Amino acid derivatives and processes for their preparation

Abstract

The present disclosure relates to a non-natural amino acid and a method of making the same.

Description

[0001] Cross-references to related applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 551,273, filed February 8, 2024, the entire contents of which are incorporated herein by reference. Background Technology

[0002] Antibody-drug conjugates (ADCs) have applications in many areas of medical and biological research, while antibody-drug conjugates (ADCs) can be used in novel therapies for cancer treatment or other therapies. Conjugation with antibodies can result in random conjugations with multiple amino acid side chains, leading to a heterogeneous mixture of conjugated antibodies. In contrast, site-specific conjugation with antibodies produces a homogeneous population of antibody-drug conjugates with consistent and / or improved pharmacological properties compared to randomly conjugated ADCs. Therefore, new methods for preparing site-specific antibody-drug conjugates remain to be developed. Summary of the Invention

[0003] This disclosure provides non-natural amino acid analogs of formula (I) as described herein, or their enantiomers, diastereomers, or pharmaceutically acceptable salts. A mutant protein is also provided comprising one or more compounds of formula (I) as non-natural amino acids as described herein.

[0004] In one respect, this paper provides a compound of formula (I): Or its enantiomers, diastereomers or pharmaceutically acceptable salts, wherein: R 1 H, C1-C6 alkyl, C3-C 10 Cycloalkyl or benzyl, wherein each C1-C6 alkyl group, C3-C6 alkyl group, and C3-C6 alkyl group are respectively cycloalkyl or benzyl. 10 Cycloalkyl and benzyl groups are optionally surrounded by one or two R groups. 9 replace; It is a 4-, 5-, 6-, or 7-membered ring, composed of C4-C7 cycloalkyl and heteroalkyl groups, wherein the heteroalkyl group has 4-6 carbon atoms and one or two selected from -NH-, -NR- 2 Composed of heteroatoms or groups of -, -O-, and -S-, wherein the 4-, 5-, 6-, or 7-membered ring is optionally surrounded by one or two R- atoms. 9 replace; R 2 It is a C1-C6 alkyl, C1-C6 haloalkyl, or -C1-C6 alkoxy group. The linker is #-(L 3 ) o -(L 2 ) p -(L 1 )q -; # indicates AND The connection; Each L 1 L 2 and L 3 Independently, it can be a bond, -O-, -NH-, C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C1-C6 heteroalkylene, C2-C6 alkenyl, C2-C6 alkyneyl, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C 10 Cycloalkylene, 3- to 10-membered heterocycloalkylene, C6-C 10 aryl or 5- to 10-membered heteroarylene, wherein each of the following is a C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C1-C6 heteroalkylene, C2-C6 alkenyl, C2-C6 alkyneyl, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C 10 Cycloalkylene, 3- to 10-membered heterocycloalkylene, C6-C 10 Aryl and 5- to 10-membered heteroaryl groups are optionally bounded by 1, 2, or 3 R groups. 9 replace; Each o and p is an independent integer between 0 and 6; q is an integer from 1 to 6; and R 9 Independently, it is a halogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, C1-C4 heteroalkyl, -C(O)H, -C(O)OH, -CN, C3-C 10 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C6-C 10aryl, 5- to 10-membered heteroaryl, -C(O)(C1-C4 alkyl), -C(O)O(C1-C4 alkyl), -C(O)NH2, -C(O)NH(C1-C4 alkyl), -C(O)N(C1-C4 alkyl)2, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, -NH(C2-C4 alkylene)-OH, -NH(C2-C4 alkylene)-O-(C1-C4 alkyl), -OH, -O(C 1-C4 alkyl), -O(C1-C4 haloalkyl), -O(C2-C4 alkylene)-NH2, -O(C2-C4 alkylene)-NH-(C1-C4 alkyl), -O(C2-C4 alkylene)-N-(C1-C4 alkyl)2, -O(C1-C4 alkylene)-C(O)OH, -O(C1-C4 alkylene)-C(O)O-(C1-C4 alkyl), -O(C2-C4 alkenyl), -O(C1-C4 alkylene)-(C6-C 10 aryl), -O(C1-C4 alkylene)-(5- to 10-heteroaryl), - O(C6-C 10 Aryl), - SH, S(O)2OH, -S(O)2 (C1-C4 alkyl), -S(O)2NH2, -S(O)2NH (C1-C4 alkyl) or -S(O)2N (C1-C4 alkyl)2; or two R 9 Together with the atoms they are attached to, they form a C3-C 10 Cycloalkyl or 3- to 10-membered heterocyclic alkyl rings.

[0005] In another aspect, this document provides a mutant protein comprising one or more non-natural amino acids, each of which is independently a compound disclosed herein.

[0006] In another aspect, this document provides the use of the compounds disclosed herein in the manufacture of a mutant protein comprising one or more non-natural amino acids, wherein at least one of the one or more non-natural amino acids is the compound.

[0007] Other aspects and advantages of this disclosure will readily become apparent to those skilled in the art from the following detailed description, in which only exemplary embodiments of the disclosure are shown and described. As will be appreciated, the disclosure may have other and different embodiments, and certain details thereof may be modified in various obvious respects without departing from the disclosure. Therefore, the drawings and descriptions should be considered illustrative in nature and not restrictive.

[0008] Incorporation All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference to the extent that each individual publication, patent, or patent application is expressly and individually indicated to be incorporated by reference. If any publication, patent, or patent application incorporated by reference contradicts the disclosure contained in this specification, this specification is intended to supersede and / or give precedence to any such contradictory content. Detailed Implementation

[0009] While various embodiments of this disclosure have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Many variations, modifications, and substitutions will occur to those skilled in the art without departing from this disclosure. It should be understood that various alternatives to the embodiments of this disclosure described herein may be employed.

[0010] Compounds are generally described herein using standard nomenclature. For compounds with asymmetric centers, it should be understood that all optical isomers and mixtures thereof are included (unless otherwise stated). Furthermore, compounds containing carbon-carbon double bonds may exist in both Z- and E-forms, and unless otherwise stated, this invention covers all isomeric forms of such compounds. When a compound exists in multiple tautomeric forms, the listed compounds are not limited to any particular tautomer, but are intended to cover all tautomeric forms.

[0011] definition As used herein, unless the context clearly indicates otherwise, the singular forms “a” and “the” include plural indicators.

[0012] When a numerical range is provided, it should be understood that every intermediate value between the upper and lower limits of the range, as well as any other specified value or intermediate value within the specified range, is included within the scope of this disclosure. When a specified range includes an upper or lower limit, the range excluding either of the included limits is also included in this disclosure. When the range is used herein for physical properties (such as molecular weight) or chemical properties (such as chemical formula), it is intended to include all combinations and sub-combinations of the range and the specific embodiments therein.

[0013] As used herein, the terms “about” or “nearly” when referring to a numerical value or range indicate that the value or range referred to is typically within + / - 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the stated value or range.

[0014] As used herein, the term “comprising” (and related terms such as “comprise”, “comprises”, “having”, or “including”) is not intended to exclude embodiments of, for example, any material composition, composition, method, or process described herein.

[0015] As used in this specification and the appended claims, unless otherwise stated to the contrary, the following terms shall have the meanings specified below.

[0016] As used herein, the term "C1-C6 alkyl" generally refers to a straight-chain or branched hydrocarbon chain having 1 to 6 carbon atoms, which is connected to the rest of the molecule by a single bond. Similarly, an alkyl group containing up to 3 carbon atoms is a C1-C3 alkyl group, and an alkyl group containing up to 4 carbon atoms is a C1-C4 alkyl group. Examples of C1-C6 alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3-dimethylbutyl, and 2-ethylbutyl. In some cases, the substituents of the alkyl group will be specifically specified. For example, "cyanoalkyl" refers to an alkyl group that is substituted by at least one cyano substituent.

[0017] C1-C6 alkyl groups may optionally be substituted with C1-C3 alkoxy groups. Examples include, but are not limited to: methoxyethyl, methoxypropyl, methoxyisopropyl, ethoxyethyl, ethoxypropyl, ethoxyisopropyl, propoxyethyl, propoxypropyl, and propoxyisopropyl.

[0018] The C1-C6 alkyl groups may optionally be replaced by C3-C6 cycloalkyl groups. Examples include, but are not limited to, 1-methylcyclopropyl, 1-methylcyclobutyl, and 1-methylcyclohexyl.

[0019] As used herein, the term "C1-C6 alkoxy" generally refers to a group of the formula -OR, where R is a C1-C6 alkyl group as defined above. Similarly, an alkoxy group containing up to 3 carbon atoms is a C1-C3 alkoxy group. Examples include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy, isopentoxy, 2-methylbutoxy, neopentoxy, 1-ethylpropoxy, n-hexyloxy, isohexyloxy, 4-methylpentoxy, 3-methylpentoxy, 2-methylpentoxy, 1-methylpentoxy, 3,3-dimethylbutoxy, 2,2-dimethylbutoxy, 1,1-dimethylbutoxy, 1,2-dimethylbutoxy, 1,3-dimethylbutoxy, 2,3-dimethylbutoxy, and 2-ethylbutoxy.

[0020] The C1-C3 alkoxy groups may optionally be substituted with C1-C3 alkoxy groups. Examples include, but are not limited to: methoxymethoxy, methoxyethoxy, methoxypropoxy, methoxyisopropoxy, ethoxymethoxy, ethoxyethoxy, ethoxypropoxy, ethoxyisopropoxy, propoxymethoxy, propoxyethoxy, propoxypropoxy, and propoxyisopropoxy.

[0021] As used herein, the term "C3-C6 cycloalkylamino" refers to, for example, aziridine, pyrrolidinyl, piperidinyl, or hexamethyleneimino.

[0022] As used herein, the term “C3-C6 cycloalkyl” generally refers to a monocyclic nonaromatic group having 3 to 6 ring atoms, wherein each atom constituting the ring (i.e., the skeletal atom) is a carbon atom. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. As used herein, the term “cycloalkenyl” generally refers to a group comprising one or more unsaturated rings, wherein all ring members are carbon atoms. Polycyclic cycloalkyl or carbocyclic groups include, for example, adamantyl, norbornyl, decahydronaphthyl, bicyclo[3.3.0]octane, bicyclo[4.3.0]nonane, cis-decahydronaphthalene, trans-decahydronaphthalene, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, and bicyclo[3.3.2]decane, as well as 7,7-dimethyl-bicyclo[2.2.1]heptyl. Unless otherwise expressly specified in the instructions, cycloalkyl groups may optionally be substituted.

[0023] As used herein, the term "alkenyl" generally refers to a straight-chain or branched olefinic group containing at least one unsaturated carbon-carbon double bond. Alkenyl groups include C... 2-8 alkenyl, C 2-6 alkenyl and C 2-4Alkenyl groups, having 2 to 8, 2 to 6, or 2 to 4 carbon atoms, include, for example, vinyl, allyl, or isopropenyl. As used herein, the term "alkynyl" generally refers to a straight-chain or branched alkyne group having one or more unsaturated carbon-carbon bonds, at least one of which is a triple bond. Alkyne groups include C... 2-8 alkynyl group, C 2-6 alkynyl group and C 2-4 The alkynyl group has 2 to 8, 2 to 6, or 2 to 4 carbon atoms, respectively.

[0024] As used herein, the term "halogen" or "halide" generally refers to fluorine, chlorine, bromine, and iodine. The term "haloalkyl" as used herein generally refers to an alkyl group substituted with one or more independently selected halogens (e.g., a "C1-C6 haloalkyl" group having 1 to 6 carbon atoms and at least one halogen). Examples of haloalkyl groups include, but are not limited to: mono-, di-, or tri-fluoromethyl; mono-, di-, or tri-chloromethyl; mono-, di-, tri-, tetra-, or penta-fluoroethyl; mono-, di-, tri-, tetra-, or penta-chloroethyl; 2,2,2-trifluoroethyl; 1,2-difluoroethyl; 3-bromo-2-fluoropropyl; 1,2-dibromoethyl; and 1,2,2,2-tetrafluoro-l-trifluoromethyl-ethyl.

[0025] As used herein, the term "heteroalkyl" generally refers to an alkyl group in which one or more skeletal atoms are selected from atoms other than carbon, such as oxygen, nitrogen (e.g., -NH-, -N(alkyl)-), sulfur, or combinations thereof. In some cases, the heteroalkyl group is attached to the remainder of the molecule at one carbon atom of the heteroalkyl group. In one aspect, the heteroalkyl group is a C1-C6 heteroalkyl group, wherein the heteroalkyl group consists of 1 to 6 carbon atoms and one or more non-carbon atoms (e.g., oxygen, nitrogen, sulfur, or combinations thereof). In some cases, the carbon atoms or heteroatoms are optionally oxidized (e.g., -C(O)OCH2-, -CH2OCH2-, -CH2S(O)2NHCH2-, -NHC(O)NHCH2-, -CH2NHC(O)CH2-). Further examples of such heteroalkyl groups include, for example, -CH2OCH3, -CH2CH2OCH3, -CH2CH2OCH2CH2OCH3, or -CH(CH3)OCH3. Unless otherwise expressly stated in the specification, the heteroalkyl group is optionally substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc. In some embodiments, the heteroalkyl group is optionally substituted with oxo, halogen, methyl, ethyl, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, the heteroalkyl group is optionally substituted with oxo, halogen, methyl, ethyl, -CN, -CF3, -OH, or -OMe.

[0026] As used herein, the term "heteroaryl" generally refers to a monocyclic aryl group comprising one or more cyclic heteroatoms selected from nitrogen, oxygen, and sulfur. Examples include, but are not limited to: pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furanyl, thiopheneyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrroleyl, pyridazinyl, triazinyl, oxadiazolyl, thiazolyl, and furazolyl.

[0027] As used herein, the terms “heterocyclic,” “heterocyclic,” “heterocyclic group,” “cycloheteroalkyl,” or “heterocyclic alkyl” generally refer to a ring structure (monocyclic or polycyclic) containing 3-12 ring atoms (3-12-membered heterocycles), 3-8 ring atoms (3-8-membered heterocycles or 3-8-membered cycloheteroalkyl groups), 3-6 ring atoms (3-6-membered heterocycles or 3-6-membered cycloheteroalkyl groups), or 5-6 ring atoms (5-6-membered heterocycles or 5-6-membered cycloheteroalkyl groups), wherein at least one ring atom is carbon, and at least one ring atom is a heteroatom selected from N, O, and S, or a heteroatom group selected from C(=O), S(=O), and S(=O)2. Heterocyclic groups can be aromatic or non-aromatic. Piperidine and oxetane are non-limiting examples of non-aromatic heterocycles. Thiazoles and pyridines are non-limiting examples of aromatic heterocycles. Other examples of heterocycles include: aziridinyl, aziridine, oxadiazinyl, pyrrolidine, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, morpholinyl, piperazinyl, 4-piperidinoneyl, pyrrolidine, pyrazolidine, quininecycloyl, thiazolyl, thiomorpholinyl, tetrahydropyranyl, 1,1-dioxothiomorpholinyl, butyrolactam, valproic acid, caprolactam, butyrolactone, valproic acid, and caprolactone. Similarly, the term "cycloheterenyl" refers to a monocyclic or polycyclic ring structure comprising a carbon atom and a heteroatom / heteroatomic group, wherein the cycloheterenyl contains at least one C=C double bond, at least one carbon ring atom, and at least one ring atom that is a heteroatom or heteroatomic group, wherein the heteroatom is selected from N, O, and S, and the heteroatomic group is selected from C(=O), S(=O), and S(=O)2. Unless otherwise expressly stated in the specification, heterocyclic or heterocyclic alkyl groups are optionally substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocyclic alkyl, heteroaryl, etc. In some embodiments, heterocyclic alkyl groups are optionally substituted with oxo, halogen, methyl, ethyl, -CN, -CF3, -OH, -OMe, -NH2, or -NO2. In some embodiments, heterocyclic alkyl groups are optionally substituted with oxo, halogen, methyl, ethyl, -CN, -CF3, -OH, or -OMe.

[0028] As used herein, the term "aryl" generally refers to a group of 6 to 12 carbon atoms (C6, C6, C7, C8, C9 ... 6-12 aryl) or 6 to 10 carbon atoms (C6-10 Aryl (or aryl) is a monocyclic or polycyclic aromatic hydrocarbon group consisting of an all-carbon ring. Examples include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indenyl, biphenyl, and anthracene. The aryl group can be substituted or unsubstituted. Typical substituents include halogen, trihalomethyl, alkyl, hydroxyl, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, nitro, carbonyl, thiocarbonyl, C-carboxyl, O-carboxyl, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-amide, N-amide, sulfinyl, sulfonyl, amino, and -NR. X R Y , where R X and R Y The substituted alkyl group is independently selected from hydrogen, alkyl, cycloalkyl, aryl, carbonyl, acetyl, sulfonyl, trifluoromethanesulfonyl, and combinations of five- or six-membered heterocyclic rings. Exemplary substituted alkyl groups include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, aminomethyl, aminoethyl, hydroxymethyl, methoxymethyl, 2-fluoroethyl, and 2-methoxyethyl.

[0029] As used herein, the term "heteroaryl" generally refers to an aromatic group in which at least one aromatic ring contains at least one heteroatom selected from N, O, and S. Heteroaryls include, for example, 5-12-membered heteroaryls, 5-10-membered heteroaryls, 5-7-membered monocyclic structures, or 7-12-membered bicyclic structures. The number of heteroatoms in a heteroaryl group can be 1, 2, 3, 4, or more. Examples include, but are not limited to: thiophene, pyridinyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridine-2(1H)-keto, pyridin-4(1H)-keto, pyrroleyl, pyrazolyl, thiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-oxadiazolyl, imidazolyl, furanyl, tetrazolyl, isothiazolyl, oxazolyl, isoxazolyl, thiazolyl, oxadiazolyl, naphthyl, benzothiophene, indolyl, benzimidazolyl, benzothiazolyl, benzofuranyl, quinolinyl, isoquinolinyl, and quinazolinyl. The heteroaryl group may be substituted or unsubstituted. Typical substituents include halogenated, trihalomethyl, alkyl, hydroxyl, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, nitro, carbonyl, thiocarbonyl, C-carboxyl, O-carboxyl, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-amide, N-amide, sulfinyl, sulfonyl, amino, and -NR. X R Y , where R X and R Y As defined above.

[0030] As used herein, the term "amino" generally refers to the primary amino group (-NH2), the secondary amino group (-NH-), and the tertiary amino group (-NH-). ).

[0031] As used herein, the term "alkylamino" generally refers to substances having the general formula -NH-R. 1 or -N(R) 1 (R) 2 secondary or tertiary amines, wherein R 1 and R 2 Independently selected from alkyl, cycloalkyl, and (cycloalkyl)alkyl groups. Such alkylamino groups include, but are not limited to, mono- and di-(C)-amino groups. 1-6 alkyl)amino groups, wherein each C 1-6 Alkyl groups can be the same or different. In this case, the definition of "alkyl" as used in the term "alkylamino" differs from the definition of "alkyl" used for all other alkyl-containing groups in that it includes cycloalkyl and (cycloalkyl)alkyl groups.

[0032] As used herein, the term "alkylthio" generally refers to an alkyl-substituted thio group, where the term alkyl is as defined above.

[0033] As used herein, the terms “substituent” and “substituted” generally refer to a molecular moiety covalently bonded to an atom in the target molecule. For example, a cyclic substituent can be a moiety such as a halogen, alkyl group, haloalkyl group, or other group, covalently bonded to an atom that is a ring member (preferably a carbon or nitrogen atom). Substituents of aromatic groups are typically covalently bonded to a ring carbon atom. A linear substituent can be a moiety such as a halogen, alkyl group, haloalkyl group, or other group, covalently bonded to an atom that is a linear member (preferably a carbon or nitrogen atom).

[0034] As used in this article, the term "cycloalkylamine" generally refers to a ring structure in which an amino group is attached to a carbon atom, or a ring structure in which a nitrogen atom is a member of the ring.

[0035] As used herein, the term "C1-C3 alkylsulfonyl" generally refers to a sulfonyl group that is substituted with a C1-C3 alkyl group as defined above. Examples include, but are not limited to, methanesulfonyl, ethanesulfonyl, n-propanesulfonyl, and isopropanesulfonyl.

[0036] As used herein, the term "C1-C4 alkoxycarbonyl" generally refers to a carbonyl group that has been substituted with a C1-C4 alkoxy group as defined above. Examples include methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, and tert-butoxycarbonyl.

[0037] As used herein, the term “pharmaceutical acceptable” generally refers to a form of compound that is safe to administer to a target. For example, the free base, salt, solvate, hydrate, prodrug, or derivative forms of the compounds described herein (which have been approved by a competent authority or regulatory agency, such as the U.S. Food and Drug Administration (FDA), for use in mammals via oral ingestion or any other route of administration) are pharmaceutically acceptable.

[0038] Compounds of formulas (I), (II), (III), and (IV) include pharmaceutically acceptable salt forms of free base compounds. As used herein, the term "pharmaceutically acceptable salt" generally refers to a salt commonly used to form alkali metal salts or addition salts of free acids or free bases, which has been approved by regulatory agencies. Salts are formed by ion association, charge-charge interactions, covalent bonding, complexation, coordination, etc. The properties of salts are not strictly limited, as long as they are pharmaceutically acceptable.

[0039] As used herein, the term "pharmaceutically acceptable salt" refers to salts that, within reasonable medical judgment, are suitable for contact with the tissues of a subject without excessive toxicity, irritation, allergic reactions, etc., and whose benefits / risks are proportionate to a reasonable ratio. For example, Berge et al. describe pharmaceutically acceptable salts in detail in Pharmaceutical Sciences (1977) 66: 1-19. Pharmaceutically acceptable salts of the compounds presented herein include salts derived from suitable inorganic and organic acids and bases. Inorganic acids from which salts may be derived include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Organic acids from which salts may be derived include, but are not limited to, 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. Examples of pharmaceutically acceptable, non-toxic acid addition salts include: salts formed with an amino group and an inorganic acid such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, or perchloric acid, or with an organic acid such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or salts prepared by other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipates, alginates, ascorbic acid salts, aspartate salts, benzenesulfonate, besylate, benzoate, hydrogen sulfate, borates, butyrates, camphorates, camphor sulfonate, citrates, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucono-p-ethyl, glycerol phosphate, gluconate, hemisulfate, heptaate, and so on. Salts, hydroiodates, 2-hydroxyethanesulfonates, lacturonates, lactates, laurates, lauryl sulfates, malates, maleates, malonic acids, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, neopentanoates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, etc. In some embodiments, organic acids from which the salts can be derived include, for example, 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.

[0040] Pharmaceutically acceptable salts derived from suitable bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and other amine salts. Inorganic bases from which salts can be derived include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. Organic bases from which salts can be derived include, but are not limited to, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins; examples include, but are not limited to, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, pharmaceutically acceptable base addition salts are ammonium, potassium, sodium, calcium, or magnesium salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. Other pharmaceutically acceptable salts, where appropriate, include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxyl, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate ions. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, pharmaceutically acceptable base addition salts are selected from ammonium, potassium, sodium, calcium, and magnesium salts. Disalts (i.e., two counterions) and higher-order salts (e.g., three or more counterions) are both included within the meaning of pharmaceutically acceptable salts.

[0041] As used herein, the term "isomer" generally refers to different compounds having the same molecular formula, including any and all geometric isomers and stereoisomers. A "stereoisomer" is an isomer that differs only in the spatial arrangement of its atoms. For example, "isomers" include geometric double-bond cis- and trans-isomers, also known as E- and Z-isomers; R- and S-enantiomers; diastereomers, (d)-isomers and (l)-isomers, racemic mixtures thereof; and other mixtures thereof falling within the scope of this disclosure, unless otherwise stated. As used herein, the term "tautomer" is a class of isomers comprising two or more interconvertible compounds produced by at least one formal migration of hydrogen atoms and at least one change in valence (e.g., from a single bond to a double bond, from a triple bond to a single bond, or vice versa).

[0042] In some embodiments, compounds of formulas (I), (II), (III), and (IV) are administered to a therapeutic subject as a pharmaceutical composition. For this purpose, in one embodiment, the compound is combined with one or more pharmaceutically acceptable excipients, including carriers, diluents, or adjuvants, to form a suitable composition, which is described in more detail herein.

[0043] As used herein, the term “excipient” generally refers to any pharmaceutically acceptable additive, carrier, excipient or other suitable ingredient other than the active pharmaceutical ingredient (API), which is typically included for formulation and / or administration purposes.

[0044] As used herein, the term "diluent" generally refers to a reagent used as a filler to achieve the desired volume or weight of the composition. The diluent may be present in the particles of the pharmaceutical composition in the form of a single compound or a mixture of compounds. Non-limiting examples of diluents include lactose, starch, pregelatinized starch, microcrystalline cellulose, silicified microcrystalline cellulose, cellulose acetate, glucose, mannitol, sodium phosphate, potassium phosphate, calcium phosphate, fructose, maltose, sorbitol, or sucrose.

[0045] As used herein, the term "excipient" generally refers to any substance or mixture of substances that, when used in conjunction with the compounds disclosed herein, enhances the efficacy or potency of the compounds disclosed herein on their target. However, when the excipient is used alone, no pharmacological effect on the same target has been observed.

[0046] Non-natural amino acids As used herein, the term "unnatural amino acid" generally refers to an amino acid that is not one of the 20 common amino acids or of pyrrolidone or selenocysteine. Other terms that may be used synonymously with "unnatural amino acid" include "non-naturally encoded amino acid," "non-naturally occurring amino acid," and their various hyphenated and unhyphenated variations.

[0047] Compared to antibodies made from 20 naturally occurring amino acids, gene-incorporated non-natural amino acids (uAAs) enable unique orthogonal coupling strategies to yield antibody-drug conjugates (ADCs). Therefore, uAAs offer a new paradigm for the production of next-generation ADCs. The incorporation of uAAs allows for unique orthogonal biological recognition of modified antibodies via other proteins (e.g., enzymes). This recognition can trigger selective biological responses targeting antibodies containing specific uAAs.

[0048] Engineered orthogonal aaRS-tRNA synthetases (aaRS) and their respective homologous transfer RNA (tRNA) pairs can be used for site-specific incorporation of these uAAs by expanding the genetic code to encode the uAAs disclosed herein. For example, the engineered orthogonal aaRS / tRNA pair can suppress a reused nonsense codon to directly incorporate these uAAs into specific sites of desired proteins. The novel uAAs disclosed herein, with defined chemical functional groups (i.e., carbonyl groups) for coupling with other molecules, are ideal for mutant proteins. Using these defined chemical functional groups, mutant proteins containing these uAAs can be coupled with other proteins, cytotoxic agents, polymers, nucleic acids, etc. For example, the amber stop codon (UAG) can be used for the incorporation of these uAAs. The unique aaRS / tRNA pairs that serve as mechanisms for introducing these uAAs can be developed in a variety of hosts, including Escherichia coli, yeast, insect cells, and mammalian cells.

[0049] Implementation plan with numbering The following embodiments describe non-limiting arrangements of combinations of features disclosed herein. Other arrangements of combinations of features are also considered. In particular, each of these numbered embodiments is considered to be subordinate to or relating to each numbered embodiment that precedes or follows it, regardless of the order in which they are listed.

[0050] Implementation Scheme 1. A compound of formula (I): Or its enantiomers, diastereomers or pharmaceutically acceptable salts, wherein: R 1 H, C1-C6 alkyl, C3-C 10 Cycloalkyl or benzyl, wherein each C1-C6 alkyl group, C3-C6 alkyl group, and C3-C6 alkyl group are respectively cycloalkyl or benzyl. 10 Cycloalkyl and benzyl groups are optionally surrounded by one or two R groups. 9 replace; It is a 4-, 5-, 6-, or 7-membered ring, composed of C4-C7 cycloalkyl and heteroalkyl groups, wherein the heteroalkyl group has 4-6 carbon atoms and one or two selected from -NH-, -NR- 2 Composed of heteroatoms or groups of -, -O-, and -S-, wherein the 4-, 5-, 6-, or 7-membered ring is optionally surrounded by one or two R- atoms. 9 replace; R 2 It is a C1-C6 alkyl, C1-C6 haloalkyl, or -C1-C6 alkoxy group. The linker is #-(L 3 ) o -(L 2 ) p-(L 1 ) q -; # indicates AND The connection; Each L 1 L 2 and L 3 Independently, it can be a bond, -O-, -NH-, C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C1-C6 heteroalkylene, C2-C6 alkenyl, C2-C6 alkyneyl, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C 10 Cycloalkylene, 3- to 10-membered heterocycloalkylene, C6-C 10 aryl or 5- to 10-membered heteroarylene, wherein each of the following is a C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C1-C6 heteroalkylene, C2-C6 alkenyl, C2-C6 alkyneyl, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C 10 Cycloalkylene, 3- to 10-membered heterocycloalkylene, C6-C 10 Aryl and 5- to 10-membered heteroaryl groups are optionally bounded by 1, 2, or 3 R groups. 9 replace; Each o and p is an independent integer between 0 and 6; q is an integer from 1 to 6; and R 9 Independently, it is a halogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, C1-C4 heteroalkyl, -C(O)H, -C(O)OH, -CN, C3-C 10 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C6-C 10aryl, 5- to 10-membered heteroaryl, -C(O)(C1-C4 alkyl), -C(O)O(C1-C4 alkyl), -C(O)NH2, -C(O)NH(C1-C4 alkyl), -C(O)N(C1-C4 alkyl)2, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, -NH(C2-C4 alkylene)-OH, -NH(C2-C4 alkylene)-O-(C1-C4 alkyl), -OH, -O(C 1-C4 alkyl), -O(C1-C4 haloalkyl), -O(C2-C4 alkylene)-NH2, -O(C2-C4 alkylene)-NH-(C1-C4 alkyl), -O(C2-C4 alkylene)-N-(C1-C4 alkyl)2, -O(C1-C4 alkylene)-C(O)OH, -O(C1-C4 alkylene)-C(O)O-(C1-C4 alkyl), -O(C2-C4 alkenyl), -O(C1-C4 alkylene)-(C6-C 10 aryl), -O(C1-C4 alkylene)-(5- to 10-heteroaryl), - O(C6-C 10 Aryl), - SH, S(O)2OH, -S(O)2 (C1-C4 alkyl), -S(O)2NH2, -S(O)2NH (C1-C4 alkyl) or -S(O)2N (C1-C4 alkyl)2; or two R 9 Together with the atoms they are attached to, they form a C3-C 10 Cycloalkyl or 3- to 10-membered heterocyclic alkyl rings.

[0051] Implementation Scheme 2. The compound according to Implementation Scheme 1, wherein the compound is of formula (II): Or its enantiomers, diastereomers or pharmaceutically acceptable salts.

[0052] Implementation Scheme 3. The compound according to Implementation Scheme 1, wherein the compound is of formula (III): Or its enantiomers, diastereomers or pharmaceutically acceptable salts.

[0053] Implementation Scheme 4. The compound according to Implementation Scheme 1, wherein the compound is of formula (IV): Or its enantiomers, diastereomers or pharmaceutically acceptable salts.

[0054] Implementation Scheme 5. The compound according to any one of Implementation Schemes 1-4, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: R 1 For H.

[0055] Implementation Scheme 6. The compound according to any one of Implementation Schemes 1-4, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: R 1 It is a C1-C6 alkyl group.

[0056] Implementation Scheme 7. The compound according to any one of Implementation Schemes 1-6, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: It is a C4-C7 cycloalkyl group, wherein the C4-C7 cycloalkyl group is optionally surrounded by one or two R groups. 9 replace.

[0057] Implementation Scheme 8. A compound according to any one of Implementation Schemes 1-6, or an enantiomer, diastereomer, or pharmaceutically acceptable salt thereof, wherein: It is a heteroalkyl group, consisting of 3-6 carbon atoms and one or two atoms selected from -NH-, -NR-. 2 Composed of heteroatoms or groups of -, -O-, and -S-, wherein the heteroalkyl group is optionally composed of one or two R- atoms. 9 replace.

[0058] Implementation Scheme 9. A compound according to any one of Implementation Schemes 1-6, or an enantiomer, diastereomer, or pharmaceutically acceptable salt thereof, wherein for: , , , , , , , , , , , , , , , , , , , , , , , , , or , Each X and Y is independently -NH- and -NR. 2 -、-O- or -S-.

[0059] Implementation Scheme 10. A compound according to any one of Implementation Schemes 1-6, or an enantiomer, diastereomer, or pharmaceutically acceptable salt thereof, wherein for: , , , , , , , , , , or , Where X is -NH- or -NR 2 -、-O- or -S-.

[0060] Implementation Scheme 11. A compound according to any one of Implementation Schemes 1-6, or an enantiomer, diastereomer, or pharmaceutically acceptable salt thereof, wherein for: , , , , , , , , , , or , Where X is -O-.

[0061] Implementation Scheme 12. The compound according to any one of Implementation Schemes 1-11, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: The linker is #-(L 3 ) o -(L 2 ) p -(L 1 ) q -; # indicates AND The connection; Each L 1 L 2 and L 3Independently, it is a bond, -O-, -NH-, C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C1-C6 heteroalkylene, alkoxycarbonyl, C3-C 10 Cycloalkylene or 3- to 10-membered heterocyclic alkylene, wherein each C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C1-C6 heteroalkylene, alkoxycarbonyl, C3-C 10 Cycloalkylene or 3- to 10-membered heterocyclic alkylene, optionally surrounded by 1, 2, or 3 Rs. 9 replace; Each o and p is an independent integer from 0 to 6; and q is an integer from 1 to 6.

[0062] Implementation Scheme 13. The compound according to any one of Implementation Schemes 1-11, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: The linker is methylene, -CF2-, -CH(CH3)-, ethylene, #-CH(CH3)-CH2-, #-CH2-CH(CH3)-, n-propylene, -CH2-CH(CH3)-CH2-, #-CH(CH3)-CH2-CH2-, #-CH2-CH2-CH(CH3)-, n-butylene, n-pentylene, -CH2-O-CH2-, #-CH2-O-CH2-CH2-, #-CH2-CH2-O-CH2-, -CH2-O-CH2-O-CH2-, or its enantiomers or diastereomers; and # indicates AND The connection.

[0063] Implementation Scheme 14. The compound according to any one of Implementation Schemes 1-11, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: The linker group is methylene, -CH(CH3)-, ethylene, #-CH(CH3)-CH2-, #-CH2-CH(CH3)-, n-propylene, -CH2-CH(CH3)-CH2-, #-CH(CH3)-CH2-CH2-, #-CH2-CH2-CH(CH3)-, n-butylene, n-pentylene, -CH2-O-CH2-, #-CH2-O-CH2-CH2-, #-CH2-CH2-O-CH2-, or their enantiomers or diastereomers; and # indicates AND The connection.

[0064] Implementation Scheme 15. The compound according to any one of Implementation Schemes 1-11, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: The linker is n-butylene, #-CH2-O-CH2-CH2-, #-CH2-CH2-O-CH2-, or its enantiomers or diastereomers; and # indicates AND The connection.

[0065] Implementation Scheme 16. The compound according to Implementation Scheme 1, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein the compound is: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

[0066] Implementation Scheme 17. The compound or a pharmaceutically acceptable salt thereof according to Implementation Scheme 1, wherein the compound is: , , , , , , , , , , , , , , or .

[0067] Implementation Scheme 18. The compound or a pharmaceutically acceptable salt thereof according to Implementation Scheme 1, wherein the compound is: , , , , , , , , , , , , , , or .

[0068] Implementation Scheme 19. The compound or a pharmaceutically acceptable salt thereof according to Implementation Scheme 1, wherein the compound is: , , , , , , , , , , , , , , or .

[0069] Implementation Scheme 20. The compound or a pharmaceutically acceptable salt thereof according to Implementation Scheme 1, wherein the compound is: , , , , , , , , , , , , , , or .

[0070] Implementation Scheme 21. The compound or a pharmaceutically acceptable salt thereof according to Implementation Scheme 1, wherein the compound is: , , , , , , , , , , , , , , or .

[0071] Implementation Scheme 22. The compound or a pharmaceutically acceptable salt thereof according to Implementation Scheme 1, wherein the compound is: , , , , , , , , , , , , , , or .

[0072] Implementation Scheme 23. A compound selected from the following: , , , , and , Or its enantiomers, diastereomers or pharmaceutically acceptable salts.

[0073] Implementation Scheme 24. The compound according to Implementation Scheme 23, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein the compound is: , , or .

[0074] Implementation Scheme 25. The compound or a pharmaceutically acceptable salt thereof according to Implementation Scheme 23, wherein the compound is: , , or .

[0075] Implementation Scheme 26. The compound or a pharmaceutically acceptable salt thereof according to Implementation Scheme 23, wherein the compound is: , or .

[0076] Implementation Scheme 27. A mutant protein comprising one or more non-natural amino acids, wherein each of the one or more non-natural amino acids is independently a compound according to any one of Implementation Schemes 1-26.

[0077] Implementation Scheme 28. The mutant protein according to Implementation Scheme 27, wherein the mutant protein comprises a compound according to any one of Implementation Schemes 1-26.

[0078] Implementation Scheme 29. The mutant protein according to Implementation Scheme 27, wherein the mutant protein comprises two or more non-natural amino acids, and wherein each of the two or more non-natural amino acids is the same compound according to any one of Implementation Schemes 1-26.

[0079] Implementation Scheme 30. Use of the compound according to any one of embodiments 1-26 in the manufacture of a mutant protein comprising one or more non-natural amino acids, wherein at least one of the one or more non-natural amino acids is a compound according to any one of embodiments 1-26.

[0080] Implementation Scheme 31. The use according to Implementation Scheme 30, wherein at least two of the one or more non-natural amino acids are compounds according to any one of Implementation Schemes 1-26.

[0081] Implementation Scheme 32. A mutant protein comprising one or more compounds as non-natural amino acids according to any one of Implementation Schemes 1-26.

[0082] Implementation Scheme 33. The mutant protein according to Implementation Scheme 32 further comprises one or more portions selected from proteins, antibodies, cytotoxic agents, drugs, and polymers, wherein said one or more portions are coupled to the non-natural amino acid.

[0083] Implementation Scheme 34. The mutant protein according to Implementation Scheme 33, wherein the polymer comprises a polyethylene glycol portion.

[0084] Implementation Scheme 35. An antibody comprising, in each heavy chain and / or light chain, one or more compounds as non-natural amino acids according to any one of Implementation Schemes 1-26.

[0085] Implementation Scheme 36. The antibody according to Implementation Scheme 35, wherein the Fc region comprises one or more of the compounds.

[0086] Example Chemical synthesis The methods of the present invention may include the use of at least one compound of formulas (I), (II), (III), and (IV) that inhibits programmed necrosis in regulating the repair and / or functional performance of various cells, tissues, and organs, and have therapeutic and cosmetic applications, encompassing the regulation of nervous tissue, bone and cartilage formation and repair, regulation of spermatogenesis, regulation of smooth muscle, regulation of the lungs, liver, and other organs derived from the primitive intestine, regulation of hematopoietic function, regulation of skin and hair growth, etc. Therefore, the methods and compositions of the present invention include using the aforementioned inhibitors in all such applications, wherever an inhibitor of programmed necrosis may be involved. Furthermore, the aforementioned methods may be performed on cells provided in a culture medium (in vitro) or on cells of a whole animal (in vivo).

[0087] The examples and preparation instructions provided below illustrate the compounds described herein and methods for preparing such compounds. Generally, the compounds described herein can be prepared using methods known in the general field of chemistry.

[0088] The compounds of the present invention can be prepared from commercially available materials using a variety of synthetic routes (including those described below). The starting materials of the present invention are known, commercially available, or can be synthesized by reference to or according to methods known in the art. Many starting materials can be prepared according to known methods, particularly those described in the examples. In some cases, when synthesizing starting materials, functional groups are protected with suitable protecting groups, if necessary. Functional groups can be removed according to steps known in the art.

[0089] Protection of functional groups by protecting groups, the protecting groups themselves, and their removal reactions (often referred to as "deprotection") are described in, for example, standard reference works such as J.F.W. McOmie, Protective Groups in Organic Chemistry, Plenum Press, London and New York (1973), in T.W. Greene, Protective Groups in Organic Synthesis, Wiley, New York (1981), in The Peptides, Vol. 3, edited by E. Gross and J. Meienhofer, Academic Press, London and New York (1981).

[0090] All synthetic steps described herein can be carried out under known reaction conditions, preferably under the conditions described herein, or in the absence or presence of (typically) solvents or diluents.

[0091] This invention also covers "intermediate" compounds, including structures produced by the synthetic steps prior to obtaining the final desired compound, regardless of whether they have been isolated. Structures obtained from transient starting materials through the steps, structures obtained at any stage deviating from the method, and structures that form the starting materials under the reaction conditions are all "intermediates" covered by this invention. Furthermore, structures produced from starting materials using reactive derivatives or salts, structures produced from compounds obtainable by the method described in this invention, and structures obtained by in-situ treatment of the compounds of this invention are also within the scope of this invention.

[0092] Novel starting materials and / or intermediates, as well as methods for their preparation, are also the subject of this invention. In selected embodiments, such starting materials are used and reaction conditions are chosen to obtain the desired compound.

[0093] The starting materials of this invention are known, commercially available, or synthesized by reference to or according to methods known in the art. Many starting materials can be prepared according to known methods, and in particular, by the methods described in the examples. In some cases, when synthesizing the starting materials, the functional groups are protected with suitable protecting groups, if necessary. Protecting groups, their introduction, and removal are described above.

[0094] Unless otherwise stated, all reagents and solvents are commercially available. Unless otherwise stated, all commercially available reagents and solvents are used without purification. If necessary, some reagents and solvents are purified using standard techniques. For example, tetrahydrofuran can be purified by sodium distillation.

[0095] The reaction progress was monitored by reversed-phase HPLC and / or thin-layer chromatography (TLC). Liquid chromatography-mass spectrometry (LC-MS) was performed using a Waters or Shimadzu 2010EV LCMS instrument with water and acetonitrile or methanol doped with 0.1% formic acid. TLC was performed on 0.25 mm plates pre-coated with silica gel 60 F254. Rapid chromatography was performed using silica gel (particle size 32-63 μm). Chromatographic purification was performed using either silica gel (m), activated basic alumina (approximately 150 mesh), or alumina (activated, basic, approximately 150 mesh). Automated chromatographic purification was performed using pre-packed silica gel or C18 columns (from RediSep and Luknova) and eluted using an ISCO Companion system. Reversed-phase purification was performed using water and acetonitrile or methanol doped with 0.1% formic acid. All final product compounds were purified using one of these two chromatographic methods. The purity and characterization of the compounds were determined by combined TLC, LC-MS, and NMR analysis. 1 H and 13 CNMR spectra were obtained at 400 MHz and 101 MHz on a Joel 400 spectrometer. Chemical shifts are reported as δ (ppm), with the deuterated solvent signal as an internal reference.

[0096] The size and scale of the synthetic method will vary depending on the amount of the desired end product. It should be understood that although specific reactants and amounts are provided in the examples, those skilled in the art will recognize other alternative and equally feasible combinations of reactants that will yield the same compound. Therefore, when using general oxidizing agents, reducing agents, and solvents of various properties (aprotic, nonpolar, polar, etc.), equivalents will be known in the art and considered herein for use in this method.

[0097] The following steps describe various post-treatments following reaction termination. One post-treatment typically involves quenching the reaction to terminate any residual catalytic activity and starting reagents. An organic solvent is then typically added, and the aqueous layer is separated from the organic layer. The product is usually obtained from the organic layer, while unused reactants and other byproducts and unwanted chemicals are typically retained in the aqueous layer and discarded. Post-treatments in standard organic synthesis steps seen in the literature typically involve drying the product by exposure to a desiccant such as anhydrous Na₂SO₄ to remove any residual excess water or aqueous byproducts partially dissolved in the organic layer and to concentrate the remaining organic layer. Concentration of the product dissolved in the solvent can be achieved by any known method, such as pressure evaporation, temperature-pressure evaporation, etc. Such concentration can be achieved using standard laboratory equipment, such as rotary evaporator distillation, etc. Optionally, one or more purification steps are then performed, which may include, but are not limited to: rapid column chromatography, filtration through various media, and / or other preparative methods known in the art, and / or crystallization / recrystallization. (See, for example, Addison Ault, “Techniques and Experiments for Organic Chemistry”, 6th ed., University Science Books, Sausalito, Calif., 1998, edited by Ann B. McGuire, pp. 45–59).

[0098] General synthetic route There are many routes available for synthesizing compounds of formulas (I), (II), (III), and (IV). The following are synthetic routes for obtaining specific compounds of formulas (I), (II), (III), and (IV). The synthetic routes shown are not limiting but are examples among many feasible routes.

[0099] In one embodiment of obtaining compound (I), as shown in Scheme 1, the ketal group of compound A-1 is protected as a ketal under acid catalysis to give ester compound A-2 without affecting the stereochemistry at the α-carbon. The ester of compound A-2 is hydrolyzed under basic conditions to give the carboxylic acid in compound A-3. The unmasked carboxylic acid is converted to an activated ester compound A-4, which reacts with amine compound A-4A to give amide compound A-5. Ring A in compound A-5 is as defined in formula (I). The ketal protecting group on compound A-5 is hydrolyzed (treated with TsOH in acetone) to demask the cyclic ketone in compound A-6, and further deprotected using TFA to remove the tert-butyloxycarbonyl (Boc) group to give compound A-7 of formula (I). Alternatively, the free carboxylic acid in compound A-6 reacts with alcohol compound A-6A (R 1The reaction of OH (e.g., azirmonotriazole tetramethyluranium hexafluorophosphate (HATU) or diisopropylethylamine (DIEA) in acetonitrile) yields α-amino acid ester compound A-8. The tert-butoxycarbonyl (Boc) group on compound A-8 is removed using TFA to give compound A-9 of formula (I).

[0100] Scheme 1: Synthesis of compounds A-7 and A-9 The coupling reaction between compound A-6 and compound A-6A can be carried out under different conditions, including but not limited to: (1) HATU, DIEA, dimethylformamide (DMF), at room temperature; (2) benzotriazole tetramethylureonium hexafluorophosphate (HBTU), DIEA, DMF, at room temperature; (3) 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), hydroxybenzotriazole (HOBt), DIEA, DMF, at room temperature; (4) benzotriazole-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP), TEA, DMF, at room temperature; and (5) benzotriazole-1-yloxytripyrrolidinylphosphonium hexafluorophosphate (PyBOP), TEA, DMF, at room temperature. Other conditions may also be used.

[0101] As shown in Scheme 1, the chirality of the amino acid moiety can be controlled by the chirality of the amine compound A-4A. The chirality of ring A can be controlled by the chirality of compound A-1. There are multiple synthetic methods for chiral α-amino acids. See (1) Y. Zou et al., Asymmetric Synthesis of Tailor-Made Amino Acids Using Chiral Ni(II) Complexes of Schiff Bases. An Update of the Recent Literature. Molecules , 2020; 25(12):2739; (2) AS Saghyan and P. Langer, Asymmetric Synthesis of Non-proteinogenic Amino Acids (Wiley-VCH, 2016); (3) C. Nájera and JM Sansano, Catalytic asymmetric synthesis of α-amino acids. Chem. Rev.2007; 107, 4584-4671; (3) Y.-P. Xue et al., Enzymatic Asymmetric Synthesis of Chiral Amino Acids. Chem. Soc. Rev. , 2018, 47: 1516-1561. Racemic amino acid products can be resolved by: (1) forming diastereomer salts with pure chiral acids or bases; or (2) enzymatic identification using enzymes such as aminoacylases during amide hydrolysis. Some commonly used chiral resolving agents for amino acid resolution include, but are not limited to, tartaric acid, camphor sulfonic acid, strychnine, strychnine, and various synthetic chiral amines. In general, there are also many chemical or enzymatic methods or methods for synthesizing chiral compounds by resolving racemic products.

[0102] The synthetic route shown in Scheme 1 can be modified to synthesize the uAA compounds disclosed herein, particularly those uAAs shown in Schemes 16 to 26.

[0103] Compounds A-7 and A-9 can also be synthesized via other routes. For example, as shown in Scheme 2, the primary amine of compound A-20 is coupled with a carboxylic acid compound A-21 (e.g., tetramethyluranium hexafluorophosphate azirmonazine (HATU), diisopropylethylamine in acetonitrile (DIEA)) to give amide compound A-22. Ring A in compound A-4 is as defined in formula (I). Acidification (H3PO4, toluene) removes the tert-butyl ester, and hydrogenation (H2 via Pd / C) removes the Cbz protecting group from compound A-22 to give compound A-7. Alternatively, the tert-butyl ester in compound A-21 is selectively removed in the presence of the Boc protecting group, followed by unmasked carboxylic acid and R 1 The coupling reaction between OH (compound A-6A) and the hydrogenation reaction yielded α-amino acid ester compound A-9.

[0104] Option 2: Another synthesis of compounds A-7 and A-9 Similarly, other protecting groups, different from tert-butyl ester and Cbz group, can be used to react the primary amine in compound A-20 with compound A-21. The ketone group in compound A-21 can be protected as a ketal, similar to the strategy used in Scheme 1, and then the route in Scheme 2 is completed by adding several deprotection steps. See Scheme 3 below, for example.

[0105] Option 3: Another synthesis of compounds A-7 and A-9 Example 1: Synthesis of LUX 102-6 Option 4: Step 1: LUX 102-2 1,2-Ethylene glycol (1.2 equivalents) and TsOH (0.05 equivalents) were dissolved in benzene, and the solution was stirred under reflux using a Dean-Stark apparatus to remove H₂O from the reagents. After 1.5 h, LUX 102-1 (1 equivalent) was added, and stirring was continued under reflux for another 2.5 h. After cooling to room temperature, the solution was diluted with EtOAc, poured into a saturated NaHCO₃ solution, and the two phases were separated. The organic layer was washed with brine, dried over anhydrous Na₂SO₄, and filtered. The solvent was evaporated, and the residue was then purified by silica gel chromatography to obtain LUX 102-2.

[0106] Step 2: LUX 102-3 LUX 102-2 (1.0 equivalent) was stirred overnight in a mixture of THF / water (10:1, v / v) and NaOH (0.05 equivalent). The mixture was quenched with aqueous NH4Cl solution, the pH was adjusted to approximately 2 with hydrochloric acid (1 M), and the mixture was extracted with EtOAc (3×). The combined organic layers were washed with brine, dried over anhydrous MgSO4, filtered, and concentrated. The crude product was purified by silica gel chromatography to give product LUX 102-3.

[0107] Step 3: LUX 102-4 A solution of LUX 102-3 (1.0 equivalent), Et3N (1.1 equivalent), DMAP (0.1 equivalent), and N,N'-disuccinimidyl carbonate (DSC, 1.1 equivalent) in DCM was stirred at ambient temperature for more than two hours. Subsequently, the reaction mixture was mixed with EtOAc, hydrochloric acid (0.1 N), water, a saturated aqueous solution of NaHCO3, and brine. The mixture was dried over anhydrous MgSO4, filtered, concentrated, and purified by silica gel chromatography to obtain the product LUX 102-4.

[0108] Step 4: LUX 102-6 A mixture of LUX 102-4 (1.0 equivalent), LUX 102-5 (1.0 equivalent), and Et3N (1.1 equivalent) in DMF was stirred overnight. The mixture was concentrated. The residue was partitioned between EtOAc and hydrochloric acid (0.1 N). The organic layer was separated, washed with water, aqueous NaHCO3 solution, and brine, then dried, filtered, and concentrated. The crude product was purified by silica gel chromatography to obtain the desired LUX102-6.

[0109] Step 5: LUX 102-7 A mixture of LUX 102-6 (1.0 equivalent) and TsOH (0.05 equivalent) in acetone was stirred overnight. The mixture was concentrated. The residue was partitioned between EtOAc and an aqueous solution of NaHCO3. The organic layer was separated, washed with brine, dried, filtered, and concentrated. The crude product was purified by silica gel chromatography to give the desired LUX 102-7.

[0110] Step 6: LUX 102-8 The mixture of LUX 102-7 in TFA / DCM (1:1, v / v) was stirred for 4 hours. The mixture was concentrated. The residue was partitioned between EtOAc and water. The organic layer was separated, washed with aqueous NaHCO3 solution and brine, then dried, filtered, and concentrated. The crude product was purified by silica gel chromatography to obtain the desired LUX 102-8.

[0111] Other compounds of formula (I) can be prepared using a similar method. For example, according to scheme 6, another compound, LUX 103-8, was prepared. Analytical analysis / data confirmed the structure of LUX 102-8.

[0112] Option 5: The ketal group of LUX 103-1 is protected as a ketal under acid catalysis to give the ester LUX 103-2 without affecting the stereochemistry at the α-carbon. The ester of LUX 103-2 is hydrolyzed under basic conditions to give the carboxylic acid in LUX 103-3. The unmasked carboxylic acid is converted to the activated ester LUX 103-4, which reacts with the amine LUX 102-5 to give the amide LUX 103-6. The ketal protecting group on LUX 103-6 is hydrolyzed (treated with TsOH in acetone) to demask the cyclic ketone in LUX 103-7. Further deprotection is performed using TFA to remove the tert-butyloxycarbonyl (Boc) group, giving LUX 103-8 of formula (I). Analytical analysis / data confirm the structure of LUX 103-8.

[0113] While preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. The invention is not intended to be limited to the specific examples provided in the specification. Although the invention has been described with reference to the foregoing specification, the description and illustration of embodiments herein are not intended to be construed in a limiting sense. Many variations, alterations, and substitutions will now occur to those skilled in the art without departing from the invention. Furthermore, it should be understood that all aspects of the invention are not limited to the specific depictions, configurations, or relative proportions described herein, but depend on various conditions and variables. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in carrying out the invention. Therefore, the invention is also intended to cover any such alternatives, modifications, variations, or equivalents. The following claims are intended to define the scope of the invention and thereby cover the methods and structures within the scope of these claims and their equivalents.

Claims

1. A compound of formula (I): Or its enantiomers, diastereomers or pharmaceutically acceptable salts, wherein: R 1 H, C1-C6 alkyl, C3-C 10 Cycloalkyl or benzyl, wherein each C1-C6 alkyl group, C3-C6 alkyl group, and C3-C6 alkyl group are respectively cycloalkyl or benzyl. 10 Cycloalkyl and benzyl groups are optionally surrounded by one or two R groups. 9 replace; It is a 4-, 5-, 6-, or 7-membered ring, composed of C4-C7 cycloalkyl and heteroalkyl groups, wherein the heteroalkyl group has 4-6 carbon atoms and one or two selected from -NH-, -NR- 2 Composed of heteroatoms or groups of -, -O-, and -S-, wherein the 4-, 5-, 6-, or 7-membered ring is optionally surrounded by one or two R- atoms. 9 replace; R 2 It is a C1-C6 alkyl, C1-C6 haloalkyl, or -C1-C6 alkoxy group. The linker is #-(L 3 ) o -(L 2 ) p -(L 1 ) q -; # indicates AND The connection; Each L 1 L 2 and L 3 Independently, it can be a bond, -O-, -NH-, C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C1-C6 heteroalkylene, C2-C6 alkenyl, C2-C6 alkyneyl, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C 10 Cycloalkylene, 3- to 10-membered heterocycloalkylene, C6-C 10 aryl or 5- to 10-membered heteroarylene, wherein each of the following is a C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C1-C6 heteroalkylene, C2-C6 alkenyl, C2-C6 alkyneyl, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C 10 Cycloalkylene, 3- to 10-membered heterocycloalkylene, C6-C 10 Aryl and 5- to 10-membered heteroaryl groups are optionally bounded by 1, 2, or 3 R groups. 9 replace; Each o and p is an independent integer between 0 and 6; q is an integer from 1 to 6; and R 9 Independently, it is a halogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, C1-C4 heteroalkyl, -C(O)H, -C(O)OH, -CN, C3-C 10 Cycloalkyl, 3- to 10-membered heterocycloalkyl, C6-C 10 aryl, 5- to 10-membered heteroaryl, -C(O)(C1-C4 alkyl), -C(O)O(C1-C4 alkyl), -C(O)NH2, -C(O)NH(C1-C4 alkyl), -C(O)N(C1-C4 alkyl)2, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, -NH(C2-C4 alkylene)-OH, -NH(C2-C4 alkylene)-O-(C1-C4 alkyl), -OH, -O(C 1-C4 alkyl), -O(C1-C4 haloalkyl), -O(C2-C4 alkylene)-NH2, -O(C2-C4 alkylene)-NH-(C1-C4 alkyl), -O(C2-C4 alkylene)-N-(C1-C4 alkyl)2, -O(C1-C4 alkylene)-C(O)OH, -O(C1-C4 alkylene)-C(O)O-(C1-C4 alkyl), -O(C2-C4 alkenyl), -O(C1-C4 alkylene)-(C6-C 10 aryl), -O(C1-C4 alkylene)-(5- to 10-membered heteroaryl), - O(C6-C 10 Aryl), - SH, S(O)2OH, -S(O)2 (C1-C4 alkyl), -S(O)2NH2, -S(O)2NH (C1-C4 alkyl) or -S(O)2N (C1-C4 alkyl)2; or two R 9 Together with the atoms they are attached to, they form a C3-C. 10 Cycloalkyl or 3- to 10-membered heterocyclic alkyl rings.

2. The compound according to claim 1, wherein the compound is of formula (II): Or its enantiomers, diastereomers or pharmaceutically acceptable salts.

3. The compound according to claim 1, wherein the compound is of formula (III): Or its enantiomers, diastereomers or pharmaceutically acceptable salts.

4. The compound according to claim 1, wherein the compound is of formula (IV): Or its enantiomers, diastereomers or pharmaceutically acceptable salts.

5. The compound according to any one of claims 1-4, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: R 1 For H.

6. The compound according to any one of claims 1-4, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: R 1 It is a C1-C6 alkyl group.

7. The compound according to any one of claims 1-6, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: It is a C4-C7 cycloalkyl group, wherein the C4-C7 cycloalkyl group is optionally surrounded by one or two R groups. 9 replace.

8. The compound according to any one of claims 1-6, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein: It is a heteroalkyl group, consisting of 3-6 carbon atoms and one or two atoms selected from -NH-, -NR-. 2 Composed of heteroatoms or groups of -, -O-, and -S-, wherein the heteroalkyl group is optionally composed of one or two R- atoms. 9 replace.

9. The compound according to any one of claims 1-6, or its enantiomer, diastereomer, or pharmaceutically acceptable salt, wherein... for: , , , , , , , , , , , , , , , , , , , , , , , ; , or Each X and Y is independently -NH-, -NR. 2 -、-O- or -S-.

10. The compound according to any one of claims 1-6, or its enantiomer, diastereomer, or pharmaceutically acceptable salt, wherein... for: , , , , , , , , , , or , Where X is -NH- or -NR 2 -、-O- or -S-.

11. The compound according to any one of claims 1-6, or its enantiomers, diastereomers, or pharmaceutically acceptable salts, wherein... for: , , , , , , , , , , or , Where X is -O-.

12. The compound according to any one of claims 1-11, or its enantiomer, diastereomer, or pharmaceutically acceptable salt, wherein: The linker is #-(L 3 ) o -(L 2 ) p -(L 1 ) q -; # indicates AND The connection; Each L 1 L 2 and L 3 Independently, it is a bond, -O-, -NH-, C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C1-C6 heteroalkylene, alkoxycarbonyl, C3-C 10 Cycloalkylene or 3- to 10-membered heterocyclic alkylene, wherein each C1-C6 alkylene, C1-C6 haloalkylene, C1-C6 alkoxy, C1-C6 heteroalkylene, alkoxycarbonyl, C3-C 10 Cycloalkylene or 3- to 10-membered heterocycloalkylene, optionally surrounded by 1, 2, or 3 Rs. 9 replace; Each o and p is an independent integer from 0 to 6; and q is an integer from 1 to 6.

13. The compound according to any one of claims 1-11, or its enantiomer, diastereomer, or pharmaceutically acceptable salt, wherein: The linker group is methylene, ss-CF2-, -CH(CH3)-, ethylene, #-CH(CH3)-CH2-, #-CH2-CH(CH3)-, n-propylene, -CH2-CH(CH3)-CH2-, #-CH(CH3)-CH2-CH2-, #-CH2-CH2-CH(CH3)-, n-butylene, n-pentylene, -CH2-O-CH2-, #-CH2-O-CH2-CH2-, #-CH2-CH2-O-CH2-, -CH2-O-CH2-O-CH2-, or its enantiomers or diastereomers; and # indicates AND The connection.

14. The compound according to any one of claims 1-11, or its enantiomer, diastereomer, or pharmaceutically acceptable salt, wherein: The linker group is methylene, -CH(CH3)-, ethylene, #-CH(CH3)-CH2-, #-CH2-CH(CH3)-, n-propylene, -CH2-CH(CH3)-CH2-, #-CH(CH3)-CH2-CH2-, #-CH2-CH2-CH(CH3)-, n-butylene, n-pentylene, -CH2-O-CH2-, #-CH2-O-CH2-CH2-, #-CH2-CH2-O-CH2-, or their enantiomers or diastereomers; and # indicates AND The connection.

15. The compound according to any one of claims 1-11, or its enantiomer, diastereomer, or pharmaceutically acceptable salt, wherein: The linker is n-butylene, #-CH2-O-CH2-CH2-, #-CH2-CH2-O-CH2-, or its enantiomers or diastereomers; and # indicates AND The connection.

16. The compound of claim 1, or its enantiomers, diastereomers, or pharmaceutically acceptable salts thereof, wherein the compound is: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

17. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is: , , , , , , , , , , , , , , or .

18. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is: , , , , , , , , , , , , , , or .

19. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is: , , , , , , , , , , , , , , or .

20. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is: , , , , , , , , , , , , , , or .

21. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is: , , , , , , , , , , , , , , or .

22. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is: , , , , , , , , , , , , , , or .

23. A compound selected from the following: , , , , and or its enantiomers, diastereomers or pharmaceutically acceptable salts.

24. The compound of claim 23, or its enantiomers, diastereomers, or pharmaceutically acceptable salts thereof, wherein the compound is: , , or .

25. The compound of claim 23 or a pharmaceutically acceptable salt thereof, wherein the compound is: , , or .

26. The compound of claim 23 or a pharmaceutically acceptable salt thereof, wherein the compound is: , or .

27. A mutant protein comprising one or more non-natural amino acids, wherein each of the one or more non-natural amino acids is independently a compound according to any one of claims 1-26.

28. The mutant protein of claim 27, wherein the mutant protein comprises a compound according to any one of claims 1-26.

29. The mutant protein of claim 27, wherein the mutant protein comprises two or more non-natural amino acids, and wherein each of the two or more non-natural amino acids is the same compound as any one of claims 1-26.

30. Use of the compound according to any one of claims 1-26 in the manufacture of a mutant protein comprising one or more non-natural amino acids, wherein at least one of the one or more non-natural amino acids is a compound according to any one of claims 1-26.

31. The use according to claim 30, wherein at least two of the one or more non-natural amino acids are independently compounds according to any one of claims 1-26.

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