Prodrugs of itaconate esters and methyl itaconate

Abstract

Disclosed are itaconic acid and prodrugs of 1-methylitaconic acid and 4-methylitaconic acid and their use for treating diseases, disorders or conditions associated with inflammation.

Description

[0001] Federally Sponsored Research or Development

[0002] This invention was made with government support under Grants AR068280 and AR064297 awarded by the National Institute of Health. The government has certain rights in this invention.

[0003] background

[0004] Inflammatory macrophages show a large accumulation of itaconate, which has been shown to exert significant anti-inflammatory activity by inhibiting succinate dehydrogenase and inducing electrophilic stress that activates the NRF2-dependent antioxidant response. However, given its charged nature, itaconate itself does not exhibit good pharmacokinetic or cell permeability properties. Therefore, the therapeutic potential of administering exogenous itaconate has not been clearly established.

[0005] Overview

[0006] The presently disclosed subject matter provides itaconic acid and prodrugs of 1-methylitaconic acid and 4-methylitaconic acid and their use for treating diseases, disorders or conditions associated with inflammation.

[0007] More particularly, in some aspects, the presently disclosed subject matter provides compounds of formula (I):

[0008]

[0009] in:

[0010] R1 and R2 may be the same or different and are each independently selected from one or more of the following and combinations thereof:

[0011] (a) -OR3, wherein R3 is H or a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group;

[0012] (b) wherein n is an integer selected from 1, 2, 3 and 4; R4 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group or -OR5, wherein R5 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group;

[0013] (c) wherein m is an integer selected from 1, 2, 3 and 4; p is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20; and R6 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group;

[0014] (d) Wherein R7 is selected from:

[0015] (i) -C(=O)-O-R8, wherein R8 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group;

[0016] (ii) Wherein R9 is H or C1-C4 straight or branched chain unsubstituted or substituted hydrocarbon group; R 10 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group; R 11 and R 12 are each independently H or a protecting group; and R 13 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group;

[0017] (iii) wherein q is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; R 11 and R 12 Each independently is H or a protecting group; R 14 is H or a C1-C4 straight or branched unsubstituted or substituted hydrocarbon group; and R 15 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group; and

[0018] (iv) Where R 16 and R 17 R is independently selected from H, C1-C4 straight or branched unsubstituted or substituted hydrocarbon group and protecting group; 18 is an aromatic group;

[0019] (e) Where R 19 is a C1-C4 straight or branched unsubstituted or substituted hydrocarbon group;

[0020] (f) wherein u is an integer selected from 1, 2, 3 and 4; R 20 is H or a C1-C4 straight or branched unsubstituted or substituted hydrocarbon group; and R 21 Yes-OR 22 , where R 22 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group or -NR 23 R 24 , where R 23 and R 24 Each is independently H or a C1-C4 straight or branched unsubstituted or substituted hydrocarbon group;

[0021] The condition is that R1 and R2 cannot both be -OH or both be -OR3;

[0022] and pharmaceutically acceptable salts thereof.

[0023] In other aspects, the presently disclosed subject matter provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0024] In yet other aspects, the presently disclosed subject matter provides methods for treating a disease, disorder, or condition associated with inflammation, comprising administering a compound of formula (I) or a pharmaceutical composition thereof to a subject in need of such treatment.

[0025] Having set forth above certain aspects of the presently disclosed subject matter, which in whole or in part are presented by the presently disclosed subject matter, other aspects will become apparent as the description proceeds when considered in conjunction with the accompanying embodiments and drawings which are best described below. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] This patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0028] Having thus generally described the presently disclosed subject matter, reference will now be made to the accompanying drawings, which are not necessarily drawn to scale, and in which:

[0029] Figure 1 Mouse plasma stability of representative prodrugs is shown;

[0030] Figure 2 shows the release of active monomethyl itaconate (in plasma) from a representative prodrug; and

[0031] Figure 3 The release of active itaconic acid (in plasma) from representative prodrugs is shown.

[0032] Details

[0033] The subject matter disclosed in the present invention will now be described more fully below with reference to the accompanying drawings, in which some but not all embodiments of the present invention are shown. The same numbers refer to the same elements throughout the text. The subject matter disclosed in the present invention can be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein; on the contrary, these embodiments are provided so that the present disclosure will satisfy applicable legal requirements. In fact, a person skilled in the art to which the subject matter disclosed in the present invention belongs will think of many modifications and other embodiments of the subject matter disclosed in the present invention set forth herein, with the benefit of the teachings presented in the previous description and the related drawings. Therefore, it should be understood that the subject matter disclosed in the present invention is not limited to the specific embodiments disclosed and modifications and other embodiments are intended to be included within the scope of the appended claims.

[0034] I. Novel Prodrugs of Itaconate Esters and Methyl Itaconate

[0035] Inflammatory macrophages show a large accumulation of itaconate, which has been shown to exert significant anti-inflammatory activity by inhibiting succinate dehydrogenase and inducing electrophilic stress that activates the NRF2-dependent antioxidant response. However, given the charged nature of itaconate, itaconate itself does not exhibit good pharmacokinetic properties or cell permeability properties. In view of the significant therapeutic potential of administering exogenous itaconate, the subject matter disclosed in the present invention provides prodrugs of itaconate and 1-methyl itaconate and 4-methyl itaconate, which have the ability to achieve enhanced cell penetration and release active itaconate and the corresponding itaconate methyl ester after oral, systemic or topical / local administration. The prodrugs disclosed in the present invention also exert anti-inflammatory effects on keratinocytes.

[0036] A. Representative compounds of formula (I)

[0037] In some embodiments, the presently disclosed subject matter provides compounds of formula (I):

[0038]

[0039] in:

[0040] R1 and R2 may be the same or different and are each independently selected from one or more of the following and combinations thereof:

[0041] (a) -OR3, wherein R3 is H or a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group;

[0042] (b) wherein n is an integer selected from 1, 2, 3 and 4; R4 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group or -OR5, wherein R5 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group;

[0043] (c) wherein m is an integer selected from 1, 2, 3 and 4; p is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20; and R6 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group;

[0044] (d) Wherein R7 is selected from:

[0045] (i) -C(=O)-O-R8, wherein R8 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group;

[0046] (ii) Wherein R9 is H or C1-C4 straight or branched chain unsubstituted or substituted hydrocarbon group; R 10 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group; R 11 and R 12 are each independently H or a protecting group; and R 13 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group;

[0047] (iii) wherein q is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; R 11 and R 12 Each independently is H or a protecting group; R 14 is H or a C1-C4 straight or branched unsubstituted or substituted hydrocarbon group; and R 15 is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group; and

[0048] (iv) Where R 16 and R 17 R is independently selected from H, C1-C4 straight or branched unsubstituted or substituted hydrocarbon group and protecting group; 18 is an aromatic group;

[0049] (e) Where R 19 is a C1-C4 straight or branched unsubstituted or substituted hydrocarbon group;

[0050] (f) wherein u is an integer selected from 1, 2, 3 and 4; R 20 is H or a C1-C4 straight or branched unsubstituted or substituted hydrocarbon group; and R 21 Yes-OR 22 , where R 22is a C1-C6 straight or branched unsubstituted or substituted hydrocarbon group or -NR 23 R 24 , where R 23 and R 24 Each is independently H or a C1-C4 straight or branched unsubstituted or substituted hydrocarbon group;

[0051] The condition is that R1 and R2 cannot both be -OH or both be -OR3;

[0052] and pharmaceutically acceptable salts thereof.

[0053] In certain embodiments of compounds of Formula (I):

[0054] (a) R1 is -OR3 and R2 is selected from -OR3, wherein R7 is selected from -C(=O)-O-R8,

[0055] (b) R1 is and R2 is selected from -OR3, wherein R7 is selected from -C(=O)-O-R8,

[0056] (c) R1 is and R2 is selected from -OR3, wherein R7 is selected from -C(=O)-O-R8,

[0057] (d) R1 is and R2 is selected from -OR3, wherein R7 is selected from -C(=O)-O-R8,

[0058] (e)R1 is and R2 is selected from -OR3, wherein R7 is selected from -C(=O)-O-R8,

[0059] (f) R1 is wherein R7 is -C(=O)-O-R8 and R2 is selected from -OR3, wherein R7 is selected from -C(=O)-O-R8,

[0060] (g) R1 is Where R7 is and R2 is selected from -OR3, wherein R7 is selected from -C(=O)-O-R8,

[0061] (h) R1 is Where R7 is and R2 is selected from -OR3, wherein R7 is selected from -C(=O)-O-R8, as well as

[0062] (i) R1 is Where R7 is and R2 is selected from -OR3, wherein R7 is selected from -C(=O)-O-R8,

[0063] The condition is that R1 and R2 cannot both be -OH or both be -OR3;

[0064] and pharmaceutically acceptable salts thereof.

[0065] In certain other embodiments of the compound of Formula (I):

[0066] (ai) R1 is -OH and R2 is selected from:

[0067] wherein R7 is -C(=O)-O-R8;

[0068] (a-ii) R1 is -OR3 and R2 is selected from:

[0069] Wherein R7 is selected from

[0070] -C(=O)-O-R8,

[0071] (b) R1 is and R2 is selected from:

[0072] -OR3,

[0073] (c) R1 is and R2 is -OR3;

[0074] (d) R1 is and R2 is selected from: -OR3 and

[0075] (e)R1 is wherein R7 is -C(=O)-O-R8 and R2 is -OR3;

[0076] (f) R1 is Where R7 is and R2 is -OR3;

[0077] (g) R1 is Where R7 is and R2 is -OR3;

[0078] (h) R1 is Where R7 is and R2 is -OR3; and

[0079] (i) R1 is and R2 is -OR3.

[0080] In some embodiments of the compound of Formula (I), R3, R4, R5, R6, R8, R9, R 10 , R 13 , R 15 and R 22 Each may independently be a C1, C2, C3, C4, C5 or C6 straight or branched unsubstituted or substituted hydrocarbon group selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl, n-hexyl, 2-methylpentane, 3-methylpentane, 2,2-dimethylbutane and 2,3-dimethylbutane.

[0081] In some embodiments of the compound of Formula (I), R9, R 14 , R 16 , R 17 , R 19 , R 20 , R 23 and R 24 Each may independently be a C1, C2, C3 or C4 straight or branched unsubstituted or substituted hydrocarbon group selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl.

[0082] Representative substituent groups include, but are not limited to, hydrocarbyl, substituted hydrocarbyl, halogen, aryl, substituted aryl, hydrocarbyloxy, hydroxy, nitro, amino, hydrocarbylamino, dihydrocarbylamino, sulfate, cyano, mercapto, and hydrocarbylthio.

[0083] In certain embodiments of the compound of formula (I), the protecting group is selected from tert-butyloxycarbonyl (boc), benzyloxycarbonyl (Cbz), p-methoxybenzylcarbonyl (Moz or MeOZ), 9-fluorenylmethoxycarbonyl (Fmoc) group, benzyl (Bn), p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-toluenesulfonyl (Ts), Troc (trichloroethyl chloroformate), (4-nitrophenyl)sulfonyl (Nosyl) and nitrophenylthio (Nps). In a specific embodiment, the protecting group is tert-butyloxycarbonyl (boc).

[0084] In some embodiments of the compound of Formula (I), R 18 Selected from phenyl.

[0085] In certain embodiments of compounds of Formula (I):

[0086] (ai) R1 is -OH and R2 is selected from:

[0087]

[0088] (a-ii) R1 is selected from -OCH3, -OCH(CH3)2 and -OC(CH3)3 and R2 is selected from:

[0089]

[0090] (b) R1 is and R2 is -OCH3(5,6) or

[0091] (c) R1 is and R2 is -OCH3(12, 13);

[0092] (d) R1 is and R2 is selected from: -OCH3(11) and

[0093] (e)R1 is and R2 is -OCH3(14);

[0094] (f) R1 is and R2 is -OCH3(17, 19);

[0095] (g) R1 is and R2 is -OCH3(15, 16);

[0096] (h) R1 is and R2 is -OCH3(18, 20); and

[0097] (i) R1 is And R2 is -OCH3(3,4).

[0098] In certain embodiments of the compounds of formula (I), the compound is selected from:

[0099]

[0100]

[0101]

[0102] In certain embodiments, the compound of formula (I) includes a pharmaceutically acceptable salt. In some embodiments, the pharmaceutically acceptable salt is an acidic salt. In a specific embodiment, the pharmaceutically acceptable salt is trifluoroacetate (TFA).

[0103] Representative compounds of formula (I) and the active agent released by each corresponding prodrug are provided in Table 1.

[0104]

[0105]

[0106]

[0107]

[0108]

[0109] 4-MI refers to 4-methylitaconate; 1-MI refers to 1-methylitaconate

[0110] In yet other embodiments, the presently disclosed subject matter provides methods for treating a disease, disorder, or condition associated with inflammation, comprising administering a compound of formula (I) or a pharmaceutical composition thereof to a subject in need of such treatment.

[0111] As used herein, the term "treating" may include reversing, alleviating, inhibiting the progression of, preventing or reducing the likelihood of the disease, disorder or condition to which such term is applied, or one or more symptoms or manifestations of such disease, disorder or condition. Prevention refers to preventing the disease, disorder, condition or its symptoms or manifestations from occurring, or preventing it from occurring with a worsening of severity. Thus, the compounds disclosed herein may be administered prophylactically to prevent or reduce the occurrence or recurrence of a disease, disorder or condition.

[0112] The "subject" treated by the methods disclosed herein in many of its embodiments is desirably a human subject, however it should be understood that the methods described herein are effective with respect to all vertebrate species, all of which are intended to be included in the term "subject". Thus, a "subject" may include a human subject for medical purposes, such as for treatment of an existing condition or disease or for prophylactic treatment to prevent the onset of a condition or disease, or an animal subject for medical purposes, veterinary purposes, or development purposes. Suitable animal subjects include mammals, including, but not limited to, primates, such as humans, monkeys, apes, etc.; bovines, such as cattle, bulls, etc.; ovines, such as sheep, etc.; caprines, such as goats, etc.; porcines, such as pigs, livestock pigs, etc.; equines, such as horses, donkeys, zebras, etc.; felines, including wild and domestic cats; canines, including dogs; lagomorphs, including rabbits, hares, etc.; and rodents, including mice, rats, etc. The animal can be a transgenic animal. In certain embodiments, the subject is a human, including, but not limited to, a fetus, a newborn, an infant, a juvenile, and an adult subject. In addition, a "subject" can include a patient suffering from or suspected of suffering from a condition or disease. Therefore, the terms "subject" and "patient" are used interchangeably herein. The term "subject" also refers to an organism, tissue, cell, or cell collection from a subject.

[0113] Typically, an "effective amount" of an active agent or drug delivery device refers to the amount necessary to elicit a desired biological response. As will be appreciated by one of ordinary skill in the art, the effective amount of an agent or device may vary depending on factors such as the desired biological endpoint, the agent to be delivered, the composition of the pharmaceutical composition, the target tissue, etc.

[0114] The term "combination" is used in its broadest sense, and means that the subject is administered at least two agents, more particularly, a compound of formula (I) and at least one β-lactam antibiotic, and optionally one or more antibacterial agents. More particularly, the term "combination" refers to the simultaneous administration of two (or more) active agents for the treatment of, for example, a single disease state. As used herein, the active agent can be combined and administered in a single dosage form, can be administered at the same time as a separate dosage form, or can be administered as a separate dosage form administered alternately or sequentially on the same or different days. In one embodiment of the subject matter disclosed in the present invention, the active agent is combined and administered in a single dosage form. In another embodiment, the active agent is administered in a separate dosage form (e.g., where it is desirable to vary the amount of one active agent, while the amount of another active agent does not vary). A single dosage form may include additional active agents for the treatment of a disease state.

[0115] In addition, the compounds of Formula (I) described herein can be administered alone or in combination with adjuvants that enhance the stability of the compounds of Formula (I), administered alone or in combination with one or more antibacterial agents, in certain embodiments facilitate administration of pharmaceutical compositions containing them (including other active ingredients), provide increased dissolution or dispersion, increase inhibitory activity, provide adjunctive therapy, etc. Advantageously, such combination therapies utilize lower doses of conventional therapeutic agents, thereby avoiding possible toxicity and adverse side effects when those agents are used as monotherapy.

[0116] The compound of formula (I) and at least one other therapeutic agent can be applied at different times (i.e., on the same day or on different days, in any order in sequence) to receive the compound of formula (I) and at least one other therapeutic agent, as long as the effect of the combination of two agents is achieved in the subject.

[0117] When administered sequentially, the doses can be administered within 1 minute, 5 minutes, 10 minutes, 30 minutes, 60 minutes, 120 minutes, 180 minutes, 240 minutes or longer of each other. In other embodiments, the doses administered sequentially can be administered within 1 day, 5 days, 10 days, 15 days, 20 days or more of each other. When the compound of formula (I) and at least one other therapeutic agent are administered simultaneously, they can be administered to the subject as a separate pharmaceutical composition (each pharmaceutical composition comprises a compound of formula (I) or at least one other therapeutic agent), or they can be administered to the subject as a single pharmaceutical composition comprising two doses.

[0118] When administered in combination, the effective concentration of each of the agents that elicit a particular biological response can be less than the effective concentration of each agent when administered alone, thereby allowing a reduction in the dose of one or more of the agents relative to the dose that would be required if the agent were administered as a single agent. The effects of multiple agents can, but need not be, additive or synergistic. An agent can be administered multiple times.

[0119] In some embodiments, when combined, two or more agents may have a synergistic effect. As used herein, the terms "synergy", "synergistic", "synergistically" and derivatives thereof, such as in "synergistic effect" or "synergistic combination" or "synergistic composition", refer to a situation where the biological activity of the combination of a compound of formula (I) and at least one other therapeutic agent is greater than the sum of the biological activities of the corresponding agents when administered alone.

[0120] Synergy can be expressed in terms of a "synergy index (SI)", which can generally be determined by the method described in FC Kull et al., Applied Microbiology 9, 538 (1961) from the ratio determined as follows:

[0121] Q a / Q A +Q b / Q B =Synergy Index (SI)

[0122] in:

[0123] Q A is the concentration of component A acting alone that produces an endpoint associated with component A;

[0124] Q a is the concentration of component A in the mixture that produces the endpoint;

[0125] Q B is the concentration of Component B, acting alone, which produces an endpoint associated with Component B; and

[0126] Q b is the concentration of component B in the mixture that produces the endpoint.

[0127] Usually, when Q a / Q A and Q b / Q B When the sum of the SI is greater than 1, antagonism is indicated. When the sum is equal to 1, an addition is indicated. When the sum is less than 1, a synergistic effect is indicated. The lower the SI, the greater the synergy shown by the particular mixture. Thus, a "synergistic combination" has an activity that is higher than what would be expected based on the activity observed for the individual components when used alone. In addition, a "synergistically effective amount" of a component refers to the amount of the component necessary to elicit a synergistic effect of, for example, another therapeutic agent present in the composition.

[0128] In another embodiment, the subject matter disclosed herein provides a pharmaceutical composition comprising a single compound of formula (I) or a combination of a compound of formula (I) and one or more additional therapeutic agents, mixed with a pharmaceutically acceptable excipient. Those skilled in the art will recognize that the pharmaceutical composition includes pharmaceutically acceptable salts of the compounds described above. Pharmaceutically acceptable salts are generally well known to those of ordinary skill in the art and include salts of active compounds prepared from relatively nontoxic acids or bases, depending on the specific substituent moieties present on the compounds described herein. When the compounds of the present disclosure contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds in a neutral form with a sufficient amount of the desired base, either pure or in a suitable inert solvent, or by ion exchange, whereby one basic counterion (base) in the ionic complex is replaced by another. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts, or similar salts.

[0129] When the compounds of the present disclosure contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent, or by ion exchange, whereby one acidic counterion (acid) in the ionic complex is replaced by another. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, and the like, and salts derived from relatively nontoxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like. Also included are salts of amino acids such as arginine salts, and salts of organic acids such as glucuronic acid or galacturonic acid, and the like (see, e.g., Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present disclosure contain both basic and acidic functional groups, which allow the compounds to be converted into either base addition salts or acid addition salts.

[0130] Thus, pharmaceutically acceptable salts suitable for use with the presently disclosed subject matter include, by way of example, but are not limited to, acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camphorsulfonate, carbonate, citrate, edetate, edisylate, estolate, esylate, fumarate, glucoheptonate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, methanesulfonate, mucate, naphthylate, nitrate, pamoate (pamoate), pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, or theoclate. Other pharmaceutically acceptable salts can be found, for example, in Remington: The Science and Practice of Pharmacy (20th ed.) Lippincott, Williams & Wilkins (2000).

[0131] In therapeutic and / or diagnostic applications, the compounds of the present disclosure can be formulated for a variety of modes of administration, including systemic administration and topical or local administration. Techniques and formulations generally can be found in Remington: The Science and Practice of Pharmacy (20th Edition) Lippincott, Williams & Wilkins (2000).

[0132] Depending on the specific conditions being treated, such agents can be formulated into liquid or solid dosage forms and are administered systemically or topically. Agents can be delivered, for example, in a timed slow release form known to those skilled in the art or in a sustained slow release form. The technology used for preparation and administration can be found in Remington: The Science and Practice of Pharmacy (20th edition) Lippincott, Williams & Wilkins (2000). Suitable routes can include oral, buccal, by inhalation spray, sublingual, rectal, transdermal, vaginal, transmucosal, nasal or intestinal administration; parenteral delivery, including intramuscular, subcutaneous, intramedullary injection, and intrathecal, direct intraventricular, intravenous, intraarticular, intrasternal, intrasynovial, intrahepatic, intralesional, intracranial, intraperitoneal, intranasal or intraocular injection or other delivery modes.

[0133] For injection, the agents of the present disclosure can be formulated and diluted in aqueous solutions, such as in physiologically compatible buffers such as Hank's solution, Ringer's solution, or saline buffer. For such transmucosal administration, penetrants suitable for the barrier to be penetrated are used in the formulation. Such penetrants are generally known in the art.

[0134] It is within the scope of the present disclosure to use a pharmaceutically acceptable inert carrier to prepare the compounds disclosed herein for practicing the present disclosure into a dosage suitable for systemic administration. In the case of appropriate selection of carriers and suitable manufacturing practices, the compositions of the present disclosure, particularly those formulated as solutions, can be administered parenterally, such as by intravenous injection. The compound can be easily formulated into a dosage suitable for oral administration using pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the present disclosure to be formulated into tablets, pills, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral intake by a subject to be treated (e.g., a patient).

[0135] For nasal or inhaled delivery, the agents of the present disclosure may also be formulated by methods known to those skilled in the art, and may include, for example, but not limited to, examples of solubilizing, diluting or dispersing substances, such as saline; preservatives, such as benzyl alcohol; absorption enhancers and fluorocarbons.

[0136] Pharmaceutical compositions suitable for use in the present disclosure include compositions in which the active ingredient is contained in an effective amount to achieve its intended purpose. The determination of the effective amount is fully within the capabilities of those skilled in the art, particularly in light of the detailed disclosure provided herein. Typically, compounds according to the present disclosure are effective over a wide dosage range. For example, in the treatment of adults, dosages from 0.01 mg to 1000 mg, from 0.5 mg to 100 mg, from 1 mg to 50 mg, and from 5 mg to 40 mg per day are examples of dosages that can be used. A non-limiting dose is 10 mg to 30 mg per day. The exact dosage will depend on the route of administration, the form in which the compound is administered, the subject to be treated, the body weight of the subject to be treated, the bioavailability of the compound, the adsorption, distribution, metabolism and excretion (ADME) toxicity of the compound, and the preference and experience of the attending physician.

[0137] In addition to the active ingredients, these pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers including excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Preparations formulated for oral administration may be in the form of tablets, dragees, capsules or solutions.

[0138] Pharmaceutical preparations for oral use can be obtained by combining the active compound with a solid excipient, optionally grinding the resulting mixture and, if desired, processing the granulated mixture after adding suitable auxiliaries to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulose preparations, for example, corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose (CMC) and / or polyvinylpyrrolidone (PVP: povidone). If desired, disintegrants such as cross-linked polyvinylpyrrolidone, agar or alginic acid or a salt thereof such as sodium alginate may be added.

[0139] The dragee core is provided with a suitable coating. For this purpose, concentrated sugar solutions can be used, which can optionally contain gum arabic, talcum powder, polyvinyl pyrrolidone, carbomer gel, polyethylene glycol (PEG) and / or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes or pigments can be added to tablets or dragee coatings for identification or characterization of the different combinations of active compound dosages.

[0140] Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, and soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. The push-fit capsules may contain the active ingredient mixed with a filler such as lactose, a binder such as starch and / or a lubricant such as talc or magnesium stearate and optionally a stabilizer. In soft capsules, the active compound may be dissolved or suspended in a suitable liquid, such as a fatty oil, liquid paraffin or a liquid polyethylene glycol (PEG). In addition, a stabilizer may be added.

[0141] In other embodiments, the compound of formula (I) disclosed in the present invention can be formulated into a viscous lotion, cream, ointment, suspension, paste, gel, oil, spray or aerosol and applied superficially. Such a viscous lotion, cream or ointment can be based on water, and can also contain oil (water-in-oil or oil-in-water), such as liquid paraffin or vegetable oil, such as peanut oil or castor oil, or solvent, and can contain one or more other components, including but not limited to penetration enhancers, such as ethanol and propylene glycol, wetting agents, including but not limited to glycerin and / or glycerol, thickeners and / or gelling agents, including but not limited to soft paraffin, aluminum stearate, cetearyl alcohol, polyethylene glycol, lanolin, beeswax, carboxypolyethylene and cellulose derivatives and / or glyceryl monostearate and / or nonionic emulsifiers, stabilizers, dispersants and suspending agents. By way of example only, a typical liquid formulation may contain between about 10% to about 60% water, between about 10% to about 70% ethanol, between about 5% to about 10% propylene glycol, and between about 2% to about 5% humectant.

[0142] II. Definitions

[0143] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter described in this invention belongs.

[0144] Although it is believed that the following terms related to compounds of formula (I) are well understood by those of ordinary skill in the art, the following definitions are proposed to help explain the subject matter disclosed in the present invention. These definitions are intended to supplement and illustrate rather than exclude definitions that will be obvious to those of ordinary skill in the art after reviewing the present disclosure.

[0145] As used herein, the term substituted (whether or not preceded by the term "optionally"), and substituents, refers to the ability to change one functional group on a molecule to another functional group, as understood by those skilled in the art, provided that the valence of all atoms is maintained. When more than one position in any given structure can be substituted by more than one substituent selected from a specified group, the substituents can be the same or different at each position. Substituents can also be further substituted (e.g., an aryl group substituent can have another substituent away from it, such as another aryl group, which is further substituted at one or more positions).

[0146] Where substituent groups or linking groups are specified by their conventional chemical formula written from left to right, they also encompass chemically identical substituents that would result from writing the structure from right to left, for example, -CH2O- is equivalent to -OCH2-; -C(=O)O- is equivalent to -OC(=O)-; -OC(=O)NR- is equivalent to -NRC(=O)O-, and so forth.

[0147] When the term "independently selected" is used, the substituents being referred to (e.g., R groups, such as groups R1, R2, and the like, or variables, such as "m" and "n") can be the same or different. For example, both R1 and R2 can be substituted hydrocarbyl, or R1 can be hydrogen and R2 can be substituted hydrocarbyl, etc.

[0148] When used with respect to a group of substituents herein, the term "a", "an" or "a(n)" means at least one. For example, in the case where a compound is substituted with "an" hydrocarbyl or aryl, the compound is optionally substituted with at least one hydrocarbyl and / or at least one aryl. In addition, in the case where a portion is substituted with an R substituent, the group may be referred to as "R-substituted". In the case where a portion is R-substituted, the portion is substituted with at least one R substituent and each R substituent is optionally different.

[0149] A named "R" or group will generally have a structure recognized in the art as corresponding to the group having that name, unless otherwise specified herein. For illustrative purposes, certain representative "R" groups as set forth above are defined below.

[0150] The description of the compounds of the present disclosure is subject to the principles of chemical bonding known to those skilled in the art. Thus, where a group may be substituted by one or more of a number of substituents, such substitutions are selected so as to comply with the principles of chemical bonding and give compounds that are not inherently unstable and / or would be considered by one of ordinary skill in the art to be likely to be unstable under environmental conditions such as aqueous, neutral, and several known physiological conditions. For example, a heterocycloalkyl or heteroaryl group is attached to the remainder of the molecule via a ring heteroatom in accordance with the principles of chemical bonding known to those skilled in the art, thereby avoiding inherently unstable compounds.

[0151] Unless expressly defined otherwise, a "substituent group" as used herein includes a functional group selected from one or more of the following moieties defined herein:

[0152] As used herein, the term hydrocarbon refers to any chemical group comprising hydrogen and carbon. Hydrocarbons can be substituted or unsubstituted. As will be known to those skilled in the art, all valences must be satisfied when any substitution is made. Hydrocarbons can be unsaturated, saturated, branched, unbranched, cyclic, polycyclic or heterocyclic. Illustrative hydrocarbons are further defined herein below and include, for example, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, allyl, vinyl, n-butyl, tert-butyl, ethynyl, cyclohexyl and similar hydrocarbons.

[0153] Unless otherwise indicated, the term "alkyl" by itself or as part of another substituent means a straight chain (i.e., unbranched) or branched, acyclic or cyclic hydrocarbon group, or combinations thereof, which may be fully saturated, monounsaturated or polyunsaturated and may include divalent and multivalent groups, having a specified number of carbon atoms (i.e., C 1-10 In certain embodiments, the term "hydrocarbyl" refers to a C alkyl group derived from a hydrocarbon moiety containing between one and twenty carbon atoms by removing a single hydrogen atom. 1-20 and 20 inclusive, including linear (i.e., "straight chain"), branched or cyclic, saturated or at least partially unsaturated, and in some cases fully unsaturated (i.e., alkenyl and alkynyl) hydrocarbon radicals of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20 carbon atoms.

[0154] Representative saturated hydrocarbon groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, isopentyl, neopentyl, n-hexyl, sec-hexyl, n-heptyl, n-octyl, n-decyl, n-undecyl, dodecyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, and homologs and isomers thereof.

[0155] "Branched" refers to a hydrocarbyl group in which a lower hydrocarbyl group such as methyl, ethyl or propyl is attached to a linear hydrocarbyl chain. "Lower hydrocarbyl" refers to a hydrocarbyl group having 1 to about 8 carbon atoms, e.g., 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms (i.e., C 1-8"Higher alkyl" refers to an alkyl group having from about 10 to about 20 carbon atoms, such as 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms. In certain embodiments, "alkyl" specifically refers to C 1-8 In other embodiments, "alkyl" refers specifically to C 1-8 Branched chain hydrocarbon.

[0156] The hydrocarbyl group may be optionally substituted with one or more hydrocarbyl substituents ("substituted hydrocarbyl") which may be the same or different. The term "hydrocarbyl substituent" includes, but is not limited to, hydrocarbyl, substituted hydrocarbyl, halogen, arylamino, acyl, hydroxyl, aryloxy, hydrocarbyloxy, hydrocarbylthio, arylthio, arylalkyloxy, arylalkylthio, carboxyl, hydrocarbyloxycarbonyl, oxo, and cycloalkyl. Along the hydrocarbyl chain there may be one or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms optionally inserted, wherein the nitrogen substituent is hydrogen, a lower hydrocarbyl (also referred to herein as "hydrocarbylaminohydrocarbyl"), or an aryl.

[0157] Thus, as used herein, the term "substituted hydrocarbyl" includes hydrocarbyl groups as defined herein, wherein one or more atoms or functional groups in the hydrocarbyl group are replaced by additional atoms or functional groups, including, for example, hydrocarbyl, substituted hydrocarbyl, halogen, aryl, substituted aryl, hydrocarbyloxy, hydroxyl, nitro, amino, hydrocarbylamino, dihydrocarbylamino, sulfate, cyano, and mercapto.

[0158] Unless otherwise stated, the term "heteroalkyl" alone or in combination with another term means a stable straight or branched chain or cyclic hydrocarbon group having from 1 to 20 carbon atoms or heteroatoms, or having from 3 to 10 carbon atoms or heteroatoms, or a combination thereof, consisting of at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, Si and S, and wherein the nitrogen, phosphorus and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. The heteroatoms O, N, P and S and Si may be located at any interior position of the heteroalkyl group or at the position where the hydrocarbon group is attached to the remainder of the molecule. Examples include, but are not limited to, -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2-S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, O-CH3, -O-CH2-CH3, and -CN. Up to two or three heteroatoms may be consecutive, such as, for example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3.

[0159] As described above, heteroalkyl groups as used herein include those groups attached to the remainder of the molecule through a heteroatom, such as -C(O)NR', ​​-NR'R", -OR', -SR, -S(O)R and / or -S(O2)R'. Where "heteroalkyl" is recited followed by a specific heteroalkyl group (e.g., -NR'R" or the like), it will be understood that the terms heteroalkyl and -NR'R" are not redundant or mutually exclusive. Rather, the specific heteroalkyl groups are recited for added clarity. Thus, the term "heteroalkyl" should not be construed herein to exclude specific heteroalkyl groups, such as -NR'R" or the like.

[0160] "Cyclic" and "cycloalkyl" refer to non-aromatic monocyclic or polycyclic ring systems having from about 3 to about 10 carbon atoms (e.g., 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms). Cycloalkyl groups may optionally be partially unsaturated. Cycloalkyl groups may also optionally be substituted with alkyl group substituents, oxo and / or alkylene groups as defined herein. Along the cycloalkyl chain there may be one or more oxygen, sulfur or substituted or unsubstituted nitrogen atoms optionally inserted, wherein the nitrogen substituent is hydrogen, unsubstituted alkyl, substituted alkyl, aryl or substituted aryl, thereby providing a heterocyclic group. Representative monocyclic cycloalkyl rings include cyclopentyl, cyclohexyl and cycloheptyl. Polycyclic cycloalkyl rings include adamantyl, octahydronaphthyl, decalin, camphor, camphane, and noradamantyl as well as fused ring systems such as dihydronaphthalene and tetrahydronaphthalene and the like.

[0161] Unless otherwise stated, the terms "cycloalkyl" and "heterocycloalkyl" alone or in combination with other terms represent the cyclic forms of "alkyl" and "heterocycloalkyl", respectively. In addition, for heterocycloalkyl, heteroatoms can occupy the position where the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and similar groups. Examples of heterocycloalkyl include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidyl, 2-piperidyl, 3-piperidyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophene-2-yl, tetrahydrothiophene-3-yl, 1-piperazinyl, 2-piperazinyl, and similar groups. The terms "cycloalkylene" and "heterocycloalkylene" refer to divalent derivatives of cycloalkyl and heterocycloalkyl, respectively.

[0162] Unsaturated hydrocarbons have one or more double bonds or triple bonds. Examples of unsaturated hydrocarbon groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1-propynyl and 3-propynyl, 3-butynyl and higher homologues and isomers. Alkyl groups limited to hydrocarbon groups are referred to as "homoalkyl".

[0163] More specifically, the term "alkenyl" as used herein refers to a C-1-alkylene group having at least one carbon-carbon double bond. 2-20(2 and 20 inclusive) a monovalent group derived from a straight or branched hydrocarbon moiety by removing a single hydrogen molecule. Alkenyl groups include, for example, ethenyl (i.e., vinyl), propenyl, butenyl, 1-methyl-2-butene-1-yl, pentenyl, hexenyl, octenyl, propadienyl and butadienyl.

[0164] As used herein, the term "cycloalkenyl" refers to a cyclic hydrocarbon containing at least one carbon-carbon double bond. Examples of cycloalkenyl groups include cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadiene, cyclohexenyl, 1,3-cyclohexadiene, cycloheptenyl, cycloheptatrienyl, and cyclooctenyl.

[0165] As used herein, the term "alkynyl" refers to a linear or branched C-chain radical having a designated number of carbon atoms and containing at least one carbon-carbon triple bond. 2-20 A hydrocarbon-derived monovalent group. Examples of "alkynyl" include ethynyl, 2-propynyl (propargyl), 1-propynyl, pentynyl, hexynyl and heptynyl groups and the like.

[0166] The term "alkylene" alone or as part of another substituent refers to a straight or branched divalent aliphatic hydrocarbon group derived from an alkyl group having from 1 to about 20 carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms). The alkylene group can be straight chain, branched or cyclic. The alkylene group can also be optionally unsaturated and / or substituted with one or more "alkyl group substituents". Along the alkylene group there can be one or more oxygen, sulfur or substituted or unsubstituted nitrogen atoms (also referred to herein as "alkylaminoalkyl"), optionally inserted, wherein the nitrogen substituent is an alkyl group as previously described. Exemplary alkylene groups include methylene (-CH2-); ethylene (-CH2-CH2-); propylene (-(CH2)3-); cyclohexylene (-C6H 10 -); -CH=CH-CH=CH-; -CH=CH-CH2-; -CH2CH2CH2CH2-, -CH2CH=CHCH2-, -CH2CsCCH2-, -CH2CH2CH(CH2CH2CH3)CH2-, -(CH2) q -N(R)-(CH2) r-, wherein q and r are each independently an integer from 0 to about 20, e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, and R is hydrogen or a lower alkyl; methylenedioxy (-O-CH2-O-); and ethylenedioxy (-O-(CH2)2-O-). The alkylene group may have from about 2 to about 3 carbon atoms and may also have 6-20 carbons. Typically, the alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with those groups having 10 or fewer carbon atoms being some embodiments of the present disclosure. "Lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, typically having eight or fewer carbon atoms.

[0167] The term "heteroalkylene" alone or as part of another substituent means a divalent group derived from a heteroalkyl group (such as, but not limited to, -CH2-CH2-S-CH2-CH2- and -CH2-S-CH2-CH2-NH-CH2-). For heteroalkylene groups, heteroatoms may also occupy one or both of the chain ends (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, and the like). Still further, for alkylene and heteroalkylene linking groups, the direction in which the formula of the linking group is written does not imply the orientation of the linking group. For example, the formula -C(O)OR'- represents both -C(O)OR'- and -R'OC(O)-.

[0168] Unless otherwise indicated, the term "aryl" means an aromatic hydrocarbon substituent which may be a single ring or multiple rings (such as from 1 to 3 rings) fused together or covalently linked. The term "heteroaryl" refers to an aryl group (or ring) containing from one to four heteroatoms (in each individual ring in the case of multiple rings) selected from N, O and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. The heteroaryl group may be attached to the remainder of the molecule through a carbon or heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl and 6-quinolyl. Substituents for each of the above aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. The terms "arylene" and "heteroarylene" refer to the divalent forms of aryl and heteroaryl, respectively.

[0169] For brevity, the term "aryl" when used in combination with other terms (e.g., aryloxy, arylthio, arylalkyl) includes both aryl and heteroaryl rings as defined above. Thus, the terms "arylalkyl" and "heteroarylalkyl" are intended to include those groups in which an aryl or heteroaryl group is attached to an alkyl group (e.g., benzyl, phenethyl, pyridylmethyl, furanylmethyl, and the like), including those alkyl groups in which a carbon atom (e.g., a methylene group) has been replaced, for example, with an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, and the like). However, the term "haloaryl" as used herein is intended to include only aryl groups substituted with one or more halogens.

[0170] In cases where the heteroalkyl, heterocycloalkyl, or heteroaryl includes a specific number of members (eg, "3- to 7-membered"), the term "member" refers to a carbon or heteroatom.

[0171] Furthermore, as used herein, structures generally represented by the formula:

[0172]

[0173] Refers to a ring structure containing a substituent R group, such as, but not limited to, 3-carbon, 4-carbon, 5-carbon, 6-carbon, 7-carbon, etc., aliphatic and / or aromatic cyclic compounds, including saturated ring structures, partially saturated ring structures and unsaturated ring structures, wherein the R group may be present or absent, and when present, one or more R groups may be substituted on one or more available carbon atoms of the ring structure. The presence or absence of the R group and the number of the R group are determined by the value of the variable "n", which is an integer generally having a value in the range of from 0 to the number of carbon atoms on the ring that can be used for substitution. Each R group, if more than one, is substituted on the available carbon of the ring structure rather than on another R group. For example, the above structure wherein n is 0 to 2 will include a compound group, which includes, but is not limited to:

[0174]

[0175] and the like.

[0176] A dotted line representing a bond in a cyclic ring structure indicates that the bond may or may not be present in the ring. That is, a dotted line representing a bond in a cyclic ring structure indicates that the ring structure is selected from the group consisting of a saturated ring structure, a partially saturated ring structure, and an unsaturated ring structure.

[0177] symbol Represents the point of attachment of the moiety to the rest of the molecule.

[0178] When a named atom of an aromatic or heterocyclic aromatic ring is defined as being "absent," the named atom is replaced by a direct bond.

[0179] As used herein, the term "acyl" refers to an organic acid group in which the -OH of the carboxyl group has been replaced by another substituent and has the general formula RC(=O)-, wherein R is an alkyl, alkenyl, alkynyl, aryl, carbocyclic, heterocyclic or aromatic heterocyclic group as defined herein. Therefore, the term "acyl" specifically includes aryl acyl groups, such as 2-(furan-2-yl)acetyl groups and 2-phenylacetyl groups. Specific examples of acyl groups include acetyl and benzoyl. Acyl groups are also intended to include amides, -RC(=O)NR', ​​esters, -RC(=O)OR', ketones, -RC(=O)R' and aldehydes, -RC(=O)H.

[0180] The terms "alkoxyl" or "alkoxy" are used interchangeably herein and refer to a saturated (i.e., alkyl-O-) or unsaturated (i.e., alkenyl-O- and alkynyl-O-) group attached to the parent molecular moiety through an oxygen atom, wherein the terms "alkyl," "alkenyl," and "alkynyl" are as previously described and may include C 1-20 (Inclusive of 1 and 20) linear, branched or cyclic, saturated or unsaturated oxyhydrocarbon chains including, for example, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy and n-pentoxy, neopentoxy, n-hexoxy and the like.

[0181] As used herein, the term "hydrocarbyloxyhydrocarbyl" refers to a hydrocarbyl-O-hydrocarbyl ether, for example, a methoxyethyl or ethoxymethyl group.

[0182] "Aryloxy" refers to an aryl-O-group, wherein the aryl group is as previously described, including substituted aryl. As used herein, the term "aryloxy" may refer to a phenoxy or hexyloxy group, and a phenoxy or hexyloxy group substituted with a hydrocarbyl, a substituted hydrocarbyl, a halogen, or a hydrocarbyloxy group.

[0183] "Arylalkyl" refers to an aryl-alkyl-group in which the aryl and alkyl are as previously described, and includes substituted aryl and substituted alkyl. Exemplary aralkyl groups include benzyl, phenylethyl, and naphthylmethyl.

[0184] "Aryloxy" refers to an aryl-O- group, wherein the aryl group is as previously described. An exemplary aryloxy group is benzyloxy, i.e., C6H5-CH2-O-. The aryloxy group may be optionally substituted.

[0185] "Hydroxycarbonyl" refers to a hydrocarbyl-OC(=O)- group. Exemplary hydrocarbyloxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl, butoxycarbonyl, and tert-butoxycarbonyl.

[0186] "Aryloxycarbonyl" refers to an aryl-OC(=O)- group. Exemplary aryloxycarbonyl groups include phenoxy-carbonyl and naphthoxy-carbonyl.

[0187] "Aryloxycarbonyl" refers to an arene-OC(=O)- group. An exemplary areneoxycarbonyl group is benzyloxycarbonyl.

[0188] "Carbamoyl" refers to an amide group of the formula -C(=O)NH2. "Alkylcarbamoyl" refers to an R'RN-C(=O)- group, wherein one of R and R' is hydrogen and the other of R and R' is a hydrocarbyl and / or substituted hydrocarbyl as previously described. "Dihydrocarbylcarbamoyl" refers to an R'RN-C(=O)- group, wherein each of R and R' is independently a hydrocarbyl and / or substituted hydrocarbyl as previously described.

[0189] The term carbonyldioxy, as used herein, refers to a carbonate group of the formula -OC(=O)-OR.

[0190] "Acyloxy" means an acyl-O- group in which the acyl group is as previously described.

[0191] The term "amino" refers to the -NH2 group and also refers to nitrogen-containing groups derived from ammonia by replacing one or more hydrogen groups with organic groups, as known in the art. For example, the terms "acylamino" and "hydrocarbylamino" refer to specific N-substituted organic groups having acyl and hydrocarbyl substituents, respectively.

[0192] As used herein, "aminoalkyl" refers to an amino group covalently bonded to an alkylene linker. More specifically, the terms alkylamino, dialkylamino and trialkylamino, as used herein, refer to one, two or three alkyl groups as previously defined, respectively, attached to the parent molecular moiety via a nitrogen atom. The term alkylamino refers to a group having the structure -NHR', wherein R' is a alkyl group as previously defined; while the term dialkylamino refers to a group having the structure -NR'R", wherein R' and R" are each independently selected from the group consisting of alkyl groups. The term trialkylamino refers to a group having the structure -NR'R"R"', wherein R', R" and R"' are each independently selected from the group consisting of alkyl groups. In addition, R', R" and / or R"' taken together may optionally be -(CH2) k -, wherein k is an integer from 2 to 6. Examples include, but are not limited to, methylamino, dimethylamino, ethylamino, diethylamino, diethylaminocarbonyl, methylethylamino, isopropylamino, piperidino, trimethylamino, and propylamino.

[0193] An amino group is -NR'R", wherein R' and R" are typically selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0194] The terms alkyl thioether and thioalkyloxy refer to saturated (i.e., alkyl-S-) or unsaturated (i.e., alkenyl-S- and alkynyl-S-) groups attached to the parent molecular moiety through a sulfur atom. Examples of thioalkyloxy moieties include, but are not limited to, methylthio, ethylthio, propylthio, isopropylthio, n-butylthio and the like.

[0195] "Acylamino" refers to an acyl-NH- group in which the acyl group is as previously described. "Aroylamino" refers to an aroyl-NH- group in which the aroyl group is as previously described.

[0196] The term "carbonyl" refers to a -C(=O)- group, and may include an aldehyde group represented by the general formula RC(=O)H.

[0197] The term "carboxyl" refers to a -COOH group. Such groups are also referred to herein as "carboxylic acid" moieties.

[0198] The term "cyano" refers to a -C≡N group.

[0199] As used herein, the terms "halo", "halide" or "halogen" refer to fluorine, chlorine, bromine and iodine groups. In addition, terms such as "haloalkyl" are intended to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C 1-4 )alkyl" is intended to include, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0200] The term "hydroxy" refers to an -OH group.

[0201] The term "hydroxyalkyl" refers to an alkyl group substituted with an -OH group.

[0202] The term "mercapto" refers to a -SH group.

[0203] The term "oxo" as used herein means an oxygen atom double-bonded to a carbon atom or another element.

[0204] The term "nitro" refers to the -NO2 group.

[0205] The term "thio" refers to compounds previously described herein wherein a carbon atom or an oxygen atom is replaced by a sulfur atom.

[0206] The term "sulfate" refers to the -SO4 group.

[0207] As used herein, the term thiol or thiol refers to a group of formula -SH.

[0208] More particularly, the term "sulfide" refers to a compound having a group of formula -SR.

[0209] The term "sulfone" refers to a compound having a sulfonyl group -S(O2)R.

[0210] The term "sulfoxide" refers to a compound having a sulfinyl group -S(O)R.

[0211] The term urea refers to a urea group of the formula -NH-CO-NH2.

[0212] Throughout the specification and claims, a given chemical formula or name will encompass all tautomers, congeners and optical and stereoisomers, as well as racemic mixtures where such isomers and mixtures exist.

[0213] Certain compounds of the present disclosure may have asymmetric carbon atoms (optical or chiral centers) or double bonds; enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomeric forms, and individual isomers that may be defined in terms of absolute stereochemistry as (R)- or (S)-, or as D- or L- for amino acids are included within the scope of the present disclosure. The present disclosure is intended to include compounds in racemic, non-racemic (scalemic) and optically pure forms. Optically active (R)- and (S)-isomers or D- and L-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein contain olefinic bonds or other centers of geometric asymmetry, and unless otherwise specified, it is intended that the compounds include both E and Z geometric isomers.

[0214] Unless otherwise stated, structures depicted herein are also intended to include all stereochemical forms of the structure; that is, the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the present disclosure.

[0215] It will be apparent to those skilled in the art that certain compounds of the present disclosure may exist in tautomeric forms, and all such tautomeric forms of the compounds are within the scope of the present disclosure. As used herein, the term "tautomer" refers to one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to another isomeric form.

[0216] Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, by replacement of hydrogen by deuterium or tritium or by 13 C or 14 Compounds having the present structures in which C-enriched carbon replaces carbon are within the scope of this disclosure.

[0217] The compounds of the present disclosure may also contain unnatural proportions of atomic isotopes at one or more of the atoms that make up such compounds. For example, the compounds may be treated with, for example, tritium ( 3 H), iodine-125 ( 125 I) or carbon-14 ( 14 C) is radiolabeled with a radioactive isotope. All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are included within the scope of the present disclosure.

[0218] Some compounds of the present disclosure may exist in unsolvated forms as well as solvated forms (including hydrated forms). Typically, the solvated forms are equivalent to the unsolvated forms and are included within the scope of the present disclosure. Some compounds of the present disclosure may exist in multiple crystalline or amorphous forms. Typically, all physical forms are equivalent for the intended uses of the present disclosure and are intended to be within the scope of the present disclosure.

[0219] The term "blocking group" refers to some or all of the reactive parts of a blocking compound and prevents such part from participating in a chemical reaction until the chemical part of the blocking group is removed, for example, those parts listed and described below: TW Greene, PGM Wuts, Protective Groups in Organic Synthesis, the 3rd edition John Wiley & Sons (1999). In the case where different blocking groups are used, it can be advantageous that each (different) blocking group is removable by different means. The blocking group cracked under completely different reaction conditions allows the removal of differences of such blocking groups. For example, blocking groups can be removed by acid, alkali and hydrogenolysis. Groups such as trityl, dimethoxytrityl, acetal and tert-butyldimethylsilyl are acid unstable, and can be used to protect carboxyl and hydroxyl reactive parts in the presence of amino groups protected with Cbz groups (which are removable by hydrogenolysis) and Fmoc groups (which are base unstable). Carboxylic acid and hydroxy reactive moieties can be blocked with base labile groups such as, but not limited to, methyl, ethyl and acetyl in the presence of amines blocked with acid labile groups such as tert-butyl carbamate or with carbamates that are stable to both acid and base but removable by hydrolysis.

[0220] Carboxylic acid and hydroxyl reactive moieties may also be blocked with hydrolytically removable protecting groups such as benzyl groups, while amine groups capable of acid hydrogen bonding may be blocked with base labile groups such as Fmoc. Carboxylic acid reactive moieties may be blocked with oxidatively removable protecting groups such as 2,4-dimethoxybenzyl, while coexisting amino groups may be blocked with fluoride-labile silyl carbamates.

[0221] Allyl blocking groups are useful in the presence of acid and base protecting groups because the former are stable and can be subsequently removed by metal or pi-acid catalysts. For example, allyl blocked carboxylic acids can be deprotected by palladium (0)-catalyzed reactions in the presence of acid-labile tert-butyl carbamate or base-labile acetate amine protecting groups. Another form of protecting groups is a resin to which a compound or intermediate can be attached. As long as the residue is attached to the resin, the functional group is blocked and cannot react. After being released from the resin, the functional group can be used for reaction.

[0222] Typical blocking / protecting groups include, but are not limited to, the following moieties: p-methoxybenzylcarbonyl (Moz or MeOZ), 3,4-dimethoxybenzyl (DMPM), Troc (trichloroethyl chloroformate), (4-nitrophenyl)sulfonyl (Nosyl), and nitrophenylsulfonyl (Nps), as well as

[0223]

[0224] Following long-standing patent law convention, the terms "a," "an," and "the" when used in this application, including the claims, mean "one or more." Thus, for example, reference to "a subject" includes more than one subject unless the context clearly indicates otherwise (e.g., more than one subject), etc.

[0225] Throughout the specification and claims, the terms "comprise," "comprises," and "comprising" are used in a non-exclusive sense, unless the context requires otherwise. Likewise, the term "include" and its grammatical variations are intended to be non-limiting, such that recitation of items in a list is not to the exclusion of other like items that can be substituted or added to the listed items.

[0226] For the purpose of this specification and the appended claims, unless otherwise indicated, all numbers used in the specification and claims to indicate amounts, sizes, dimensions, ratios, shapes, formulas, parameters, percentages, quantities, characteristics and other numerical values ​​should be understood to be modified by the term "about" in all cases, even if the term "about" may not be explicitly presented with the value, amount or range. Therefore, unless otherwise indicated, the numerical parameters listed in the following specification and the appended claims are not and need not be precise, but may be close and / or larger or smaller as desired, reflecting tolerances, conversion factors, rounding, measurement errors, etc., and other factors known to those skilled in the art, depending on the desired properties sought to be obtained by the subject matter disclosed in the present invention. For example, when referring to a value, the term "about" may be intended to include differences of ±100% in some embodiments, ±50% in some embodiments, ±20% in some embodiments, ±10% in some embodiments, ±5% in some embodiments, ±1% in some embodiments, ±0.5% in some embodiments, and ±0.1% in some embodiments from the specified amount, because such differences are appropriate for carrying out the disclosed method or employing the disclosed composition.

[0227] In addition, when used in conjunction with one or more numerical values ​​or numerical ranges, the term "about" should be understood to refer to all such values, including all values ​​within the range and to modify the range by extending the boundaries to above and below the listed values. Numerical ranges stated by endpoints include all values, such as all integers, included within the range, including fractions thereof (e.g., a statement of 1 to 5 includes 1, 2, 3, 4 and 5, and fractions thereof, such as 1.5, 2.25, 3.75, 4.1, etc.) and any range within the range. Example

[0228] The following examples have been included to provide guidance to those of ordinary skill in the art for practicing representative embodiments of the subject matter disclosed herein. Based on the present disclosure and the general level of skill in the art, it will be appreciated by the skilled person that the following examples are intended to be exemplary only, and that many variations, modifications, and changes may be employed without departing from the scope of the subject matter disclosed herein. The following descriptions and specific examples of the synthesis are intended only for illustrative purposes and are not to be construed as limiting in any way the compounds of the disclosure prepared by other methods.

[0229] Example 1

[0230] Stability of the prodrug in mouse skin homogenate and plasma

[0231] Mouse skin homogenate stability was performed using washed mouse skin or CD1 mouse plasma diluted 10-fold in 0.1M potassium phosphate buffer and homogenized using a tissue homogenizer. To evaluate the stability of the intact prodrug, 1 mL aliquots of skin homogenate or plasma were spiked with the prodrug to a final assay concentration of 20 μM. Spiked skin homogenate and plasma samples were incubated at 37°C in an orbital shaker for 1 h, and then the reaction was quenched in triplicate with three volumes of acetonitrile containing an internal standard (IS; losartan: 0.5 μM). The samples were vortexed for 30 s and centrifuged at 10,000 × g for 10 min at 4°C. 50 microliters of the supernatant were diluted with 50 μL of water and transferred to a 250 μL polypropylene vial sealed with a Teflon cap. The release of itaconic acid or methyl itaconate was measured by liquid chromatography tandem mass spectrometry (LC-MS / MS).

[0232] Example 2

[0233] Pharmacokinetics in mice

[0234] Male CD1 mice (25 g-30 g) were obtained from Harlan and maintained on a 12 h light-dark cycle with adlibitum access to food and water. IS-100-142, MK939 and MK941 were administered via oral gavage at a molar equivalent dose of 10 mg / kg monomethyl itaconate. Blood samples were collected at 0.25 h and 1 h after dosing (n=3 for each time point). Mice were euthanized with carbon dioxide at designated time points after drug administration, and blood samples (~0.8 mL) were collected in heparinized microtubes by cardiac puncture. Blood samples were centrifuged at 3000×g for 10 min at a temperature of 4°C. Plasma samples (~300 μL) were collected in polypropylene tubes and stored at -80°C until bioanalysis. The 1:1 ratio of the 1:1 ratio was 1:1. The 1:1 ratio was 1:1. The 1:1 ratio was 1:1. The 1:1 ratio was 1:1. The 1:1 ratio was 1:1. The plasma of mice was used to prepare calibration standards. Itaconate monomethyl ester standards and samples were extracted from plasma by one-step protein precipitation using methanol (100% v / v) containing internal standard methyl succinate (5 μM). Aliquots of the supernatant (100 μL) were diluted with water (100 μL) and transferred to 250 μL polypropylene vials sealed with Teflon caps and analyzed via LC-MS / MS.

[0235] Example 3

[0236] Treatment of human keratinocytes with itaconate prodrugs

[0237] Neonatal human epidermal keratinocytes (NHEK) isolated from neonatal foreskin were seeded at a density of 100,000 cells per well and maintained in KGM (KGM-GOLD Bullet Kit, #192060) supplemented with growth factors. Prodrugs were reconstituted in DMSO. NHEKs were pretreated with vehicle (0.1% DMSO) or prodrugs. After 2 days, NHEKs were treated with 50 μg / μl poly(I:C) for 24 h.

[0238] Example 4

[0239] RNA isolation and quantitative real-time PCR

[0240] Total RNA was isolated and purified from cultured NHEK using RNeasy Mini kit (Qiagen, Valencia, CA, #74106). After RNA purity and concentration were assessed using NanoDrop 2000UV-Vis spectrophotometer, RNA was converted to cDNA using reverse transcription kit and random hexamer primers (Applied Biosystems, #4368814). mRNA expression was determined by qRT-PCR using gene-specific, fluorophore-based TaqMan probes and universal master mix (Applied Biosystems, #4366072). qRT-PCR reactions were multiplexed using target probes and reference gene (RPLP0) probes. The relative mRNA change multiples were then quantified using the ΔΔCt method.

[0241] Example 5

[0242] Representative compounds

[0243] (S)-4-((1-ethoxy-1-oxo-3-phenylpropan-2-yl)amino)-2-methylene-4-oxobutanoic acid (IS-100-127)

[0244]

[0245] Itaconic anhydride (50 mg, 0.45 mmol) was dissolved in anhydrous THF (5 mL), and solid potassium carbonate (0.19 g, 1.34 mmol) was added followed by L-phenylalanine ethyl ester hydrochloride (0.1 g, 0.45 mmol). The reaction mixture was stirred at room temperature for 16 hours. The volatiles were then evaporated, the residue was redissolved in DCM (30 mL), and extracted with 1 M aqueous HCl (10 mL) and brine (10 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to final purification on reverse phase HPLC to provide 100 mg (73%) of the desired compound as a colorless semi-solid.

[0246] 1 H NMR (401 MHz, DMSO-d6): δ H 1.25 (t, J=7.1Hz, 3H), 3.13 (dd, J=13.9, 5.9Hz, 2H), 3.28 (s, 2H), 4.18 (q, J=7.2Hz, 2H), 4.86 (dt, J =7.8, 6.0Hz, 1H), 5.90 (s, 1H), 6.46 (s, 1H), 6.65 (d, J = 8.0Hz, 1H), 7.06-7.38 (m, 5H), 9.98 (s, 1H).

[0247] ESI MS: 304.1 ([MH] + ).

[0248] HRMS(ESI): For C 16 H 18 Calculated value of O5N: 304.11905. Measured value: 304.11910.

[0249] (S)-3-((1-ethoxy-1-oxo-3-phenylpropan-2-yl)carbamoyl)but-3-enoic acid methyl ester (IS-100-128)

[0250]

[0251] β-Methylitaconate (50 mg, 0.35 mmol) and HATU (0.13 g, 0.34 mmol) were dissolved in anhydrous DMF (3 mL), N,N-diisopropylethylamine (0.12 mL, 0.69 mmol) was added and the mixture was stirred at room temperature for 5 minutes. L-phenylalanine ethyl ester hydrochloride (80 mg, 0.35 mmol) was added and the reaction mixture was stirred at room temperature for 3 hours. The volatiles were then evaporated, the residue was redissolved in DCM (30 mL) and extracted with 1 M aqueous HCl (10 mL) and brine (10 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: cyclohexane / ethyl acetate 2:1) to provide 100 mg (90%) of compound IS-100-128 as a colorless semisolid.

[0252] 1 H NMR (401 MHz, DMSO-d6): δ H 1.27 (t, J=7.1Hz, 3H), 3.19 (dd, J=5.7, 4.6Hz, 2H), 3.37 (d, J=1.1Hz, 2H), 3.69 (s, 3H), 4.20 (q, J=7.1Hz, 2H), 4.90 (d t, J=7.6, 5.7Hz, 1H), 5.50 (t, J=1.2Hz, 1H), 5.74 (s, 1H), 6.60 (d, J=7.6Hz, 1H), 7.09-7.19 (m, 2H), 7.21-7.36 (m, 3H).

[0253] 13 C NMR (101 MHz, CDCl3): δ C 12.76, 36.46, 36.53, 50.77, 52.04, 60.20, 120.63, 125.72, 127.13, 128.06, 134.53, 136.59, 165.52, 169.86, 170.01.

[0254] ESI MS: 342.1 ([M+Na] + ).

[0255] HRMS(ESI): For C 17 H 22 Calculated value of O5N: 320.14925. Measured value: 320.14892.

[0256] (S)-3-((1-amino-1-oxo-3-phenylpropan-2-yl)carbamoyl)but-3-enoic acid methyl ester (IS-100-129)

[0257]

[0258] β-Methylitaconate (70 mg, 0.49 mmol) and HATU (0.18 g, 0.49 mmol) were dissolved in anhydrous DMF (5 mL), N,N-diisopropylethylamine (0.17 mL, 0.97 mmol) was added and the mixture was stirred at room temperature for 5 minutes. L-phenylalanine amide hydrochloride (97 mg, 0.49 mmol) was added and the reaction mixture was stirred at room temperature for 3 hours. The volatiles were then evaporated, the residue was redissolved in DCM (30 mL) and extracted with 1 M aqueous HCl (10 mL) and brine (10 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: dichloromethane / methanol 20:1) to provide 120 mg (85%) of compound IS-100-129 as a colorless semi-solid.

[0259] 1 H NMR (401 MHz, DMSO-d6): δ H 3.14-3.31 (m, 3H), 3.46-3.56 (m, 1H), 3.68 (s, 3H), 4.77 (dt, J=8.0, 6.8Hz, 1H), 5.43 (s, 1 H), 5.57-5.63 (m, 1H), 5.67 (s, 1H), 6.38 (s, 1H), 6.67 (d, J=8.0Hz, 1H), 7.21-7.38 (m, 5H).

[0260] 13 C NMR (101 MHz, CDCl3): δ C 37.45, 38.05, 52.24, 54.01, 121.42, 127.11, 128.76, 129.32, 136.60, 138.22, 167.87, 171.74, 173.19.

[0261] ESI MS: 313.1 ([M+Na] + ).

[0262] HRMS(ESI): For C 15 H 18 Calculated value for O4N2Na: 313.11588. Found: 313.11545.

[0263] 1-(((Isopropoxycarbonyl)oxy)methyl)4-methyl 2-methylene succinate (IS-100-142)

[0264]

[0265] β-Methyl itaconate (0.2 g, 1.39 mmol), chloromethyl isopropyl carbonate (0.22 mL, 1.66 mmol) and potassium carbonate (0.29 g, 2.08 mmol) were dissolved in anhydrous MeCN (5 mL), and the mixture was stirred at room temperature for 16 hours. EtOAc (60 mL) was added and the mixture was washed with brine (20 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: cyclohexane / ethyl acetate 5: 1) to provide 90 mg (25%) of compound IS-100-142 as a colorless oil.

[0266] 1 H NMR (401 MHz, DMSO-d6): δ H 1.34 (d, J=6.3Hz, 6H), 3.38 (s, 2H), 3.72 (s, 3H), 4.86-5.02 (m, 1H), 5.82-5.88 (m, 3H), 6.46 (s, 1H).

[0267] 13 C NMR (101 MHz, CDCl3): δ C 21.64, 37.20, 52.12, 73.10, 82.20, 130.50, 132.76, 153.29, 164.52, 170.77.

[0268] ESI MS: 283.1 ([M+Na] + ).

[0269] HRMS(ESI): For C1 x H 16 Calculated value for O7Na: 283.07882. Found: 283.07855.

[0270] 4-Methyl 1-((pivaloyloxy)methyl)2-methylene succinate (IS-100-143)

[0271]

[0272] β-Methylitaconate (0.2 g, 1.39 mmol), chloromethyl pivalate (0.26 mL, 1.8 mmol), sodium iodide (50 mg, 0.33 mmol) and potassium carbonate (0.29 g, 2.08 mmol) were dissolved in anhydrous MeCN (5 mL), and the mixture was stirred at 40° C. for 16 hours. EtOAc (60 mL) was added and the mixture was washed with brine (20 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: cyclohexane / ethyl acetate 5:1) to provide 0.33 g (92%) of compound IS-100-143 as a colorless oil.

[0273] 1 H NMR (401 MHz, DMSO-d6): δ H 1.20 (s, 9H), 3.34 (s, 2H), 3.68 (s, 3H), 5.79 (d, J=1.1Hz, 1H), 5.82 (s, 2H), 6.39 (s, 1H).

[0274] 13 C NMR (101 MHz, CDCl3): δ C 26.95, 37.38, 38.90, 52.21, 79.91, 130.30, 133.06, 164.82, 170.90, 177.19.

[0275] ESI MS: 281.1 ([M+Na] + ).

[0276] HRMS(ESI): For C 12 H 18 Calculated value for O6Na: 281.09956. Found: 281.09921.

[0277] 1-(tert-Butyl) 4-((pivaloyloxy)methyl) 2-methylene succinate (IS-100-146)

[0278]

[0279] 3-(tert-Butoxycarbonyl)but-3-enoic acid (0.2 g, 1.07 mmol), chloromethyl pivalate (0.2 mL, 1.4 mmol), sodium iodide (30 mg, 0.21 mmol) and potassium carbonate (0.22 g, 1.61 mmol) were dissolved in anhydrous MeCN (4 mL), and the mixture was stirred at 45° C. for 16 hours. EtOAc (60 mL) was added and the mixture was washed with brine (20 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: cyclohexane / ethyl acetate 6:1) to provide 0.23 g (71%) of compound IS-100-143 as a colorless oil.

[0280] 1 H NMR (401 MHz, DMSO-d6): δ H 1.21 (s, 9H), 1.48 (s, 9H), 3.33 (d, J=1.2Hz, 2H), 5.63 (d, J=1.2Hz, 1H), 5.76 (s, 2H), 6.25 (d, J=1.0Hz, 1H).

[0281] 13 C NMR (101 MHz, CDCl3): δ C 27.00, 28.10, 37.72, 38.90, 79.88, 81.44, 127.92, 134.86, 165.13, 169.79, 177.23.

[0282] ESI MS: 323.2 ([M+Na] + ).

[0283] HR ESI MS: For C 15 H 24 Calculated value of O6Na: 323.14651. Found: 323.14622.

[0284] 2-Methylene-4-oxo-4-((pivaloyloxy)methoxy)butanoic acid (IS-100-147)

[0285]

[0286] 1-(tert-Butyl) 4-((pivaloyloxy)methyl) 2-methylene succinate (0.14 g, 0.47 mmol) was dissolved in anhydrous DCM (0.5 mL) and trifluoroacetic acid (4 mL), and the mixture was stirred at room temperature for 2 hours. The volatiles were evaporated, and the residue was dissolved in DCM (3×15 mL) and evaporated 3 times to provide 0.11 g (97%) of compound IS-100-147 as a colorless oil.

[0287] 1 H NMR (401 MHz, DMSO-d6): δ H 1.21 (s, 9H), 3.37 (s, 2H), 5.77 (s, 2H), 5.87 (s, 1H), 6.49 (s, 1H), 11.12 (s, 1H).

[0288] 13 C NMR (101 MHz, CDCl3): δ C 26.81, 36.97, 38.76, 79.66, 131.36, 132.54, 169.22, 171.11, 177.13.

[0289] ESI MS: 267.1 ([M+Na] + ).

[0290] HRMS(ESI): For C 11 H 16 Calculated value for O6Na: 267.08391. Found: 267.08375.

[0291] Bis((pivaloyloxy)methyl)2-methylenesuccinate (IS-100-148)

[0292]

[0293] Itaconic acid (0.2 g, 1.53 mmol), chloromethyl pivalate (0.55 mL, 3.84 mmol), sodium iodide (50 mg, 0.3 mmol) and potassium carbonate (0.64 g, 4.60 mmol) were dissolved in anhydrous MeCN (5 mL), and the mixture was stirred at 45° C. for 16 hours. EtOAc (60 mL) was added and the mixture was washed with brine (20 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: cyclohexane / ethyl acetate 5:1) to provide 0.20 g (36%) of compound IS-100-148 as a colorless oil.

[0294] 1H NMR (401 MHz, DMSO-d6): δ H 1.20 (s, 18H), 3.38 (d, J=1.0Hz, 2H), 5.74 (s, 2H), 5.82 (s, 3H), 6.41 (s, 1H).

[0295] 13 C NMR (101 MHz, CDCl3): δ C 26.83, 26.84, 37.12, 38.75, 38.78, 79.70, 79.82, 130.52, 132.33, 164.45, 169.12, 177.07.

[0296] ESI MS: 381.2 ([M+Na] + ).

[0297] HRMS(ESI): For C 17 H 26 Calculated value for O8Na: 381.15199. Found: 381.15158.

[0298] Bis(((isopropoxycarbonyl)oxy)methyl)2-methylenesuccinate (IS-100-149)

[0299]

[0300] Itaconic acid (0.2 g, 1.53 mmol), chloromethyl isopropyl carbonate (0.51 mL, 3.84 mmol), sodium iodide (50 mg, 0.3 mmol) and potassium carbonate (0.64 g, 4.60 mmol) were dissolved in anhydrous MeCN (5 mL), and the mixture was stirred at 45° C. for 16 hours. EtOAc (60 mL) was added and the mixture was washed with brine (20 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: cyclohexane / ethyl acetate 5:1) to provide 80 mg (14%) of compound IS-100-149 as a colorless oil.

[0301] 1 H NMR (401 MHz, DMSO-d6): δ H 1.31 (d, J=6.3Hz, 12H), 3.41 (s, 2H), 4.82-4.98 (m, 2H), 5.75 (s, 2H), 5.82 (s, 2H), 5.85 (s, 1H), 6.46 (s, 1H).

[0302] 13C NMR (101 MHz, CDCl3): δ C 21.63, 36.95, 73.13, 82.00, 82.22, 131.05, 131.99, 153.26, 153.29, 164.28, 168.95.

[0303] ESI MS: 385.1 ([M+Na] + ).

[0304] HRMS(ESI): For C 15 H 22 O 10 Calculated value of Na: 385.11052. Found: 385.11069.

[0305] 4-Methyl 1-((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl 2-methylene succinate (LTP1025)

[0306]

[0307] β-Methylitaconate (200 mg, 1.39 mmol) was dissolved in anhydrous MeCN (5 mL), KCO (384 mg, 2.78 mmol, 2 eq) and NaI (416 mg, 2.78 mmol, 2 eq) were added and the resulting mixture was heated to 40° C. for 10 min under an inert atmosphere. (4-Chloromethyl)-5-methyl-1,3-dioxol-2-one (412 mg, 303 μL, 2.78 mmol, 2 eq) was finally added and the mixture was heated to 40° C. for 48 h. MeCN was evaporated, EtOAc (50 mL) was added and the organic phase was washed with 10% NaS0 (50 mL), distilled H0 (50 mL) and saturated NaCl (2×50 mL). The organic phase was dried over MgSO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: cyclohexane / EtOAc, 2:1) to provide 320 mg (90%) of compound LTP1025 as a light yellow oil.

[0308] 1 H NMR (401 MHz, CDCl3): δ H 2.15(s, 3H), 3.31(s, 2H), 3.65(s, 3H), 4.89(s, 2H), 5.75(s, 1H), 6.33(s, 1H).

[0309] 13 C NMR (101 MHz, CDCl3): δ C9.36, 37.36, 52.12, 54.28, 129.98, 132.99, 133.36, 140.30, 152.11, 165.51, 170.91.

[0310] ESI MS: 279.1 ([M+Na] + ).

[0311] HRMS(ESI): For C 11 H 12 Calculated value for O7Na: 279.04752. Found: 279.04757.

[0312] Alkyl β-methyl itaconate

[0313]

[0314] R = alkyl, aryl, alkyl salicyl, Boc-tyrosyl alkyl amide, Boc-tyrosyl ester, (di)peptide with terminal tyrosyl ester, (di)peptide with terminal tyrosyl alkyl amide

[0315]

[0316] Examples of R-OH of alkyl β-methyl itaconate include, but are not limited to:

[0317] The general procedure was performed according to a modified method described in: Boschert, D.; Schneider-Chaabane, A.; Himmelsbach, A.; Eickenscheidt, A.; Lienkamp, ​​K. Synthesis and Bioactivity of Polymer-Based Synthetic Mimics of Antimicrobial Peptides (SMAMPs) Made from Asymmetrically Disubstituted Itaconates. Chem. Eur. J. 2018, 24, 8217-8227.

[0318] L-Tyrosyl alkyl amides were synthesized according to McKenna, CE; Kashemirov, BA; Krylov, IS; Zakharova, VM Method to improve antiviral activity of nucleotide analogue drugs. US9550803 B2, January 24, 2017; and Hidaka, K.; Gohda, K.; Teno, N.; Wanaka, K.; Tsuda, Y. Active site-directed plasmin inhibitors: Extension on the P2 residue. Bioorg. Med. Chem. 2016, 24, 545-553.

[0319] β-Methyl itaconate (1 mmol) was dissolved in dry dichloromethane (3 mL). Appropriate hydroxy derivatives (1.2 mmol) and DMAP (1.5 mmol) were added, and the solution was cooled to 0 ° C. A solution of DCC (1.5 mmol) in dichloromethane (2 mL) was added dropwise under nitrogen, and the entire mixture was stirred at 0 ° C for 1 h, and then stirred at room temperature overnight. The mixture was filtered, and the solution was extracted with 10% aqueous KHSO (3 × 5 mL) and saturated NaHCO (5 mL). The organic phase was dried over sodium sulfate, evaporated, and the residue was chromatographed on a silica gel column in the system described below.

[0320] 1-(2-(Hexadecyloxy)ethyl)4-methyl 2-methylene succinate (MK-933)

[0321]

[0322] Chromatography in cyclohexane-ethyl acetate (80:15) system. Yield: 253 mg (61%) of colorless liquid.

[0323] 1 H NMR (CDCl3, ppm) δ: 0.88 (t, 3H, J CH3，CH2 =7.1, CH3(18′)), 1.38-1.20(m, 26H, 13xCH2(5′-17′)), 1.57(m, 2H, CH2(4′)), 3.35(d, 2H, J 2，＝C H =1.1, CH2(2)), 3.46(t, 2H, J 3′，4′=6.7, OCH2(3′)), 3.65(m, 2H, OCH2(2′)), 3.69(s, 3H, OCH3), 4.30(m, 2H, OCH2(1′)), 5.73(q, 1H, J gem =J H，2 =1.1, =C H a ), 6.37 (m, 1H, =C H b ).

[0324] 13 C NMR (CDCl3, ppm) δ: 14.11 (C-18′), 22.68 (C-17′), 26.05 (C-5′), 29.35, 29.46, 29.61 and 29.68 (C-4′, C-6′ -C-15′), 31.91(C-16′), 37.48(C-2), 52.03(OCH3), 64.29(C-1′), 68.39(C-2′), 71.46(C-3′), 128.74(= C H2), 133.64(C-3), 166.08(C-4), 171.11(C-1).

[0325] CI MS: 413.3 (MH) + (60).

[0326] HRMS(CI): For C 24 H 45 O5(MH) + Calculated value: 413.3267; measured value: 413.3264.

[0327] 4-Methyl 1-(4-(tetradecyloxy)butyl) 2-methylene succinate (MK-937)

[0328]

[0329] Chromatography in cyclohexane-ethyl acetate (7:1) system. Yield: 259 mg (63%) of colorless liquid.

[0330] 1 H NMR (CDCl3, ppm) δ: 0.87 (t, 3H, J CH3，CH2 =7.1, CH3(18′)), 1.37-1.20(m, 22H, 11xCH2(7′-17′)), 1.55(m, 2H, CH2(6′)), 1.64(m, 2H, CH2(3′)), 1.74(m, 2H, CH2(2′)), 3.33(d, 2H, J 2，＝C H =1.1, CH2(2)), 3.38(t, 2H, J 5′，6′ =6.7, OCH2(5′)), 3.42(t, 2H, J 4′，3′ =6.4, OCH2(4′)), 3.69(s, 3H, OCH3), 4.18(t, 2H, J 1′，2′ =6.6, OCH2(1′)), 5.70(q, 1H, J gem =J Ha，2 =1.1, =C H a ), 6.32 (m, 1H, =C H b ).

[0331] 13 C NMR (CDCl3, ppm) δ: 14.10 (C-18′), 22.67 (C-17′), 25.46 (C-2′), 26.17 and 26.21 (C-3′, C-7′), 29.34, 29.49, 29.60, 29.65 and 29.73 (C-6′ and C-8′-C-15′), 31.90 (C-16′), 37.53 (C-2), 52.01 (OCH3), 64.93 (C-1′), 70.10 (C-4′), 71.05 (C-5′), 128.36 (= C H2), 133.89(C-3), 166.11(C-4), 171.15(C-1).

[0332] CI MS: 412.3(M) + (6).

[0333] HRMS(CI): For C 24 H 44 O5(M) + Calculated value: 412.3189; measured value: 412.3187.

[0334] 1-(2-(Butoxycarbonyl)phenyl)4-methyl 2-methylene succinate (MK-939)

[0335]

[0336] Chromatography in cyclohexane-ethyl acetate (6:1) system. Yield: 230 mg (72%) of colorless liquid.

[0337] 1 H NMR (CDCl3, ppm) δ: 0.94 (t, 3H, CH3 (4′)), 1.42 (m, 2H, CH2 (3′)), 1.67 (m, 2H, CH2 (2′)), 3.48 (d, 2H, JCH2，H =1.1, C H 2-C=O), 3.73(s, 3H, OCH3), 4.25(t, 2H, J 1′，2′ =6.7, OCH2(1′)), 5.94(q, 1H, J gem =J Ha，CH2 =1.1, =C H a ), 6.62 (m, 1H, =C H b ), 7.14(ddd, 1H, J 3，4 =8.1, J 3，5 =1.2, J 3，6 =0.6, H-3), 7.32 (ddd, 1H, J 5，6 =7.9, J 5，4 =7.4, J 5，3 =1.2, H-5), 7.56 (ddd, 1H, J 4，3 =8.1, J 4，5 =7.4, J 4，6 =1.7, H-4), 8.02 (ddd, 1H, J 6，5 =7.9, J 6，4 =1.7, J 6，3 =0.3, H-6).

[0338] 13 C NMR (CDCl3, ppm) δ: 13.70 (C-4′), 19.16 (C-3′), 30.60 (C-2′), 37.43CH2C=O), 5 2.09(OCH3), 65.05(C-1′), 123.70(C-1), 123.80(C-3), 126.06(C-5), 130.41(= C H2), 131.74(C-6), 133.18( C =CH2), 133.64(C-4), 150.43(C-2), 164.52( C OOBu), 164.70( C OOPh), 170.95( C OOMe).

[0339] ESI MS: 343.1 (M+Na) + (100).

[0340] HRMS(ESI): For C 17 H 20 O6Na(M+Na) + Calculated value: 343.11521; measured value: 343.11487.

[0341] (S)-1-(4-(2-(tert-Butyloxycarbonyl)amino)-3-(octylamino)-3-oxopropyl)phenyl)4-methyl 2-methylene succinate (MK-940)

[0342]

[0343] Chromatography in a cyclohexane-ethyl acetate (2:1) system followed by additional chromatography in a CHCl3 system containing 1.5% MeOH. Yield: 325 mg (63%) of a white amorphous solid.

[0344] 1 HNMR (CDCl3, ppm) δ: 0.86 (t, 3H, J CH3，CH2 =7.1, CH3(11′)), 1,14-1.31 (m, 10H, 5xCH2(6′-10′)), 1.36 (m, 2H, CH2(5′)), 1.41 (s, 9H, CH3(t-Bu)), 3.02 and 3.06 (m, 2H, CH2(tyrosine)), 3.14 (m, 2H, CH2(4′)), 3.44 (s, 2H, CH2 -C=O), 3.71(s, 3H, OCH3), 4.25(m, 1H, CH -NH(2′)), 5.09(bs, 1H, 2′- NH ), 5.83 (bs, 1H, 3′- NH ), 5.88 (m, 1H, =CH a ), 6.53 (s, 1H, =CH b ), 7.04(d, 2H, J 2，3 =8.4, H-2 (aromatic), 7.21 (d, 2H, J 3，2 =8.4, H-3 (aromatic)).

[0345] 13 C NMR (CDCl3, ppm) δ: 14.04 (C-11′), 22.58 (C-10′), 26.75 (C-6′), 28.24 ( C H3 (t-Bu), 29.10, 29.13 and 29.32 (C-5′, C-7′, C-8′), 31.74 (C-9′), 37.48 ( C H2C=O), 37.91(C-1′), 39.49(C-4′), 52.11(O C H3), 55.90(C-2′), 80.16( C (CH3)3), 121.59(C-2), 130.08(=C H2), 130.29(C-3), 133.36( C =CH2), 134.51(C-4), 149.61(C-1), 155.38(NH- C OO), 164.60( C OOPh), 170.75 and 170.91 (C-3′, CH3O- C =O).

[0346] ESI MS: 1059.6 (2M+Na) + (10), 541.3 (M+Na) + (100).

[0347] HRMS(ESI): For C 28 H 42 O7N2Na(M+Na) + Calculated value: 541.28842; measured value: 541.28735.

[0348] 1-(4-((S)-2-((S)-2-((tert-Butoxycarbonyl)amino)-3-methylbutyrylamino)-3-isopropoxy-3-oxopropyl)phenyl)4-methyl 2-methylene succinate (MK-942)

[0349]

[0350] Chromatography in cyclohexane-acetone (4:1) system. Yield: 420 mg (77%) of white solid.

[0351] 1 H NMR (DMSO-d6, ppm) δ: 0.77-0.80 (m, 6H, CH( CH3 )2), 1.05 and 1.13 (2x d, 6H, J CH3，CH =6.3, OCH( CH3 )2), 1.37(s, 9H, C( CH3 )3), 1.85 (m, 1H, CH (CH3)2), 2.94 (dd, 1H, J 3′a，2′ =8.5, J gem =13.9, H-3′a), 3.00(dd, 1H, J 3′b，2′ =6.4, J gem =13.9, H-3′b), 3.50(s, 2H, CH 2-COOMe), 3.62(s, 3H, OCH3), 3.80(dd, 1H, J 2″，NH =9.2, J2″，3″ =7.1, H-2″), 4.42 (m, 1H, H-2′), 4.81 (septet, 1H, J CH，CH3 =6.3,O CH (CH3)2), 6.02 and 6.42 (2x d, 2H, J gem =1.1, =CH2), 6.60(d, 1H, J NH，2″ =9.2, NH -2″), 7.00 (m, 2H, H-aromatic (2)), 7.28 (m, 2H, H-aromatic (3)), 8.29 (d, J NH，2′ =7.5, NH-2′).

[0352] 13 C NMR (DMSO-d6, ppm) δ: 18.39 and 19.35 (C-4″), 21.54 and 21.69 (O—CH( C H3)2), 28.38(C( C H3)3), 30.73(C-3″), 36.24(C-3′), 37.16( C H2-COOMe), 52.01(OCH3), 53.70(C-2′), 59.61(C-2″), 68.23(O- CH (CH3)2), 78.16( C (CH3)3), 121.43(C-2), 130.47(C-3), 130.88(=CH2), 133.59( C =CH2), 135.00(C-4), 149.24(C-1), 155.50(NH- C OO), 164.61( C OO-Ph), 170.96 and 171.02 (C-1′, C OOMe), 171.63(C-1″).

[0353] ESI MS: 571.3 (M+Na) + (100), 549.3 (MH) + (2).

[0354] HRMS(ESI): For C 28 H 40 O9N2Na(M+Na) + Calculated value: 571.26260; measured value: 571.26247.

[0355] (S)-1-(4-(3-benzyloxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropyl)phenyl)4-methyl 2-methylene succinate (MK-943)

[0356]

[0357] Chromatography in cyclohexane-acetone (4:1) system. Yield: 439 mg (88%) of colorless syrup.

[0358] 1 H NMR (DMSO-d6, ppm) δ: 1.32 (s, 9H, C ( CH3 )3), 2.90(dd, 1H, J 3′a，2′ =10.0, J gem =13.8, H-3′a), 3.02(dd, 1H, Jx ′b，2′ =5.4, J gem =13.8, H-3′b), 3.51(s, 2H, CH2 -COOMe), 3.63(s, 3H, OCH3), 4.21(ddd, 1H, J 2′，3′ =10.0 and 5.4, J 2′，NH =8.0, (H-2′), 5.10(s, 2H, O- CH2 Ph), 6.04 and 6.43 (2x d, 2H, J gem =1.1, =CH2), 7.02 (m, 2H, H-2 (aromatic)), 7.27-7.37 (m, 7H, H-3, H-2″, H-3″, H-4″ (aromatic)), 7.41 (bd, 1H, J NH，CH =8.0, NH).

[0359] 13 C NMR (DMSO-d6, ppm) δ: 28.33 (C ( C H3)3), 35.85(C-3′), 37.18( C H2-COOMe), 52.04(O C H3), 55.60(C-2′), 66.17(O- C H2Ph), 78.60( C (CH3)3), 121.51(C-2), 128.05(C-2″), 128.24(C-4″), 128.59(C-3″), 130.46(C-3), 130.95(=CH2), 133.59( C=CH2), 135.48(C-4), 136.06(C-1″), 149.21(C-1), 155.68(NH- C O), 164.68( C OO-C(1)), 171.04( C OO-Me), 172.19 (C-1′).

[0360] ESI MS: 1017.5 (2M+Na) + (15), 520.2 (M+Na) + (100).

[0361] HRMS(ESI): For C 27 H 31 O8NNa(M+Na) + Calculated value: 520.19419; measured value: 520.19394.

[0362] Deprotection of tert-butoxycarbonyl (Boc) group. General procedure.

[0363] A mixture of dichloromethane and trifluoroacetic acid (1: 1, 16 mL) was added to an appropriate Boc derivative (0.8 mmol). The solution was stirred at room temperature for 20 min and evaporated. The residue was crystallized (MK-944), or chromatographically separated on a silica gel column (100 mL) in the system described below, then purified on reversed-phase HPLC and freeze-dried (compound MK-941, MK-945).

[0364] The following compounds were synthesized:

[0365] (S)-1-(4-(2-amino-3-(octylamino)-3-oxopropyl)phenyl)4-methyl 2-methylene succinate trifluoroacetate (MK-941)

[0366]

[0367] Chromatography in ethyl acetate-methanol (15:2) system gave 183 mg (43%) of yellow foam which was subjected to final purification on reverse phase HPLC (gradient 20% to 60% CH3CN in 0.1% TFA / H2O over 40 min, retention time 36.8 min). Yield: 124 mg (29%) of white solid.

[0368] 1 H NMR (DMSO-d6, ppm) δ: 0.85 (t, 3H, J CH3，CH2=7.0, CH3), 1.11-1.32(m, 12H, 6x CH2(2″-7″)), 2.94(m, 1H, NH- CH a ), 3.00(m, 2H, Ph- CH2 ), 3.11 (m, 1H, NH- CH b ), 3.51(s, 2H, CH2 COOMe), 3.63(s, 3H, OCH3), 3.91(m, 1H, CH -NH3 + ), 6.05 and 6.42 (2x d, J gem =1.1, =CH2), 7.08 (m, 2H, H-2 (aromatic)), 7.26 (m, 2H, H-3 (aromatic)), 8.22 (bs, 3H, NH3 + ), 8.31(bt, 1H, J NH，1″ =5.6, NH).

[0369] 13 C NMR (DMSO-d6, ppm) δ: 14.44 (CH3(8")), 22.57 (CH2(7")), 26.76 (CH2(3"), 29.07, 29.13 and 29.15 (CH2(2", 4", 5")), 31.73 (CH2(6"), 36.93 (Ph- C H2), 37.37( C H2COOMe), 39.12(NH- C H2), 52.26(O C H3), 53.96( C H-NH3 + ), 122.04 (C-2 (aromatic)), 131.05 (C-3 (aromatic)), 131.21 (= C H2), 133.12(C-4(aromatic)), 133.80( C =CH2), 149.98 (C-1 (aromatic)), 164.80 (COO), 167.89 (NH- C =0), 171.26( C OOMe).

[0370] ESI MS: 441.2 (M+Na) + (100), 419.3 (MH) + 35).

[0371] HRMS(ESI): For C 23 H 34 O5N2Na(M+Na)+ Calculated value: 441.23599; measured value: 441.23569. 23 H 35 O5N2(MH) + Calculated value: 419.25405; measured value: 419.25385.

[0372] 1-(4-((S)-2-((S)-2-amino-3-methylbutyrylamino)-3-isopropoxy-3-oxopropyl)phenyl)4-methyl 2-methylene succinate trifluoroacetate (MK-944)

[0373]

[0374] Crystallization from ethyl acetate to which diethyl ether was added. Yield: 324 mg (72%) of white crystals.

[0375] 1 H NMR (DMSO-d6, ppm) δ: 0.93 and 0.97 (2x d, 6H, J CH3，CH =6.9, CH( CH3 )2), 1.07 and 1.15 (2x d, 6H, J CH3，CH =6.2, O-CH( CH3 )2), 2.13 (m, 1H, CH (CH3)2), 2.99 (dd, 1H, J 3′a，2′ =8.3, J gem =14.1, H-3′a), 3.04(dd, 1H, J 3′b，2′ =6.3, J gem =14.1, H-3′b), 3.51(s, 2H, CH2 -COOMe), 3.63(s, 3H, OCH3), 3.65(d, 1H, J 2″，3″ =5.1, H-2″), 4.50 (m, 1H, H-2′), 4.85 (septet, 1H, J CH，CH3 =6.2,O- CH (CH3)2), 6.04 and 6.43 (2x m, 2H, =CH2), 7.05 (m, 2H, H-2), 7.32 (m, 2H, H-3), 8.09 (bs, 3H, NH3 + ), 8.90(d, 1H, J NH，2′ =7.0, NH).

[0376] 13 C NMR (DMSO-d6, ppm) δ: 17.44 and 18.56 (C-4″), 21.52 and 21.69 (O—CH(C H3)2), 30.10(C-3″), 36.03(C-3′), 37.16( C H2-COOMe), 52.03(OCH3), 54.27(C-2′), 57.24(C-2″), 68.66(O- CH (CH3)2), 121.66(C-2), 130.53(C-3), 131.04(= C H2), 133.54( C =CH2), 134.68(C-4), 149.41(C-1), 164.71( C OO-Ph), 168.41(NH- C O), 170.56( C OO-iPr), 171.07( C OOMe).

[0377] ESI MS: 471.2 (M+Na) + (58), 449.2 (MH) + (100).

[0378] HRMS(ESI): For C 23 H 32 O7N2Na(M+Na) + Calculated value: 471.21017; Measured value: 471.20999. 23 H 33 O7N2(MH) + Calculated value: 449.22823; measured value: 449.22815.

[0379] (S)-1-(4-(2-amino-3-(benzyloxy)-3-oxopropyl)phenyl)4-methyl 2-methylene succinate trifluoroacetate (MK-945)

[0380]

[0381] Chromatography in a dichloromethane-methanol (50:2) system gave 270 mg (68%) of a yellow syrup which was subjected to final purification on reverse phase HPLC (gradient 15% to 50% CH3CN in 1% TFA / H2O over 40 min, retention time 37.1 min). Yield: 85 mg (21%) of a viscous semisolid.

[0382] 1 H NMR (DMSO-d6, ppm) δ: 3.08 (dd, 1H, J 3′a，2′ =7.7, Jgem =14.1, H-3′a), 3.16(dd, 1H, J 3′b，2′ =6.0, J gem =14.1, H-3′b), 3.53(s, 2H, CH2 -COOMe), 3.63(s, 3H, OCH3), 4.40(dd, 1H, J 2′，3′ =7.6 and 6.1, H-2'), 5.13 and 5.17 (2x d, 2H, J gem =12.3,O- CH2 Ph), 6.06 and 6.45 (2xd, 2H, J gem =1.1, =CH2), 7.05 (m, 2H, H-2 (aromatic)), 7.24 (m, 2H, H-3 (aromatic)), 7.26 (m, 2H, H-2″), 7.34-7.38 (m, 3H, H-3″, H-4″ (aromatic)), 8.58 (bs, 3H, NH3 + ).

[0383] 13 C NMR (DMSO-d6, ppm) δ: 35.68 (C-3′), 37.21 ( C H2-COOMe), 52.07(O C H3), 53.35(C-2′), 67.44(O- C H2Ph), 121.97(C-2), 128.05(C-2″), 128.67-128.69(C-2″, C-3″, C-4″), 130.87(C-3), 131.13(= C H2), 132.48(C-4), 133.56( C =CH2), 135.02(C-1″), 149.86(C-1), 164.62(CH2- C OO), 169.20( C OO-Bn), 171.11( C OO-Me),

[0384] ESI MS: 420.1 (M+Na) + (38), 398.2 (MH) + (100).

[0385] HRMS(ESI): For C 22 H 23 O6NNa(M+Na) + Calculated value: 420.14176; measured value: 420.14120.22 H 24 O6N(MH) + Calculated value: 398.15981; measured value: 398.15939.

[0386] Alkyl α-methyl itaconate

[0387]

[0388] R = alkyl, aryl, alkyl salicyl, Boc-tyrosyl alkyl amide, Boc-tyrosyl ester, (di)peptide with terminal tyrosyl ester, (di)peptide with terminal tyrosyl alkyl amide

[0389] Examples of R-OH of alkyl alpha-methyl itaconate include, but are not limited to:

[0390]

[0391] Alpha-methyl itaconate (1 mmol) was dissolved in dry dichloromethane (3 mL). Appropriate hydroxy derivatives (1.2 mmol) and DMAP (1.5 mmol) were added, and the solution was cooled to 0 ° C. A solution of DCC (1.5 mmol) in dichloromethane (2 mL) was added dropwise under nitrogen, and the entire mixture was stirred at 0 ° C for 1 h, and then stirred at room temperature overnight. The mixture was filtered, and the solution was extracted with 10% aqueous KHSO (3 × 5 mL) and saturated NaHCO (5 mL). The organic phase was dried over sodium sulfate, evaporated, and the residue was chromatographed on a silica gel column in the system described below.

[0392] 1-Methyl 4-(4-(tetradecyloxy)butyl) 2-methylene succinate (MK-956)

[0393]

[0394] Chromatography in cyclohexane-ethyl acetate (10:1) system. Yield: 244 mg (59%) of colorless liquid.

[0395] 1 H NMR (CDCl3, ppm) δ: 0.87 (t, 3H, J CH3，CH2 =7.0, CH3(14)), 1.24-1.33(m, 22H, 11xCH2(3-13)), 1.54(m, 2H, CH2(2)), 1.62(m, 2H, COO(CH2)2 CH2 CH2O), 1.69 (m, 2H, COO-CH2 CH2 ), 3.33(d, 2H, J CH2，C＝CH2 =1.2, CH2 -COO), 3.38 (t, 2H, J 1，2 =6.7, CH2(1)), 3.41(t, 2H, J CH2，CH2 =6.4, COO-(CH2)3 CH2 O), 3.76(s, 3H, OCH3), 4.12(t, 2H, J CH2，CH2 =6.5, COO-CH2), 5.70(q, 1H, J Ha，CH2 =J gem 1.1, =CH a ), 6.32(d, 1H, J gem 1.1, =CH b ).

[0396] 13 C NMR (CDCl3, ppm) δ: 14.11 (C-14), 22.67 (C-13), 25.42 (COO-CH2- C H2), 26.14 and 26.17 (C-3, COO(CH2)2 C H2CH2O), 29.34-29.73(m, C-2, C-4-C-11), 31.91(C-12), 37.75( C H2COO), 52.10(OCH3), 64.81(COO- C H2), 70.10(COO-(CH2)3 C H2O), 71.05(C-1), 128.44(C= C H2), 133.75( C =CH2), 166.65( C OO-CH3), 170.72(CH2- C OO).

[0397] ESI MS: 847.6 (2M+Na) + (10), 435.3 (M+Na) + (100).

[0398] HRMS(ESI): For C 24 H 44 O5Na(M+Na) + Calculated value: 435.30810; measured value: 435.30841.

[0399] 4-(3-(Hexadecyloxy)propyl)1-methyl 2-methylene succinate (MK-957)

[0400]

[0401] Chromatography in cyclohexane-ethyl acetate (80:15) system. Yield: 120 mg (28%) of colorless liquid.

[0402] 1 H NMR (CDCl3, ppm) δ: 0.87 (t, 3H, J CH3，CH2 =7.0, CH3(16)), 1.23-1.33(m, 26H, 13xCH2(3-15)), 1.54(m, 2H, CH2(2)), 1.88(p, 2H, OCH2- CH2 -CH2O), 3.33 (d, 2H, J CH2，C＝CH2 =1.2, CH2 -COO), 3.38 (t, 2H, J 1，2 =6.7,OCH2(1)),3.45(t,2H,J CH2，CH2 =6.3, COO-(CH2)2 CH2 O), 3.76(s, 3H, OCH3), 4.19(t, 2H, J CH2，CH2 =6.5, COO- CH2 ), 5.70(q, 1H, J Ha，CH2 =J gem 1.1, =CH a ), 6.32(d, 1H, J gem 1.1, =CH b ).

[0403] 13 C NMR (CDCl3, ppm) δ: 14.10 (C-16), 22.67 (C-15), 26.14 (C-3), 28.98 (OCH2- C H2-CH2O), 29.34-29.69(m, C-2, C-4-C-13), 31.91(C-14), 37.72( C H2COO), 52.10(OCH3), 62.27(COO-CH2), 66.99(COO-(CH2)2 C H2O), 71.16(C-1), 128.44(C= C H2), 133.73( C =CH2), 166.63( C OO-CH3), 170.64(CH2- C OO).

[0404] ESI MS: 875.7 (2M+Na) +(5), 449.3 (M+Na) + (100).

[0405] HRMS(ESI): For C 25 H 46 O5Na(M+Na) + Calculated value: 449.32375; measured value: 449.32382.

[0406] 4-(2-(Butoxycarbonyl)phenyl)1-methyl 2-methylene succinate (MK-961)

[0407]

[0408] Chromatography in cyclohexane-ethyl acetate (6:1) system. Yield: 63 mg (20%) of colorless liquid.

[0409] 1 H NMR (CDCl3, ppm) δ: 0.97 (t, 3H, J CH3，CH2 =7.4, CH3(4′)), 1.45(m, 2H, CH2(3′)), 1.72(m, 2H, CH2(2′)), 3.67(d, 2H, J CH2，C＝CH2 =1.1, C H2 -C=O), 3.81(s, 3H, OCH3), 4.27(t, 2H, J 1′，2′ =6.7, OCH2(1′)), 5.89(q, 1H, J gem =J Ha，CH2 =1.1, =C H a ), 6.42(d, 1H, J gem =1.1, =C H b ), 7.11(dddd, 1H, J 3，4 =8.1, J 3，5 =1.1, H-3), 7.30 (m, 1H, H-5), 7.54 (ddd, 1H, J 4，3 =8.2, J 4，5 =7.4, J 4，6 =1.7, H-4), 8.00 (dd, 1H, J 6，5 =7.9, J 6，4 =1.7, H-6).

[0410] 13 C NMR (CDCl3, ppm) δ: 13.74 (C-4′), 19.20 (C-3′), 30.68 (C-2′), 37.50 CH2C=O), 52.20(OCH3), 64.97(C-1′), 123.43(C-1), 123.79(C-3), 126.04(C-5), 129.30(= C H2), 131.59(C-6), 133.19( C =CH2), 133.67(C-4), 150.56(C-2), 164.38( C OOBu), 166.59( C OOCH3), 169.37(CH2- C OO).

[0411] ESI MS: 343.1 (M+Na) + (100).

[0412] HRMS(ESI): For C 17 H 20 O6Na(M+Na) + Calculated value: 343.11521; measured value: 343.11532.

[0413] 4-(4-((S)-2-((S)-2-((tert-Butoxycarbonyl)amino)-3-methylbutyrylamino)-3-isopropoxy-3-oxopropyl)phenyl)1-methyl 2-methylene succinate (MK-962)

[0414]

[0415] Chromatography in cyclohexane-acetone (4:1) system. Yield: 255 mg (47%) of white solid.

[0416] 1 H NMR (DMSO-d6, ppm) δ: 0.77-0.79 (m, 6H, CH( CH3 )2), 1.04 and 1.12 (2x d, 6H, Jx H3，CH =6.2, OCH( CH3 )2), 1.37(s, 9H, C( CH3 )3), 1.85 (m, 1H, CH (CH3)2), 2.93 (dd, 1H, J 3′a，2′ =8.5, J gem =14.0, H-3′a), 2.99(dd, 1H, J 3′b，2′ =6.6, J gem =14.0, H-3′b), 3.62(s, 2H, CH2-COOMe), 3.72(s, 3H, OCH3), 3.80(m, 1H, H-2″), 4.42(q, 1H, J 2′，3′ =J 2′，NH =7.4, H-2′), 4.81 (septet, 1H, J CH，CH3 =6.2,O CH (CH3)2), 5.96 and 6.28 (2x s, 2H, =CH2), 6.57 (d, 1H, J NH，2″ =9.2, NH -2″), 6.97 (m, 2H, H-aromatic (2)), 7.26 (m, 2H, H-aromatic (3)), 8.26 (d, J NH，2′ =7.4, NH-2′).

[0417] 13 C NMR (DMSO-d6, ppm) δ: 18.35 and 19.31 (C-4″), 21.50 and 21.65 (O—CH( C H3)2), 28.36(C( C H3)3), 30.71(C-3″), 36.24(C-3′), 37.38( C H2-COOMe), 52.29(OCH3), 53.66(C-2′), 59.61(C-2″), 68.19(O- CH (CH3)2), 78.16( C (CH3)3), 121.42(C-2), 129.66(=CH2), 130.41(C-3), 133.62( C =CH2), 134.89(C-4), 149.24(C-1), 155.48(NH- C OO), 166.27( C OO-Me), 169.38( C OO-Ph), 170.94(C-1′), 171.59(C-1″).

[0418] ESI MS: 571.3 (M+Na )+ (100).

[0419] HRMS(ESI): For C 28 H 40 O9N2Na(M+Na )+ Calculated value: 571.26260; measured value: 571.26280.

[0420] (S)-4-(4-(3-benzyloxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropyl)phenyl)1-methyl 2-methylene succinate (MK-963)

[0421]

[0422] Chromatography in cyclohexane-acetone (4:1) system. Yield: 223 mg (45%) of amorphous solid.

[0423] 1 H NMR (CDCl3, Ppm) δ: 1.41 (s, 9H, C ( CH3 )3), 3.04 (dd, 1H, J 3′a，2′ =6.0, J gem =14.0, H-3′a), 3.09(dd, 1H, J 3′b，2′ =6.0, J gem =13.9, H-3′b), 3.56(d, 2H, J CH2，C＝CH2 =1.1, CH2 -COOMe), 3.80(s, 3H, OCH3), 4.60(m, 1H, CH -NH), 4.99(d, J NH，CH = 8.3, NH), 5.09 and 5.17 (2x d, 2H, J gem =12.2,O- CH2 Ph), 5.82(q, 1H, J CH2，＝CHa =J gem =1.1, =CH a ), 6.40(d, 1H, J gem =0.8, =CH b ), 6.95 (m, 2H, H-2 (aromatic)), 7.02 (m, 2H, H-3 (aromatic)), 7.28-7.38 (m, 5H, H-2″, H-3″, H-4″ (aromatic)).

[0424] 13 C NMR (CDCl3, ppm) δ: 28.25 (C ( C H3)3), 37.51(C-3′), 37.86( C H2-COOMe), 52.23(O C H3), 54.31(C-2′), 67.15(O- C H2Ph), 79.97( C(CH3)3), 121.40 (C-2), 128.48 (C-4"), 128.58 and 128.60 (C-2", C-3"), 129.00 (=CH2), 130.28 (C-3), 133.29 ( C =CH2), 133.50(C-4), 135.06(C-1″), 149.61(C-1), 155.02(NH- C O), 166.47( C OO-Me), 169.07(CH2- C OO), 171.51 (C-1′).

[0425] ESI MS: 1017.7 (2M+Na) + (3), 520.3 (M+Na) + (100).

[0426] HRMS(ESI): For C 27 H 31 O8NNa(M+Na) + Calculated value: 520.19419; measured value: 520.19324. 27 H 32 O8N(MH) + Calculated value: 498.21224; measured value: 498.21161.

[0427] Deprotection of tert-butoxycarbonyl (Boc) group. General procedure.

[0428] A mixture of dichloromethane and trifluoroacetic acid (1:1, 16 mL) was added to the appropriate Boc derivative (0.8 mmol). The solution was stirred at room temperature for 20 min and evaporated. The residue was worked up as described below.

[0429] 4-(4-((S)-2-((S)-2-amino-3-methylbutyrylamino)-3-isopropoxy-3-oxopropyl)phenyl)1-methyl 2-methylene succinate trifluoroacetate (MK-964)

[0430]

[0431] The residue was crystallized from a mixture of ethyl acetate and diethyl ether. Yield: 340 mg (75%) of white crystals.

[0432] 1 H NMR (DMSO-d6, ppm) δ: 0.93 and 0.97 (2x d, 6H, CH( CH3)2), 1.06 and 1.14 (2x d, 6H, J CH3，CH =6.2, O-CH( CH3 )2), 2.12 (septet d, 1H, J 3″，4″ =6.9, J 3″，2″ =5.0, H-3″), 2.98(dd, 1H, J 3′a，2′ =8.2, J gem =14.2, H-3′a), 3.03(dd, 1H, J 3′b，2′ =6.6, J gem =14.2, H-3′b), 3.64(m, 2H, CH2 -COOMe), 3.65 (d, 1H, J 2″，3″ =5.1, H-2″), 3.73 (s, 3H, OCH3), 4.49 (m, 1H, H-2′), 4.84 (septet, 1H, J CH，CH3 =6.2,O- CH (CH3)2), 5.96 (q, 1H, J CH2，＝CHa =J gem =1.2, =CH a ), 6.28(d, 1H, J gem =1.2, =CH b ), 7.02(m, 2H, H-2), 7.31(m, 2H, H-3), 8.09(bs, 3H, NH3 + ), 8.89(d, 1H, J NH，2′ =7.0, NH).

[0433] 13 C NMR (DMSO-d6, ppm) δ: 17.43 and 18.51 (C-4″), 21.47 and 21.65 (O—CH( C H3)2), 30.07(C-3″), 36.03(C-3′), 37.40( C H2-COOMe), 52.32(OCH3), 54.23(C-2′), 57.23(C-2″), 68.60(O- CH (CH3)2), 121.62(C-2), 129.72(= C H2), 130.47(C-3), 133.60( C =CH2), 134.56(C-4), 149.40(C-1), 166.30( C OO-Me), 168.37(NH- C O), 169.51 (CH2- COO), 170.53( C OO-iPr).

[0434] ESI MS: 471.2 (M+Na) + (20), 449.2 (MH) + (100).

[0435] HRMS(ESI): For C 23 H 33 O7N2(MH) + Calculated value: 449.22823; measured value: 449.22745.

[0436] (S)-4-(4-(2-amino-3-(benzyloxy)-3-oxopropyl)phenyl)1-methyl 2-methylene succinate trifluoroacetate (MK-965)

[0437]

[0438] The residue was chromatographed on a silica gel column (60 mL) with a cyclohexane-acetone (1:1 to 1:4) gradient followed by chloroform-methanol (1:1) system to give 380 mg (93%) of a white viscous foam.

[0439] 1 H NMR (DMSO-d6, ppm) δ: 3.07 (dd, 1H, J 3′a，2′ =7.5, J gem =14.1, H-3′a), 3.15(dd, 1H, J 3′b，2′ =6.1, J gem =14.1, H-3′b), 3.66(d, 2H, J CH2，C＝CH2 =1.2, CH2 -COOMe), 3.73(s, 3H, OCH3), 4.37(dd, 1H, J 2′，3′ =7.6 and 6.1, H-2'), 5.13 and 5.17 (2x d, 2H, J gem =12.3,O- CH2 Ph), 5.98 (q, 1H, J gem =J CH2，＝CHa =1.2, =CH a ), 6.30(d, 1H, J gem =1.2, =CH b), 7.02 (m, 2H, H-2 (aromatic)), 7.22 (m, 2H, H-3 (aromatic)), 7.26 (m, 2H, H-2″ (aromatic)), 7.34-7.37 (m, 3H, H-3″, H-4″ (aromatic)), 8.48 (bs, 3H, NH3 + ).

[0440] 13 C NMR (DMSO-d6, ppm) δ: 35.71 (C-3′), 34.73 ( C H2-COOMe), 52.33(O C H3), 53.35(C-2′), 67.35(O- C H2Ph), 121.88(C-2), 128.60(C-2″, C-3″), 128.63(C-4″), 129.72(= C H2), 130.80(C-3′), 132.42(C-4), 133.62( C =CH2), 135.01(C-1″), 149.82(C-1), 166.32( C OO-Me), 169.25( C OO-Bn), 169.40(CH2- C OO).

[0441] ESI MS: 420.1 (M+Na) + (15), 398.2 (MH) + (100).

[0442] HRMS(ESI): For C 22 H 23 O6NNa(M+Na) + Calculated value: 420.14176; measured value: 420.14105. 22 H 24 O6N(MH) + Calculated value: 398.15981; measured value: 398.15930.

[0443] 4-(((Isopropoxycarbonyl)oxy)methyl)1-methyl 2-methylene succinate (IS-101-088)(21)

[0444]

[0445] α-Methyl itaconate (0.2 g, 1.39 mmol), chloromethyl isopropyl carbonate (0.22 mL, 1.66 mmol), sodium iodide (50 mg, 0.33 mmol) and potassium carbonate (0.29 g, 2.08 mmol) were dissolved in anhydrous MeCN (5 mL), and the mixture was stirred at 50° C. for 16 hours. EtOAc (60 mL) was added and the mixture was washed with brine (20 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: cyclohexane / ethyl acetate 5:1) to provide 0.34 g (93%) of compound IS-101-088 (21) as a colorless oil.

[0446] 1 H NMR (401 MHz, CDCl3): δ H 1.31 (d, J=6.2Hz, 6H), 3.37-3.43 (m, 2H), 3.76 (d, J=0.6Hz, 3H), 4.85-4.97 (m, 1H), 5.72-5.78 (m, 3H), 6.36 (d, J=0.8Hz, 1H).

[0447] 13 C NMR (101 MHz, CDCl3): δ C 21.6, 21.7, 37.3, 52.2, 73.1, 81.9, 129.2, 132.9, 153.3, 166.3, 169.2.

[0448] ESI MS: 283.1 ([M+Na] + ).

[0449] HRMS(ESI): For C 11 H 16 Calculated value for O7Na: 283.07882. Found: 283.07925.

[0450] 1-Methyl 4-((pivaloyloxy)methyl)2-methylene succinate (IS-101-089)(23)

[0451]

[0452] α-Methyl itaconate (0.2 g, 1.39 mmol), chloromethyl pivalate (0.26 mL, 1.8 mmol), sodium iodide (50 mg, 0.33 mmol) and potassium carbonate (0.29 g, 2.08 mmol) were dissolved in anhydrous MeCN (5 mL), and the mixture was stirred at 40° C. for 16 hours. EtOAc (60 mL) was added and the mixture was washed with brine (20 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: cyclohexane / ethyl acetate 5:1) to provide 0.33 g (92%) of compound IS-101-089 (23) as a colorless oil.

[0453] 1 H NMR (401 MHz, CDCl3): δ H 1.21 (s, 9H), 3.38 (d, J=1.2Hz, 2H), 3.76 (s, 3H), 5.74 (d, J=1.1Hz, 1H), 5.76 (s, 2H), 6.35 (d, J=0.8Hz, 1H).

[0454] 13 C NMR (101 MHz, CDCl3): δ C 26.9, 37.4, 38.8, 52.2, 79.7, 129.0, 133.0, 166.3, 169.4, 177.1.

[0455] ESI MS: 281.1 ([M+Na] + ).

[0456] HRMS(ESI): For C 12 H 18 Calculated value for O6Na: 281.09956. Found: 281.09993.

[0457] 1-(((tert-Butoxycarbonyl)oxy)methyl)4-methyl 2-methylene succinate (IS-102-081)

[0458]

[0459] β-Methylitaconate (1.00 g, 6.94 mmol), tert-butyl (chloromethyl) carbonate (1.27 g, 7.60 mmol), sodium iodide (0.20 g, 1.33 mmol) and potassium carbonate (1.44 g, 10.4 mmol) were dissolved in anhydrous MeCN (20 mL), and the mixture was stirred at 50° C. for 16 hours. The volatiles were then evaporated, the residue was redissolved in EtOAc (60 mL), and the mixture was washed with a saturated solution of sodium thiosulfate (20 mL) and brine (70 mL). The organic phase was dried over Na2SO4, the volatiles were evaporated, and the residue was subjected to flash column chromatography (silica gel 60 mesh 70-230, solvent: cyclohexane / ethyl acetate 5:1) to provide 1.65 g (87%) of compound IS-102-081 as a colorless oil.

[0460] 1 H NMR (401 MHz, CDCl3): δ H 1.49 (s, 9H), 3.34 (s, 2H), 3.68 (s, 3H), 5.78 (s, 2H), 5.81 (s, 1H), 6.42 (s, 1H).

[0461] 13 C NMR (101 MHz, CDCl3): δ C 27.7, 37.3, 52.2, 82.0, 83.7, 130.5, 133.0, 152.0, 164.7, 170.9.

[0462] ESI MS: 297.1 ([M+Na] + ).

[0463] HRMS(ESI): For C 12 H 18 Calculated value for O7Na: 297.09447. Found: 297.09465.

[0464] Scheme 1. Structural numbering for NMR assignment (alkyl β-methyl itaconate)

[0465]

[0466] Scheme 2. Structural numbers for NMR assignments (alkyl α-methyl itaconate)

[0467]

[0468] Example 6

[0469] In vitro data - mouse plasma stability

[0470] To evaluate the mouse plasma stability of the intact prodrug over time, the prodrug was spiked into mouse plasma to a final assay concentration of 10 μM. The spiked samples were incubated at 37°C in an orbital shaker for 1 h, and the reaction was then quenched with three volumes of acetonitrile containing an internal standard (IS; losartan: 0.5 μM). The samples were vortexed and centrifuged at 16,000 g for 5 min at 4°C. 50 microliters of the supernatant was diluted with 50 μL of water and transferred to a 250 μL polypropylene vial sealed with a Teflon cap. The disappearance of the prodrug over time was monitored using liquid chromatography-mass spectrometry (LC-MS). Representative results for the prodrugs disclosed in the present invention are presented in Figure 1 The release of active monomethyl itaconate (in plasma) from a representative prodrug is shown in Figure 2 The release of active itaconic acid (in plasma) from a representative prodrug is shown in Figure 3 Shown in.

[0471] References

[0472] All publications, patent applications, patents and other references mentioned in the specification are indicative of the level of those skilled in the art to which the subject matter disclosed herein belongs. All publications, patent applications, patents and other references are hereby incorporated by reference to the same extent as if each individual publication, patent application, patent and other reference was specifically and individually indicated to be incorporated by reference. It will be understood that although many patent applications, patents and other references are referred to herein, such references do not constitute an admission that any of these documents form part of the common general knowledge in the art.

[0473] International PCT Patent Application Publication No. WO2017142855, Artyomov et al., Immunomodulatory Agents and Methods of Use Thereof, published on August 24, 2017;

[0474] International PCT Patent Application Publication No. WO2019036509, Artyomov et al., Methods and Compositions for the Treatment of Diseases Associated with Cancer, Inflammation, or Immune Response, published on February 21, 2019;

[0475] O'Neill, LAJ and Artyomov, MN, Itaconate: the poster child of metabolic reprogramming in macrophage function, Nature Reviews: Immunology, 19, 273-281 (2019).

[0476] Boschert, D.; Schneider-Chaabane, A.; Himmelsbach, A.; Eickenscheidt, A.; Lienkamp, ​​K. Synthesis and Bioactivity of Polymer-Based Synthetic Mimics of Antimicrobial Peptides (SMAMPs) Made from Asymmetrically Disubstituted Itaconates. Chem. Eur. J. 2018, 24, 8217-8227.

[0477] McKenna, CE; Kashemirov, BA; Krylov, IS; Zakharova, VMM. Method to improve antiviral activity of nucleotide analogue drugs. US9550803B2, January 24, 2017.

[0478] Hidaka, K.; Gohda, K.; Teno, N.; Wanaka, K.; Tsuda, Y. Active site-directed plasmin inhibitors: Extension on the P2 residue. Bioorg. Med. Chem. 2016, 24, 545-553.

[0479] Although the foregoing subject matter has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be understood by those skilled in the art that certain changes and modifications may be practiced within the scope of the appended claims.

Claims

1. A compound selected from the following or a pharmaceutically acceptable salt thereof: 、 、 、 、 、 , or 。 2. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

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