Prodrugs, prodrug compositions, and related methods

Abstract

Embodiments of the present invention generally relate to prodrugs, and more specifically to prodrugs that can target brain-related diseases.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the benefit and priority of U.S. Provisional Patent Application No. 63 / 491,873, filed on 23 March 2023, the entire disclosure thereof being incorporated by reference.

[0002] Embodiments of the present invention generally relate to prodrugs, and more specifically to prodrugs that can target brain-related diseases. [Background technology]

[0003] The blood-brain barrier (BBB) ​​is a systemic membrane barrier that prevents the absorption of circulating drugs, protein therapies, RNAi drugs, and gene therapies into the brain. To deliver drugs or genes to the human brain for the treatment of serious brain diseases, delivery can be done in two ways: (a) by directly injecting the drug or gene into the brain to bypass the BBB, or (b) by injecting the drug or gene into the bloodstream so that it enters the brain via a vascular route across the BBB. Drug administration into the brain is highly invasive and not very effective. Vascular delivery is non-invasive and has the potential to distribute drugs more broadly to target cells in the brain. However, this latter approach requires the ability to undergo transport across the BBB, which has been a difficult barrier to cross safely.

[0004] The development of prodrugs has become one strategy to improve the physicochemical, pharmacokinetic, and / or pharmacodynamic properties of pharmacologically active substances, thereby enhancing their efficacy and / or reducing side effects. For example, prodrugs offer the potential to overcome various barriers in drug formulation and delivery, such as low water solubility, physical and / or chemical instability, poor absorption, rapid pre-systemic metabolism, poor brain penetration, toxicity, and / or local irritation. Prodrugs may also be effective in improving blood-brain barrier (BBB) ​​transport, drug targeting, and duration of action. Prodrugs are typically inactive derivatives of drug molecules that require chemical or enzymatic bioconversion to release the active parent drug in the body. Therefore, prodrugs need to be efficiently converted to the parent drug so that they exert a significant effect as soon as the drug target is achieved.

[0005] Therefore, there is a need for novel prodrug chemistry that enables efficient drug delivery across the blood-brain barrier (BBB). Furthermore, there is a need for novel prodrug chemistry that specifically targets target cells and / or target sites in the brain while minimizing toxicity. [Overview of the project]

[0006] The following summary is for illustrative purposes only and is not intended to limit it in any way. Further embodiments, exemplary designs, and features will become apparent by referring to the detailed description below, in addition to the exemplary aspects, exemplary designs, and features described.

[0007] In some aspects of the present invention, compounds having the structure of formula (I), or pharmaceutically acceptable salts thereof, are provided. Formula (I) is, [ka] And, In the formula, Z represents the therapeutic moiety. R 1 is either O or S, R 2is, independently at each occurrence, hydrogen or a C1-C3 alkyl group, R 3 is hydrogen or a C1-C3 alkyl group, R 4 is a C1-C6 alkyl group, a C6-C 10 aryl group, or a C5-C 10 heteroaryl group.

[0008] In some embodiments of the present invention, pharmaceutical compositions are presented. The pharmaceutical composition includes a compound of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, or cocrystal thereof, and a pharmaceutical carrier, diluent, or excipient.

[0009] In some embodiments of the present invention, a method of treating a brain disease is presented. This method includes administering to a patient a pharmaceutical composition comprising an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, or cocrystal thereof.

Embodiments for Carrying out the Invention

[0010] Hereinafter, various embodiments will be described in more detail with reference to the accompanying drawings showing only some embodiments. The specific structures and functions disclosed herein are representative only for the purpose of explaining examples of embodiments. However, the example embodiments can be implemented in many alternative forms and should not be construed as limited to only the example embodiments described herein. On the contrary, the example embodiments cover all modifications, equivalents, and alternatives thereof.

[0011] The terms used herein are solely for the purpose of describing specific embodiments and are not limiting. Unless otherwise defined, all terms used herein (including technical and scientific terms) have the same meaning as those generally understood by those skilled in the art in the field to which the example embodiments belong. Furthermore, terms defined in commonly used dictionaries, etc., should be interpreted as having the meaning consistent with their meaning in the context of the relevant art, and it will be understood that they should not be interpreted in an idealized or overly formal sense unless expressly defined herein.

[0012] Where used herein, the singular forms “a,” “an,” and “the” also include the plural forms unless the context clearly indicates otherwise. Where used herein, the terms “and / or” and “at least one” include any combination of one or more of the relevant list items. Furthermore, where used herein, the terms “comprises,” “comprising,” “includes,” and / or “including” identify the presence of the described features, integers, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof.

[0013] The approximation language used throughout this specification and the claims may be applied to modify any quantitative expressions that may change to an acceptable extent without altering the underlying functionalities. Therefore, values ​​modified with terms such as “approximately” are not limited to the specified exact values. Unless otherwise noted, all numerical values ​​used in this specification and the claims, representing quantities of components, properties such as molecular weight, reaction conditions, etc., are understood in all cases to be modified with the term “approximately.” Therefore, unless otherwise indicated, the numerical parameters described in the following specification and the appended claims are approximations and may vary depending on the desired properties sought to be obtained by the present invention. At a minimum, each numerical parameter should be interpreted using the usual rounding method, taking into account the reported number of significant digits.

[0014] To more clearly and concisely describe and point out the subject matter of the claimed invention, the following definitions are provided for the specific terms used in the following description and the appended claims. Definitions of specific functional groups and chemical terms are described in detail below. For the purposes of the present invention, chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed., on the inside of the cover, and specific functional groups are defined in general as described therein. Unless otherwise specified, structures shown herein include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structure of the present invention, including substitution of hydrogen with deuterium or tritium, substitution of carbon with 13C- or 14-C enriched carbon, or substitution of fluorine with 18F-enriched fluorine, are within the scope of the present invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents according to the present invention.

[0015] Unless otherwise specified, the structures shown herein include all isomers of the structure (e.g., enantiomers, diastereomers, and geometric isomers (or conformational isomers)), such as the R and S configurations of each chiral center, the Z and E double bond isomers, and the Z and E conformational isomers. Therefore, single stereochemical isomers of the compounds of the present invention, as well as enantiomers, diastereomers, and geometric (or conformational) mixtures, are within the scope of the present invention. Unless otherwise specified, all tautomers of the compounds of the present invention are within the scope of the present invention. Therefore, unless otherwise specified, the structures shown herein include compounds that differ only in the presence of one or more isotopically enriched atoms.

[0016] Where a particular enantiomer is preferred, in some embodiments the compound is provided substantially free of the corresponding enantiomer and is also referred to as “optically concentrated.” As used herein, “optically concentrated” means that the compound is composed of a significantly higher proportion of one enantiomer. In certain embodiments, the compound is made of at least about 90% by weight of the preferred enantiomer. In other embodiments, the compound is made of at least about 95%, 98%, or 99% by weight of the preferred enantiomer. The preferred enantiomer can be isolated from the racemic mixture by any method known to those skilled in the art, including chiral high-pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts, or it can be prepared by asymmetric synthesis.

[0017] As used herein, the terms "L-isomer" and "D-isomer" refer to enantiomers of amino acids. Amino acids, with the exception of glycine which lacks a chiral center, exist in nature in two forms (L and D optical isomers). L and D enantiomers differ in the orientation of the four substituents attached to the α-carbon and are also known as non-superimposed mirror images.

[0018] As used herein, the terms "syn isomer" and "anti isomer" refer to the diastereomers of the compounds of the present invention. Diastereomers are defined as non-mirror image, non-identical stereoisomers. In the present specification, these occur when two or more stereoisomers of a compound have different arrangements at one or more (but not all) of the equivalent (relevant) stereocenters and are not mirror images of each other. The prefixes "syn" and "anti" are used to denote geometric isomerism. The alternative name "syn isomer" indicates that two functional groups are present on the same side of a double bond (e.g., the N=C bond). The alternative name "anti isomer" indicates that two functional groups are present on opposite sides of a double bond (e.g., the N=C bond).

[0019] As used herein, the term "alkyl group" refers to a saturated monovalent group consisting of a non-cyclic, straight-chain or branched arrangement of atoms. An alkyl group is defined to contain at least one carbon atom and is represented by the formula C n H 2n+1 . The arrangement of atoms contained in an alkyl group may consist of only carbon and hydrogen. For example, the term "C1-C 10 alkyl group" contains at least 1 and no more than 10 carbon atoms. The methyl group (i.e., CH3-) is an example of a monovalent C1 alkyl group. The decyl group (i.e., CH3(CH2)9-) is an example of a monovalent C 10 alkyl group. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0020] As used herein, the term "aryl group" refers to a monovalent aromatic hydrocarbon radical derived by removing one hydrogen atom from one carbon atom of a parent aromatic ring system. Aryl groups include 5- and 6-membered carbocyclic aromatic rings (e.g., benzene), bicyclic ring systems (where at least one ring is carbocyclic and aromatic) (e.g., naphthalene, indane, tetralin), and tricyclic ring systems (where at least one ring is carbocyclic and aromatic) (e.g., fluorene).

[0021] The term "aryl group" also encompasses multiple ring systems having at least one carbocyclic aromatic ring, cycloalkyl ring, or heterocycloalkyl ring fused with at least one carbocyclic aromatic ring. For example, aryl groups include 5 and 6-membered carbocyclic aromatic rings fused with 5- to 7-membered heterocycloalkyl rings containing one or more heteroatoms selected from N, O, and S. In such fused bicyclic ring systems (where only one of the rings is a carbocyclic aromatic ring), the attachment site may be on the carbocyclic aromatic ring or the heterocycloalkyl ring. Examples of aryl groups include, but are not limited to, groups derived from acetantrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene, fluorantene, fluorene, hexacene, hexaphene, hexalene, as-indacene, s-indacene, indan, indene, naphthalene, octacene, octafen, octalene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene, rubicene, triphenylene, trinaphthalene, etc. In certain embodiments, the aryl group may contain 5 to 20 carbon atoms, and in certain embodiments, it may contain 5 to 12 carbon atoms.

[0022] As used herein, the term “heteroaryl group” refers to a monovalent heteroaromatic radical induced by removing one hydrogen atom from one atom of a parent heteroaromatic ring system. Heteroaryl groups encompass multiple ring systems in which at least one aromatic ring is fused to at least one other ring, which may be aromatic or nonaromatic, with at least one ring atom being a heteroatom. Heteroaryl groups encompass 5-12 membered aromatic monocyclic rings (such as 5-7 membered rings) containing one or more heteroatoms selected from N, O, and S, e.g., 1-4, or in certain embodiments, 1-3, with the remaining ring atoms being carbon, as well as bicyclic heterocycloalkyl rings containing one or more heteroatoms selected from N, O, and S, e.g., 1-4, or in certain embodiments, 1-3, with the remaining ring atoms being carbon, and at least one heteroatom present in the aromatic ring. For example, heteroaryls include 5-7 membered heteroaromatic rings fused to 5-7 membered cycloalkyl rings. In the case of fused bicyclic heteroaryl ring systems in which one or more heteroatoms are contained in only one of the rings, the attachment sites may be on heteroaromatic rings or cycloalkyl rings. In certain embodiments, if the total number of N, S, and O atoms in the heteroaryl group is greater than 1, the heteroatoms are not adjacent to each other. In certain embodiments, the total number of N, S, and O atoms in the heteroaryl group is 2 or less. In certain embodiments, the total number of N, S, and O atoms in the aromatic heteroring is 1 or less.

[0023] Examples of heteroaryl groups include, but are not limited to, groups derived from acridine, arsindol, carbazole, beta-carbolin, chroman, chromene, cinnoline, furan, furazan, imidazole, indazole, indole, indoline, indidine, isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, perimidine, phenantholidine, phenanthroline, phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolidine, quinazoline, quinoline, quinolidine, quinoxaline, tetrazoline, thiadiazole, thiazole, thiophene, triazole, xanthene, and others.

[0024] As used herein, the term “amino acid” refers to both naturally occurring and unnaturally occurring amino acids. Therefore, the term “amino acid” includes naturally occurring proteolytic L-amino acids, D-amino acids, chemically modified amino acids, such as amino acid variants and derivatives, naturally occurring unproteolytic amino acids, such as norleucine, β-alanine, and ornithine, as well as chemically synthesized compounds having properties known in the art as characteristic of amino acids.

[0025] Standard naturally occurring nonpolar (hydrophobic) amino acids include alanine (Ala), leucine (Leu), isoleucine (Ile), valine (Val), proline (Pro), phenylalanine (Phe), tryptophan (Trp), and methionine (Met). Polar neutral amino acids include glycine (Gly), serine (Ser), threonine (Thr), cysteine ​​(Cys), tyrosine (Tyr), asparagine (Asn), and glutamine (Gln). Positively charged (basic) amino acids include arginine (Arg), lysine (Lys), and histidine (His). Negatively charged (acidic) amino acids include aspartic acid (Asp) and glutamic acid (Glu). Non-standard amino acids can be formed in the body, for example, by post-translational modification. Some examples of such naturally occurring non-standard amino acids include selenocysteine ​​and pyrrolicin. Other non-limiting examples of amino acids include α-methyl amino acids (e.g., α-methylalanine), β-amino acids, histidine-like amino acids (e.g., 2-aminohistidine, β-hydroxyhistidine, homohistidine, α-fluoromethylhistidine, and α-methylhistidine), amino acids having an extra methylene group in the side chain ("homo" amino acids), and amino acids in which the carboxylic acid functional group of the side chain is replaced with a sulfonic acid group (e.g., cysteic acid).

[0026] As used herein, the term “amino acid” encompasses amino acids in which one or more amino groups in an amino acid compound may be further substituted with alkyl groups to form monoalkyl amino acids or dialkyl amino acids. For example, N,N-dimethyl-L-phenylalanine is an example of a dialkyl amino acid, and methyl-L-alanine is an example of a monoalkyl amino acid. Furthermore, in this specification, any designation of an amino acid without specifying its stereochemistry is intended to encompass either the L-form or D-form of the amino acid, or a racemic mixture thereof.

[0027] As used herein, the term “cleavable portion” refers to a portion having a cleavable bond under specific conditions of use, for example, after administration to a patient. The bond may be cleaved by enzymatic or non-enzymatic means. Cleavage may proceed spontaneously, such as by hydrolysis, or may be catalyzed or induced by another factor such as enzymes, acids, or bases, or by changes in or exposure to physical or environmental parameters such as temperature or pH. The drug may be endogenous to the conditions of use, such as enzymes present in the systemic circulation of the patient to whom the prodrug is administered, or the acidic conditions of the stomach, or the drug may be supplied exogenously. As used herein, the term “enzymatically cleavable portion” refers to a portion having a bond that can be cleaved by enzymatic means. In certain embodiments, the cleavable portion has a bond that can be cleaved by enzymatic means present in the affected organ, tissue, or cell.

[0028] As used herein, the term “specific binding” refers to a state in which one of two distinct molecules is specifically recognized by the other, but the recognition of the other molecule is significantly lower. A molecule may have regions on its surface or within its cavities that give rise to specific recognition between two molecules resulting from one or more electrostatic interactions, hydrogen bonds, or hydrophobic interactions. Specific examples of binding include, but are not limited to, antibody-antigen interactions, enzyme-substrate interactions, polynucleotide interactions, and receptor interactions. In some embodiments, specific binding refers to the binding of a compound of the present invention to one or more enzymes located within or near a target cell or target site (such as a target tissue or organ).

[0029] As used herein, the term “target” refers to a portion that is naturally expressed in greater quantities in a diseased state compared to a healthy state, or a portion that is naturally expressed in greater quantities in a target cell or target site (such as a tissue or organ). Generally, the compounds of the present invention may bind to a target via one or more individual chemical moieties of the target, or via three-dimensional structural components of the target (e.g., 3D structures resulting from peptide folding). Targets include one or more of the following: native or modified peptides, proteins (e.g., antibodies, aphibodies, aptamers, lectins), nucleic acids (e.g., polynucleotides, DNA, RNA, aptamers), polysaccharides (e.g., sugars), lipids, enzymes, enzyme substrates, ligands, receptors, antigens, and haptens. In some embodiments, a target may include one or more enzymes present in or near a target cell or target site (such as a target tissue or organ). Non-limiting examples of targets include brain amidase, fatty acid amidohydrolase (FAAH), kallikrein 6 (KLK6), acetyl-CoA synthetase 2 (ACSS2), and dipeptidyl peptidase 4 (DPP4).

[0030] As used herein, the term “prodrug” refers to a drug or derivative of a pharmacologically active substance that is administered in an inactive or completely inactive form and subsequently converted to an active form in the body. In some embodiments, the conversion releases the parent drug or pharmaceutically active substance. In some embodiments, a bioactive derivative of the parent drug or pharmaceutically active agent is produced.

[0031] As used herein, the term “therapeutic dose” means an amount of the compound that is sufficient to provide a therapeutic benefit to a patient in the treatment or management of a disease or disorder, or to delay or minimize one or more symptoms associated with the disease or disorder.

[0032] As used herein, the term “pharmaceutically acceptable salt” refers to any salt suitable for administration to a patient. Examples of salts include, but are not limited to, salts of acids, bases, organic, inorganic, amines, and alkali or alkaline earth metals (including, but not limited to, salts of calcium salts, magnesium salts, potassium salts, sodium salts, hydrochlorides, hydrobroms, sulfuric acid, nitric acid, phosphoric acid, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.).

[0033] In some embodiments, compounds having formula (I) or pharmaceutically acceptable salts thereof are presented. [ka] In the formula, Z represents the treatment area. R 1 is either O or S, R 2 In each occurrence, it is independently hydrogen or a C1-C3 alkyl group. R 3 is hydrogen or a C1-C3 alkyl group, R 4 C1-C6 alkyl groups, C6-C 10 Aryl group, or C5~C 10 It is a heteroaryl group.

[0034] R 2 Non-limiting examples include hydrogen, a methyl group, or an ethyl group. 3 Non-restrictive examples include those containing hydrogen or a methyl group. 4 Non-limiting examples include methyl, isopropyl, phenyl, or indole groups.

[0035] The compound is L and its syn isomer of formula (I). The compound is D and its syn isomer of formula (II). The compound is L and its anti isomer of formula (I). The compound is D and its syn isomer of formula (I).

[0036] In some embodiments, compounds having formula (II) or pharmaceutically acceptable salts thereof are presented. [ka] In the formula, Z represents the treatment area. R 1 is either O or S, R 2 In each occurrence, it is independently hydrogen or a C1-C3 alkyl group. R 3 is hydrogen or a C1-C3 alkyl group, R 4 C1-C6 alkyl groups, C6-C 10 Aryl group, or C5~C 10 It is a heteroaryl group.

[0037] The compound is the syn isomer of formula (II). The compound is the anti isomer of formula (II).

[0038] In some embodiments, compounds having formulas (III) to (VI) or pharmaceutically acceptable salts thereof are presented. [ka]

[0039] In some embodiments, compounds having formula (VII) or pharmaceutically acceptable salts thereof are presented. [ka] In the formula, R 2 In each instance, it is independently either hydrogen or a C1-C3 alkyl group.

[0040] As mentioned above, Z is the therapeutic portion derived from the therapeutic agent. Non-exclusive examples of therapeutic agents include ACE inhibitors, antianginergics, antiarrhythmics, antiasthmatics, anticholesterolemia drugs, anticonvulsants, antidepressants, antidiarrheals, antihistamines, antihypertensives, antiinfectives, anti-inflammatory drugs, antilipids, antimanic drugs, anti-nausea drugs, antistroke drugs, antithyroid drugs, antitumor drugs, antitussives, antiuricemia drugs, antiviral drugs, acne drugs, alkaloids, amino acid preparations, anabolic agents, analgesics, anesthetics, angiogenesis inhibitors, antacids, antiarthritis drugs, antibiotics, anticoagulants, antiemetics, antiobesity drugs, antiparasitic drugs, antipsychotics, antipyretics, antispasmodics, antithrombotic drugs, anxiolytics, appetite stimulants, appetite suppressants, beta-blockers, bronchodilators, cardiovascular agents, cerebral stimulants, chelating agents, This includes cholecystokinin antagonists, chemotherapy agents, cognitive stimulants, contraceptives, coronary artery dilators, antitussives, decongestants, deodorants, dermatological agents, antidiabetic agents, diuretics, emollients, enzymes, erythropoiesis-producing agents, expectorants, infertility treatments, antiseptics, gastrointestinal drugs, growth regulators, hormone replacement drugs, hyperglycemia drugs, hypnotics, hypoglycemia drugs, laxatives, migraine treatments, mucolytics, narcotics, nerve relaxants, neuromuscular drugs, NSAIDs, peripheral vasodilators, prostaglandins, psychotropic drugs, renin inhibitors, respiratory stimulants, steroids, stimulants, sympathomimetic agents, thyroid preparations, tranquilizers, uterine relaxants, vaginal preparations, vasoconstrictors, vasodilators, dizziness treatments, vitamins, and wound healing agents.

[0041] In some embodiments, Z is a therapeutic portion obtained from a therapeutic agent selected from the group consisting of antibiotics, anti-inflammatory agents, antiviral agents, anticancer agents, anti-infective agents, and combinations thereof. In some embodiments, Z is a therapeutic portion obtained from a corticosteroid. Non-limiting examples of corticosteroids include dexamethasone, prednisone, prednisolone, cortisone, hydrocortisone, betamethasone, and combinations thereof. In certain embodiments, Z is a dexamethasone residue. In some embodiments, Z is a therapeutic portion obtained from an antibiotic. In certain embodiments, Z is a mitramycin residue.

[0042] In some embodiments, compounds having formulas (VIII) to (X) or pharmaceutically acceptable salts thereof are presented. [ka] [ka] In the formula, R 1 is either O or S, R 2 In each occurrence, it is independently hydrogen or a C1-C3 alkyl group. R 3 is hydrogen or a C1-C3 alkyl group, R 4 C1-C6 alkyl groups, C6-C 10 Aryl group, or C5~C 10 It is a heteroaryl group.

[0043] According to embodiments of the present invention, the prodrug compounds of formulas (I) to (X) can specifically bind to a target and then release the therapeutic moiety Z, where the Z=N bond functions as a cleavable moiety. Furthermore, the prodrug compounds of formulas (I) to (X) of the present invention can effectively transport the therapeutic moiety across the blood-brain barrier.

[0044] In some embodiments, pharmaceutical compositions are presented. These pharmaceutical compositions comprise a compound described herein or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, or cocrystal thereof, and a pharmaceutical carrier, diluent, or excipient. In some embodiments, the pharmaceutical composition comprises a compound having formula (I).

[0045] The pharmaceutical composition of the present invention may be in any form that can be administered to a subject. For example, the composition may be in the form of a solid, liquid, or gas (aerosol). The pharmaceutical composition may be formulated such that, upon administration to a subject, the active ingredients contained therein become bioavailable. To further optimize the pharmacokinetic profile of the compounds of the present invention, the compounds may be administered in combination with a suitable delivery medium (e.g., lipid-based delivery systems such as microcapsules, microspheres, biodegradable polymer films, liposomes, and lipid foams, viscous infusions, and absorbable mechanical barriers) that helps maintain the required concentration of the prodrug or therapeutic agent at the disease site.

[0046] Processes for preparing pharmaceutical compositions are also presented. These processes involve mixing the compound or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, or cocrystal thereof with a pharmaceutically acceptable carrier, diluent, or excipient. In some embodiments, these processes involve mixing a compound having formula (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, or cocrystal thereof with a pharmaceutically acceptable carrier, diluent, or excipient.

[0047] In some embodiments, methods are also presented for treating or alleviating the symptoms of a particular disease by administering the compounds of the present invention. The compounds or their derivatives can be administered in amounts effective to treat the disorder to any host, including humans, non-human animals, and mammals.

[0048] In some embodiments, a method for treating brain diseases is presented. This method involves administering to a patient a pharmaceutical composition comprising an effective amount of the compound of the present invention or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, or cocrystal thereof. In some embodiments, this method involves administering to a patient a pharmaceutical composition comprising an effective amount of the compound of formula (I) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, or cocrystal thereof. In some embodiments, the brain disease is glioblastoma, glioma, medulloblastoma, or metastatic brain disease. According to embodiments of the present invention, the compound of formula (I) can effectively deliver the therapeutic portion Z across the blood-brain barrier.

[0049] Pharmaceutical compositions may be administered by any suitable method known to those skilled in the art. Typical routes of administration include, but are not limited to, oral, topical, parenteral, sublingual, rectal, vaginal, ocular, and nasal. As used herein, the term “parenteral” includes intravenous, intraperitoneal, intramuscular, intradermal, and epidermal administration including subcutaneous and intradermal, application to mucosal surfaces by oral or intranasal administration using, for example, inhalation of an aerosol suspension, and application to mucosal surfaces by implantation into the muscle or other tissue of the subject. Specific Embodiments

[0050] The embodiments listed below represent several aspects of the present invention.

[0051] Embodiment 1. A compound having formula (I) or a pharmaceutically acceptable salt thereof, [ka] In the formula, Z represents the treatment area. R 1 is either O or S, R 2 In each occurrence, it is independently hydrogen or a C1-C3 alkyl group. R 3 is hydrogen or a C1-C3 alkyl group, R 4 C1-C6 alkyl groups, C6-C10 Aryl group, or C5~C 10 A compound or a pharmaceutically acceptable salt thereof that contains a heteroaryl group.

[0052] Embodiment 2. The compound according to Embodiment 1, wherein the compound is L and a syn isomer of formula (I).

[0053] Embodiment 3. The compound according to Embodiment 1, wherein the compound is L and an antiisomer of formula (I).

[0054] Embodiment 4. The compound described in Embodiment 1, wherein the compound is D and a syn isomer of formula (I).

[0055] Embodiment 5. The compound according to Embodiment 1, wherein the compound is D and an antiisomer of formula (I).

[0056] Embodiment 6. The compound according to any one of Embodiments 1 to 3, wherein the compound has formula (II) or a pharmaceutically acceptable salt thereof. [ka]

[0057] Embodiment 7. The compound according to any one of Embodiments 1 to 6, wherein the compound has formula (III) to (VI) or a pharmaceutically acceptable salt thereof. [ka]

[0058] Embodiment 8.Z is a compound according to any one of Embodiments 1 to 7, which is a therapeutic portion obtained from a therapeutic agent selected from the group consisting of antibiotics, anti-inflammatory agents, antiviral agents, anticancer agents, anti-infective agents, and combinations thereof.

[0059] Embodiment 9.Z is a compound according to any one of Embodiments 1 to 8, wherein the therapeutic portion is obtained from a corticosteroid selected from the group consisting of dexamethasone, prednisone, prednisolone, triamcinolone, cortisone, hydrocortisone, and betamethasone.

[0060] Embodiment 10.Z is a compound according to any one of Embodiments 1 to 9, wherein the compound is a dexamethasone residue.

[0061] Embodiment 11.Z is a compound according to any one of Embodiments 1 to 9, wherein Z is a mitramycin residue.

[0062] Embodiment 12. A pharmaceutical composition comprising a compound described in any one of Embodiments 1 to 11 or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, or cocrystal thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.

[0063] Embodiment 13. A method for treating a brain disease, comprising administering to a patient a pharmaceutical composition containing an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, [ka] In the formula, Z represents the treatment area. R 1 is either O or S, R 2 In each occurrence, it is independently hydrogen or a C1-C3 alkyl group. R 3 is hydrogen or a C1-C3 alkyl group, R 4 C1-C6 alkyl groups, C6-C 10 Aryl group, or C5~C 10 A method involving a heteroaryl group.

[0064] Embodiment 14. The method according to Embodiment 14, wherein the compound is L and a syn isomer of formula (I).

[0065] Embodiment 15. The method according to Embodiment 14, wherein the compound is L and an antiisomer of formula (I).

[0066] Embodiment 16. The method according to Embodiment 14, wherein the compound is D and a syn isomer of formula (I).

[0067] Embodiment 17. The method according to Embodiment 14, wherein the compound is D and an antiisomer of formula (I).

[0068] Embodiment 18. The method according to any one of Embodiments 13 to 15, wherein the compound has formula (II) or a pharmaceutically acceptable salt thereof. [ka]

[0069] Embodiment 19. The method according to any one of Embodiments 1 to 18, wherein the compound has formulas (III) to (VI) or a pharmaceutically acceptable salt thereof. [ka]

[0070] Embodiment 20. The method according to any one of Embodiments 1 to 19, wherein the brain disease includes glioblastoma, medulloblastoma, glioma, or metastatic brain disease. [Examples]

[0071] General method for synthesizing prodrugs Dexamethasone (purity 95% or higher) and mitramycin A (purity 95% or higher) were purchased commercially and used for linking with the linker. All commercially available reagents and solvents were used as received. The reaction progress, intermediates, and final product were monitored and evaluated using LC / MS, HPLC-UV, and NMR (1H and 13C). Purification of the prodrug was performed using flash column chromatography on a silica gel column, or by preparative LC as needed. 1H NMR spectra were recorded at 300 MHz with a Bruker BioSpin GmbH spectrometer. The binding constant (J) is reported in Hertz (Hz). The chemical shift (δ) is reported in ppm relative to 2.50 ppm of residual solvent (DMSO-d6) or 0.00 ppm of internal standard tetramethylsilane. The following abbreviations are used in the reported spectra. s is a singlet, d is a doublet, t is a triplet, q is a quartet, m is a multiplet, dd is one doublet out of several doublets, td is a triplet of doublets, and ddd is a doublet of one doublet out of several doublets. The purity of all prodrugs synthesized in this project was 95% or higher.

[0072] Example 1: Synthesis of Compound I [ka]

[0073] To a solution of dexamethasone (5 g, 12.73 mmol) in DMF (25 mL), imidazole (1.73 g, 25.47 mmol) and TBDMSCl (2.30 g, 0.30 mmol) were added at 0°C, and the resulting mixture was stirred at room temperature for 12 hours. The reaction process was monitored by TLC. After the reaction was complete, ice water (50 mL) was added to the reaction mixture to form a precipitate. The precipitate was filtered, washed with water, and vacuum-dried to obtain compound 2 (5.2 g, 80%) as a white solid.

[0074] A solution of compound 2 (3.0 g, 5.92 mmol) in methanol (12 mL) was stirred, and propionohydrazide (1.5 g, 17.78 mmol) and AcOH (177 mg, 2.96 mmol) were added at 0°C. The reaction mixture was stirred at ambient temperature for 12 hours. After the reaction was complete, the methanol was concentrated under reduced pressure, diluted with H2O (200 mL), and extracted with ethyl acetate (2 × 200 mL). The organic layer was washed with brine (150 mL), dried over sodium sulfate, and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by flash column chromatography to obtain compound 3b (800 mg, 23%) as a pale yellow solid.

[0075] A solution of compound 3b (0.5 g, 0.86 mmol) in THF (5 mL) was stirred, and TBAF (1 M, 1.7 mL, 1.72 mmol) was added at 0°C. The reaction mixture was stirred at ambient temperature for 2 hours. After the reaction was complete, ice water (30 mL) was added to the reaction mixture, and it was extracted with ethyl acetate (2 × 150 mL). The organic layer was washed with brine (120 mL), dried over sodium sulfate, and concentrated under reduced pressure to obtain 300 mg of the crude compound. 100 mg of the 300 mg of crude compound was purified by preparative HPLC and lyophilized. The compound was made basic with saturated NaHCO3 solution, extracted with ethyl acetate (50 mL), concentrated under reduced pressure, and lyophilized to obtain compound I (15 mg, yield 11.27%) as a white solid.

[0076] 1H NMR(400MHz,DMSO-d6):δ10.48(dd,1H),6.74(dd,1H),6.41(dd,J=17.0,10.1Hz,1H),6.22(dd,J=22.5,10.0 Hz,1H),5.13(dd,J=10.4,2.8Hz,1H),4.94(s,1H),4.68(t,J=5.8Hz,1H),4.49(dd,J=19.3,5.9Hz,1H),4.07 (dd,J=19.3,5.8Hz,2H),2.94(m,1H),2.67-2.55(m,2H),2.34-2.09(m,5H),1.71(m,1H),1.58(m,1H),1.41( d,J=5.2Hz,3H),1.29(m,2H),1.02(q,J=7.4Hz,4H),0.84(s,3H),0.78(d,J=7.2Hz,3H)ppm.LC-MS(ESI):m / z Calculated value 462, actual value 463 (M+H)+, (RT=5.1, purity 99.78%). HPLC: 99.31% (250nm), 99.49% (295nm).

[0077] Example 2: Synthesis of Compound II [ka]

[0078] To a stirred solution of dexamethasone (1.0 g, 2.54 mmol) in methanol and DMSO (10 mL), propionohydrazide (0.67 g, 7.64 mmol) and AcOH (75 mg, 1.27 mmol) were added at 0°C. The reaction mixture was stirred at 50°C for 12 hours. After the reaction was complete, methanol was evaporated under reduced pressure, diluted with well-chilled water (50 mL), and extracted with ethyl acetate (100 mL x 2). The organic layer was washed with brine solution (50 mL), dried over sodium sulfate, and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by preparative HPLC and lyophilized. The compound was made basic with saturated NaHCO3 solution, extracted with ethyl acetate (50 mL), concentrated under reduced pressure, and lyophilized to obtain compound II (26 mg, yield 2.22%) as a white solid. 1H NMR (400MHz, DMSO-d6): δ10.54(bd,1H),6.99(bd,10.1Hz,1H),6.60(bd,J=30.4,10.3Hz,1H),5.96 (d,J=24.5Hz,1H),5.04(d,J=86.4Hz,1H),4.48(d,J=19.2Hz,1H),4.09(s,2H),2.92(m,3H),2.36- 2.20(m,2H),2.17-2.04(m,2H),1.71-1.53(m,3H),1.41(d,J=6.3Hz,4H),1.40-1.35(m,1H),1.33- 1.22(m,2H),1.02(dd,J=14.9,7.5Hz,4H),0.83(s,3H),0.77(d,J=7.2Hz,3H)ppm.LC-MS(ESI):m / z Calculated value 462, actual value 463 (M+H)+, (RT=5.3, purity 99.38%). HPLC: 98.15% (250nm), 98.89% (290nm).

[0079] Example 3: Synthesis of Compounds IIIA and IIIB [ka]

[0080] To a stirred solution of (tert-butoxycarbonyl)phenylalanine (2 g, 7.53 mmol) in THF (12 mL), CDI (1.35 g, 8.27 mmol) was added at room temperature and the mixture was stirred for 1 hour. Hydrazine hydride was added to the reaction mixture at 0°C. The resulting mixture was stirred at room temperature for 12 hours and monitored by TLC. The reaction mixture was quenched with ice water (150 mL), and the compound was extracted with ethyl acetate (2 × 200 mL). The combined organic layers were washed with brine solution, dried over sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash column chromatography, and the compound was eluted with 5-6% methanol in a DCM gradient to obtain compound 3 (1.5 g, 71%) as an off-white solid.

[0081] To a solution of dexamethasone (1) (1 g, 2.55 mmol) and compound 3 (1.5 g, 5.10 mmol) in DCE (10 mL), TEA (0.55 mL, 3.82 mmol) and STAB (650 mg, 3.06 mmol) were slowly added at 0°C. The resulting mixture was stirred at ambient temperature for 12 hours and monitored by LC-MS. The reaction mixture was quenched with water (100 mL), and the compounds were extracted with DCM (2 × 150 mL). The combined organic layers were washed with brine solution, dried over sodium sulfate, and then concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash chromatography, and the compounds were eluted with a 2-4% methanol gradient in DCM to obtain compound 4 (1.2 g, 72%) as a white solid.

[0082] A solution of compound 4 (400 mg, 0.61 mmol) in DCM (6 mL) was stirred, and TFA (2 mL) was slowly added at -10°C. The resulting mixture was stirred at -10 to -5°C for 3 hours and monitored by TLC and LC-MS. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain the residue, which was ground with diethyl ether to obtain the crude product. The crude product was purified by preparative HPLC to obtain compound IIIA (P1-30.8 mg) and compound IIIB (P2-40 mg) as white solids.

[0083] Compound IIIA (Pike 1): ¹H NMR (400 MHz, DMSO-d6): δ 10.48 (s, ¹H), 7.37–7.11 (m, 6H), 6.22 (d, J = 10.1 Hz, 1H), 6.00 (s, 1H), 5.54 (d, J = 87.6 Hz, 1H), 5.29 (dd, J = 12.6, 4.2 Hz, 1H), 4.53 (d, J = 56.8 Hz, 1H), 4.35–4.05 (m, 3H), 3.51 (dd, J = 9.4, 4.0 Hz, 1H), 3.18 (s, 1H), 3.0 8 (dd, J=13.5, 4.2Hz, 1H), 2.95 (dd, J=13.4, 4.3Hz, 1H), 2.72-2.57 (m, 2H), 2.39-2.25 (m, 3H), 2.21-2.05 (m, 2H), 1.78 (s, 1H), 1.65-1.51 (m, 1H), 1.50-1.38 (m, 4H), 1.38-1.19 (m, 2H), 1.14-1.02 (m, 1H), 0.96-0.77 (m, 6H). LC-MS (ESI): m / z Calculated value 553, measured value 554 (M+H)+, (RT=4.45, and purity 99.69%). HPLC: 99.08% (230nm), 98.84% (240nm), MR: 184℃~188℃.

[0084] Compound IIIB (Pike 2): ¹H NMR (400MHz, DMSO-d6): δ 10.49 (s, ¹H), 7.37–7.15 (m, 6H), 6.23 (t, J = 9.7 Hz, ¹H), 6.00 (s, ¹H), 5.69–5.36 (m, ¹H), 5.28 (s, ¹H), 4.53 (d, J = 65.7 Hz, ¹H), 4.29 (d, J = 3.4 Hz, ¹H), 4.25–4.15 (m, ¹H), 4.10 (dd, J = 23.5, 17.2 Hz, 2H), 3.56–3.42 (m, ¹H), 3.17 (s, ¹H), 3.13–3. 0.5 (m, 1H), 2.97 (dd, J=13.2, 4.6Hz, 1H), 2.72-2.54 (m, 4H), 2.33 (s, 4H), 2.19-2.05 (m, 3H), 1.77 (s, 1H), 1.71-1.53 ​​(m, 1H), 1.45 (dd, J=22.6, 10.9Hz, 5H), 1.34 (d, J=9.2Hz, 1H), 1.23 (s, 1H), 1.07 (s, 2H), 0.96-0.84 (m, 5H), 0.81 (d, J=7.1Hz, 1H). LC-MS (ESI): m / z Calculated value 553, measured value 554 (M+H)+, (RT=4.5, and purity 99.58%). HPLC: 99.07% (230nm), 98.99% (240nm). MR:184℃~188℃.

[0085] Example 4, synthesis of compounds IVA and IVB

change

[0086] To a stirred solution of (tert-butoxycarbonyl)valine (2 g, 9.20 mmol) in THF (12 mL), CDI (1.64 g, 10.12 mmol) was added at room temperature and the mixture was stirred for 1 hour. Hydrazine hydride (0.88 g, 27.6 mmol) was added to the reaction mixture at 0°C. The resulting mixture was stirred at room temperature for 12 hours and monitored by TLC. The reaction mixture was quenched with ice water (100 mL), and the compounds were extracted with ethyl acetate (2 × 150 mL). The combined organic layers were washed with brine solution, dried over sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash column chromatography, and the compounds were eluted with 4-6% methanol in a DCM gradient to obtain compound 3 (1.2 g, 56%) as an off-white solid.

[0087] To a solution of dexamethasone (1) (1 g, 2.54 mmol) and compound 3 (590 mg, 2.54 mmol) in DCE (10 mL), TEA (0.35 mL, 2.54 mmol) and STAB (538 mg, 2.54 mmol) were slowly added at 0°C. The resulting mixture was stirred at ambient temperature for 12 hours and monitored by TLC. The reaction mixture was quenched with water (100 mL), and the compounds were extracted with DCM (2 × 150 mL). The combined organic layers were washed with brine solution, dried over sodium sulfate, and then concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash column chromatography, and the compounds were eluted with 2-4% methanol in a DCM gradient to obtain compound 4 (870 mg, 56%) as an off-white solid.

[0088] A solution of compound 4 (400 mg, 0.66 mmol) in DCM (10 mL) was stirred, and TFA (2 mL) was slowly added at 0°C. The resulting mixture was stirred at ambient temperature for 3 hours and monitored by TLC. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain the residue, which was ground with diethyl ether to obtain the crude product. The crude product was purified by preparative HPLC to obtain compound IVA (P1-28 mg) and compound IV (P2-26 mg) as white solids.

[0089] Compound IVA (Pike 1): ¹H NMR (400MHz, DMSO-d6): δ 10.84 (s, ¹H), 7.33 (d, J = 10.1 Hz, ¹H), 6.23 (d, J = 10.1 Hz, ¹H), 6.02 (s, ¹H), 5.75 (d, J = 40.7 Hz, ¹H), 5.34 (dd, J = 27.7, 4.1 Hz, ¹H), 4.64 (d, J = 30.9 Hz, ¹H), 4.31 (dd, J = 33.6, 14.9 Hz, 2H), 4.12 (s, ¹H), 3 0.09(s,1H), 2.62(d,J=5.1Hz,1H), 2.30(dd,J=24.2,6.0Hz,2H), 2.09(ddd,J=25.1,15.2,6.6Hz,3H), 1.77(s,1H), 1.64-1.45(m,5H), 1.35(dd,J=13.1,4.5Hz,1H), 1.08(d,J=8.4Hz,1H), 1.05-0.77(m,12H). LC-MS(ESI): m / z Calculated value 505, measured value 506(M+H)+, (RT=4.18, and purity 99.85%). HPLC: 99.14% (230nm), 99.52% (240nm), MR: 226℃~230℃.

[0090] Compound IVB (Pike 2): ¹H NMR (400 MHz, DMSO-d6): δ 10.88 (s, ¹H), 7.33 (d, J = 10.1 Hz, ¹H), 6.23 (dd, J = 10.1, 1.5 Hz, ¹H), 6.01 (s, ¹H), 5.80 (t, J = 41.7 Hz, ¹H), 5.34 (dd, J = 9.5, 4.3 Hz, ¹H), 4.79–4.56 (m, ¹H), 4.39–4.05 (m, 4H), 3.2 3-3.96 (m, 1H), 2.60 (d, J=13.6Hz, 1H), 2.40-2.25 (m, 2H), 2.23-2.00 (m, 3H), 1.78 (dd, J=10.7, 5.8Hz, 1H), 1.66-1.44 (m, 5H), 1.41-1.29 (m, 1H), 1.17-1.09 (m, 1H), 1.07-0.77 (m, 12H). LC-MS (ESI): m / z Calculated value 505, Detected value 506 (M+H)+, (RT=4.16, purity 99.82%). HPLC: 99.69% (230nm), 99.41% (240nm). MR: 226℃~230℃

[0091] Example 5: Synthesis of compounds VA and VB [ka]

[0092] To a stirred solution of (tert-butoxycarbonyl)valine (209 mg, 0.96 mmol) and compound-1 (500 mg, 0.96 mmol) in DMF (5 mL), TEA (0.34 mL, 2.43 mmol), HOBT (208 mg, 1.53 mmol), and DCC (317 mg, 1.53 mmol) were added at 0°C. The resulting mixture was stirred at room temperature for 12 hours and monitored by TLC. After the reaction was complete, it was quenched with cold water (100 mL) and extracted with DCM (2 × 150 mL). The combined organic layer was washed with brine solution, dried over sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash column chromatography, and the compound was eluted with 3-4% methanol in a DCM gradient to obtain compound 2 (620 mg, 89%) as an off-white solid.

[0093] To a solution of compound 2 (400 mg, 0.55 mmol) in DCM (6 mL), TFA (2 mL) was slowly added at 0°C, and the resulting mixture was stirred at room temperature for 12 hours. After the reaction was complete, the reaction was monitored by TLC. The reaction mixture was concentrated under reduced pressure to obtain the residue, which was made basic with saturated NaHCO3 solution and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were dried over sodium sulfate and then concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC to obtain compound VA (P1-14 mg) and compound VB (P2-15 mg) as off-white solids.

[0094] Compound VA (Pike 1): ¹H NMR (400MHz, DMSO-d6): δ 11.13 (s, ¹H), 6.73 (d, J = 62.1Hz, ¹H), 6.50 (dd, J = 17.5, 10.2Hz, ¹H), 6.27 (dd, J = 30.7, 11.3Hz, ¹H), 5.21–5.10 (m, ¹H), 4.95 (d, J = 2.4Hz, ¹H), 4.69 (t, J = 5.8Hz, ¹H), 4.49 (dd, J = 19.3, 5.9Hz, ¹H), 4.32 (d, J = 4.8Hz, ¹H), 4.07 (dd, J = 19.2, 5.6Hz, 2H), 3.56 (d, J = 6... .1Hz,1H),2.94(s,1H),2.61(d,J=8.9Hz,1H),2.24(d,J=8.9Hz,1H),2.13(d,J=8.7Hz,3H),2.04-1.96(m,1H),1.74(d,J=5.8Hz,1H),1.67-1. 53(m,1H),1.42(d,J=7.2Hz,4H),1.34(d,J=11.8Hz,1H),1.06(s,1H),0.99-0.86(m,6H),0.85(s,3H),0.78(d,J=7.2Hz,3H).LC-MS(ESI):m / z Calculated value 505, measured value 506 (M+H)+, (RT=4.14, 4.28 and 95.19, and purity 4.51%). HPLC: 95.64 and 3.34% (250nm), 96.88 and 2.80% (300nm), MR: >240°C.

[0095] Compound VB (peak 2): 1H NMR (400MHz, DMSO-d6): δ11.13(s,1H),6.84-6.61(m,1H),6.49(t,J=11.8Hz,1H),6.26(dd,J=29.3,10.2Hz,1H),5.16(d,J=9. 5Hz,1H),4.95(d,J=3.2Hz,1H),4.70(s,1H),4.50(dd,J=19.1,5.8Hz,1H),4.28-4.00(m,2H),3.47-3.39(m,1H),2.94(s,1H),2 .61(s,1H),2.24(d,J=9.4Hz,1H),2.13(d,J=13.9Hz,2H),2.00-1.88(m,1H),1.74(s,1H),1.61(d,J=11.0Hz,1H),1.43(d,J=7. 5Hz,4H),1.35(d,J=10.0Hz,1H),1.06(s,1H),0.96(d,J=6.9Hz,1H),0.93-0.83(m,7H),0.78(d,J=7.2Hz,3H).LC-MS(ESI):m / z Calculated value 505, found value 506 (M+H)+, (RT=4.14, 4.26 and 92.5, and purity 6.9%). HPLC: 91.71 and 5.08% (250 nm), 92.21 and 5.23% (300 nm), MR: >240°C.

[0096] Example 6: Synthesis of compounds VIA and VIB [ka]

[0097] To a solution of dexamethasone (5 g, 12.73 mmol) in DMF (25 mL), imidazole (1.73 g, 25.47 mmol) and TBDMSCl (2.30 g, 0.30 mmol) were added at 0°C, and the resulting mixture was stirred at room temperature for 12 hours. The reaction process was monitored by TLC. After the reaction was complete, well-chilled water (50 mL) was added to the reaction mixture to form a precipitate. The precipitate was filtered, washed with water, and vacuum-dried to obtain compound 2 (5.2 g, 80%) as a white solid.

[0098] Compound 2 (5 g, 9.88 mmol) was dissolved in MeOH (50 mL) and stirred. Acetic acid (0.28 mL, 2.667 mmol) and hydrazine hydrate (1.48 g, 29.64 mmol) were added at 0°C, and the resulting mixture was stirred at room temperature for 3 hours. After the reaction was complete, the solvent was removed, the resulting residue was diluted with water (250 mL), and extracted with ethyl acetate (2 × 400 mL). The combined organic layer was washed with brine solution (300 mL), dried over sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash column chromatography, and the pure compound was eluted using a 70% ethyl acetate gradient in petroleum ether.

[0099] To a stirred solution of compound 3 (500 mg, 0.96 mmol) and (tert-butoxycarbonyl)phenylalanine (255 mg, 0.96 mmol) in DMF (5 mL), TEA (0.34 mL, 2.43 mmol), HOBT (208 mg, 1.53 mmol), and DCC (317 mg, 1.53 mmol) were slowly added at 0°C. The resulting mixture was stirred at ambient temperature for 12 hours and monitored by TLC. After the reaction was complete, the mixture was quenched with cold water (50 mL), and the compound was extracted with ethyl acetate (2 × 100 mL). The combined organic layer was washed with brine solution, dried over sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash chromatography, and the compound was eluted with a 30-35% ethyl acetate gradient in petroleum ether to obtain compound 3 (480 mg, 65%) as a pale yellow solid.

[0100] To a stirred solution of compound 3 (300 mg, 0.39 mmol) in DCM (6 mL) at 0°C, TFA (2 mL) was added, and the reaction mixture was stirred at ambient temperature for 12 hours. The reaction was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated to obtain the residue. Water was added, the mixture was made basic with saturated NaHCO3 solution, and extracted with ethyl acetate. The combined organic layers were washed with brine solution. The organic layers were dried over sodium sulfate and then concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC to obtain compound VIA (P1-17 mg) and compound VIB (P2-19 mg) as white solids.

[0101] Compound VIA (Pike 1): ¹H NMR (400 MHz, DMSO-d6): δ 11.19 (s, ¹H), 7.34–7.22 (m, 6H), 6.77 (s, ¹H), 6.52 (t, J = 10.8 Hz, ¹H), 6.32–6.24 (m, ¹H), 5.17 (s, ¹H), 4.96 (d, J = 5.2 Hz, ¹H), 4.71–4.66 (m, ¹H), 4.53–4.46 (m, ¹H), 4.11–4.05 (m, 2H) ,3.14-2.93(m,4H),2.63-2.62(m,1H),2.26-2.11(m,4H),1.76-1.74(m,1H),1.66-1.57(m,1H),1.43-1.42(m,4H),1.36-1.29(m,1H),1.08-1.06(m,1H),0.85(s,3H),0.79(d,J=6.8Hz,3H). LC-MS (ESI): m / z Calculated value 553, Detected value 554 (M+H)+, (RT=4.35, purity 96.98%). HPLC: 96.86% (250nm), 97.35% (300nm). MR:>240℃.

[0102] Compound VIB (Pike 2): 1H NMR (400MHz, DMSO-d6): δ 11.17 (s, 1H), 7.35-7.24 (m, 6H), 6.81 (s, 1H), 6.54-6.27 (m, 2H), 5.17 (s, 1H), 4.95 (s, 1H), 4.71-4.68 (m, 1H), 4.61 (dd, J1=7.2Hz, J2=3.6Hz, 1H), 4.10-4. 0.4 (m, 3H), 3.05-2.88 (m, 3H), 2.67-2.60 (m, 1H), 2.33-2.10 (m, 4H), 1.76-1.73 (m, 1H), 1.65-1.56 (m, 1H), 1.43-1.17 (m, 5H), 1.07-1.05 (m, 1H), 0.85 (s, 3H), 0.78 (d, J=6.4Hz, 3H). LC-MS (ESI): m / z calculated value 553, measured value 554 (M+H)+, (RT=4.4, purity 94.83%). HPLC: 92.31% (250nm), 92.65% (300nm). MR: >240℃.

[0103] Synthesis of Compounds VIIA and VIIB [ka]

[0104] To a solution of compound 1 (5.0 g, 17.9 mmol, 1 equivalent) and dexamethasone (7.0 g, 17.9 mmol, 1 equivalent) in MeOH (20 mL), TFA (2.6 mL, 35.8 mmol, 2 equivalents) was added at room temperature. The resulting mixture was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (1:1) to obtain compound 2 (10 g, 76.91%) as a white solid. MS(ESI)(m / z):[M+1]+ Calculated value: 654.3, Measured value: 654.4.

[0105] To a solution of compound 2 (10 g, 15.3 mmol, 1 equivalent) in DCM (75 mL), TFA (25 mL) was added at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The resulting mixture was concentrated under vacuum. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel, mobile phase, MeCN (10 mmol / L NH4HCO3) in water, gradient from 20% to 60% over 30 minutes, detector: UV 254 nm. The resulting mixture was concentrated under reduced pressure to obtain the product (mixture, ratio: approximately 1:1, 2.7 g) as a white solid. The product (mixture, ratio: approximately 1:1, 2.7g) was purified by chiral preparative HPLC under the following conditions (column: CHIRAL ART Cellulose-SB, 2*25cm, 5μm, mobile phase A: MtBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 20mL / min, gradient: 7.5 min from 30%B to 30%B, wavelength: 220 / 254nm, front peak (min): 4.63, post peak (min): 6.65, sample solvent: EtOH--HPLC, injection volume: 0.7mL, number of runs: 10), yielding compound VIIA (1.0877g, 45.48%) as an off-white solid with a front peak and compound VIIB (1.0355g, 43.25%) as an off-white solid with a post peak.

[0106] Compound VIIA: ¹H NMR (300 MHz, DMSO-d6) δ 10.73 (s, ¹H), 7.32–7.14 (m, 5H), 6.50–6.40 (m, 2H), 6.26 (ddd, J = 9.1, 7.1, 1.9 Hz, ¹H), 5.20–5.12 (m, ¹H), 4.98–4.93 (m, ¹H), 4.70 (s, ¹H), 4.50 (dd, J = 19.1, 4.9 Hz, 1H), 4.11 (s, 2H), 3.61 (ddd, J = 7.9, 6.0, 2.0 Hz, 1H), 2.94 (s, 2H). 2.98-2.87(m,1H),2.68(dd,J=13.2,8.0Hz,1H),2.17(dp,J=31.4,12.0,11.3Hz,4H),1.72(s,2H),1.61(q,J=11.9Hz,1H),1.42(s,3H),1.38(d,J=14.7Hz,1H),1.07(ddd,J=11.8,8.0,4.1Hz,1H),0.85(s,3H),0.78(d,J=7.2Hz,3H)..MS(ESI)(m / z)[M+H]+ Calculated value: 554.3, Measured value: 554.3.

[0107] Compound VIIB: ¹H NMR (300 MHz, DMSO-d⁶) δ 10.66 (s, ¹H), 7.23 (ddt, J = 22.0, 14.6, 6.8 Hz, 5H), 6.50–6.40 (m, 2H), 6.26 (ddd, J = 9.3, 7.1, 1.8 Hz, 1H), 5.19–5.12 (m, 1H), 4.95 (d, J = 3.3 Hz, 1H), 4.70 (t, J = 5.8 Hz, 1H), 4.50 (dd, J = 19.2, 5.9 Hz, 1H), 4.16–4.02 (m, 2H), 3.64–3.56 (m,1H),2.93(dd,J=13.4,5.9Hz,2H),2.63(ddd,J=31.1,13.9,8.2Hz,1H),2.23(t,J=13.9Hz,1H),2.17-2.05(m,4H),1.61(q,J=11.7Hz,1H),1.42(s,3H),1.34(d,J=8.7Hz,3H),1.12-1.01(m,1H),0.85(s,3H),0.78(d,J=7.2Hz,3H).MS(ESI)(m / z)[M+H]+ Calculated value: 554.3, Measured value: 554.3.

[0108] Synthesis of compounds VIIIA and VIIIB [ka]

[0109] To a solution of compound 1 (2.0 g, 7.5 mmol, 1 equivalent) and 1-hydroxypyrrolidine-2,5-dione (0.9 g, 8.3 mmol, 1.1 equivalents) in DMF (20 mL), DCC (1.5 g, 7.5 mmol, 1.0 equivalent) was added at 0°C. The resulting mixture was stirred at 0°C for 1 hour and at room temperature for 2 hours. Hydrazine hydrate (3.7 g, 75.3 mmol, 10.0 equivalents) was added dropwise to the above mixture at room temperature. The resulting mixture was stirred at room temperature overnight. The resulting mixture was filtered, and the filter cake was washed with HCl (2 × 50 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was diluted with HCl (100 mL). The combined organic layers were washed with KHCO3 (saturated 2 × 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. As a result, compound 2 (2.1 g, 90.7%) was obtained as a white solid. MS(ESI)(m / z):[M+1] + Calculated value: 280.2, Measured value: 280.3.

[0110] To a solution of compound 2 (2.1 g, 7.5 mmol, 1 equivalent) and dexamethasone (2.9 g, 7.5 mmol, 1.00 equivalent) in MeOH (20 mL), TFA (1.1 mL, 15.0 mmol, 2.0 equivalent) was added at room temperature. The resulting mixture was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel, mobile phase, MeCN in water ((0.1% FA)), gradient from 20% to 70% over 30 minutes, detector: UV 254 nm. The resulting mixture was concentrated under reduced pressure. As a result, compound 3 (3.1 g, 61.2%) was obtained as a pale yellow solid. MS(ESI)(m / z):[M+1] + Calculated value: 654.3, Measured value: 654.4.

[0111] To a solution of compound 3 (3.0 g, 4.6 mmol, 1 equivalent) in DCM (30 mL), TFA (10 mL) was added at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The resulting mixture was concentrated under vacuum. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel, mobile phase, MeCN (10 mmol / L NH4HCO3) in water, gradient from 30% to 50% over 30 minutes, detector: UV 254 nm. The resulting mixture was concentrated under reduced pressure to obtain the product (mixture, ratio: approximately 3:2, 2.5 g) as a white solid. The product (mixture, ratio: approximately 3:2, 2.5g) was purified by chiral preparative HPLC under the following conditions (column: Lux 5um Cellulose-4 2.12×25cm, 5μm, mobile phase A: hexane (0.3% IPA amine)-HPLC, mobile phase B: EtOH-HPLC, flow rate: 20mL / min, gradient: isocratic 50, wavelength: 220 / 254nm, front peak (min): 7.689, post peak (min): 10.526, sample solvent: EtOH-HPLC, injection volume: 0.5mL, number of analyses: 36). The front peak yielded compound VIIIA (347.7mg, 12.24%) as a white solid, and the post peak yielded compound VIIIB (512.3mg, 18.69%) as a white solid.

[0112] Compound VIIIA: 1H NMR(400MHz,DMSO-d6)δ10.72(s,1H),7.35-7.13(m,5H),6.76(dd,J=10.3,1.9Hz,1H),6.63(dd,J=19.7,10.4Hz,1H),6.00(d,J=7.3Hz,1H),5.15( d,J=4.1Hz,1H),4.67(t,J=5.7Hz,1H),4.49(dd,J=19.2,5.5Hz,1H),4.25 (dd,J=8.1,4.8Hz,1H),4.13-4.02(m,2H),3.60(dd,J=7.9,5.8Hz,1H),2. 93(dt,J=11.0,6.1Hz,2H),2.66(dd,J=13.4,7.9Hz,1H),2.34-2.26(m,1H ),2.24(s,1H),2.13(t,J=9.8Hz,2H),1.69(d,J=12.2Hz,1H),1.59(t,J=1 1.5Hz,1H),1.47(d,J=16.0Hz,1H),1.43(s,3H),1.32(d,J=14.6Hz,2H),1 .17-1.02(m,1H),0.85(s,3H),0.79(d,J=7.2Hz,3H).MS(ESI)(m / z)[M+H] + Calculated value: 554.3, measured value: 554.3.

[0113] Compound VIIIB: 1H NMR(400MHz,DMSO-d6)δ10.66(s,1H),7.34-7.14(m,5H),6.50-6.35(m,2H),6.27(dd,J=10.1,1.8Hz,1H),5.21-5.11(m,1 H),4.94(d,J=2.7Hz,1H),4.68(d,J=7.1Hz,1H),4.50(dd,J=19.2,5.2Hz,1H),4.17-4.02(m,2H),3.62(dd,J=7.8,5.9Hz, 1H),2.94(ddt,J=13.5,8.5,4.4Hz,2H),2.73-2.52(m,2H),2.33-2.08(m,4H),1.74(d,J=13.1Hz,1H),1.61(q,J=11.7Hz, 1H),1.47-1.38(m,4H),1.33(d,J=11.7Hz,1H),1.10-1.02(m,1H),0.86(s,3H),0.79(d,J=7.2Hz,3H).MS(ESI)(m / z)[M+H] + Calculated value: 554.3, Measured value: 554.3.

[0114] Synthesis of compounds IXA and IXB [ka]

[0115] To a stirred solution of compound 1 (3.0 g, 16.73 mmol, 1.0 equivalent) and HCHO (10.22 mL, 100.43 mmol, 6 equivalents, 36%) in MeOH (15 mL), AcONa (1.37 g, 16.73 mmol, 1.0 equivalent) and NaBH3CN (2.10 g, 33.47 mmol, 2.0 equivalents) were added at 0°C. The resulting mixture was stirred overnight at room temperature. The resulting mixture was diluted with water (30 mL). The resulting mixture was extracted with SiO2 (2 × 50 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (5:1) to obtain compound 2 (1.6 g, 46.11%) as a white solid. MS(ESI)(m / z):[M+1]+ Calculated value: 208.2, Measured value: 208.3.

[0116] Compound 2 (1.6 g, 7.71 mmol, 1 equivalent) and hydrazine hydrate (1.55 g, 30.87 mmol, 4 equivalents) were stirred in EtOH (10 mL) at room temperature. The resulting mixture was stirred overnight at 80 °C. The resulting mixture was concentrated under vacuum. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel, mobile phase, MeCN (10 mmol / L NH4HCO3) in water, gradient from 10% to 50% over 10 minutes, detector: UV 254 nm. As a result, compound 3 (0.8 g, 50.00%) was obtained as a pale yellow oily substance. MS (ESI) (m / z): [M+1]+ Calculated value: 208.1, Measured value: 208.1.

[0117] To a solution of compound 3 (400 mg, 1.93 mmol, 1 equivalent) and dexamethasone (757.38 mg, 1.93 mmol, 1.00 equivalent) in MeOH (10.0 mL), TFA (0.29 mL, 3.86 mmol, 2.0 equivalent) was added at room temperature. The resulting mixture was stirred overnight at room temperature. The resulting mixture was concentrated under vacuum. The crude product (410 mg) was analyzed by chiral preparative HPLC under the following conditions (2#SHIMADZU(HPLC-01)): column: YMC-Actus Triart C18 ExRS, 30*150 mm, 5 μm, mobile phase: 10 mmol NH4HCO3 + 0.05% NH3H2O ​​and ACN (from 35% ACN to 65% in 8 mins), detector: UV 254 nm, and compound 4 (200 mg, 17.10%) was obtained as a white solid. MS(ESI)(m / z):[M+1]+ Calculated value: 582.3, Measured value: 582.3.

[0118] Compound 4 was purified by chiral preparative HPLC under the following conditions (column: CHIRALPAK ID, 2 × 25 cm, 5 μm; mobile phase A: hexane (0.1% FA)-HPLC; mobile phase B: IPA-HPLC; flow rate: 20 mL / min; gradient: 40% B to 40% B (33 mins); wavelength: 220 / 254 nm; front peak (min): 11.911; post peak (min): 25.866; sample solvent: IPA-HPLC; injection volume: 1 mL; number of analyses: 5). The front peak was compound IXA (23.2 mg, 10.73%) as a white solid, and the post peak was compound IXB (15.0 mg, 6.84%) as a white solid.

[0119] Compound IXA:1H NMR(400MHz,Methanol-d4)δ7.33-7.16(m,5H),6.70(d,J=10.4Hz,1H),6.26(dd,J=10.4,1.9Hz,1H),6.10(s,1H),4.6 1(dd,J=19.3,6.1Hz,1H),4.32-4.13(m,2H),3.54(s,1H),3.20-3.03(m,3H),2.81-2.69(m,1H),2.68-2.53(m,7H),2. 48-2.31(m,1H),2.29(s,2H),2.26(s,1H),2.21(td,J=11.6,8.0Hz,1H),1.83-1.65(m,2H),1.50(s,3H),1.44(dd,J=1 3.7,2.2Hz,1H),1.20(ddd,J=12.1,8.1,4.0Hz,1H),1.00(d,J=3.5Hz,3H),0.88(t,J=7.4Hz,3H).MS(ESI)(m / z)[M+H]+ Calculated value: 582.3, actual value: 582.3.

[0120] Compound IXB:1H NMR(400MHz,Methanol-d4)δ7.33-7.22(m,4H),7.26-7.15(m,1H),6.60(d,J=10.1Hz,1H),6.35(dd,J=10.1,2.1Hz,1H),6.04(s ,1H),4.64(s,1H),4.29(s,1H),4.22(d,J=10.2Hz,1H),3.55-3.47(m,1H),3.19-3.01(m,3H),2.76-2.60(m,1H),2.54(d,J=1.4H z,6H),2.40-2.28(m,1H),2.30-2.24(m,1H),2.22(dd,J=13.6,5.7Hz,2H),1.88-1.74(m,1H),1.71(d,J=11.6Hz,1H),1.49(s,3 H),1.57-1.41(m,1H),1.31(s,1H),1.22(ddd,J=12.1,8.1,4.1Hz,1H),1.00(s,3H),0.89(d,J=7.2Hz,3H).MS(ESI)(m / z)[M+H]+ Calculated value: 582.3, actual value: 582.3.

[0121] Example 9: Synthesis of Compound X [ka]

[0122] To a solution of compound 1 (100 mg, 0.35 mmol, 1 equivalent) in MeOH (20 mL), hydrazine hydrate (0.87 mL, 14.32 mmol, 2.0 equivalents, 80%) was added at 0°C. The resulting mixture was stirred at room temperature for 5 hours. The resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with RINKAN (2 × 50 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. As a result, compound 2 (1.5 g, 75.00%) was obtained as a white solid. The crude product was used directly in the next step without further purification. MS(ESI)(m / z):[M+1]+ Calculated value: 280.2, Measured value: 280.3.

[0123] To a stirred solution of compound 2 (500 mg, 1.79 mmol, 1 equivalent) in pyridine (8.0 mL), P2S5 (397.81 mg, 1.790 mmol, 1.00 equivalent) was added at room temperature. The resulting mixture was stirred overnight at 90°C. The resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with  (2 × 50 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. As a result, compound 3 (320 mg, 60.52%) was obtained as a yellow solid. The crude product was used directly in the next step without further purification. MS(ESI)(m / z):[M+1]+ Calculated value: 296.2, Measured value: 296.2.

[0124] To a solution of compound 3 (320 mg, 1.08 mmol, 1.0 equivalent) and dexamethasone (425.15 mg, 1.08 mmol, 1.0 equivalent) in MeOH (5.0 mL), TFA (0.16 mL, 2.16 mmol, 2.0 equivalent) was added at room temperature. The resulting mixture was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with PE / EA (2:1) to obtain compound 4 (200 mg, 27.56%) as a yellow solid. MS(ESI)(m / z):[M+1]+ Calculated value: 670.3, Measured value: 670.4.

[0125] To a solution of compound 4 (200 mg, 0.30 mmol, 1 equivalent) in DCM (3.0 mL), TFA (1.0 mL) was added at room temperature. The resulting mixture was stirred at room temperature for 1 hour. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography under the following conditions: column: C18 silica gel, mobile phase: MeCN in water (10 mmol / L NH4HCO3), gradient from 30% to 50% over 30 minutes, detector: UV 254 nm, to obtain compound X (27.9 mg, 16.40%) as a yellow solid.

[0126] Compound X:1H NMR(400MHz,Methanol-d4)δ7.41-7.28(m,6H),6.93(d,J=10.0Hz,1H),6.71(d,J=3.8Hz,1H),4.64-4.59(d,J=1 4.3Hz,1H),4.32-4.22(m,3H),3.39(dd,J=13.2,7.2Hz,1H),3.24-3.10(m,2H),2.79(td,J=13.7,6.2Hz,1H),2.5 2(dtd,J=29.0,11.9,4.8Hz,2H),2.27(dd,J=25.3,11.9Hz,2H),2.01-1.92(m,1H),1.81-1.75(d,J=11.9Hz,1H) ,1.61-1.48(m,5H),1.22(dt,J=14.9,7.2Hz,1H),1.03(s,3H),0.88(dd,J=7.4,2.5Hz,3H).MS(ESI)(m / z)[M+H]+ Calculated value: 570.3, actual value: 570.1.

[0127] Example 10: Synthesis of Compound XI [ka]

[0128] Mitramycin A (1 equivalent), MeOH, hydrazine (2 equivalents), and TFA (2 equivalents) were added to an 8 mL vial at room temperature. The resulting mixture was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure. The crude product was purified by preparative HPLC to obtain compound XI.

[0129] Compound XI: 1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),7.34(s,1H),7.31(d,J=6.4Hz,4H),7. 25(t,J=6.7Hz,2H),6.33(s,1H),6.24(s,1H),6.17(s,1H),5.96(s,1H),5.2 3(s,1H),5.17(d,J=9.4Hz,1H),4.96(s,2H),4.87(dd,J=13.8,8.2Hz,5H),4 .61(td,J=24.0,20.6,11.9Hz,5H),4.22-4.13(m,4H),4.03(s,1H),3.96(s,2 H),3.75(d,J=11.8Hz,3H),3.62(s,1H),3.55(s,1H),3.24(s,5H),2.95(d,J =9.3Hz,2H),2.88(s,1H),2.77(t,J=8.6Hz,5H),2.40(s,2H),2.28(d,J=8.7 Hz,1H),2.04-1.90(m,5H),1.76(d,J=12.5Hz,2H),1.67-1.57(m,2H),1.50- 1.33(m,3H),1.23(d,J=7.1Hz,5H),1.21-1.06(m,23H).MS(ESI)(m / z)[M+H]+ Calculated value: 1246.6, Measured value: 1246.6.

[0130] Example 11, Synthesis of Compound XII [ka]

[0131] Mitramycin A (1 equivalent), MeOH, hydrazine (2 equivalents), and TFA (2 equivalents) were added to an 8 mL vial at room temperature. The resulting mixture was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure. The crude product was purified by preparative HPLC to obtain compound XII.

[0132] Compound XII: 1H NMR(300MHz,DMSO-d6)δ14.80(d,J=12.4Hz,1H),10.72(s,1H),7.35(d,J=7.5Hz,2 H),7.25(t,J=6.6Hz,5H),6.61(s,1H),6.38(s,1H),6.22(s,1H),6.15(s,2H),5.2 4(d,J=9.3Hz,1H),5.06(s,1H),5.03-4.86(m,10H),4.69(d,J=6.7Hz,1H),4.42(s ,1H),4.26(s,2H),4.16(s,1H),4.04(d,J=5.5Hz,2H),3.83(s,3H),3.70(s,1H),3 .61(s,2H),3.48(s,1H),3.23(d,J=16.7Hz,1H),2.94(s,3H),2.79(d,J=10.4Hz,6 H),2.39(s,3H),2.29(d,J=8.1Hz,11H),2.08(d,J=6.6Hz,4H),1.98(s,6H),1.87( s,4H),1.76(d,J=13.1Hz,1H),1.49-1.36(m,3H),1.21(s,13H),1.30-1.11(m,22H) ),1.04(s,9H),1.04(d,J=16.4Hz,2H),0.84(d,J=6.4Hz,1H).MS(ESI)(m / z)[M+H] + Calculated value: 1274.6, Measured value: 1274.6.

[0133] In vitro and in vitro biological research After synthesizing various compounds, each compound was tested in various preclinical models to optimize the lead compounds. Initial testing included i) chemical stability at two different pH levels (4.5 and 7.4), ii) human and rodent plasma stability, iii) rat liver microsomal stability, iv) cell permeability, v) pharmacokinetic and tissue distribution analysis, and vi) biological assays such as glucocorticoid receptor (GR) binding.

[0134] In vivo PK studies were conducted in accordance with the guidelines of the Institutional Animal Care and Use Committee (IACUC). No abnormal clinical symptoms were observed in the rats throughout the experiment.

[0135] chemical stability The chemical stability of the prodrug compound was tested at pH 4.5 and pH 7.4 (pH adjusted in phosphate-buffered saline). Both pH buffers were prepared in-house. Two μL of a 500 μM stock solution of the prodrug was added to each vial containing 198 μL of PBS at pH 7.4 or pH 4.5 and mixed homogeneously. The assay was performed redundantly. The final concentration of the test compound was 5 μM. Samples were incubated at 37°C and 600 rpm. The reaction start times were staggered so that all time points (0, 30, 60, 120, 180, 240 minutes) were terminated simultaneously with 1000 μL of cold quench solution (acetonitrile containing internal standards (IS, 200 nM labetalol, 100 nM alprazolam, 2 μM ketoprofen)). Samples from separate vials were used for each different time point. The sample was vortexed for 2 minutes and centrifuged at 3,220 g for 5 minutes at 4°C. 100 μL of supernatant was transferred to a new plate. The supernatant was diluted with 100 μL or 200 μL of water depending on the LC / MS signal response and peak shape. The sample was thoroughly mixed and analyzed using LC / MS / MS. The t1 / 2 value was calculated using the parent drug residue versus reaction time.

[0136] Plasma stability Rat and human plasma were obtained from accredited vendors. 5 μL of test substance solution (500 μM) was added to pre-incubated human or rat plasma (495 μL) to a final concentration of 5 μM. The final concentration of the organic solvent was less than 0.5%. The assay was performed redundantly. Reaction samples were incubated in a 37°C water bath with shaking at approximately 60 rpm. 50 μL of reaction sample was collected at 0, 30, 60, 120, and 180 minutes. The reaction was stopped by adding 300 μL of quench solution at room temperature (acetonitrile containing internal standards (IS, 200 nM labetalol, 100 nM alprazolam, 2 μM ketoprofen)). After vortexing the samples for 5 minutes, the proteins were precipitated by centrifugation at 3,220 g for 30 minutes at room temperature. 100 μL of supernatant was transferred to a new plate and diluted with 100 μL or 200 μL of water depending on the LC / MS signal response and peak shape. The samples were thoroughly mixed and analyzed using LC / MS / MS. The t1 / 2 value was calculated using the parent drug residue versus reaction time.

[0137] Metabolic stability: Rat liver microsomes were obtained from an authorized vendor, and the metabolic stability of the test compound was determined at 1 μM. The compound was incubated at 37°C in buffer in the presence of 1 mM NADPH at a concentration of 0.5 mg microsomes / mL. 50 μL samples were taken from the reaction solution at 0, 15, 30, 45, and 60 minutes. The reaction was stopped by adding four times the volume of cold acetonitrile containing internal standards (IS, 200 nM labetalol, 100 nM alprazolam, 2 μM ketoprofen). Precipitated proteins were removed by centrifugation, and the supernatant was collected, diluted, and analyzed. Two positive controls and a negative control (without NADPH) were used in this study. All samples were analyzed redundantly. LC / MS / MS was performed for detection and quantification of the test compound. Linear regression was determined using the remaining percentage of the parent drug at each incubation time, and the t1 / 2 value was calculated.

[0138] Permeability studies using a monolayer of Caco-2 cells: The objective of this study was to evaluate the bidirectional permeability and absorption mechanisms of the test compound. CaCO-2 cells were cultured, and monolayers were developed in-house in 96-well plates over 18 days using a standard protocol. The integrity of the monolayer cells was tested before use.

[0139] The test compound was added to the Transwell insert (apical compartment) to determine the drug transport rate from the apex to the basal outward direction. To determine the drug transport rate from the basal outward direction to the apex, the test compound was added to the wells of the receiver plate (basal outward compartment). An appropriate amount of HBSS (10 mM HEPES, pH 7.4) was added to both compartments to a final volume of 200 μL. The plate was incubated at 37°C for 2 hours while shaking in a rotary shaker at 150 rpm. At the end of the transport period, 50 μL was taken from the apical and basal outward wells, and the reaction was terminated by adding four times the volume of cold acetonitrile containing internal standards (IS: 100 nM alprazolam, 200 nM labetalol, 2 μM ketoprofen) to each sample. The samples were vortexed and centrifuged at 3,800 g for 20 minutes. A 150 μL supernatant was divided and used for LC / MS / MS analysis. All samples were analyzed redundantly. Samples were analyzed by LC / MS / MS to estimate the permeability and efflux of compounds across the Caco-2 cell monolayer.

[0140] Glucocorticoid receptor binding assay The glucocorticoid receptor (GR) binding of prodrugs was determined using the TR-FRET GR competitive binding assay. Three binding points (0.1, 1, and 10 μM) were evaluated for each prodrug. Dexamethasone was used as a positive control.

[0141] Pharmacokinetic studies in rats: SD rats aged 6–8 weeks were obtained from qualified providers. The test compound was introduced into the rats by a single intravenous dose. A liquid formulation (2 mg / kg dexamethasone or an equivalent prodrug) was prepared in 20% HP-β-CD and administered via the tail vein of rats that had been fasted overnight as a prerequisite. After administration, blood and brain samples were collected from three rats (3 repeats) at three time points (15, 60, and 360 minutes). Blood samples (0.1, 0.2, or 0.5 mL) were collected in a 1 mL syringe containing the anticoagulant K2EDTA. Blood samples were centrifuged to obtain plasma for analysis. Additional blood samples were collected from the 360-minute group at 15, 30, 60, 120, and 240 minutes. After collection at each time point, brain samples were frozen at -75°C. Prior to analysis, a portion of the brain sample was weighed and homogenized with phosphate-buffered saline. The concentrations of the prodrug and dexamethasone released from the prodrug in both plasma and brain tissue were measured by LC / MS / MS. Data analysis and pharmacokinetic parameters were generated using WinNonlin software.

[0142] Table 1 shows the chemical stability of the prodrug compound according to embodiments of the present invention at two different pH values ​​(4.5 and 7.4). As shown in Table 2, the prodrug compound was also stable at pH 7.4. [Table 1]

[0143] Table 2 shows the human plasma stability and rat plasma stability of the prodrug compounds according to embodiments of the present invention. As shown in Table 2, the prodrug compounds showed stability in human plasma. [Table 2]

[0144] Table 3 shows the rat liver microsome stability data for prodrug compounds. [Table 3]

[0145] Table 4 shows the permeability data for prodrug compounds. [Table 4]

[0146] Table 5 shows the glucocorticoid receptor (GR) binding data of prodrug compounds according to embodiments of the present invention. As shown in Table 3, the prodrugs of the present invention showed significantly lower GR binding compared to dexamethasone. [Table 5]

[0147] Table 6 shows in vitro biological assay data for a mixture of prodrug isomers according to embodiments of the present invention. [Table 6]

[0148] Pharmacokinetics and tissue distribution in vivo: A mixture of compounds VIA and VIB was tested in rats, and the concentrations of the prodrug and active molecule (dexamethasone) in plasma and brain were measured at three different time points. Table 7 shows that the prodrug compounds showed higher dexamethasone concentrations in the brain compared to plasma. [Table 7-1] [Table 7-2]

[0149] Although the plasma dexamethasone concentration released from the prodrug was significantly lower than that of dexamethasone administered to rats, the brain concentration of dexamethasone was higher with the prodrug. Therefore, the data suggest that the prodrug compound of the present invention may deliver a sufficient amount of dexamethasone to the brain with less systemic exposure, thereby reducing the toxicity of dexamethasone in the body.

[0150] While this specification illustrates and describes only certain features of several embodiments, many modifications and changes are possible for those skilled in the art. Therefore, it should be understood that the appended claims are intended to cover all modifications and changes that fall within the scope of the present invention and the appended claims.

Claims

1. A compound having formula (I) or a pharmaceutically acceptable salt thereof, 【Chemistry 1】 In the formula, Z represents the treatment area. R 1 is either O or S, R 2 In each appearance, independently, hydrogen or C 1 ~C 3 It is an alkyl group, R 3 is hydrogen or C 1 ~C 3 It is an alkyl group, R 4 is a C 1 to C 6 alkyl group, a C 6 to C 10 aryl group, or a C 5 to C 10 heteroaryl group, said compound or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, wherein the compound is L and a syn isomer of formula (I).

3. The compound according to claim 1, wherein the compound is L and an antiisomer of formula (I).

4. The compound according to claim 1, wherein the compound is D and a syn isomer of formula (I).

5. The compound according to claim 1, wherein the compound is D and an antiisomer of formula (I).

6. The compound according to claim 1, wherein the compound has formula (II) or a pharmaceutically acceptable salt thereof. 【Chemistry 2】

7. The compound according to claim 1, wherein the compound has formula (III) to (VI) or a pharmaceutically acceptable salt thereof. 【Transformation 3】

8. The compound according to claim 1, wherein Z is a therapeutic portion obtained from a therapeutic agent selected from the group consisting of antibiotics, anti-inflammatory agents, antiviral agents, anticancer agents, anti-infective agents, and combinations thereof.

9. The compound according to claim 8, wherein Z is a therapeutic portion obtained from a corticosteroid selected from the group consisting of dexamethasone, prednisone, prednisolone, triamcinolone, cortisone, hydrocortisone, and betamethasone.

10. The compound according to claim 9, wherein Z is a dexamethasone residue.

11. The compound according to claim 8, wherein Z is a mitramycin residue.

12. A pharmaceutical composition, A compound according to claim 1 or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, or cocrystal thereof, Pharmaceutical carrier, diluent, or excipient The pharmaceutical composition comprising the above.

13. A method for treating a brain disease, comprising administering to a patient a pharmaceutical composition containing an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, 【Chemistry 4】 In the formula, Z represents the treatment area. R 1 is either O or S, R 2 In each appearance, independently, hydrogen or C 1 ~C 3 It is an alkyl group, R 3 is hydrogen or C 1 ~C 3 It is an alkyl group, R 4 C 1 ~C 6 alkyl group, C 6 ~C 10 Aryl group, or C 5 ~C 10 The method, wherein the group is a heteroaryl group.

14. The method according to claim 13, wherein the compound is L and a syn isomer of formula (I).

15. The method according to claim 13, wherein the compound is L and an antiisomer of formula (I).

16. The method according to claim 13, wherein the compound is D and an antiisomer of formula (I).

17. The method according to claim 13, wherein the compound is D and a syn isomer of formula (I).

18. The method according to claim 13, wherein the compound has formula (II) or a pharmaceutically acceptable salt thereof. 【Transformation 5】

19. The method according to claim 13, wherein the compound comprises formula (III) to (VI) or a pharmaceutically acceptable salt thereof. 【Transformation 6】

20. The method according to claim 13, wherein the brain disease includes glioblastoma, medulloblastoma, glioma, or metastatic brain disease.

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