concentrated.The residue was dissolved in CHCl (5 mL) and DBU (0.16 g, 0.15 mL, 3 eq, 1.0 mmol) was added. The resulting mixture was stirred at room temperature for 90 min. The mixture was diluted with CHCl (20 mL), washed with HCl (1M, 20 mL) and brine, dried over NaSO, filtered and concentrated. The crude material was purified by automated reverse phase FCC to give methyl 2-(2-(2-(4-(hex-5-ynamido)phenyl)thiazole-4-carboxamido)acrylamido)acrylate (4.5 mg, 9.6 μmol, 2.8%) as a white solid. LCMS (22010199C TFA LCMS-5 C3) RT: 1.287 min; area % (215 nm): 93.0%; m / z=467.2 [M+H]. + . 1 H NMR (400 MHz, CDCl3) δ 9.98 (s, 1H), 8.53 (s, 1H), 8.00 - 7.92 (m, 2H), 7.62 (d, J = 8.3 Hz, 2H), 7.31 (s, 1H), 6.76 (d, J = 2.2 Hz, 1H), 6.69 (s, 1H), 6.01 (d, J = 1.2 Hz, 1H), 5.48 (t, J = 1.9 Hz, 1H), 3.88 (s, 3H), 2.55 (t, J = 7.3 Hz, 2H), 2.34 (td, J = 6.7, 2.6 Hz, 2H), 2.04 - 1.92 (m, 3H).

[0303] Example 10: Compound 51 [ka] Compound 51 was prepared according to the general procedure for conjugation chemistry between azide-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 (regioisomers); Area: 95.3% (215 nm), 99.6% (ELSD) m / z = 1744.1 [M + H] +

[0304] Example 11: Compound 52 Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxamido)acrylamido)acrylate [ka]

[0305] Step 1. Methyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylate was prepared according to general experimental procedure 1. Methyl-2-bromo-5-methyl-thiazole-4-carboxylate (0.91 g, 3.8 mmol) and (4-((tert-butoxycarbonyl)amino)phenyl)boronic acid (0.86 g, 3.6 mmol) gave methyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylate (1.1 g, 3.1 mmol, 88%). LCMS (general tribasic) RT: 1.80 min; area % (214 nm): 87%; m / z = 347.2 [MH] - .

[0306] Step 2. 2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylic acid was prepared according to general experimental procedure 2. Methyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylate (1.1 g, 3.1 mmol) gave methyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylate (1.0 g, 3.1 mmol, quantitative). LCMS (general tribasic) RT: 1.17 min; area % (214 nm): 81%; m / z = 335.4 [M+H] + .

[0307] Step 3. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serinate was prepared according to general experimental procedure 3. 2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxylic acid (1.1 g, 3.2 mmol) and L-Ser-OMe.HCl (0.60 g, 1.2 equiv, 3.9 mmol) gave crude methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serinate (2.6 g), which was used in the next step without purification. LCMS (general tribasic) RT: 1.61 min; area % (214 nm): 69%; m / z=436.2 [M+H] + .

[0308] Step 4. (2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serine was prepared according to general experimental procedure 2. Crude methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serinate (2.6 g crude) gave crude (2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serine (1.8 g), which was used in the next step without purification. LCMS (general tribasic) RT: 1.06 min; area % (214 nm): 69%; m / z=422.4 [M+H] + .

[0309] Step 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to general experimental procedure 4. (2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)serine (1.4 g, crude) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (1.6 g, 4.0 mmol) gave methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.2 g, 1.5 mmol) as a clear yellow oil. LCMS (general 3 acids) RT: 1.85 min; Area% (214nm): 73%; m / z=761.6[M+H] + .

[0310] Step 6. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to general experimental procedure 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.2 g, 1.5 mmol) and acetic anhydride (0.16 mL, 1.1 equiv, 1.7 mmol) gave methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.1 g, 1.3 mmol, 87%) as a yellow viscous oil. LCMS (general tribasic) RT: 2.31 min; area % (214 nm): 86%; m / z=801.5 [M+H] + .

[0311] Step 7. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxamido)acrylamide)acrylate was prepared according to General experimental procedure 7. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.21 g, 0.26 mmol) gave methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)-5-methylthiazole-4-carboxamido)acrylamide)acrylate (11 mg, 23 μmol, 13%) as a white solid. LCMS(22010199C TFA LCMS-5 C3)RT:1.702min;Area:91.1%(215nm);m / z487.2[M+H] + . 1 H NMR (400 MHz,CDCl3) δ 10.08 (s,1H),8.51 (s,1H),7.88 - 7.80 (m,2H),7.42 (d, J =8.6 Hz,2H),6.72 - 6.66 ( m,2H),6.57 (s,1H),6.00 (d, J =1.2 Hz,1H), 5.42 (t, J =1.8 Hz,1H), 3.88 (s,3H), 2.85 (s,3H), 1.52(s,9H).

[0312] Example 12: Compound 53 Methyl (Z)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoate [ka]

[0313] Step 1. Methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl-L-threoninate was prepared according to General Experimental Procedure 3. N-(2-(4-((tert-butoxycarbonyl)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 methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl-L-threoninate (0.29 g, 0.38 mmol, 27%) as a pale yellow foam. LCMS (general tribasic) RT: 2.20 min; area % (254 nm): 81%; m / z=761.3 [M+H] + .

[0314] Step 2. Methyl O-acetyl-N-(N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl)-L-threoninate was prepared according to General Experimental Procedure 5. Methyl N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl-L-threoninate (0.29 g, 0.38 mmol) and acetic anhydride (37 μL, 1.05 equiv, 0.39 mmol) gave methyl O-acetyl-N-(N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl)-L-threoninate (0.30 g, 0.37 mmol, 98%) as a pale yellow foam.

[0315] Step 3. Methyl (Z)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)but-2-enoate was prepared according to General Experimental Procedure 7. Methyl O-acetyl-N-(N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-O-(tert-butyldiphenylsilyl)seryl)-L-threoninate (0.10 g, 0.13 mmol) gave methyl (Z)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)but-2-enoate (22 mg, 45 μmol, 36%). LCMS(22010199C TFA LCMS-5 C3)RT:1.424min;Area%(215nm):92.9%;m / z=487.2[M+H] + . 1 H NMR (400 MHz, CDCl3) δ 9.97 (s, 1H), 8.07 (s, 1H), 7.94 - 7.86 (m, 2H), 7.48 (s, 1H), 7.42 (d, J = 8.6 Hz, 2H), 6.93 (q, J = 7.2 Hz, 1H), 6.76 (d, J = 1.8 Hz, 1H), 6.59 (s, 1H), 5.53 (t, J = 1.7 Hz, 1H), 3.79 (s, 3H), 1.82 (dd, J = 7.2, 0.7 Hz, 3H), 1.51 (s, 9H).

[0316] Example 13: Compound 55 Methyl (E)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoate [ka]

[0317] Step 1. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allosthreoninate was prepared according to General Experimental Procedure 4. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) (785 mg, 1.93 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-allothreoninate (859 mg, 1.2 equiv, 2.31 mmol) gave methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreoninate (1.54 g, 2.02 mmol, quantitative) as a viscous oil. LCMS (general 3 acid) RT: 1.81 min; area % (254 nm): 96%; m / z=761.3 [M+H] + .

[0318] Step 2. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allosthreoninate was prepared according to general experimental procedure 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allosthreoninate (1.6 g, 2.1 mmol) and acetic anhydride (0.22 mL, 1.1 equiv, 2.3 mmol) gave methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allosthreoninate (1.37 g, 1.71 mmol, 81%) as a yellow solid. LCMS (general 3 acid) RT: 1.88 min; area % (254 nm): 75%; m / z=803.5 [M+H] + .

[0319] Step 3. Methyl (E)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)but-2-enoate was prepared according to General Experimental Procedure 7. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allosthreoninate (306 mg, 0.38 mmol) gave methyl methyl (E)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)but-2-enoate (4.6 mg, 9.5 μmol, 2.5%). LCMS(22010199C TFA LCMS-5 C3)RT:1.470min;Area:93.8%(215nm);m / z487.2[M+H] + . 1 H NMR (400 MHz, CDCl3) δ 9.98 (s, 1H), 8.17 (s, 1H), 8.06 (s, 1H), 7.95 - 7.88 (m, 2H), 7.44 (d, J = 8.6 Hz, 2H), 7.33 (q, J = 7.7 Hz, 2H), 6.70 (d, J = 2.0 Hz, 1H), 6.59 (s, 1H), 5.42 (s, 1H), 3.88 (s, 3H), 2.15 (d, J = 7.7 Hz, 3H), 1.52 (s, 9H).

[0320] Example 14: Compound 56 Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acetamido)acrylate [ka]

[0321] Step 1. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycinate was prepared according to general experimental procedure 3. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (intermediate 2) (0.41 g, 1 eq, 1.3 mmol) and methyl glycinate hydrochloride (0.17 g, 1 eq, 1.3 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycinate (0.37 g, 0.94 mmol, 74%) as a white foam. LCMS (general tribasic) RT: 1.63 min; Area % (254 nm): 92%; m / z=392.2 [M+H] + . 1 H NMR (400 MHz,CDCl3) δ 8.03 (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).

[0322] Step 2. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycine was prepared according to general experimental procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycinate (0.37 mg, 1 eq, 0.94 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycine (0.32 g, 0.85 mmol, 90%) as a white solid. LCMS (general tribasic) RT: 1.12 min; Area % (254 nm): 96%; m / z=378.2 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 9.69 (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 = 6.1 Hz, 2H), 1.49 (s, 9H).

[0323] Step 3. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycyl-L-serinate was prepared according to general experimental procedure 3. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycine (0.31 g, 1 eq, 0.83 mol) and methyl L-serinate hydrochloride (0.16 g, 1.2 eq, 1.0 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)glycylserinate (0.29 mg, 0.60 mmol, 72%). LCMS (general tribasic) RT: 1.47 min; area % (254 nm): 93%; m / z = 479.3 [M+H] + . 1 H NMR (400 MHz, MeOD) δ 8.07 (s, 1H), 7.90 - 7.82 (m, 2H), 7.52 - 7.44 (m, 2H), 4.56 (t, J = 4.3 Hz, 1H), 4.15 (d, J = 2.1 Hz, 2H), 3.92 - 3.76 (m, 2H), 3.71 (s, 3H), 1.49 (s, 9H).

[0324] 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)glycylserinate (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-5 C8)RT:1.382min;Area%(215nm):98.0%;m / z=461.2[M+H] + . 1 H NMR (400 MHz, CDCl3) δ 8.36 (s, 1H), 8.09 (d, J = 2.4 Hz, 1H), 8.00 (d, J = 6.5 Hz, 1H), 7.91 - 7.80 (m, 2H), 7.45 (d, J = 8.3 Hz, 2H), 6.61 (d, J = 2.5 Hz, 2H), 5.92 (d, J = 1.5 Hz, 1H), 4.24 (d, J = 5.9 Hz, 2H), 3.82 (s, 3H), 1.52 (s, 9H).

[0325] Example 15: Compound 57 Methyl (S)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)propanamido)acrylate [ka]

[0326] Step 1. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-alaninate was prepared according to general experimental procedure 3. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (intermediate 2) (0.53 g, 1.7 mmol) and H-Ala-OMe.HCl (0.27 g, 1.2 equiv, 1.9 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-alaninate (0.43 mg, 1.1 mmol, 63%). LCMS (general tribasic) RT: 1.77 min; area % (254 nm): 86%; m / z=404.2 [M+H] + .

[0327] Step 2. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-D-alanine was prepared according to general experimental procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-D-alaninate (0.43 g, 1.1 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-D-alanine (0.41 g, 1.1 mmol, quantitative) as a yellow foam. LCMS (general tribasic) RT: 1.16 min; area % (254 nm): 95%; m / z=392.2 [M+H] + .

[0328] Step 3. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-alanyl-L-serinate was prepared according to general experimental procedure 3. (2-(4-((tert-butoxycarbonyl)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 methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-alanyl-L-serinate (0.24 mg, 0.50 mmol, 47%) as a yellow foam. LCMS (general tribasic) RT: 1.54 min; Area% (254nm): 85%; m / z=493.2[M+H] + .

[0329] Step 4. Methyl (S)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)propanamide)acrylate was prepared using the following procedure. To a solution of methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-alanylserinate (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 methyl (S)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)propanamide)acrylate as a white solid. LCMS(22010199D TFA LCMS-5 C8)RT:1.453min;Area%(215nm):94.9%;m / z=475.2[M+H] + . 1H NMR (400 MHz, CDCl3) δ 8.54 (s, 1H), 8.08 (s, 1H), 7.90 - 7.80 (m, 3H), 7.49 - 7.42 (m, 2H), 6.70 (s, 1H), 6.59 (s, 1H), 5.91 (d, J = 1.4 Hz, 1H), 4.77 (p, J = 7.1 Hz, 1H), 3.81 (s, 3H), 1.56 (d, J = 7.0 Hz, 3H), 1.52 (s, 9H).

[0330] Example 16: Compound 58 Methyl (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)glycinate [ka]

[0331] Step 1. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serinate was prepared according to general experimental procedure 3. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (intermediate 2) (1.0 g, 3.2 mmol) and L-Ser-OMe.HCl (0.59 g, 1.2 equiv, 3.8 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serinate (0.88 g, 2.1 mmol, 66%). LCMS (general tribasic) RT: 1.54 min; area % (254 nm): 64%; m / z = 420.2 [MH] - .

[0332] Step 2. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) was prepared according to general experimental procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serinate (0.88 g, 2.1 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) (0.84 mg, 2.1 mmol, 99%) as a white solid.

[0333] Step 3. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl glycinate was prepared according to general experimental procedure 3. (2-(4-((tert-butoxycarbonyl)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 methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl glycinate (0.35 g, 0.72 mmol, 70%) as a yellow solid. 1 H NMR (400 MHz,MeOD) δ 8.16 (s,1H),7.98 - 7.88 (m,2H),7.59 - 7.51 (m,2H),4.71 (t, J =4.9 Hz,1H),4.04 - 3.96 (m, 3H), 3.92 (dd, J = 11.3, 4.8 Hz, 1H), 3.73 (s, 3H), 1.54 (s, 9H).

[0334] Step 4. Methyl O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl glycinate was prepared according to general experimental procedure 5. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl glycinate (0.13 mg, 0.27 mmol) and acetic anhydride (26 μL, 1 equiv, 0.27 mmol) gave methyl O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl glycinate (153 mg, quantitative). LCMS (general 3 acid) RT: 1.14 min; Area % (254 nm): 92%; m / z = 521.5 [M+H] + .

[0335] Step 5. Methyl (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)glycinate was prepared using the following procedure. To a solution of methyl (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)glycinate (51 mg, 99 μmol) in CHCl (1 mL) was added DBU (30 μL, 2 equiv, 197 μmol) and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure and the crude material was purified by automated reverse-phase FCC to give methyl (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)glycinate (12.5 mg, 27.1 μmol, 28%) as a white solid. LCMS(22010199D TFA LCMS-5 C8)RT:1.403min;Area%(215nm):93.2%;m / z=461.2[M+H] + . 1H NMR (400 MHz, CDCl3) δ 9.98 (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 = 1.7 Hz, 1H), 4.18 (d, J = 5.0 Hz, 2H), 3.79 (s, 3H), 1.52 (s, 9H).

[0336] Example 17: Compound 59 Methyl (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)-L-alaninate [ka]

[0337] Step 1. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl-L-alaninate was prepared according to general experimental procedure 3. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) (0.42 g, 1.0 mmol) and H-Ala-OMe.HCl (0.19 g, 1.3 equiv, 1.3 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl-L-alaninate (0.24 g, 0.49 mmol, 48%) as a yellow foam. LCMS (General Tribasic) RT: 1.60 min; Area % (254 nm): 81%; m / z=493.2 [M+H] + .

[0338] Step 2. Methyl O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl-L-alaninate was prepared according to General Experimental Procedure 5. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl-L-alaninate (0.20 g, 0.41 mmol) and acetic anhydride (42 μL, 0.44 mmol) gave methyl O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl-L-alaninate (0.23 g, 0.44 mmol, quantitative) as a pale yellow foam. LCMS (general tribasic) RT: 1.59 min; Area% (254nm): 86%; m / z=535.2[M+H] + .

[0339] Step 3. Methyl (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)-L-alaninate was prepared using the following procedure. To a solution of methyl O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl-L-alaninate (52 mg, 98 μmol) in CHCl (1 mL) was added DBU (30 μL, 0.20 mmol) and the resulting mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure and the crude material was purified by automated reverse phase FCC to give methyl (2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acryloyl)-L-alaninate (10.8 mg, 22.8 μmol, 23%). LCMS (22010199D TFA LCMS-5 C8) RT: 1.472 min; Area % (215 nm): 97.4%; m / z=475.2 [M+H] + . 1H NMR (400 MHz, CDCl3) δ 9.99 (s, 1H), 8.05 (s, 1H), 7.90 (d, J = 8.4 Hz, 2H), 7.43 (d, J = 8.3 Hz, 2H), 6.80 (d, J = 7.2 Hz, 1H), 6.72 (s, 1H), 6.67 (d, J = 1.8 Hz, 1H), 5.44 - 5.39 (m, 1H), 4.69 (p, J = 7.2 Hz, 1H), 3.79 (s, 3H), 1.50 (m, 12H).

[0340] Example 18: Compound 60 N3(CH2CH2O)3CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl salt [ka] N3(CH2CHO)3CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl 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(CH2CHO)3CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl salt (53 mg, 42 μmol, 17%) was isolated as a white solid. LCMS (30833 LCMS-6): RT: 2.365 min; Area: 83.7% (215 nm), 99.8% (ELSD); m / z=1160.8 [M+H] +

[0341] Example 19: Compound 61 [ka] Compound 61 was prepared according to the general procedure for conjugation chemistry between azide-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] +

[0342] Example 20: Compound 62 Methyl (E)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoate [ka] Step 1. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allosthreoninate was prepared according to General Experimental Procedure 4. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (Intermediate 4) (785 mg, 1.93 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-allothreoninate (859 mg, 1.2 equiv, 2.31 mmol) gave methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allothreoninate (1.54 g, 2.02 mmol, quantitative) as a viscous oil. LCMS (general 3 acid) RT: 1.81 min; area % (254 nm): 96%; m / z=761.3 [M+H] + .

[0343] Step 2. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allosthreoninate was prepared according to general experimental procedure 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allosthreoninate (1.6 g, 2.1 mmol) and acetic anhydride (0.22 mL, 1.1 equiv, 2.3 mmol) gave methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allosthreoninate (1.37 g, 1.71 mmol, 81%) as a yellow solid. LCMS (general 3 acid) RT: 1.88 min; area % (254 nm): 75%; m / z=803.5 [M+H] + .

[0344] Step 3. Methyl (E)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)but-2-enoate was prepared according to General Experimental Procedure 7. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)seryl)-O-(tert-butyldiphenylsilyl)-L-allosthreoninate (306 mg, 0.38 mmol) gave methyl methyl (E)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)but-2-enoate (4.6 mg, 9.5 μmol, 2.5%). LCMS(22010199C TFA LCMS-5 C3)RT:1.470min;Area:93.8%(215nm);m / z487.2[M+H] + . 1H NMR (400 MHz, CDCl3) δ 9.98 (s, 1H), 8.17 (s, 1H), 8.06 (s, 1H), 7.95 - 7.88 (m, 2H), 7.44 (d, J = 8.6 Hz, 2H), 7.33 (q, J = 7.7 Hz, 2H), 6.70 (d, J = 2.0 Hz, 1H), 6.59 (s, 1H), 5.42 (s, 1H), 3.88 (s, 3H), 2.15 (d, J = 7.7 Hz, 3H), 1.52 (s, 9H).

[0345] Example 21: Compound 63 Methyl (E)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)but-2-enoate [ka]

[0346] Step 1. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreoninate was prepared according to general experimental procedure 4. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (intermediate 2) (0.98 g, 3.1 mmol) and methyl L-allothreoninate hydrochloride (0.57 g, 1.1 equiv, 3.4 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreoninate (1.6 g, 3.2 mmol, quantitative) as a white solid.

[0347] Step 2. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonine was prepared according to general experimental procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonine (1.6 g, 3.2 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonine (1.4 g, 3.2 mmol, quantitative) as a white solid.

[0348] Step 3. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allosthreonyl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 4. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonine (1.4 g, 3.2 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (1.4 g, 1.1 equiv., 3.5 mmol) gave methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serinate (2.3 g, 3.0 mmol, 93%).

[0349] Step 4. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serinate (2.3 g, 3.0 mmol) gave methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serinate (2.2 g, 2.7 mmol, 90%).

[0350] Step 5. Methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-L-serinate was prepared using the following procedure. To a solution of methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-O-(tert-butyldiphenylsilyl)-L-serinate (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 equiv, 0.56 mmol) and the resulting mixture was stirred for 1 h. The reaction mixture was concentrated under reduced pressure and the residue was dissolved in MeCN (6 mL). Triethylamine (0.13 mL, 2.5 equiv, 0.93 mmol) and acetic anhydride (48 μL, 1.35 equiv, 0.50 mmol) were added and the resulting mixture was stirred at room temperature for 45 min. Water was added and the mixture was concentrated under reduced pressure 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 give methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allostreonyl)-L-serinate (0.16 g, 0.27 mmol, 72%) as a white solid.

[0351] Step 6. Methyl (Z)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)but-2-enamido)acrylate (E / Z stereochemistry arbitrarily assigned) was prepared according to general experimental procedure 8. Methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-allothreonyl)-L-serinate (0.15 g, 0.25 mmol) gave methyl (Z)-2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)but-2-enamido)acrylate (30 mg, 62 μmol, 25%) as a white solid. LCMS(22010199C TFA LCMS-5 C3)RT:1.387min;Area:99.6%(215nm),99.3%(306nm);m / z487.2[M+H] + . 1 H NMR (400 MHz, CDCl3) δ 8.73 (s, 1H), 8.48 (s, 1H), 8.13 (s, 1H), 7.95 - 7.88 (m, 2H), 7.48 (d, J = 8.5 Hz, 2H), 6.73 (q, J = 7.1 Hz, 1H), 6.69 (s, 1H), 6.63 (s, 1H), 5.94 (d, J = 1.4 Hz, 1H), 3.81 (s, 3H), 1.89 (d, J = 7.1 Hz, 3H), 1.54 (s, 9H).

[0352] Example 22: Compound 67 Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxamido)acrylamido)acrylate [ka]

[0353] Step 1. Ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylate was prepared according to 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 2-(4-((tert-butoxycarbonyl)ethylamino)phenyl)thiazole-5-carboxylate (1.26 g, 3.62 mmol, 43%) as a yellow solid. 1 H NMR (299 MHz,CDCl3) δ 8.39 (s,1H),7.94 (d, J =8.8 Hz,2H),7.49 (d, J = 8.6 Hz,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).

[0354] Step 2. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid was prepared according to general experimental procedure 2. Ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylate (1.2 g, 3.4 mmol) gave 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid (1.0 g, 3.1 mmol, 91%) as a white solid.

[0355] Step 3. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serinate was prepared according to general experimental procedure 4. 2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxylic acid (1.6 g, 5.0 mmol) and H-Ser-OMe.HCl (0.93 g, 1.2 equiv, 6.0 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serinate (1.4 g, 3.3 mmol, 67%) as an orange oil. LCMS (general 3 acid) RT: 0.99 min; Area % (254 nm): 88%; m / z=422.3 [M+H] + .

[0356] Step 4. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine was prepared according to general experimental procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serinate (1.4 g, 3.3 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine (0.98 g, 2.4 mmol, 72%).

[0357] Step 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 4. (2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-serine (0.98 g, 2.4 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (1.1 g, 1.2 equiv., 2.9 mmol) gave crude methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (2.25 g), which was used directly in the next step.

[0358] Step 6. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 5. Crude methyl N-((2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (2.25 g) and acetic anhydride (0.34 mL, 3.6 mmol) gave methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.40 g, 0.51 mmol, 21% in two steps). LCMS (general 3 acid) RT: 1.78 min; area % (214 nm): 100%; m / z=789.6 [M+H] + .

[0359] Step 7. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxamido)acrylamide)acrylate was prepared according to General experimental procedure 7. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.19 g, 0.24 mmol) gave methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-5-carboxamido)acrylamide)acrylate (7.8 mg, 16 μmol, 27%) as a white solid. LC-MS (22010199A TFA LCMS-5 C1): RT: 1.353 min; area 97.7% (215nm), 96.7% (304nm); m / z=473.2[M+H] + . 1H NMR (400 MHz,CDCl3) δ (ppm) 8.66 (s,1H),8.57 (s,1H),8.24 (s,1H),7.92 (d,J=8.8 Hz,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).

[0360] Example 23: Compound 68 Methyl 2-(2-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxamido)acrylamido)acrylate [ka]

[0361] Step 1. Ethyl 5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxylate was prepared according to 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 (299 MHz, CDCl3) δ 8.00 (m, 1H), 7.65 (m, 1H), 7.55 (d, J = 8.7 Hz, 2H), 7.41 (d, J = 8.5 Hz, 2H), 7.28 (s, 4H), 6.54 (s, 1H), 4.37 (q, J = 7.1 Hz, 2H), 1.55 (s, 9H), 1.40 (t, J = 7.1 Hz, 3H).

[0362] Step 2. 5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxylic acid was prepared according to General Experimental Procedure 2. Ethyl 5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxylate (1.1 g, 3.2 mmol) and 5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxylic acid (0.90 g, 2.8 mmol, 89%) as white solids. 1 H NMR (299 MHz, CD3OD) δ 8.08 (m, 1H), 7.65 (m, 1H), 7.62 - 7.52 (m, 2H), 7.48 (d, J = 8.7 Hz, 2H), 1.54 (s, 8H).

[0363] Step 3. Methyl (5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serinate was prepared according to General experimental procedure 3. 5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxylic acid (0.90 g, 2.8 mmol) and methyl L-serinate hydrochloride (0.44 g, 2.8 mmol) gave methyl (5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serinate (1.0 g, 2.4 mmol, 84%) as a yellow oil. 1 H NMR (299 MHz, CDCl3) δ 7.84 (m, 1H), 7.56 - 7.46 (m, 3H), 7.40 (d, J = 8.5 Hz, 2H), 7.03 (d, J = 7.2 Hz, 1H), 6.64 (s, 1H), 4.87 (dt, J = 7.2, 3.6 Hz, 1H), 4.39 (q, J = 7.1 Hz, 1H), 4.09 (t, J = 4.0 Hz, 2H), 3.85 (s, 3H), 1.55 (s, 9H).

[0364] Step 4. (5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serine was prepared according to general experimental procedure 2. Methyl (5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serinate (1.0 g, 0.24 mmol) gave (5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serine (0.90 g, 2.2 mmol, 93%) as a yellow solid. 1 H NMR (299 MHz, CD3OD) δ 9.05 (s, 1H), 8.25 (d, J = 8.0 Hz, 0H), 8.04 (d, J = 1.4 Hz, 1H), 7.74 (d, J = 1.4 Hz, 1H), 7.63 - 7.52 (m, 2H), 7.48 (d, J = 8.6 Hz, 2H), 4.76 - 4.67 (m, 1H), 4.10 - 3.92 (m, 2H), 1.55 (s, 9H).

[0365] Step 5. Methyl N-((5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to general experimental procedure 4. (5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-serine (0.20 g, 0.49 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (0.23 g, 1.2 equiv., 0.59 mmol) gave crude methyl N-((5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (233 mg), which was used in the next step without purification.

[0366] Step 6. Methyl N-(O-acetyl-N-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 5. From crude methyl N-((5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (233 mg), methyl N-(O-acetyl-N-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.10 g, 0.13 mmol, 26% in two steps).

[0367] Step 7. Methyl 2-(2-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxamido)acrylamide)acrylate was prepared according to General experimental procedure 7. Methyl N-(O-acetyl-N-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (50 mg, 63 μmol) gave methyl 2-(2-(5-(4-((tert-butoxycarbonyl)amino)phenyl)thiophene-3-carboxamido)acrylamide)acrylate (3.9 mg, 8.3 μmol, 31%) as a white solid. LC-MS (22010199A TFA LCMS-5 C1): RT 1.483 min; area 84.0% (215nm), 88.7% (304nm); m / z=472.0[M+H] +

[0368] Example 24: Compound 69 Methyl 2-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxamido)acrylamido)acrylate [ka]

[0369] Step 1. Methyl 4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxylate was prepared using the following procedure. To a solution of methyl 4'-amino-[1,1'-biphenyl]-4-carboxylate (0.50 g, 2.2 mmol) in CHCl (20 mL) was added DMAP (0.54 g, 2 eq, 4.4 mmol) and BocO (0.48 g, 1 eq, 2.2 mmol) and the resulting mixture was stirred at room temperature for 16 h. The mixture was filtered and HCl (1 M, aq. 50 mL) was added. The mixture was extracted with EtOAc and the organic layer was dried over NaSO, filtered and concentrated. The crude material was purified by FCC to give methyl 4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxylate (0.30 g, 0.92 mmol, 42%). 1 H NMR (299 MHz, CDCl3) δ 8.10 (m, 2H), 7.62 (m, 5H), 7.53 - 7.44 (m, 2H), 6.70 (s, 1H), 3.95 (s, 3H), 1.55 (s, 9H).

[0370] Step 2. 4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxylic acid was prepared according to general experimental procedure 2. Methyl 4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxylate (0.52 g, 1.6 mmol) gave 4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxylic acid (0.34 g, 1.1 mmol, 66%). 1 HNMR (299 MHz, CD3OD) δ 8.13 - 8.00 (m, 2H), 7.77 - 7.58 (m, 4H), 7.53 (m, 2H), 1.55 (s, 9H).

[0371] Step 3. Methyl (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serinate was prepared according to general experimental procedure 4. 4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxylic acid (90 mg, 0.29 mmol) and methyl L-serinate hydrochloride (90 mg, 2 equiv, 0.57 mmol) gave methyl (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serinate (34 mg, 82 μmol, 29%). LCMS (general 3 acid) RT: 1.07 min; Area % (254 nm): 100%; m / z = 415.1 [M+H] + .

[0372] Step 4. (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serine was prepared according to general experimental procedure 2. Methyl (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serinate (34 mg, 82 μmol) gave crude (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serine (77 mg), which was used directly in the next step. LCMS (general 3 acid) RT: 0.95 min; area % (254 nm): 82%; m / z = 401.3 [M+H] + .

[0373] Step 5. Methyl N-((4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 4. (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-serine (100 mg, 0.25 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (118 mg, 0.30 mmol) gave methyl N-((4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (66 mg, 89 μmol, 36%). LCMS (general 3 acid) RT: 1.72 min; area % (254 nm): 77%; m / z=740.6 [M+H] + .

[0374] Step 6. Methyl N-(O-acetyl-N-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to general experimental procedure 5. Methyl N-((4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (66 mg, 89 μmol) gave methyl N-(O-acetyl-N-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (44 mg, 56 μmol, 63%). LCMS (general 3 acid) RT: 1.81 min; area % (254 nm): 100%; m / z=740.6 [M+H] + .

[0375] Step 7. Methyl 2-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxamido)acrylamide)acrylate was prepared according to General Experimental Procedure 7. Methyl N-(O-acetyl-N-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (42 mg, 54 μmol) gave methyl 2-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-4-carboxamido)acrylamide)acrylate (5.0 mg, 11 μmol, 20%). LC-MS (22010199A TFA LCMS-5 C1): rt 1.483 min; area 78.4% (215nm), 83.7% (304nm), m / z=[M+H] + =466.2. 1 H NMR (CDCl3) δ (ppm) 8.92 (s, 1H), 8.60 (s, 1H), 7.92 (d, J = 8.3 Hz, 2H), 7.67 (d, J = 8.1 Hz, 2H), 7.57 (d, J = 8.4 Hz, 2H), 7.47 (d, J = 8.4 Hz, 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).

[0376] Example 25: Compound 70 Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxamido)acrylamido)acrylate [ka]

[0377] Step 1. Ethyl 2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylate was prepared according to 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%). 1 H NMR (400 MHz, CDCl3) δ 8.20 (t, J = 8.2 Hz, 1H), 8.10 (s, 1H), 7.79 (dd, J = 11.8, 2.0 Hz, 1H), 7.68 (d, J = 8.6 Hz, 1H), 6.82 (s, 1H), 4.43 (q, J = 7.1 Hz, 2H), 1.52 (s, 9H), 1.41 (t, J = 7.1 Hz, 3H).

[0378] Step 2. 2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylic acid was prepared according to general experimental procedure 2. Ethyl 2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylate (0.71 g, 1.9 mmol) gave 2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylic acid (0.63 g, 1.9 mmol, 97%). 1 H NMR (400 MHz, CDCl3) δ 8.25 (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).

[0379] Step 3. Methyl (2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serinate was prepared according to General Experimental Procedure 4. 2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxylic acid (0.63 g, 1.9 mmol) and methyl L-serinate hydrochloride (0.32 g, 1.1 equiv, 2.1 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serinate (0.87 g, 90% pure, 1.8 mmol, 96%).

[0380] Step 4. (2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serine was prepared according to general experimental procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serinate (0.87 g, purity 90%, 1.8 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serine (0.85 g, purity: 90%, 1.8 mmol, quantitative) as an off-white solid.

[0381] Step 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinathene was prepared according to General Experimental Procedure 4. (2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)serine (0.76 g, 1.8 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (0.78 g, 1.1 equiv, 2.0 mmol) gave methyl N-((2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.2 g, 1.5 mmol, 84%).

[0382] Step 6. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to general experimental procedure 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.2 g, 1.5 mmol) gave methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.1 g, 1.4 mmol, 93%). 1 H NMR (400 MHz, CDCl3) δ 8.21 (t, J = 8.2 Hz, 1H), 8.09 (d, J = 7.7 Hz, 1H), 8.04 (s, 1H), 7.72 (dd, J = 11.7, 2.0 Hz, 1H), 7.63 (d, J = 8.4 Hz, 1H), 7.61 - 7.50 (m, 5H), 7.44 - 7.26 (m, 7H), 7.12 (d, J = 8.1 Hz, 1H), 6.83 (d, J = 3.5 Hz, 1H), 4.95 - 4.85 (m, 1H), 4.67 (dt, J = 8.2, 2.9 Hz, 1H), 4.52 (dd, J = 11.3, 5.6 Hz, 1H), 4.39 (dd, J = 11.3, 6.2 Hz, 1H), 4.11 (dd, J = 10.3, 2.7 Hz, 2H), 3.88 (dd, J = 10.3, 3.0 Hz, 1H), 3.73 (s, 3H), 2.08 (s, 3H), 1.53 (s, 9H), 0.96 (s, 9H).

[0383] Step 7. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxamido)acrylamide)acrylate was prepared according to 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-serinate (0.30 g, 0.37 mmol) gave methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)-3-fluorophenyl)thiazole-4-carboxamido)acrylamide)acrylate (30 mg, 59 μmol, 16%) as a white solid. LCMS (22010199A TFA LCMS-5 C1): RT: 1.815 min; Area: 98.1% (215nm), 96.8% (306nm); m / z=491.2[M+H] + . 1 H NMR (400 MHz, CDCl3) δ 9.97 (s, 1H), 8.55 (s, 1H), 8.23 ​​(t, J = 8.3 Hz, 1H), 8.12 (s, 1H), 7.81 (dd, J = 11.7, 2.0 Hz, 1H), 7.68 (dd, J = 8.6, 1.3 Hz, 1H), 6.86 (m, 1H), 6.77 (d, J = 2.2 Hz, 1H), 6.72 (s, 1H), 6.04 (d, J = 1.4 Hz, 1H), 5.51 (t, J = 1.9 Hz, 1H), f3.90 (s, 3H), 1.57 - 1.49 (m, 9H).

[0384] Example 26: Compound 71 Methyl 2-(2-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamido)acrylate [ka]

[0385] Step 1. Ethyl 2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylate was prepared according to 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. 1 H NMR (400 MHz, CDCl3) δ 8.13 (s, 1H), 7.94 (t, J = 2.0 Hz, 1H), 7.64 (ddd, J = 7.7, 1.7, 1.0 Hz, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.35 (t, J = 8.0 Hz, 1H), 6.56 (s, 1H), 4.43 (q, J = 7.1 Hz, 2H), 1.51 (s, 9H), 1.41 (t, J = 7.1 Hz, 3H).

[0386] Step 2. 2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid was prepared according to general experimental procedure 2. Ethyl 2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylate (1.4 g, 4.0 mmol) gave 2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (1.3 g, 3.8 mmol, 95%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 8.22 (s, 1H), 8.05 (s, 1H), 7.59 (d, J = 7.6 Hz, 1H), 7.46 (d, J = 8.2 Hz, 1H), 7.37 (t, J = 7.9 Hz, 1H), 1.53 (s, 9H).

[0387] Step 3. Methyl (2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serinate was prepared according to general experimental procedure 4. 2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxylic acid (1.3 g, 3.8 mmol) and methyl L-serinate hydrochloride (0.65 g, 1.1 equiv, 4.2 mmol) gave methyl (2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serinate (1.5 g, 3.5 mmol, 93%). 1 H NMR (299 MHz, CDCl3) δ 8.27 (d, J = 7.6 Hz, 1H), 8.14 (d, J = 1.5 Hz, 1H), 7.98 (d, J = 1.9 Hz, 1H), 7.61 (m, 2H), 7.41 (t, J = 8.0 Hz, 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).

[0388] Step 4. (2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine was prepared according to general experimental procedure 2. Methyl (2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serinate (1.5 g, 3.5 mmol) gave crude (2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (1.5 g, 3.8 mmol, quantitative), which was used directly in the next step.

[0389] Step 5. Methyl N-((2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to general experimental procedure 4. (2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)serine (0.80 g, 2.0 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (0.75 g, 1.9 mmol) gave methyl N-((2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.2 g, 1.6 mmol, 79%).

[0390] Step 6. Methyl N-(O-acetyl-N-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 5. Methyl N-((2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.2 g, 1.6 mmol) gave methyl N-(O-acetyl-N-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (1.1 g, 1.4 mmol, 89%). 1H NMR (400 MHz, CDCl3) δ 8.11 (d, J = 7.8 Hz, 1H), 8.07 (s, 1H), 7.90 (t, J = 2.0 Hz, 1H), 7.62 - 7.50 (m, 6H), 7.44 - 7.26 (m, 8H), 7.13 (d, J = 8.2 Hz, 1H), 6.57 (s, 1H), 4.92 (dt, J = 7.9, 5.9 Hz, 1H), 4.67 (dt, J = 8.1, 2.8 Hz, 1H), 4.51 (dd, J = 11.3, 5.7 Hz, 1H), 4.39 (dd, J = 11.3, 6.2 Hz, 1H), 4.11 (dd, J = 10.3, 2.7 Hz, 1H), 3.87 (dd, J = 10.3, 3.0 Hz, 1H), 3.73 (s, 3H), 2.08 (s, 3H), 1.52 (s, 9H), 0.96 (s, 9H).

[0391] Step 7. Methyl 2-(2-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)acrylate was prepared according to General experimental procedure 7. Methyl N-(O-acetyl-N-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.30 g, 0.38 mmol) gave methyl 2-(2-(2-(3-((tert-butoxycarbonyl)amino)phenyl)thiazole-4-carboxamido)acrylamide)acrylate (51 mg, 0.11 mmol, 57%) as a white solid. LCMS (22010199A TFA LCMS-5 C1): RT: 1.704 min; Area: 98.9% (215nm), 98.6% (306nm); m / z473.2[M+H] + . 1H NMR (400 MHz, CDCl3) δ 10.00 (s, 1H), 8.55 (s, 1H), 8.15 (s, 1H), 7.93 (t, J = 2.0 Hz, 1H), 7.67 (d, J = 7.8 Hz, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.40 (t, J = 8.0 Hz, 1H), 6.78 (d, J = 2.2 Hz, 1H), 6.71 (s, 1H), 6.66 (s, 1H), 6.03 (d, J = 1.3 Hz, 1H), 5.51 (t, J = 2.0 Hz, 1H), 3.91 (s, 3H), 1.55 (s, 9H).

[0392] Example 27: Compound 72 Methyl 2-(2-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxamido)acrylamido)acrylate [ka]

[0393] Step 1. Ethyl 2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylate was prepared according to 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 (400 MHz, CDCl3) δ 9.15 (s, 2H), 8.62 (s, 1H), 8.18 (s, 1H), 4.43 (q, J = 7.1 Hz, 2H), 1.63 (s, 1H), 1.56 (s, 9H), 1.41 (t, J = 7.1 Hz, 3H).

[0394] Step 2. 2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylic acid was prepared according to general experimental procedure 2. Ethyl 2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylate (2.1 g, 5.9 mmol) gave crude 2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylic acid (2.0 g), which was used in the next step without purification.

[0395] Step 3. Methyl (2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serinate was prepared according to General Experimental Procedure 4. 2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxylic acid (0.64 g, 2.0 mmol) and methyl L-serinate hydrochloride (0.34 g, 1.1 equiv, 2.2 mmol) gave methyl (2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serinate (0.93 g, 2.0 mmol, quantitative).

[0396] Step 4. (2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serine was prepared according to general experimental procedure 2. Methyl (2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serinate (0.84 g, 2.0 mmol) gave (2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serine (0.55 g, 1.3 mmol, 67%). 1 H NMR (400 MHz, DMSO) δ 10.48 (s, 1H), 9.19 (s, 1H), 8.41 (s, 1H), 8.37 (d, J = 8.1 Hz, 1H), 4.49 (dt, J = 8.3, 4.2 Hz, 1H), 3.84 (m, 2H), 1.48 (s, 9H).

[0397] Step 5. Methyl N-((2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General experimental procedure 4. (2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)serine (0.54 g, 1.3 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (0.57 g, 1.1 equiv, 1.4 mmol) gave methyl N-((2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.42 g, 0.56 mmol, 43%) as an off-white solid. LCMS (general 3 acid) RT: 1.53 min; area % (254 nm): 76%; m / z=749.7 [M+H] + .

[0398] Step 6. Methyl N-(O-acetyl-N-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to general experimental procedure 5. Methyl N-((2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.40 g, 0.53 mmol) gave methyl N-(O-acetyl-N-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.24 g, 0.30 mmol, 57%). LCMS (general 3 acid) RT: 1.63 min; area % (254 nm): 96%; m / z=691.5 [M-Boc+H] + .

[0399] Step 7. Methyl O-acetyl-N-(O-acetyl-N-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-L-serinate was prepared using the following procedure. To a solution of methyl N-(O-acetyl-N-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (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 equiv, 0.45 mmol) and the resulting mixture was stirred for 1 h. The mixture was concentrated and the residue was taken up in MeCN (5 mL). Triethylamine (0.10 mL, 2.4 equiv, 0.72 mmol) and acetic anhydride (38 μL, 1.3 equiv, 0.40 mmol) were added and the resulting mixture was stirred at room temperature for 45 min. 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 give methyl O-acetyl-N-(O-acetyl-N-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-L-serinate (87 mg, 0.15 mmol, 48%).

[0400] Step 8. Methyl 2-(2-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxamide)acrylamide)acrylate was prepared using the following procedure. To a solution of methyl O-acetyl-N-(O-acetyl-N-(2-(2-((tert-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carbonyl)-L-seryl)-L-serinate (86 mg, 0.14 mmol) in CHCl (1.5 mL) and THF (1.5 mL) was added DBU (87 μL, 4 equiv, 0.58 mmol) at 0 °C and the mixture was stirred at 0 °C for 2 h. Water was added and the aqueous phase was adjusted to pH 4 by addition of 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-butoxycarbonyl)amino)pyrimidin-5-yl)thiazole-4-carboxamido)acrylamido)acrylate (27 mg, 57 μmol, 39%). LCMS (22010199A TFA LCMS-5 C1): RT: 1.460 min; Area: 96.8% (215 nm), 96.2% (306 nm); m / z 473.0 [MH] - . 1 H NMR (400 MHz, CDCl3) δ 10.01 (s, 1H), 9.17 (s, 2H), 8.54 (s, 1H), 8.20 (s, 1H), 7.96 (s, 1H), 6.77 (d, J = 2.3 Hz, 1H), 6.70 (s, 1H), 6.03 (d, J = 1.3 Hz, 1H), 5.51 (t, J = 1.9 Hz, 1H), 3.90 (s, 3H), 1.57 (s, 9H).

[0401] Example 28: Compound 75 Methyl 2-(2-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carboxamido)acrylamido)acrylate [ka]

[0402] 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 (2 M aqueous solution, 10 equiv, 42 mmol) and palladium tetrakis (0.24 g, 0.05 equiv, 0.21 mmol) and the resulting mixture was stirred at 80° C. for 16 h. The mixture was cooled to room temperature, diluted with ethyl acetate and washed with water. The aqueous phase was acidified with 1 M aqueous HCl 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% pure, 1.4 mmol, 35%) as a mixture with the decarboxylated product. The mixture was used directly in the next step.

[0403] Step 2. Methyl (4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serinate was prepared according to General Experimental Procedure 4. Crude 4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carboxylic acid (1.1 g, 43% purity, 1.4 mmol) and methyl L-serinate hydrochloride (0.26 g, 1.2 equiv., 1.7 mmol) gave methyl (4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serinate (0.61 g, 1.45 mmol, quantitative).

[0404] Step 3. (4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serine was prepared according to General Experimental Procedure 2. Methyl (4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serinate (0.60 g, 1.4 mmol) gave (4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serine (0.59 g, 1.4 mmol, quantitative).

[0405] Step 4. Methyl (4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl-L-serinate was prepared according to general experimental procedure 4. (4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-serine (0.58 g, 1.4 mmol) and methyl L-serinate hydrochloride (0.27 g, 1.2 equiv, 1.7 mmol) gave methyl (4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl-L-serinate (0.52 g, 1.0 mmol, 72%). LCMS (general 3 acid) RT: 0.94 min; area % (254 nm): 91%; m / z = 509.4 [M+H] + .

[0406] Step 5. Methyl O-acetyl-N-(O-acetyl-N-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl)-L-serinate was prepared according to the following general experimental procedure 5. Methyl (4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl-L-serinate (0.52 g, 1.0 mmol) and acetic anhydride (0.25 mL, 4 eq, 2.7 mmol) gave methyl O-acetyl-N-(O-acetyl-N-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl)-L-serinate (0.33 g, 0.56 mmol, 55%).

[0407] Step 6. Methyl 2-(2-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carboxamido)acrylamide)acrylate was prepared according to General experimental procedure 8. Methyl O-acetyl-N-(O-acetyl-N-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carbonyl)-L-seryl)-L-serinate (0.35 g, 0.59 mmol) gave methyl 2-(2-(4-(4-((tert-butoxycarbonyl)amino)phenyl)thiazole-2-carboxamido)acrylamide)acrylate (85 mg, 0.18 mmol, 30%) as a white solid. LC-MS (22010199A TFA LCMS-5 C1): RT: 1.731 min; area 98.6% (215nm), 97.8% (304nm); m / z=473.2[M+H] + . 1 H NMR (400 MHz, CDCl3) δ (ppm) 9.92 (s, 1H), 8.55 (s, 1H), 7.89 (d, J = 8.7 Hz, 2H), 7.67 (s, 1H), 7.45 (d, J = 8.6 Hz, 2H), 6.76 (d, J = 2.4 Hz, 1H), 6.71 (s, 1H), 6.59 (s, 1H), 6.04 (d, J = 1.0 Hz, 1H), 5.54 (dd, J = 2.3 Hz, J = 1.6 Hz, 1H), 3.90 (s, 3H), 1.54 (s, 9H).

[0408] Example 29: Compound 76 Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxamido)acrylamido)acrylate [ka]

[0409] Step 1. Ethyl 2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxylate was prepared according to 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 3 acid) RT: 1.58 min; Area % (254 nm): 96%; m / z=383.2 [M+H] + .

[0410] Step 2. 2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxylic acid was prepared according to general experimental procedure 2. Ethyl 2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxylate (0.38 g, 0.99 mmol) gave 2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxylic acid (0.30 g, 0.84 mmol, 85%). 1 H NMR (299 MHz, CDCl3) δ 8.36 (s, 1H), 8.25 (d, J = 8.9 Hz, 1H), 7.82 (s, 1H), 7.31 - 7.22 (m, 2H), 6.78 (s, 1H), 1.56 (s, 9H).

[0411] Step 3. Methyl (2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serinate was prepared according to general experimental procedure 4. 2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxylic acid (0.44 g, 1.2 mmol) and methyl L-serinate hydrochloride (0.23 g, 1.2 equiv, 1.5 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serinate (0.42 g, 0.92 mmol, 75%). LCMS (general 3 acid) RT: 1.23 min; Area % (254 nm): 93%; m / z = 456.3 [M+H] + .

[0412] Step 4. (2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serine was prepared according to general experimental procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serinate (0.42 g, 0.92 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serine (0.33 g, 0.75 mmol, 81%) as an orange solid.

[0413] Step 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to General Experimental Procedure 4. 2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-serine (0.33 g, 0.75 mmol) and methyl O-(tert-butyldiphenylsilyl)-L-serinate hydrochloride (0.35 g, 1.2 equiv., 0.90 mmol) gave methyl N-((2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.37 g, 0.48 mmol, 64%).

[0414] Step 6. Methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate was prepared according to general experimental procedure 5. Methyl N-((2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.37 g, 0.48 mmol) and acetic anhydride (68 μL, 1.5 equiv, 0.72 mmol) gave methyl N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carbonyl)-L-seryl)-O-(tert-butyldiphenylsilyl)-L-serinate (0.22 g, 0.27 mmol, 57%). 1H NMR (299 MHz, CDCl3) δ 8.28 - 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 = 6.3 Hz, 1H), 4.70 (d, J = 8.1 Hz, 1H), 4.56 (dd, J = 11.3, 5.7 Hz, 1H), 4.43 (dd, J = 11.3, 6.2 Hz, 1H), 4.14 (d, J = 8.1 Hz, 1H), 3.96 - 3.86 (m, 1H), 3.77 (d, J = 1.2 Hz, 3H), 2.11 (s, 3H), 1.56 (s, 9H), 1.00 (s, 9H).

[0415] Step 7. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxamido)acrylamide)acrylate was prepared according to 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-serinate (0.22 g, 0.27 mmol) gave methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)-2-chlorophenyl)thiazole-4-carboxamido)acrylamide)acrylate (15 mg, 30 μmol, 29%) as a white solid. LCMS (22010199A TFA LCMS-5 C1):rt 1.880 min;Area 96.5%(215nm),95.6%(304nm);m / z=507.2[M+H] + . 1H NMR (400 MHz, CDCl3) δ (ppm) 10.02 (s, 1H), 8.55 (s, 1H), 8.31 (d,J= 8.7 Hz, 1H), 8.23 ​​(s, 1H), 7.84 (d,J= 1.8 Hz, 1H), 7.21 (dd,J= 8.7 Hz,J= 2.3 Hz, 1H), 6.78 (d,J= 2.2 Hz, 1H), 6.70 (s, 1H), 6.66 (s, 1H), 6.02 (d,J= 1.1 Hz, 1H), 5.48 (t,J= 1.8 Hz, 1H), 3.90 (s, 3H), 1.54 (s, 9H).

[0416] Example 30: Compound 77 Methyl 2-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxamido)acrylamido)acrylate [ka]

[0417] Step 1. 4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxylic acid was prepared using the following procedure. To a solution of methyl 4'-amino-[1,1'-biphenyl]-3-carboxylate (0.50 g, 2.2 mmol) in CHCl (20 mL) was added DMAP (0.54 g, 2 eq, 4.4 mmol). Then BocO (0.72 g, 1.5 eq, 3.3 mmol) was added in portions and the mixture was stirred at room temperature for 16 h. The mixture was washed with water and brine, dried over NaSO and concentrated under reduced pressure. The crude material was dissolved in THF (20 mL) and NaOH solution (1 M aq., 7 eq, 15.4 mL, 15 mmol) was added and the resulting mixture was stirred for 64 h. The mixture was quenched with 1 M HCl and extracted with EtOAc. The organic phase was dried over Na2SO4, filtered and concentrated under reduced pressure to give 4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxylic acid (0.47 g, 1.5 mmol, 68%) as a yellow solid.

[0418] Step 2. Methyl (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serinate was prepared according to General Experimental Procedure 4. 4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxylic acid (0.45 g, 1.4 mmol) and methyl L-serinate hydrochloride (0.27 g, 1.2 equiv, 1.7 mmol) gave crude methyl (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serinate (0.60 g), which was used directly in the next step.

[0419] Step 3. (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serine was prepared according to general experimental procedure 2. Methyl (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serinate (0.58 g, 1.4 mmol) gave (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serine (0.37 g, 0.92 mmol, 66% for 2 steps) as a yellow oil. LCMS (general 3 acid) RT: 0.97 min; Area % (254 nm): 90%; m / z = 401.3 [M+H] + .

[0420] Step 4. Methyl (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl-L-serinate was prepared according to General Experimental Procedure 4. (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-serine (0.20 g, 0.50 mmol) and methyl L-serinate hydrochloride (0.93 mg, 1.2 equiv., 0.60 mmol) gave crude methyl (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl-L-serinate (0.16 g), which was used directly in the next step. LCMS (general 3 acids) RT: 0.95 min; Area% (254nm): 84%; m / z=502.4[M+H] + .

[0421] Step 5. Methyl O-acetyl-N-(O-acetyl-N-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl)-L-serinate was prepared according to General Experimental Procedure 5 below. Crude methyl (4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl-L-serinate (0.16 g) and acetic anhydride (0.12 mL, 2.6 equiv., 1.3 mmol) gave methyl O-acetyl-N-(O-acetyl-N-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl)-L-serinate (130 mg, 0.22 mmol, 44%) and the mono-elimination product methyl O-acetyl-N-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxamido)acryloyl)-L-serinate (0.10 g, 0.19 mmol, 38%).

[0422] Step 6. Methyl 2-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxamido)acrylamido)acrylate was prepared according to General Experimental Procedure 8. A mixture of methyl O-acetyl-N-(O-acetyl-N-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carbonyl)-L-seryl)-L-serinate (130 mg, 0.22 mmol) prepared in step 5 and methyl O-acetyl-N-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxamido)acryloyl)-L-serinate (0.10 g, 0.19 mmol) gave methyl 2-(2-(4'-((tert-butoxycarbonyl)amino)-[1,1'-biphenyl]-3-carboxamido)acrylamide)acrylate (6.6 mg, 14 μmol) as a white solid. LCMS (22010199A TFA LCMS-5 C1): RT: 1.468 min; area 98.0% (215nm), 97.2% (304nm); m / z=466.2[M+H] + . 1 H NMR (400 MHz, CDCl3) δ 8.95 (s, 1H), 8.61 (s, 1H), 8.08 (t, J = 1.9 Hz, 1H), 7.78 (ddt, J = 15.4, 7.9, 1.3 Hz, 2H), 7.62 - 7.45 (m, 6H), 6.85 (d, J = 2.3 Hz, 1H), 6.68 (s, 1H), 6.60 (s, 1H), 6.05 (d, J = 1.4 Hz, 1H), 5.53 (t, J = 1.8 Hz, 1H), 3.93 (s, 3H), 1.56 (s, 9H).

[0423] Example 31: Compound 78 Methyl 2-(2-(2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxamido)acrylamido)acrylate [ka]

[0424] Step 1. tert-Butyl (3-carbamoylbicyclo[1.1.1]pentan-1-yl)carbamate was prepared according to general experimental procedure 9. 3-((tert-butoxycarbonyl)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.

[0425] Step 2. tert-Butyl (3-carbamoylbicyclo[1.1.1]pentan-1-yl)carbamate was prepared according to 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-carbamoylbicyclo[1.1.1]pentan-1-yl)carbamate (1.5 g, 6.4 mmol, 78%). LCMS (general 3 acid) RT: 0.79 min; Area % (254 nm): 100%

[0426] Step 3. Ethyl 2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylate was prepared according to general experimental procedure 11. tert-Butyl (3-carbamothioylbicyclo[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 (400 MHz, CDCl3) δ 8.06 (s, 1H), 5.00 (s, 1H), 4.41 (q, J = 7.1 Hz, 2H), 2.49 (s, 6H), 1.46 (s, 9H), 1.39 (t, J = 7.1 Hz, 3H).

[0427] Step 4. 2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylic acid was prepared according to general experimental procedure 2. Ethyl 2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylate (1.8 g, 5.4 mmol) gave 2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylic acid (1.7 g, 5.4 mmol, quantitative).

[0428] Step 5. Methyl (2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serinate was prepared according to general experimental procedure 4. 2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxylic acid (1.7 g, 5.4 mmol) and methyl L-serinate hydrochloride (0.92 g, 1.1 equiv, 5.9 mmol) gave methyl (2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serinate (1.6 g, 90% pure, 3.6 mmol, 67%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 8.07 (d, J = 7.7 Hz, 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).

[0429] Step 6. (2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serine was prepared according to General Experimental Procedure 2. Methyl (2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serinate (1.6 g, 90% purity, 3.6 mmol) gave (2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serine (1.5 g, 85% purity, 3.2 mmol, 89%).

[0430] Step 7. Methyl (2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-seryl-L-serinate was prepared according to general experimental procedure 4. (2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-serine (1.5 g, 3.2 mmol) and methyl L-serinate hydrochloride (0.70 g, 1.4 equiv, 4.5 mmol) gave methyl (2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-seryl-L-serinate (1.6 g, 3.2 mmol, 98%).

[0431] Step 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-serinate was prepared according to General Experimental Procedure 5 below. Methyl (2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carbonyl)-L-seryl-L-serinate (1.6 g, 3.2 mmol) gave 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-serinate (1.2 g, 2.1 mmol, 68%) as a white solid.

[0432] Step 9. Methyl 2-(2-(2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxamide)acrylamide)acrylate was prepared according to 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-serinate (0.17 g, 0.32 mmol) gave methyl 2-(2-(2-(3-((tert-butoxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)thiazole-4-carboxamide)acrylamide)acrylate (85 mg, 0.18 mmol, 58%) as a white solid. LCMS (22010199A TFA LCMS-5 C1):rt 1.319 min;Area 99.0%(215nm),98.7%(304nm);m / z=463.2[M+H] + . 1H NMR (400 MHz, CDCl3) δ 9.81 (s, 1H), 8.51 (s, 1H), 8.05 (s, 1H), 6.73 (d, J = 2.1 Hz, 1H), 6.69 (s, 1H), 6.01 (d, J = 1.3 Hz, 1H), 5.47 (t, J = 1.9 Hz, 1H), 5.02 (s, 1H), 3.90 (s, 3H), 2.48 (s, 6H), 1.47 (s, 9H).

[0433] Example 32: Compound 79 Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxamido)acrylamido)acrylate [ka]

[0434] Step 1. tert-Butyl (trans-4-carbamoylcyclohexyl)carbamate was prepared according to General Experimental Procedure 9. trans-4-((tert-butoxycarbonyl)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.

[0435] Step 2. tert-Butyl (4-carbamothioylcyclohexyl)carbamate was prepared according to general experimental procedure 10. tert-Butyl (trans-4-carbamoylcyclohexyl)carbamate (1.9 g, 7.7 mmol) gave tert-butyl (4-carbamothioylcyclohexyl)carbamate (1.2 g, 4.7 mmol, 61%, trans / cis=97:3) as an off-white solid.

[0436] Step 3. Ethyl 2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylate was prepared according to General Experimental Procedure 11. tert-Butyl (4-carbamothioylcyclohexyl)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).

[0437] Step 4. 2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylic acid was prepared according to general experimental procedure 2. Ethyl 2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylate (1.1 g, 3.0 mmol) gave 2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylic acid (0.97 g, 3.0 mmol, 99%, trans / cis=9:1).

[0438] Step 5. Methyl (2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serinate was prepared according to General Experimental Procedure 4. 2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxylic acid (0.97 g, 3.0 mmol) and methyl L-serinate hydrochloride (0.51 g, 1.1 equiv, 3.3 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serinate (1.1 g, 89% pure, 2.2 mmol, 75%).

[0439] Step 6. (2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serine was prepared according to General Experimental Procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serinate (1.1 g, 89% purity, 2.2 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-serine (1.0 g, 88% purity, 2.1 mmol, 97%).

[0440] Step 7. Methyl (2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl-L-serinate was prepared according to General Experimental Procedure 4. (2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl-L-serine (1.0 g, 88% purity, 2.1 mmol) and methyl L-serinate hydrochloride (0.36 g, 1.1 equiv, 2.3 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl-L-serinate (0.55 g, 90% purity, 0.96 mmol, 45%).

[0441] Step 8. Methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl)-L-serinate was prepared according to General Experimental Procedure 5 below. Methyl (2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl-L-serinate (0.55 g, 90% purity, 0.96 mmol) gave methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl)-L-serinate (0.46 g, 0.77 mmol, 80%, trans / cis=87:13).

[0442] Step 9. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxamido)acrylamide)acrylate was prepared according to General Experimental Procedure 8. Methyl O-acetyl-N-(O-acetyl-N-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carbonyl)-L-seryl)-L-serinate (0.20 g, 0.33 mol) gave two batches of methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)cyclohexyl)thiazole-4-carboxamido)acrylamide)acrylate 79-1 and 79-2.

[0443] 79-1 (81 mg, 0.17 mmol, 51%, trans / cis=89:11) LCMS (22010199A TFA LCMS-5 C1): RT: 1.492 min; Area: 86.2% (215 nm), 88.4% (306 nm); m / z 479.2(M-H+);RT:1.520min;Area:10.80%(215nm), 11.61%(306nm);m / z=479.2[M+H] + 1 H-NMR (major isomer) (400 MHz, CDCl3) δ 9.90 (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, 3.6 Hz, 1H), 2.25 (d, J = 13.4 Hz, 2H), 2.17 (d, J = 12.7 Hz, 2H), 1.66 (qd, J = 13.1, 3.2 Hz, 2H), 1.46 (s, 9H), 1.27 (qd, J = 12.7, 3.4 Hz, 2H).

[0444] 79-2 14mg, 29μmol, 8.7%, trans / cis=63:37. LCMS-5(22010199A TFA LCMS-5 C1):RT:1.493min;Area:55.3%(215nm), 54.5%(306nm);m / z479.2(M-H+);RT:1.526min;Area:42.9%(215nm),43.6%(306nm);m / z=379.2[M-Boc+H] + . 1 H NMR (400 MHz, CDCl3) δ 9.99 (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 = 3.7 Hz, 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).

[0445] Example 33: Compound 80 Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxamido)acrylamido)acrylate [ka]

[0446] Step 1. tert-Butyl (4-carbamoylbicyclo[2.2.2]octan-1-yl)carbamate was prepared according to General Experimental Procedure 9. 4-((tert-butoxycarbonyl)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% pure, 3.5 mmol, 95%).

[0447] Step 2. tert-Butyl (4-carbamoylbicyclo[2.2.2]octan-1-yl)carbamate was prepared according to general experimental procedure 10. tert-Butyl (4-carbamoylbicyclo[2.2.2]octan-1-yl)carbamate (1.2 g, 82% purity, 3.5 mmol) gave tert-butyl (4-carbamoylbicyclo[2.2.2]octan-1-yl)carbamate (0.48 g, 1.7 mmol, 47%) as a white solid.

[0448] Step 3. Ethyl 2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylate was prepared according to general experimental procedure 11. tert-Butyl (4-carbamothioylbicyclo[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 (400 MHz, CDCl3) δ 8.00 (s, 1H), 4.37 (q, J = 7.1 Hz, 2H), 2.12 - 1.99 (m, 6H), 1.99 - 1.91 (m, 6H), 1.41 (s, 9H), 1.36 (t, J = 7.1 Hz, 3H).

[0449] Step 4. 2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylic acid was prepared according to general experimental procedure 2. Ethyl 2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylate (0.31 g, 0.82 mmol) gave 2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylic acid (0.32 g, 90% pure, 0.82 mmol, quantitative).

[0450] Step 5. Methyl (2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serinate was prepared according to general experimental procedure 4. 2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxylic acid (0.32 g, 90% purity, 0.82 mmol) and methyl L-serinate hydrochloride (0.14 g, 1.1 equiv, 0.90 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serinate (0.33, 88% purity, 0.64 mmol, 78%).

[0451] Step 6. (2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serine was prepared according to General Experimental Procedure 2. Methyl (2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serine (0.33 g, 0.64 mmol) gave (2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serine (0.30 g, 0.66 mmol, quantitative).

[0452] Step 7. Methyl (2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-seryl-D-serinate was prepared according to general experimental procedure 4. (2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-serine (0.30 g, 0.66 mmol) and methyl L-serinate hydrochloride (0.11 g, 1.1 equiv, 0.73 mmol) gave methyl (2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-seryl-D-serinate (0.30 g, 0.55 mmol, 83%). 1 H NMR (400 MHz, CDCl3) δ 8.25 (d, J = 7.4 Hz, 1H), 7.96 (s, 1H), 7.67 (d, J = 7.9 Hz, 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).

[0453] Step 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-serinate was prepared according to general experimental procedure 5. Methyl (2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carbonyl)-L-seryl-L-serinate (0.30 g, 0.55 mmol) and acetic anhydride (0.13 mL, 2.5 equiv, 1.4 mmol) gave 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-serinate (0.26 g, 0.41 mmol, 76%).

[0454] Step 9. Methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxamide)acrylamide)acrylate was prepared according to 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-serinate (0.15 g, 0.23 mmol) gave methyl 2-(2-(2-(4-((tert-butoxycarbonyl)amino)bicyclo[2.2.2]octan-1-yl)thiazole-4-carboxamide)acrylamide)acrylate (65 mg, 0.13 mmol, 55%) as a white solid. LCMS (22010199A TFA LCMS-5 C1): RT: 1.783 min; Area: 99.9% (215nm), 99.6% (306nm), m / z=505.2[M+H] +

[0455] General Procedure for Solid Phase Peptide Synthesis Fmoc Rink Amide AM resin (0.70 mmol / g) was swollen by washing with CHCl (2×1 min, 10 mL / gram resin). The resin was washed with DMF (3×1 min, 10 mL / gram resin) and treated with 20% piperidine in DMF (10 mL / gram resin) for 30 min. The resin was washed with DMF (3×1 min, 10 mL / gram resin). The resin was treated with a solution of Fmoc-AA(PG)-OH (3 eq.), HATU (3 eq.), and DiPEA (3 eq.) for 2-3 h or 16 h. The resin was washed with DMF (3×1 min, 10 mL / gram resin). The wash-deprotect-wash-coupling cycle was repeated for all amino acid couplings. After completion of the coupling cycle, the resin was treated with TFA:TIPS:water (95:2.5:2.5, 10 mL / gram resin) for 2 h. The resin was removed by filtration and the filtrate was precipitated with MTBE:heptane (1:1 v / v). After centrifugation (5 min, 3000 rpm), the supernatant was discarded and the residue was resuspended in MTBE:heptane (1:1 v / v) and centrifuged again (5 min, 3000 rpm). The supernatant was discarded and the pellet was taken up in water / MeCN (1:1). The solution was partially concentrated and lyophilized. The crude peptide was purified by automated reversed-phase FCC.

[0456] Example 34: Compound 81 [ka] 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 using HCl buffer afforded 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 (30833 LCMS-6): RT: 2.403 min; Area: 78.0% (215 nm), 99.6% (ELSD); m / z = 1084.7 [M+H] +

[0457] Example 35: Compound 82 N3(CH2CH2O)6CH2CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl salt [ka] N3(CH2CHO)6CH2CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl 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(CH2CHO)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 (30833 LCMS-6): RT: 2.365 min; Area: 97.8% (215 nm), 99.6% (ELSD); m / z=1306.8 [M+H] +

[0458] Example 36: Compound 83 N3(CH2CH2O)9CH2CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl salt [ka] N3(CH2CHO)9CH2CH2C(O)-Cha-D-Arg-Cha-D-Arg-Cha-D-Arg-NH2HCl 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(CH2CHO)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 (30833 LCMS-6): RT: 2.415 min; Area: 98.4% (215 nm), 99.8% (ELSD); m / z=1438.9 [M+H] +

[0459] General procedure for the conjugation chemistry of azido-peptides with compound 49 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.

[0460] Example 37: Compound 84 Compound 49 bound to compound 82 [ka] Compound 84 was prepared following the general procedure for the conjugation chemistry of azido-peptides with 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] +

[0461] Example 38: Compound 85 Compound 49 bound to compound 83 [ka] Compound 85 was prepared according to the general procedure for conjugation chemistry with azide-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+

[0462] Example 39: Hypothetical Assay To examine the mechanism of binding and explore the binding target, cell treatment and Western blotting are performed as described in Cunniff et al. 2015. The covalent cross-linking of the enzyme can be detected by protein Western blotting using specific antibodies.

[0463] Briefly, human tumor cell lines (HMESO cell line derived from a malignant mesothelioma patient) are cultured in appropriate medium and treated with various concentrations of test compounds (0.1 μM to 100 μM) for 24 h. After 24 h exposure to test compounds, cell lysates are generated in standard lysis buffer (RIPA buffer). Protein abundance is quantified and equal protein concentrations are separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Protein Western blotting is performed using antibodies specific for proteins such as PRX1, PRX2, PRX3, and PRX4. Covalent cross-linking modifications are detectable on protein Western blots by the presence of an antibody-reactive species of approximately 46 kD.

[0464] Cell death assays are performed as described in Nelson et al. 2021. Briefly, human tumor cell lines (HMESO cell lines derived from malignant mesothelioma patients) are cultured in 96-well plates and incubated with test compounds for 48 hours. The amount of remaining cellular material is stained with crystal violet and a total cell count is performed to determine % cell viability.

[0465] Example 40: Cell viability studies Cell lines 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, followed by incubation for 48 hours. After incubation, cells were washed with PBS (Corning Cellgro, Manassas, VA, USA), fixed with 3.0% formaldehyde in PBS (Fisher BioReagents, Fair Lawn, NJ, USA), and stained with 0.1% crystal violet in water (Acros Organics, Fair Lawn, NJ, USA) for 30 minutes. The crystal violet stain was removed, and the plates were 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). Effective cytotoxic concentrations (IC 50 To determine the α-associated β ...

[0466] Data from cell viability assays on HMESO mesothelioma cells treated with compound (1)-HCl or (5)-HCl are summarized in Table 2 below. IC 50 (concentration of drug required to kill 50% of the cells) is given in μM concentration. [Table 10]

[0467] Example 41: Cross-linking test Malignant mesothelioma (MM) cells (H-MESO cell line) were seeded in complete tissue culture medium in 6-well plates. Cells were allowed to adhere for 24 h and then treated with the indicated concentrations of thiostrepton (TS), (1) or (5) (DMSO stock) for 24 h. 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 reducing SDS-polyacrylamide gel electrophoresis. Proteins were transferred to PVDF membranes, blocked with 5% bovine serum albumin (BSA) for 1 h, and incubated with PRX3 primary antibody in 1X Tris-buffered saline with Tween (TBST) overnight at 4 °C. Membranes were washed three times with 1X TBST and incubated with horseradish peroxidase-conjugated (HRP) secondary antibody for 1 h at room temperature. Membranes were washed three times with 1× TBST, and the HRP signal was developed using enhanced chemiluminescence and visualized with a GE digital imager.

[0468] 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 in cells treated with (5) (Figure 3). No PRX3-X-PRX3 cross-linking was observed in cells treated with (1) (also Figure 3).

[0469] Example 42: Cytotoxic activity of test compounds in malignant mesothelioma cell lines 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 followed by incubation for 48 h (in technical replicates). After incubation, cells were washed with PBS (Corning Cellgro, Manassas, VA, USA), fixed with 3.0% formaldehyde in PBS (Fisher BioReagents, Fair Lawn, NJ, USA), and stained with 0.1% crystal violet in water (Acros Organics, Fair Lawn, NJ, USA) for 30 min. Crystal violet stain was removed and plates were 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). Effective cytotoxic concentrations (EC 50 To determine the α-associated β ...

[0470] Example 43: Covalent cross-linking of recombinant peroxiredoxin 3 (rPRX3) with test compounds The master mix reagents in Table 3 were combined for one reaction in an Eppendorf tube on ice. Reactions were scaled by the number of test compounds to be tested. 16 μL of master mix was added to a new Eppendorf tube containing 1 μL of test compound (10 mM stock diluted in DMSO) and mixed by gentle finger flicking and rapid centrifugation at 1,000 RPM. Reactions were incubated at 37° C. for 18 hours. Reactions were removed from incubation and quenched by adding 2 μL of Laemmli buffer containing 0.2 M dithiothreitol (DTT) and 10% sodium dodecyl sulfate. Samples were boiled at 98° C. for 5 minutes. Samples were separated by polyacrylamide gel electrophoresis, transferred to PVDF membranes, and subjected to protein western blotting using anti-PRX3 antibody (AbFrontier, LF-PA0255). The membrane was incubated with ECL reagent (ThermoScientific, Rockford, IL, USA) and visualized using a GE Amersham Imager chemiluminescence detection system. Unmodified rPRX3 is visualized as a single band at approximately 23 kDa, and rPRX3 covalently modified by the test compound is analyzed as a band at approximately 45 kDa. This is a qualitative assay that evaluates the presence or absence of the approximately 45 kDA band. The qualitative results are tabulated in Figure 5. [Table 11]

[0471] Example 44: Covalent cross-linking of peroxiredoxin 3 (PRX3) in malignant mesothelioma cells by test compounds 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 using RIPA buffer (50 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 0.25% sodium deoxycholate, 0.1% sodium dodecyl sulfate in deionized (DI) water) to reduce samples analyzed by reducing SDS-PAGE. Protein concentration was measured by Bradford assay (ThermoScientific, Rockford, IL, USA). Lysates (15 μg protein / well) were separated by SDS-PAGE under reducing conditions on 4–12% gradient Bis-Tris Midi gels (Invitrogen, Carlsbad, CA, USA) at a constant 200 V for 50 m. Gels were transferred to PVDF membranes at a constant 1 A for 50 min, blocked for a minimum of 1 h with 5% BSA diluted in 1× Tris-buffered saline containing 1% Tween-20 (TBS-T), and incubated with anti-PRX3 antibodies in 5% BSA TBS-T overnight at 4 °C. Membranes were washed for 1 h with 1× TBS-T, incubated for 1 h with the appropriate secondary antibodies, and washed again for 1 h with 1× TBS-T. Membranes were incubated with ECL reagent (ThermoScientific, Rockford, IL, USA) and visualized using a GE Amersham Imager chemiluminescence detection system. Qualitative results are tabulated in Figure 5.

[0472] Incorporation by Reference All publications and patents mentioned herein are herein 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 case of conflict, the present application, including any definitions herein, will control.

[0473] Equivalent Moreover, the present invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the enumerated claims are introduced into another claim. For example, any claim that is dependent on another claim may be amended to include one or more limitations found in any other claim that is dependent on the same base claim. When elements are presented as a list, e.g., in Markush group format, each subgroup of elements is also disclosed, and any element(s) may be removed from the group. In general, when the invention, or aspects of the invention, are described as comprising certain elements and / or features, it is to be understood that a particular embodiment of the invention or aspect of the invention consists of or consists essentially of such elements and / or features. For purposes of brevity, those embodiments have not been specifically described verbatim herein. It should also be noted that the terms "comprising" and "containing" are open-ended, allowing for the inclusion of additional elements or steps. When ranges are given, the endpoints are included. Furthermore, unless otherwise indicated or otherwise apparent from the context and the understanding of one of ordinary skill in the art, values ​​expressed as ranges should be understood to be able to take any specific value or subrange within the stated ranges of different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0474] The foregoing written specification is believed to be sufficient to enable one skilled in the art to practice the invention. The present invention is not limited in scope by the examples provided, which are intended to be merely illustrative of one aspect of the invention, and other functionally equivalent embodiments are also included within the scope of the present invention. Various modifications of the present invention in addition to those shown and described herein will become apparent to those skilled in the art from the above description. These modifications are intended to be included within the scope of the appended "claims". The advantages and objectives of the present invention are not necessarily encompassed in each embodiment of the present invention.

Claims

1. A compound having the structure of formula (IA): 【Chemistry 1】 or a pharmaceutically acceptable salt thereof, wherein: R １ is -NH ２ or —O—CH 3 , R ２ is =CH2, R ３ is =CH2, R ５ is —C(O)—R 1, Ring A is cycloalkyl; Ring B is aryl; R ４ is hydrogen or —C(O)—CH ３ and 【Chemistry 2】 are double bonds, The compound or a pharmaceutically acceptable salt thereof, wherein any hydrogen atom is optionally replaced with deuterium.

2. R １ is -OCH ３ 2. The compound of claim 1, wherein:

3. R １ is -NH ２ 2. The compound of claim 1, wherein:

4. 10. The compound of claim 1, wherein Ring A is a 5-membered cycloalkyl or a 5-membered cycloalkyl fused to a second ring.

5. The compound of claim 1 , wherein Ring A is a 5-membered cycloalkyl.

6. The compound of claim 5 , wherein Ring A is cyclopentyl.

7. Ring A is 【Transformation 3】 7. The compound of claim 6, wherein * indicates a bond to ring B.

8. The compound of claim 1 , wherein Ring A is a bridged bicyclic cycloalkyl.

9. 9. The compound of claim 8, wherein ring A is bicyclo[2.1.1]hexyl.

10. Ring A is 【Chemistry 4】 10. The compound of claim 9, wherein * indicates a bond to ring B.

11. The compound of claim 1 , wherein Ring A is a polycyclic cycloalkyl.

12. The compound of any one of claims 1 to 11, wherein Ring B is phenyl.

13. The compound of any one of claims 1 to 11, wherein Ring B is unsubstituted phenyl.

14. The compound according to any one of claims 1 to 11, wherein ring B is a halogen-substituted phenyl.

15. Ring B is 【Transformation 5】 12. The compound of any one of claims 1 to 11, wherein Z is halo and ** indicates a bond to ring A.

16. R ４ The compound according to any one of claims 1 to 11, wherein is hydrogen. 【Request Item 17】 【Chemistry 6】 2. The compound of claim 1 selected from:

18. 10. A pharmaceutically acceptable composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier.

19. 20. Use of a compound of claim 1 or a composition of claim 18 in the manufacture of a cancer therapeutic agent.

20. The use of claim 19, wherein the cancer is a solid tumor or a blood cancer.

21. 20. The use of claim 19, wherein the cancer is selected from lung, breast, prostate, melanoma, esophageal, leukemia, cervical, liver, colon, stomach, colorectal, glioblastoma, head and neck, pancreatic, mesothelioma, and ovarian cancer.

22. 20. The use of claim 19, wherein the cancer is selected from mesothelioma, lung, ovarian, and breast cancer.

23. The composition of claim 18 for use in the treatment of cancer.

24. The composition described in claim 23, wherein the cancer is a solid tumor or a blood cancer.

25. The composition of claim 23, wherein the cancer is selected from lung, breast, prostate, melanoma, esophageal, leukemia, cervical, liver, colon, stomach, colorectal, glioblastoma, head and neck, pancreatic, mesothelioma, and ovarian cancer.

26. The composition described in claim 23, wherein the cancer is selected from mesothelioma, lung, ovarian, and breast cancer.

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