Process for preparing functionalized cyclooctene

Abstract

The present disclosure relates to processes for preparing functionalized cyclooctenes and synthetic intermediates prepared thereby.

Description

[0001] This application is a divisional application of a patent application with application number 201980080633.9, application date October 10, 2019, and name “Process for Preparing Functionalized Cyclooctene”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 62 / 744,041, filed on October 10, 2018, the entire contents of which are incorporated herein by reference. Technical Field

[0004] The present disclosure relates to processes for preparing functionalized cyclooctenes and synthetic intermediates prepared thereby. Background Art

[0005] Bioorthogonal conjugation or click reactions are selective and orthogonal (non-interacting) functions found in biological systems and have found use in a variety of applications in chemistry, chemical biology, molecular diagnostics, and medicine, where they can be used to facilitate the selective manipulation of molecules, cells, particles, and surfaces in vitro and in vivo, as well as the labeling and tracking of biomolecules. These reactions include Staudinger ligation, azide-cyclooctyne cycloaddition, and inverse electron demand Diels-Alder reactions. The present disclosure provides methods for preparing functionalized payload compositions for such reactions for delivery of therapeutic agents to subjects, the functionalized payload compositions having improved water solubility. Summary of the invention

[0006] Compared to payloads linked to unsubstituted cyclooctene, functionalized payloads according to the present disclosure have improved water solubility and are therefore easier to formulate and administer.

[0007] The present invention provides a process for preparing a compound of formula I or a salt thereof:

[0008]

[0009] in

[0010] R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety;

[0011] n is 0, 1, 2, 3 or 4;

[0012] Each R 2 Independently for C 1-4 Alkyl, C1-4 Haloalkyl and C 1-4 Alkoxy;

[0013] D is the payload portion; and

[0014] R 4 is hydrogen or C 1-4 alkyl;

[0015] This includes making a compound of formula II:

[0016]

[0017] Contact HO-R 4 , an optionally substituted heterocyclyl or an amino acid moiety thereof or a salt thereof, and a compound of formula III:

[0018]

[0019] Among them, each R 3 In the presence of a base in an organic solvent, independently C 1-4 alkyl.

[0020] Also provided is a process for preparing a compound of formula X or a salt thereof:

[0021]

[0022] in

[0023] R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety;

[0024] n is 0, 1, 2, 3 or 4;

[0025] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 alkoxy; and

[0026] R 4 is hydrogen or C 1-4 alkyl;

[0027] This includes making a compound of formula IV:

[0028]

[0029] Contact HO-R 4 , an optionally substituted heterocyclyl or an amino acid moiety thereof or a salt thereof, and a compound of formula III:

[0030]

[0031] Among them, each R 3 In the presence of a base in an organic solvent, independently C 1-4 alkyl.

[0032] Also provided is a process for preparing a compound of formula I or a salt thereof:

[0033]

[0034] in

[0035] R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety;

[0036] n is 0, 1, 2, 3 or 4;

[0037] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 Alkoxy;

[0038] D is the payload portion; and

[0039] R 4 is hydrogen or C 1-4 alkyl;

[0040] The method comprises making a compound of formula X or a salt thereof:

[0041]

[0042] The payload moiety or a salt thereof is contacted in the presence of a base in an organic solvent.

[0043] Also provided is a process for preparing a composition comprising a compound of formula IV or a salt thereof:

[0044]

[0045] in

[0046] n is 0, 1, 2, 3 or 4; and

[0047] Each R 2 are independently hydrogen or C 1-4 alkyl;

[0048] This includes making one equivalent of a compound of formula V or a salt thereof:

[0049]

[0050] 3-5 equivalents of N,N'-disuccinimidyl carbonate are contacted in 15-25 volumes of anhydrous organic solvent in the presence of a base at a temperature of 15-30°C for 5-10 hours.

[0051] Also provided is a process for resolving a composition comprising one or more stereoisomers of a compound of Formula VI or a salt thereof:

[0052]

[0053] in

[0054] n is 0, 1, 2, 3 or 4;

[0055] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 Alkoxy;

[0056] The following steps are involved:

[0057] (a) contacting a composition comprising one or more stereoisomers of Formula VI with a chiral base to provide a chiral salt of a compound of Formula VI;

[0058] (b) cooling the composition of step (a);

[0059] (c) isolating a chiral salt of the compound of Formula VI; and

[0060] (d) hydrolyzing a chiral salt of a compound of Formula VI to provide an enantiomerically enriched composition comprising a compound of Formula VI or a salt thereof.

[0061] In certain embodiments, the process comprises repeating steps (b) and (c) prior to step (d).

[0062] Also provided is a process for preparing an enantiomerically enriched composition comprising a compound of formula VII or a salt thereof:

[0063]

[0064] in

[0065] n is 0, 1, 2, 3 or 4;

[0066] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 alkoxy; and

[0067] R 4 are independently hydrogen or C 1-4 alkyl;

[0068] Compositions comprising enantiomerically enriching a compound of Formula VI or a salt thereof:

[0069]

[0070] contacting in sequence: (i) a solution comprising KI and I2, (ii) a base, and (iii) a solution comprising a base and a 4 -OH to provide an enantiomerically enriched composition of a compound of Formula VII or a salt thereof.

[0071] Also provided is a process for resolving a composition comprising one or more stereoisomers of a compound of Formula VIIA or a salt thereof:

[0072]

[0073] in

[0074] R 5 is hydrogen or C 1-4 Alkyl, comprising the following steps:

[0075] (a) contacting a composition comprising one or more stereoisomers of Formula VIIA with a chiral base to provide a chiral salt of a compound of Formula VIIA;

[0076] (b) cooling the composition of step (a);

[0077] (c) isolating a chiral salt of a compound of formula VIIA; and

[0078] (d) hydrolyzing a chiral salt of a compound of Formula VIIA to provide an enantiomerically enriched composition comprising a compound of Formula VIIA or a salt thereof.

[0079] Also provided is a process for preparing an enantiomerically enriched composition comprising a compound of formula VB or a salt thereof:

[0080]

[0081] in

[0082] R 4 is hydrogen or C 1-4 Alkyl; and

[0083] R 5 is hydrogen or C 1-4 alkyl;

[0084] An enantiomerically enriched composition comprising an isomerized compound of formula VIIB or a salt thereof:

[0085]

[0086] Also provided is an enantiomerically enriched composition comprising a compound selected from Table 1 or a salt thereof. DETAILED DESCRIPTION

[0087] 1. Definition

[0088] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art. In the event of a conflict, this document (including definitions) shall prevail. Preferred methods and materials are as follows, but methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods and examples disclosed herein are illustrative only and are not intended to be limiting.

[0089] As used herein, the terms "comprise(s)", "include(s)", "having", "has", "can", "contain(s)" and variations thereof are intended to be open-ended transitional phrases, terms or words that do not exclude the possible presence of additional acts or structures. The singular forms "a", "an", and "the" include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates additional embodiments "comprising", "consisting of", and "consisting essentially of" embodiments or elements set forth herein, whether or not explicitly set forth.

[0090] The modifier "about" used in conjunction with an amount is inclusive of the stated value and has the meaning dictated by the context (e.g., it includes at least the degree of error associated with the measurement of a particular amount). The modifier "about" should also be considered to disclose a range defined by the absolute values ​​of the two endpoints. For example, the expression "about 2 to about 4" also discloses a range of "2 to 4". The term "about" can refer to plus or minus 10% of the indicated number. For example, "about 10%" can mean a range of 9% to 11%, and "about 1" can mean 0.9-1.1. Other meanings of "about" are also apparent from the context, such as rounding, so that, for example, "about 1" can also mean 0.5 to 1.4.

[0091] The conjunction "or" includes any and all combinations of one or more listed elements associated with the conjunction. For example, the phrase "a device comprising A or B" may refer to a device comprising A without B, a device comprising B without A, or a device with both A and B present. The phrase "at least one of A, B, ... and N" or "at least one of A, B, ... and N or a combination thereof" is defined in the broadest sense to refer to one or more elements selected from the group comprising A, B, ... and N, that is, any combination of one or more of elements A, B, ... or N, including any one element alone, or in combination with one or more of other elements, which may also include additional elements not listed in the combination.

[0092] The definitions of specific functional groups and chemical terms are described in more detail below. For the purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition, inside cover, and specific functional groups are generally defined as described herein. In addition, general principles of organic chemistry as well as specific functional moieties and reactivities are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March, March's Advanced Organic Chemistry, 5th edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; Carruthers, Some Modern Methods of Organic Synthesis, 3rd edition, Cambridge University Press, Cambridge, 1987; the entire contents of each of the above references are incorporated herein by reference.

[0093] As used herein, the term "enantiomerically enriched" refers to a composition of a chiral substance having an enantiomeric ratio greater than 50:50 but less than 100:0. In certain embodiments, the % ee of the enantiomerically enriched composition is greater than about 25%, or greater than about 30%, or greater than about 35%, or greater than about 40%, or about 45%, or about 50%, or about 55%, or greater than about 60%, or greater than about 65%, or greater than about 70%, or greater than about 75%, or greater than about 80%, or greater than about 85%, or greater than about 90%, or greater than about 95%, or greater than about 97%, or greater than about 99%, or about 25%, or about 30%, or about 35%, or about 40%, or about 45%, or about 50%, or about 55%, or about 60%, or about 65%, or about 70%, or about 75%, or about 80%, or about 85%, or about 90%, or about 95%, or about 97%, or about 99%, or about 100.

[0094] As used herein, the term "alkyl" refers to a straight or branched saturated hydrocarbon chain containing 1 to 30 carbon atoms. The term "lower alkyl" or "C1-C6 alkyl" refers to a straight or branched hydrocarbon containing 1 to 6 carbon atoms. The term "C1-C3 alkyl" refers to a straight or branched hydrocarbon containing 1 to 3 carbon atoms. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl and n-decyl.

[0095] As used herein, the term "alkoxy" refers to an alkyl group, as defined herein, attached to the parent molecular moiety through an oxygen atom. Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, and tert-butoxy.

[0096] As used herein, the term "alkenyl" refers to a hydrocarbon chain containing 2 to 30 carbon atoms and having at least one carbon-carbon double bond. The alkenyl group may be substituted or unsubstituted. For example, the alkenyl group may be substituted by an aryl group such as a phenyl group.

[0097] As used herein, the term "alkynyl" refers to a straight or branched monovalent hydrocarbon radical having 2 to 30 carbon atoms (such as 2 to 20 or 2 to 10 carbon atoms) and having at least one triple bond unsaturated site. The term "alkyne" also includes non-aromatic cycloalkyls having 5 to 20 carbon atoms (such as 5 to 10 carbon atoms) having a monocyclic or multiple rings and having at least one triple bond. Examples of such alkynyls include, but are not limited to, ethynyl (-C≡CH) and propargyl (-CH2C≡CH) and cycloalkynyl moieties, such as, but not limited to, substituted or unsubstituted cyclooctyne moieties.

[0098] The term "alkoxyalkyl" as used herein, means an alkoxy group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein.

[0099] As used herein, the term "alkylene" refers to a divalent group derived from a straight or branched hydrocarbon having 1 to 30 carbon atoms, for example, 2 to 10 carbon atoms. Representative examples of alkylene include, but are not limited to, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, and -CH2CH2CH2CH2CH2-.

[0100] As used herein, the term "aryl" refers to a phenyl or bicyclic aryl or tricyclic aryl fused (fused) ring system. Examples of bicyclic fused ring systems include phenyls attached to the parent molecular moiety and fused to the phenyl. Examples of tricyclic fused ring systems include phenyls attached to the parent molecular moiety and fused to two other phenyls. Representative examples of bicyclic aryls include, but are not limited to, naphthyl. Representative examples of tricyclic aryls include, but are not limited to, anthracenyl. Monocyclic, bicyclic, and tricyclic aryls are connected to the parent molecular moiety by any carbon atom contained in the ring, and can be unsubstituted or substituted.

[0101] As used herein, the term "cycloalkyl" refers to a carbocyclic ring system containing three to ten carbon atoms, zero heteroatoms and zero double bonds. Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl and cyclodecyl. "Cycloalkyl" also includes carbocyclic ring systems in which the cycloalkyl is attached to the parent molecular moiety and fused to an aryl as defined herein, a heteroaryl as defined herein, or a heterocycle as defined herein.

[0102] As used herein, the term "cycloalkenyl" refers to a non-aromatic monocyclic or polycyclic ring system containing at least one carbon-carbon double bond and preferably having 5-10 carbon atoms per ring. Exemplary monocyclic cycloalkenyl rings include, but are not limited to, cyclopentenyl, cyclohexenyl, or cycloheptenyl.

[0103] As used herein, the term "cyclooctene" refers to a substituted or unsubstituted non-aromatic cyclic alkyl group having 8 carbon atoms, which has a single ring with a double bond. Examples of such cyclooctenyl groups include, but are not limited to, substituted or unsubstituted trans-cyclooctene (TCO).

[0104] As used herein, the term "fluoroalkyl" refers to an alkyl group as defined herein, wherein one, two, three, four, five, six, seven or eight hydrogen atoms are substituted by fluorine. Representative examples of fluoroalkyl groups include, but are not limited to, 2-fluoroethyl, 2,2,2-trifluoroethyl, trifluoromethyl, difluoromethyl, pentafluoroethyl and trifluoropropyl, such as 3,3,3-trifluoropropyl.

[0105] The term "alkoxyfluoroalkyl" as used herein, means an alkoxy group, as defined herein, appended to the parent molecular moiety through a fluoroalkyl group, as defined herein.

[0106] The term "fluoroalkoxy" as used herein, refers to at least one fluoroalkyl group, as defined herein, attached to the parent molecular moiety through an oxygen atom. Representative examples of fluoroalkoxy include, but are not limited to, difluoromethoxy, trifluoromethoxy, and 2,2,2-trifluoroethoxy.

[0107] As used herein, the term "halogen" or "halo" refers to Cl, Br, I or F.

[0108] As used herein, the term "haloalkyl" refers to an alkyl group as defined herein wherein one, two, three, four, five, six, seven or eight hydrogen atoms are replaced by halogen.

[0109] The term "haloalkoxy" as used herein, means at least one haloalkyl group, as defined herein, attached to the parent molecular moiety through an oxygen atom.

[0110] As used herein, the term "heteroalkyl" refers to an alkyl group as defined herein, wherein one or more carbon atoms have been replaced by a heteroatom selected from S, Si, O, P and N. The heteroatom may be oxidized. Representative examples of heteroalkyl groups include, but are not limited to, alkyl ethers, secondary and tertiary alkyl amines, and alkyl sulfides.

[0111] As used herein, the term "heteroaryl" refers to an aromatic monocyclic or aromatic bicyclic ring system or an aromatic tricyclic ring system. An aromatic monocyclic ring is a five-membered or six-membered ring containing at least one heteroatom independently selected from the group consisting of N, O and S (e.g., 1, 2, 3 or 4 heteroatoms independently selected from O, S and N). A five-membered aromatic monocyclic ring has two double bonds, and a six-membered six-membered aromatic monocyclic ring has three double bonds. Examples of bicyclic heteroaryls include a monocyclic heteroaryl ring attached to a parent molecular moiety and fused to a monocyclic cycloalkyl as defined herein, a monocyclic aryl as defined herein, a monocyclic heteroaryl as defined herein, or a monocyclic heterocyclic ring as defined herein. Examples of tricyclic heteroaryls include a monocyclic heteroaryl ring attached to a parent molecular moiety and fused to two monocyclic cycloalkyls as defined herein, a monocyclic aryl as defined herein, a monocyclic heteroaryl as defined herein, or a monocyclic heterocyclic ring as defined herein. Representative examples of monocyclic heteroaryls include, but are not limited to, pyridyl (including pyridin-2-yl, pyridin-3-yl, pyridin-4-yl), pyrimidinyl, pyrazinyl, thienyl, furanyl, thiazolyl, thiadiazolyl, isoxazolyl, pyrazolyl, and 2-oxo-1,2-dihydropyridinyl. Representative examples of bicyclic heteroaryls include, but are not limited to, chromenyl, benzothienyl, benzodioxolyl, benzotriazolyl, quinolinyl, thienopyrrolyl, thienothiphenylyl, imidazothiazolyl, benzothiazolyl, benzofuranyl, indolyl, quinolinyl, imidazopyridinyl, benzooxadiazolyl, and benzopyrazolyl. Representative examples of tricyclic heteroaryls include, but are not limited to, dibenzofuranyl and dibenzothienyl. The monocyclic, bicyclic and tricyclic heteroaryl groups are attached to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the rings, and can be unsubstituted or substituted.

[0112] As used herein, the term "heterocycle" or "heterocyclic" refers to a monocyclic heterocycle, a bicyclic heterocycle or a tricyclic heterocycle. The monocyclic heterocycle is a three-, four-, five-, six-, seven- or eight-membered ring comprising at least one heteroatom independently selected from the group consisting of O, N and S. The three- or four-membered ring comprises zero or one double bond and a heteroatom selected from the group consisting of O, N and S. The five-membered ring comprises zero or one double bond and one, two or three heteroatoms selected from the group consisting of O, N and S. The six-membered ring comprises zero, one or two double bonds and one, two or three heteroatoms selected from the group consisting of O, N and S. The seven- and eight-membered ring comprises zero, one, two or three double bonds and one, two or three heteroatoms selected from the group consisting of O, N and S. Representative examples of monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiolanyl, 1,3-dithianyl, 1,3-dimethylpyrimidine-2,4(1H,3H)-dione, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl , oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, oxetanyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydrothiophenyl, thiadiazolinyl, thiadiazolidinyl, 1,2-thiazinyl, 1,3-thiazinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1-dioxothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, and trithianyl. The bicyclic heterocycle is a monocyclic heterocycle fused to a phenyl group, or a monocyclic heterocycle fused to a monocyclic cycloalkyl group, or a monocyclic heterocycle fused to a monocyclic cycloalkenyl group, or a monocyclic heterocycle fused to a monocyclic heterocycle, or a spiroheterocyclic group, or a bridged monocyclic heterocycle system in which two non-adjacent atoms of the ring are linked by an alkylene bridge of 1, 2, 3 or 4 carbon atoms or an alkenylene bridge of two, three or four carbon atoms. Representative examples of bicyclic heterocycles include, but are not limited to, benzopyranyl, benzothiopyranyl, chromanyl, 2,3-dihydrobenzofuranyl, 2,3-dihydrobenzothiophenyl, 2,3-dihydroisoquinolinyl, 2-azaspiro[3.3]hept-2-yl, azabicyclo[2.2.1]hept-2-yl (including 2-azabicyclo[2.2.1]hept-2-yl), 2,3-dihydro-1H-indolyl, isoindolinyl, octahydrocyclopenta[c]pyrrolyl, octahydropyrrolopyridinyl, and tetrahydroisoquinolinyl.Examples of tricyclic heterocycles include bicyclic heterocycles fused to phenyl, or bicyclic heterocycles fused to monocyclic cycloalkyl, or bicyclic heterocycles fused to monocyclic cycloalkenyl, or bicyclic heterocycles fused to monocyclic heterocycles or bicyclic heterocycles wherein two non-adjacent atoms of the bicyclic ring are linked by an alkylene bridge of 1, 2, 3 or 4 carbon atoms or an alkenylene bridge of 2, 3 or 4 carbon atoms. Examples of tricyclic heterocycles include, but are not limited to, octahydro-2,5-epoxypentalene, hexahydro-2H-2,5-methanol cyclopenta[b]furan, hexahydro-1H-1,4-methanol cyclopenta[c]furan, aza-adamantane (1-azatricyclo[3.3.1.13,7]decane) and oxadamantane (2-oxatricyclo[3.3.1.13,7]decane). The monocyclic, bicyclic and tricyclic heterocycles are attached to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the rings, and can be unsubstituted or substituted.

[0113] As used herein, the term "hydroxy" refers to an -OH group.

[0114] As used herein, the term "hydroxyalkyl" refers to an alkyl group as defined herein wherein one, two, three, four, five, six, seven or eight hydrogen atoms are replaced by hydroxy groups.

[0115] In some cases, the number of carbon atoms in a hydrocarbyl substituent (e.g., alkyl or cycloalkyl) is indicated by the prefix "C x -C y -" or "C x-y " means, where x is the minimum number of carbon atoms in the substituent and y is the maximum number of carbon atoms in the substituent. Thus, for example, "C1-C3-alkyl" and "C 1-3 "Alkyl" refers to an alkyl substituent containing 1 to 3 carbon atoms. x -C y -" and "C x-y ” These two conventions can be used interchangeably and have the same meaning.

[0116] The term "substituted" refers to a group that can be further substituted by one or more non-hydrogen substituents. Substituents include, but are not limited to, halogen, =O, =S, cyano, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic radical, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxyalkyl, alkylene, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, -COOH, keto, amide, carbamate and acyl.

[0117] The term "therapeutic agent" refers to an agent capable of treating and / or alleviating a condition or disease, or one or more symptoms thereof, in a subject. Therapeutic agents of the present disclosure also include prodrug forms of the therapeutic agents.

[0118] The term "diagnostic agent" refers to an agent that helps diagnose a condition or disease. Representative diagnostic agents include imaging agents, such as paramagnetic agents, optical probes, radionuclides, and the like. Paramagnetic agents are imaging agents that have magnetism under an externally applied field. Examples of paramagnetic agents include, but are not limited to, iron particles, including iron nanoparticles and iron microparticles. Optical probes are fluorescent compounds that can be detected by excitation at one wavelength of radiation and detection at a second, different wavelength of radiation. Optical probes disclosed herein include, but are not limited to, Cy5.5, Alexa 680, Cy5, DiD (1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine perchlorate) and DiR (1,1'-dioctadecyl 3,3,3',3'-tetramethylindotricarbocyanine iodide). Other optical probes include quantum dots. Radionuclides are elements that undergo detectable radioactive decay. Radionuclides that can be used in embodiments of the present disclosure include, but are not limited to 3 H. 11 C. 13 N. 18 F. 19 F. 60 Co. 64 Cu, 67 Cu, 68 Ga, 82 Rb, 90 Sr. 90 Y. 99 Tc, 99m Tc, 111 In, 123 I. 124 I. 125 I. 129 I. 131 I. 137 Cs, 177 Lu, 186 Re, 188 Re, 211 At, Rn, Ra, Th, U, Pu and 241 Am.

[0119] The term "contacting" or "contact" refers to the process of bringing at least two different species into contact so that they can interact with each other, such as in a non-covalent or covalent binding interaction or binding reaction. However, it should be understood that the resulting complex or reaction product can be produced directly from the interaction or reaction between the added reagents, or from one or more added reagents or moieties of intermediates that can be produced in the contact mixture.

[0120] The term "leaving group" refers to an atom (or group of atoms) with electron-withdrawing ability that can be replaced with a stable species along with the bonding electrons. Examples of suitable leaving groups include halides (e.g., Br, Cl, I), sulfonates (e.g., triflate, mesylate, tosylate, and brosylate), and nitrophenols.

[0121] The term "physiological conditions" is meant to encompass those conditions that are compatible with living cells, for example, predominantly aqueous conditions of temperature, pH, salinity, etc. that are compatible with living cells.

[0122] The term "chiral base" refers to a chiral compound having at least one stereogenic center and at least one nitrogen atom. Chiral bases can be synthesized using methods known in the art or purchased from commercial sources (e.g., Sigma Aldrich). Exemplary chiral bases include, but are not limited to, L-(-)-α-amino-ε-caprolactam hydrochloride, (R)-(-)-1-amino-2-propanol, (S)-(+)-1-amino-2-propanol, L-aspartic acid, cis-(1S,2R)-(-)-2-(benzylamino)cyclohexanemethanol, (S)-N-benzyl-1-(1-naphthyl)ethylamine hydrochloride, (-)-1,4-bis-O-(4-chlorobenzyl)-L-threitol, (1R,2R)-1,2-bis(2-hydroxyphenyl)ethylenediamine, (1S,2S)-1,2-bis(2-hydroxyphenyl)ethylenediamine, (+) -Bis[(R)-1-phenylethyl]amine hydrochloride, N,N-bis[(S)-(-)-1-phenylethyl] o-aminobenzoic acid, (R)-(+)-1-(4-bromophenyl)ethylamine, (S)-(-)-1-(4-bromophenyl)ethylamine, (R)-4-chloro-α-methylbenzylamine, (S)-4-chloro-α-methylbenzylamine, cinchonidine, (+)-cinchonine, (R)-(+)-N,α-dimethylbenzylamine, (S)-(-)-N,α-dimethylbenzylamine, (R)-(+)-N,N-dimethyl-1-phenylethylamine, (S)-(-)-N,N-dimethyl-1-phenyl Ethylamine, (R)-5,5-dimethyl-6-phenyl-3,4,5,6-tetrahydropyrimidine, (S)-5,5-dimethyl-6-phenyl-3,4,5,6-tetrahydropyrimidine, (R)-(-)-3,5-dinitro-N-(1-phenylethyl)benzamide, (S)-(+)-3,5-dinitro-N-(1-phenylethyl)benzamide, (1R,2S)-(-)-ephedrine, D-glutamic acid, L-glutamic acid, (R)-(+)-α-methylbenzylamine, (S)-(-)-α-methylbenzylamine, (R)-α-methyl-4-nitrobenzylamine hydrochloride, (S)-α -Methyl-4-nitrobenzylamine hydrochloride, (R)-(+)-α-methyl-4-pyridinemethanol, (S)-(-)-α-methyl-4-pyridinemethanol, (S)-(-)-1-(2-naphthyl)ethylamine, (S)-(-)-N-[1-(1-naphthyl)ethyl]succinic acid, (R)-(+)-N-(1-phenylethyl)-o-carbamoylbenzoic acid, (S)-(-)-N-(1-phenylethyl)-o-carbamoylbenzoic acid, (R)-(+)-N-(1-phenylethyl)succinic acid, (S)-(-)-N-(1-phenylethyl)succinic acid, quinine, D-valine and L-valine.

[0123] For the compounds described herein, groups and substituents may be selected based on the allowed valences of atoms and substituents such that the selections and substituents result in stable compounds, e.g., compounds that do not spontaneously transform (such as by rearrangement, cyclization, elimination, etc.).

[0124] Where a numerical range is provided, it will be understood that, unless the context clearly specifies otherwise, each intermediate value to one tenth of a unit of the lower limit between the upper and lower limits of the range or any other specified value or intermediate value within the specified range is encompassed within the present invention. The upper and lower limits of these smaller ranges may be independently included in the smaller ranges and are also encompassed within the present invention, but any explicitly excluded limits within the specified ranges are to be observed. Where a specified range includes one or two limits, the range excluding one or both of those included limits is also encompassed within the present invention.

[0125] For the purpose of reciting the numerical ranges herein, each intervening number with the same degree of precision therebetween is expressly contemplated. For example, for the range of 6-9, in addition to 6 and 9, the numbers 7 and 8 are also contemplated, and for the range of 6.0-7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly contemplated.

[0126] It should be understood that certain features of the invention described in the context of separate embodiments for clarity may also be provided in combination in a single embodiment. Conversely, for brevity, various features of the invention described in the context of a single embodiment may also be provided individually or in any suitable subcombination. All combinations of embodiments related to the present invention are expressly included in the present invention and disclosed herein, just as each combination is individually and expressly disclosed, to the extent that such combinations include, for example, the subject matter of compounds that are stable compounds (i.e., compounds that can be prepared, isolated, characterized, and tested for biological activity). In addition, all subcombinations of various embodiments and elements thereof (i.e., elements of the chemical groups listed in the embodiments describing such variables) are expressly included in the present invention and disclosed herein, just as each such subcombination is individually and expressly disclosed.

[0127] 2. Craftsmanship

[0128] The processes described herein provide functionalized payloads with improved water solubility compared to payloads attached to unsubstituted cyclooctenes. Functionalized payloads can be used as reagents in bioorthogonal conjugation or click reactions and have been used in a variety of applications in chemistry, chemical biology, molecular diagnostics, and medicine, where they can be used to facilitate selective manipulation of molecules, cells, particles, and surfaces in vitro and in vivo, as well as labeling and tracking of biomolecules. The processes described herein are performed using appropriate reaction conditions and optionally one or more protecting groups as desired.

[0129] The term "reaction conditions" is intended to refer to the physical and / or environmental conditions under which a chemical reaction is carried out. Examples of reaction conditions include, but are not limited to, one or more of the following: reaction temperature, solvent, pH, pressure, reaction time, the molar ratio of reactants, the presence of a base or acid, one or more protecting groups or catalysts, radiation, etc. Reaction conditions can be named after a specific chemical reaction in which the conditions are employed, such as coupling conditions, hydrogenation conditions, acylation conditions, reduction conditions, etc. The reaction conditions of most reactions are generally known to those skilled in the art, or can be easily obtained from the literature. In the full text, particularly in the following examples, exemplary reaction conditions sufficient to perform chemical transformations provided herein can be found. It can also be expected that reaction conditions can also include reagents other than those listed in the specific reaction.

[0130] The term "protecting group" refers to those groups intended to protect a given atom or functional group from undesirable reactions during a synthetic procedure, and includes, but is not limited to, silyl ethers, such as 2-(trimethylsilyl)ethoxymethyl (SEM) ether, or alkoxymethyl ethers, such as methoxymethyl (MOM) ether, tert-butoxymethyl (BUM) ether, benzyloxymethyl (BOM) ether, or methoxyethoxymethyl (MEM) ether. Additional protecting groups include tert-butyl, acetyl, benzyl, benzyloxycarbonyl (carbonylbenzyloxy, CBZ), p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, tert-butoxycarbonyl (BOC), trifluoroacetyl, and the like. Certain protecting groups may be preferred over other protecting groups due to their convenience or relative ease of removal or due to their stereospecific effects in subsequent steps of the process. Additional suitable amino protecting groups are taught in TW Greene and PGM Wuts, Protecting Groups in Organic Synthesis, 5th ed., Wiley, New York, 2014, and all references cited therein are incorporated herein by reference in their entirety.

[0131] In one embodiment, the present disclosure provides a process for preparing a compound of formula I or a salt thereof:

[0132]

[0133] in

[0134] R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety;

[0135] n is 0, 1, 2, 3 or 4;

[0136] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 Alkoxy;

[0137] D is the payload portion; and

[0138] R 4 is hydrogen or C 1-4 alkyl;

[0139] This includes making a compound of formula II:

[0140]

[0141] Contact HO-R 4 , an optionally substituted heterocyclyl or an amino acid moiety thereof or a salt thereof, and a compound of formula III:

[0142]

[0143] Among them, each R 3 In the presence of a base in an organic solvent, independently C 1-4 alkyl.

[0144] In one embodiment, the present disclosure provides a process for preparing a compound of formula I or a salt thereof:

[0145]

[0146] in

[0147] R 1 is the amino acid part;

[0148] n is 0, 1, 2, 3 or 4;

[0149] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 alkoxy; and

[0150] D is the effective load part;

[0151] This includes making a compound of formula II:

[0152]

[0153] contacting an amino acid moiety or a salt thereof, and a compound of formula III:

[0154]

[0155] Among them, each R 3 In the presence of a base in an organic solvent, independently C 1-4 alkyl.

[0156] In one embodiment, the present disclosure provides a process for preparing a compound of formula I or a salt thereof:

[0157]

[0158] where

[0159] R 1 Selected from the group consisting of: G 1 , OH, -NR 1c -C 1-4 Alkylene-G 1 ,

[0160] -NR 1c -C 1-4 Alkylene-N(R 1d )2、-N(R 1c )CHR 1e CO2H, -N(R 1c )-C 1-6 Alkylene-CO2H, -N(R 1f )-C 2-4 Alkylene-

[0161] (N(C 1-4 Alkylene-CO2H)-C 2-4 Alkylene) m -N(C 1-4 Alkylene -CO2H)2, -N(R 1c )CHR 1e C(O)OC 1-6 alkyl,

[0162] -N(R 1c )-C 1-6 Alkylene-C(O)OC 1-6 Alkyl, and -N(R 1f )-C 2-4 Alkylene-(N(C 1-4 Alkylene-C(O)OC1-6 Alkyl)-C 2-4 Alkylene) m -N(C 1-4 Alkylene-C(O)OC 1-6 Alkyl)2;

[0163] Each time it appears, R 1c and R 1d are independently hydrogen or C 1-4 alkyl;

[0164] R 1e For -C 1-4 Alkylene -CO2H, -C 1-4 Alkylene -CONH2 or -C 1-4 Alkylene-OH;

[0165] R 1f For hydrogen, C 1-6 Alkyl or C 1-4 Alkylene-CO2H;

[0166] G 1 is an optionally substituted heterocyclic group.

[0167] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 Alkoxy;

[0168] D is the effective load part;

[0169] n is 0, 1, 2, 3 or 4; and

[0170] m is 0, 1, 2 or 3;

[0171] This includes making a compound of formula II:

[0172]

[0173] Suitable reagents and compounds for contacting formula III:

[0174]

[0175] Among them, each R 3 In the presence of a base in an organic solvent, independently C 1-4 alkyl.

[0176] In certain embodiments, R 1 Selected from the group consisting of: G 1 ,OH,

[0177] -NR 1c -C1-4 Alkylene-G 1 、-NR 1c -C 1-4 Alkylene-N(R 1d )2、-N(R 1c )CHR 1e CO2H, -N(R 1c )CH2CO2H and -N(R 1f )-CH2CH2-(N(CH2CO2H)CH2CH2) m -N(CH2CO2H)2;

[0178] R 1e For -C 1-4 Alkylene-CO2H;

[0179] R 1f is hydrogen or -C 1-4 Alkylene-CO2H;

[0180] G 1 is a 4- to 8-membered monocyclic heterocyclic group comprising a first nitrogen atom and optionally one further heteroatom selected from nitrogen, oxygen and sulfur, G 1 is attached to the first nitrogen atom and is optionally substituted with 1-4 substituents independently selected from the group consisting of:

[0181] C 1-4 Alkyl, C 1-4 Haloalkyl, halo, cyano, OH, -OC 1-4 alkyl and oxo; and

[0182] m is 0, 1 or 2;

[0183] In certain embodiments, R 1 The amino acid part.

[0184] In certain embodiments, the compound of Formula III is N,O-bis(trimethylsilyl)acetamide.

[0185] In certain embodiments, the organic solvent comprises DMF, DCM or a mixture thereof.

[0186] In certain embodiments, the base comprises an organic base. In certain embodiments, the base comprises an amine base. In certain embodiments, the base comprises DIPEA.

[0187] In certain embodiments, the contacting comprises stirring at room temperature for about 24 hours.

[0188] In certain embodiments, the process further comprises preparing a compound of formula II by reacting a compound of formula IV:

[0189]

[0190] The payload moiety or a salt thereof is contacted in the presence of a base in an organic solvent.

[0191] In certain embodiments, the organic solvent comprises DMF, DCM or a mixture thereof.

[0192] In certain embodiments, the base comprises an organic base. In certain embodiments, the base comprises an amine base. In certain embodiments, the base comprises DIPEA.

[0193] In certain embodiments, the contacting comprises stirring at room temperature for about 1 to 2 hours.

[0194] In certain embodiments, the compound of Formula I is represented by Formula IA:

[0195]

[0196] in

[0197] R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety;

[0198] R 4 is hydrogen or C 1-4 alkyl;

[0199] R 5 is hydrogen or C 1-4 Alkyl; and

[0200] D is the effective load part;

[0201] And the compound of formula II is represented by formula IIA:

[0202]

[0203] In certain embodiments, the compound of Formula I is represented by Formula IA:

[0204]

[0205] in

[0206] R 1 is the amino acid part;

[0207] R 5 is hydrogen or C 1-4 Alkyl; and

[0208] D is the effective load part;

[0209] And the compound of formula II is represented by formula IIA:

[0210]

[0211] In certain embodiments, the compound of Formula IV is represented by Formula IVA:

[0212]

[0213] The present invention also provides a process for preparing a compound of formula X or a salt thereof:

[0214]

[0215] in

[0216] R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety;

[0217] n is 0, 1, 2, 3 or 4;

[0218] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 alkoxy; and

[0219] R 4 is hydrogen or C 1-4 alkyl;

[0220] This includes making a compound of formula IV:

[0221]

[0222] Contact HO-R 4 , an optionally substituted heterocyclyl or an amino acid moiety thereof or a salt thereof, and a compound of formula III:

[0223]

[0224] Among them, each R 3 In the presence of a base in an organic solvent, independently C 1-4 alkyl.

[0225] The present invention also provides a process for preparing a compound of formula X or a salt thereof:

[0226]

[0227] in

[0228] R 1 is the amino acid part;

[0229] n is 0, 1, 2, 3 or 4;

[0230] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 alkoxy; and

[0231] This includes making a compound of formula IV:

[0232]

[0233] contacting an amino acid or a salt thereof and a compound of formula III:

[0234]

[0235] Among them, each R 3 In the presence of a base in an organic solvent, independently C 1-4 alkyl.

[0236] In certain embodiments, the compound of Formula III is N,O-bis(trimethylsilyl)acetamide.

[0237] In certain embodiments, the organic solvent comprises DMF, DCM or a mixture thereof.

[0238] In certain embodiments, the base comprises an organic base. In certain embodiments, the base comprises an amine base. In certain embodiments, the base comprises DIPEA.

[0239] In certain embodiments, the contacting comprises stirring at room temperature for about 24 hours.

[0240] In certain embodiments, the compound of Formula X is represented by Formula XA:

[0241]

[0242] And the compound of formula IV is represented by formula IVA:

[0243]

[0244] Among them, R 1 As defined herein, and R 5 is hydrogen or C 1-4 alkyl.

[0245] In certain embodiments, a process for preparing a compound of formula IV or a salt thereof is provided, wherein R 2 and n are as defined herein, the process comprising treating a compound of formula V or a salt thereof:

[0246]

[0247] The N,N'-disuccinimidyl carbonate is contacted in the presence of a base in an organic solvent.

[0248] In certain embodiments, the organic solvent comprises acetonitrile. In certain embodiments, the organic solvent comprises dry acetonitrile.

[0249] In certain embodiments, the base comprises an organic base. In certain embodiments, the base comprises an amine base. In certain embodiments, the base comprises DIPEA.

[0250] In certain embodiments, the contacting comprises stirring at room temperature.

[0251] In certain embodiments, the compound of Formula V is represented by Formula VA:

[0252]

[0253] Among them, R 4 is hydrogen or C 1-4 Alkyl and R 5 is hydrogen or C 1-4 alkyl.

[0254] Also provided is a process for preparing a compound of formula I or a salt thereof:

[0255]

[0256] in

[0257] R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety;

[0258] n is 0, 1, 2, 3 or 4;

[0259] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 Alkoxy;

[0260] D is the payload portion; and

[0261] R 4 is hydrogen or C 1-4 alkyl;

[0262] The method comprises making a compound of formula X or a salt thereof:

[0263]

[0264] The payload moiety or a salt thereof is contacted in the presence of a base in an organic solvent.

[0265] Also provided is a process for preparing a compound of formula I or a salt thereof:

[0266]

[0267] in

[0268] R 1 is the amino acid part;

[0269] n is 0, 1, 2, 3 or 4;

[0270] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 alkoxy; and

[0271] D is the effective load part;

[0272] The method comprises making a compound of formula X or a salt thereof:

[0273]

[0274] The payload moiety or a salt thereof is contacted in the presence of a base in an organic solvent.

[0275] In certain embodiments, the organic solvent comprises DMF, DCM or a mixture thereof.

[0276] In certain embodiments, the base comprises an organic base. In certain embodiments, the base comprises an amine base. In certain embodiments, the base comprises DIPEA.

[0277] In certain embodiments, the contacting comprises stirring at room temperature for about 1 to 2 hours.

[0278] In certain embodiments, the compound of Formula I is represented by Formula IA:

[0279]

[0280] And the compound of formula X is represented by formula XA:

[0281]

[0282] Among them, R 5 is hydrogen or C 1-4 alkyl.

[0283] Also provided herein is a process for preparing a composition comprising a compound of formula IV or a salt thereof:

[0284]

[0285] in

[0286] n is 0, 1, 2, 3 or 4; and

[0287] Each R 2 are independently hydrogen or C 1-4 alkyl;

[0288] This includes making one equivalent of a compound of formula V or a salt thereof:

[0289]

[0290] 3-5 equivalents of N,N'-disuccinimidyl carbonate are contacted in 15-25 volumes of anhydrous organic solvent in the presence of a base at a temperature of 15-30°C for 5-10 hours.

[0291] In certain embodiments, the process further comprises adding the composition comprising the compound of Formula IV to 40-60 volumes of water at room temperature to form an aqueous product mixture.

[0292] In certain embodiments, the process further comprises separating solids formed in the aqueous product mixture.

[0293] In certain embodiments, the process further comprises grinding the solid in acetonitrile at a temperature of about 30°C to about 50°C to form a ground solid.

[0294] In certain embodiments, the process further comprises separating the ground solids.

[0295] In certain embodiments, the anhydrous organic solvent is anhydrous acetonitrile.

[0296] In certain embodiments, the base is a trialkylamine base.

[0297] In certain embodiments, the base is diisopropylethylamine.

[0298] In certain embodiments, the compound of Formula IV is represented by Formula IVA:

[0299]

[0300] And the compound of formula V is represented by formula VA:

[0301]

[0302] Among them, R 4 is hydrogen or C 1-4 Alkyl and R 5 is hydrogen or C1-4 alkyl.

[0303] Also provided herein is a process for resolving a composition comprising one or more stereoisomers of a compound of Formula VI or a salt thereof:

[0304]

[0305] in

[0306] n is 0, 1, 2, 3 or 4;

[0307] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 Alkoxy;

[0308] The following steps are involved:

[0309] (a) contacting a composition comprising one or more stereoisomers of Formula VI with a chiral base to provide a chiral salt of a compound of Formula VI;

[0310] (b) cooling the composition of step (a);

[0311] (c) isolating a chiral salt of the compound of Formula VI; and

[0312] (d) hydrolyzing a chiral salt of a compound of Formula VI to provide an enantiomerically enriched composition comprising a compound of Formula VI or a salt thereof.

[0313] In certain embodiments, the chiral base is cinchonidine.

[0314] In certain embodiments, the contacting of step (a) is performed in a solvent selected from acetone / water or acetone / isopropanol.

[0315] In certain embodiments, the cooling of step (b) is performed at a temperature at or below about 15°C.

[0316] In certain embodiments, the cooling of step (b) is at a temperature of about 10°C to about 15°C.

[0317] In certain embodiments, the cooling of step (b) is maintained for at least about 8 hours.

[0318] In certain embodiments, the separating of step (c) is via filtration.

[0319] In certain embodiments, the process further comprises dissolving the chiral salt of the compound of Formula VI obtained from step (c) in a suitable solvent, and repeating steps (b) and (c) before step (d).

[0320] In certain embodiments, the process further comprises the steps of dissolving the chiral salt of the compound of Formula VI obtained from step (c) in a suitable solvent, and repeating steps (b) and (c) two, three, four, five or six times before step (d).

[0321] In certain embodiments, the compound of Formula VI is represented by Formula VIA:

[0322]

[0323] Among them, R 5 is hydrogen or C 1-4 alkyl.

[0324] Also provided herein is a process for preparing an enantiomerically enriched composition comprising a compound of Formula VII or a salt thereof:

[0325]

[0326] in

[0327] n is 0, 1, 2, 3 or 4;

[0328] Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 alkoxy; and

[0329] R 4 are independently hydrogen or C 1-4 alkyl;

[0330] Compositions comprising enantiomerically enriching a compound of Formula VI or a salt thereof:

[0331]

[0332] contacting in sequence: (i) a solution comprising KI and I2, (ii) a base, and (iii) a solution comprising a base and a 4 -OH to provide an enantiomerically enriched composition of a compound of Formula VII or a salt thereof.

[0333] Also provided herein is a process as claimed in claim 34, wherein the compound of formula VII is represented by formula VIIA:

[0334]

[0335] And the compound of formula VI is represented by formula VIA:

[0336]

[0337] Among them, R 5 is hydrogen or C 1-4 alkyl.

[0338] Also provided herein is a process for preparing an enantiomerically enriched composition comprising a compound of Formula VIIB or a salt thereof:

[0339]

[0340] Among them, R 4 is hydrogen or C 1-4 Alkyl, R 5 is hydrogen or C 1-4 Alkyl, comprising an enantiomerically enriched composition of a hydrolyzed compound of Formula IXB or a salt thereof:

[0341]

[0342] Using a base and a formula R 4 The hydrolysis composition of compounds having -OH is hydrolyzed to provide an enantiomerically enriched composition of a compound of Formula VIIB or a salt thereof.

[0343] Also provided herein is a process for resolving a composition comprising one or more stereoisomers of a compound of Formula VIIA or a salt thereof:

[0344]

[0345] in

[0346] R 5 is hydrogen or C 1-4 Alkyl, comprising the following steps:

[0347] (a) contacting a composition comprising one or more stereoisomers of Formula VIIA with a chiral base to provide a chiral salt of a compound of Formula VIIA;

[0348] (b) cooling the composition of step (a);

[0349] (c) isolating a chiral salt of a compound of formula VIIA; and

[0350] (d) hydrolyzing a chiral salt of a compound of Formula VIIA to provide an enantiomerically enriched composition comprising a compound of Formula VIIA or a salt thereof.

[0351] In some embodiments, the chiral base is (R)-1-amino-2-propanol. In some embodiments, the chiral base is L-phenylalaninol. In some embodiments, the chiral base is (S)-phenylglycinol. In some embodiments, the chiral base is (S)-diphenyl-2-pyrrolidinemethanol.

[0352] Also provided herein is a process for preparing an enantiomerically enriched composition comprising a compound of Formula IXB or a salt thereof:

[0353]

[0354] Among them, R 5 is hydrogen or C 1-4 Alkyl, including compositions that enantiomerically enrich a compound of Formula VIIIB or a salt thereof:

[0355]

[0356] Contacting with a base provides an enantiomerically enriched composition of the compound of Formula IXB or a salt thereof.

[0357] Also provided herein is a process for preparing an enantiomerically enriched composition comprising a compound of Formula VIIIB or a salt thereof:

[0358]

[0359] Among them, R 5 is hydrogen or C 1-4 Alkyl, including compositions that enantiomerically enrich a compound of Formula VIB or a salt thereof:

[0360]

[0361] Contacting a solution comprising KI and I2 to provide an enantiomerically enriched composition of the compound of Formula VIIIB or a salt thereof.

[0362] Also provided herein is a process for preparing an enantiomerically enriched composition comprising a compound of Formula VIIB or a salt thereof:

[0363]

[0364] in

[0365] R 4 is hydrogen or C 1-4 Alkyl; and

[0366] R 5 is hydrogen or C 1-4 alkyl;

[0367] The following steps are involved:

[0368] (a) A composition which enriches the enantiomer of a compound of formula VIB or a salt thereof:

[0369]

[0370] contacting a solution comprising KI and I2 to provide an enantiomerically enriched composition of a compound of Formula VIIIB or a salt thereof:

[0371]

[0372] (b) contacting the enantiomerically enriched composition of the compound of Formula VIIIB with a base to provide an enantiomerically enriched composition of the compound of Formula IXB or a salt thereof:

[0373]

[0374] (c) using a compound comprising a base and a compound of formula R 4 The hydrolysis composition of compounds having -OH hydrolyzes an enantiomerically enriched composition of a compound of Formula IXB or a salt thereof to provide an enantiomerically enriched composition of a compound of Formula VIIB or a salt thereof.

[0375] Also provided herein is a process for preparing an enantiomerically enriched composition comprising a compound of Formula VB or a salt thereof:

[0376]

[0377] in

[0378] R 4 is hydrogen or C 1-4 Alkyl; and

[0379] R 5 is hydrogen or C 1-4 alkyl;

[0380] An enantiomerically enriched composition comprising an isomerized compound of formula VIIB or a salt thereof:

[0381]

[0382] In certain embodiments, the isomerizing comprises exposing the compound of Formula VA to UV light.

[0383] In certain embodiments, when R 4 C 1-4 When alkyl, the process further comprises a hydrolysis step to convert R 4 Converted into hydrogen.

[0384] Also provided herein is a process for preparing an enantiomerically enriched composition comprising a compound of Formula IVB or a salt thereof:

[0385]

[0386] Among them, R 5 is hydrogen or C 1-4Alkyl, including compositions that enantiomerically enrich a compound of formula VB or a salt thereof:

[0387]

[0388] and contacting with N,N'-disuccinimidyl carbonate (DSC) under conditions to provide an enantiomerically enriched composition comprising a compound of Formula IVB.

[0389] In certain embodiments of any of the formulas described herein, R 1 In certain embodiments of any of the formulas described herein, R 1 For glycine.

[0390] In certain embodiments of any of the formulas described herein, n is 1.

[0391] In certain embodiments of any of the formulas described herein, R 2 C 1-4 In certain embodiments of any of the formulae described herein, R 2 It is methyl.

[0392] In certain embodiments of any of the formulas described herein, R 5 It is methyl.

[0393] Amino Acid Part

[0394] The term "amino acid" refers to both natural and non-natural amino acids. In certain embodiments, the amino acid moiety is a natural amino acid. Natural amino acids include 20 proteinogenic amino acids directly encoded by triplet codons in the genetic code, and include alanine (ala, A), arginine (arg, R), asparagine (asn, N), aspartic acid (asp, D), cysteine ​​(cys, C), glutamine (gln, Q), glutamic acid (glu, E), glycine (gly, G), histidine (his, H), isoleucine (ile, I), leucine (leu, L), lysine (lys, K), methionine (met, M), phenylalanine (phe, F), proline (pro, P), serine (ser, S), threonine (thr, T), tryptophan (trp, W), tyrosine (tyr, Y) and valine (val, V).

[0395] The amino acid moiety may also include unnatural amino acids. In the broadest sense, the term "amino acid moiety" includes any organic compound having amine (-NH2) and carboxylic acid (-CO2H) functional groups.

[0396] In certain embodiments, the amino acid moiety is selected from the group consisting of: -NR1c -C 1-4 Alkylene-optionally substituted heterocyclyl, -NR 1c -C 1-4 Alkylene-N(R 1d )2.

[0397] -N(R 1c )CHR 1e CO2H, -N(R 1c )-C 1-6 Alkylene-CO2H, -N(R 1f )-C 2-4 Alkylene-(N(C 1-4 Alkylene-CO2H)-C 2-4 Alkylene) m -N(C 1-4 Alkylene -CO2H)2, -N(R 1c )CHR 1e C(O)OC 1-6 Alkyl, -N(R 1c )-C 1-6 Alkylene-C(O)OC 1-6 Alkyl and -N(R 1f )-C 2-4 Alkylene-(N(C 1-4 Alkylene-C(O)OC 1-6 Alkyl)-C 2-4 Alkylene) m -N(C 1-4 Alkylene-C(O)OC 1-6 Alkyl)2; wherein,

[0398] R 1c and R 1d are independently hydrogen or C 1-4 alkyl;

[0399] R 1e For -C 1-4 Alkylene -CO2H, -C 1-4 Alkylene -CONH2 or -C 1-4 Alkylene –OH;

[0400] R 1f Hydrogen, -C 1-6 Alkyl or -C 1-4 Alkylene –CO2H; and

[0401] m is 0, 1, 2 or 3.

[0402] In certain embodiments, the amino acid moiety is glycine. In certain embodiments, the amino acid moiety is alanine.

[0403] Payload

[0404] As used herein, "payload moiety" refers to the payload D minus the nucleophilic group attached to the linker, such as NH, NC 1-4 alkyl, O or S, or minus its electrophilic group attached to the linker, such as C(O), i.e., the remainder of the payload. Generally, the term "payload" refers to an agent for delivery to a target site in a subject, and includes, but is not limited to, therapeutic agents, diagnostic agents, targeting agents, and the like.

[0405] In some embodiments, the effective load is a therapeutic agent, such as an antibiotic, an antifungal, an antiviral, an anticancer, a cardiovascular, a CNS, an anti-inflammatory / anti-arthritic, an anti-TB / anti-leprosy, an antihistamine / respiratory agent, a corticosteroid, an immunosuppressant, or an anti-ulcer agent. Specific therapeutic drugs include paclitaxel, doxorubicin, daunorubicin, etoposide, irinotecan, SN-38, docetaxel, gemcitabine, podophyllotoxin, carmustine, ixabepilone, patupirone, cyclosporin A, rapamycin, amphotericin, vancomycin, daptomycin, doxycycline, ceftriaxone, trimethoprim, sulfamethoxazole, acyclovir, nystatin, amphotericin B, flucytosine, emtricitabine, gentamicin, colistin, levodopa, oseltamivir, cephalexin, 5-aminolevulinic acid, cysteine, celecoxib, and nimodipine.

[0406] In certain embodiments, the payload moiety is an anthracycline moiety, an auristatin moiety, a glycopeptide antibiotic moiety, or a lipopeptide antibiotic moiety.

[0407] In certain embodiments, the payload moiety is a doxorubicin moiety, a daunorubicin moiety, a monomethyl auristatin E moiety, a vancomycin moiety, or a daptomycin moiety.

[0408] In certain embodiments, D is an antibiotic agent, an antifungal agent, an antiviral agent, an anticancer agent, a cardiovascular agent, a CNS agent, an anti-inflammatory / anti-arthritic agent, an anti-TB / anti-leprosy agent, an antihistamine / respiratory disease agent, a corticosteroid agent, an immunosuppressant, or an anti-ulcer agent.

[0409] In certain embodiments, D is an antibiotic. Suitable antibiotics include, but are not limited to, β-lactams, including penicillins and cephalosporins, such as thiamycin, monobactam, β-lactamase inhibitors, and methicillin; aminoglycosides, including streptomycin, gentamicin, kanamycin, tobramycin, amikacin, neomycin, ribosomycin, succinomimecin, and astamicin; tetracyclines, including tetracycline, oxytetracycline, chlortetracycline, and doxycycline; chloramphenicol, including chloramphenicol and thiamphenicol; macrolides, including erythromycin, leucomycin, erythromycin etoposide, and erythromycin sirolimus. , erythromycin ethylsuccinate, azithromycin, acetylspiramycin, midecamycin and josamycin; other antibiotics that act on gram-positive bacteria, such as lincomycin, clindamycin, vancomycin and bacitracin; other antibiotics that act on gram-positive bacteria, such as polymyxin, fosfomycin, schizotypin, cycloserine and rifampicin; antifungal antibiotics, such as griseofulvin; anticancer antibiotics, such as mitomycin, actinomycin D, bleomycin and doxorubicin; and immunosuppressive antibiotics, such as cyclosporine.

[0410] In some embodiments, D is an anticancer drug, an anticoagulant, a microbial immunosuppressant or an anti-restenotic drug. The anticancer drug can be selected from one or more of methotrexate, purine, pyrimidine, plant alkaloids, epothilone, triptolide compounds, antibiotics (particularly actinomycin D), hormones and antibodies. Among plant alkaloids, paclitaxel, adriamycin, maytansin, auristatin, calicheamicin, duocarmycin, tubulysin and camptothecin can be mentioned in particular. The anticoagulant can be selected from one or more of heparin, aspirin, hirudin, colchicine and platelet GPIIb / IIIa receptor antagonists.

[0411] The platelet GPIIb / IIIa receptor antagonist may be one or more selected from tirofiban, abciximab and eptifibatide. The microbial immunosuppressant may be one or more selected from the following: cyclosporin A, tacrolimus and its analogs, despergualin, mycophenolate mofetil, rapamycin and its derivatives, FR-900520 substance from Streptomyces strains, FR-900523 substance from Streptomyces strains, daclizumab, valeramide, kanglemycin C, spergualin, prodigiotensin-25C, tranilast, myriocin, cyclosporin C, brevitacillin, mycophenolic acid, brefeldin A and steroids. The anti-restenosis drug can be one or more selected from the following: batimastat, metalloproteinase inhibitors, 17β-estradiol, NO donor, 2-chlorodeoxyadenosine, 2-deoxycoformycin, fingolimod, mycophenolate sodium, ISATX247 (cyclosporine A derivative), esabucol, daclizumab, basiliximab, antithymocyte globulin, everolimus, methotrexate, neoadjuvant, cyclophosphamide, brequinar sodium, leflunomide and imidazolin.

[0412] In certain embodiments, D is an anticancer drug. Exemplary anticancer drugs include, but are not limited to, abiraterone acetate, Abitrexate (methotrexate), Abraxane (paclitaxel albumin-stable nanoparticle formulation), ABVD, ABVE, ABVE-PC, AC, AC-T, Adcetris (Bentuximab Restatin), ADE, Ado-Trastuzumab Emtansine, Adriamycin (doxorubicin hydrochloride), Adrucil (fluorouracil), Afatinib maleate, Afinitor (everolimus), Aiteler (imiquimod), Aldesleukin, Alemtuzumab, Eninda (pemetrexed disodium), Alexi (palonosetron hydrochloride), Ambochlorin (chlorambucil), Am boclorin (chlorambucil), aminolevulinic acid, anastrozole, aprepitant, Aredia (pamidronate disodium), Arimidex (anastrozole), Aromasin (exemestane), Arranon (nelarabine), arsenic trioxide, Arzerra (ofatumumab), asparaginase erwinia, Avastin (bevacizumab), axitinib, azacitidine, BEACOPP, bendamustine hydrochloride, BEP, bevacizumab, bexarotene, bexar (tositumomab and I131 iodotositumomab), bicalutamide, bleomycin, bortezomib, Bosulif (bosutinib), bosutinib, bentuximab, busulfan, busulfan ( busulfan), cabazitaxel, cabozantinib-S-malate, CAF, Campath (alemtuzumab), Camptosar (irinotecan hydrochloride), capecitabine, CAPOX, carboplatin, carboplatin-paclitaxel, carfilzomib, Casodex (bicalutamide), CeeNU (lomustine), Cerubidine (daunorubicin hydrochloride), Cervarix (recombinant HPV bivalent vaccine), cetuximab, chlorambucil, chlorambucil-prednisone, CHOP, cisplatin, Clafen (cyclophosphamide), clofarabine, Clofarex (clofarabine), Clolar (clofarabine), CMF, Cometriq (cabozantinib-S-malate), CO PP, COPP-ABV, Cosmegen (dactinomycin), crizotinib, CVP, cyclophosphamide, Cyfos (ifosfamide), cytarabine, cytarabine, liposomal cytarabine, Cydesa-U (cytarabine), Cytoxan (cyclophosphamide), dabrafenib, dacarbazine, Dakogin (decitabine), dactinomycin, dasatinib, daunorubicin hydrochloride, decitabine, degarelix, denileukin, denosumab, DepoCyt (liposomal cytarabine), DepoFoam (liposomal cytarabine), dexrazoxane hydrochloride, docetaxel, Dox (liposomal doxorubicin hydrochloride), doxorubicin hydrochloride, Dox-SL (liposomal doxorubicin hydrochloride),DTIC-Dome (dacarbazine), Efudex (fluorouracil), Ellite (rasburicase), Ellence (epirubicin hydrochloride), Eloxatin (oxaliplatin), Idrapolamine, Emend (aprepitant), Enzalutamide, Epirubicin hydrochloride, EPOCH, Erbitux (cetuximab), Eribulin mesylate, Erivedge (vismodegib), Erlotinib hydrochloride, Erwinaze (asparaginase), Etopoph os (etoposide phosphate), etoposide, etoposide phosphate, Evacet (liposomal doxorubicin hydrochloride), everolimus, Evacet (raloxifene hydrochloride), exemestane, Faleton (toremifene), Faslodex (fulvestrant), FEC, Femara (letrozole), filgrastim, Fludara (fludarabine phosphate), fludarabine phosphate, Fluoroplex (fluorouracil), fluorouracil, Folex (methotrexate), Folex PFS (methotrexate), Folfiri, Folfiri-bevacizumab, Folfiri-cetuximab, Folfirinox, Folfox (leucovorin, fluorouracil, oxaliplatin), Folotyn (pralatrexate), FU-LV, Fulvestrant, Gardasil (recombinant HPV quadrivalent vaccine), Gazyva (obitutuzumab), gefitinib, gemcitabine hydrochloride, gemcitabine-cisplatin, gemcitabine-oxaliplatin , Gemtuzumab uzumab, Gemzar (gemcitabine hydrochloride), Gitler (afatinib maleate), Gleevec (imatinib mesylate), Carboxypeptidase, Goserelin acetate, Halaven (eribulin mesylate), Herceptin (trastuzumab), HPV bivalent vaccine, HPV quadrivalent vaccine recombinant, recombinant, Hycamtin (topotecan hydrochloride), Hyper-CVAD, Ibritumomab tiuxetan, Ibrutinib, ICE, Iclusig (ponatinib hydrochloride ), Ifex (ifosfamide), ifosfamide, Ifosfamidum (ifosfamide), imatinib mesylate, Imbruvica (ibrutinib), imiquimod, Inlyta (axitinib), Intron A (recombinant interferon alpha-2b), iodine-131 tositumomab and tositumomab, ipilimumab, Iressa (gefitinib), irinotecan hydrochloride, Istodax (romidepsin), ixabepilone, Ixempra (i Sapiron), Jakafi (ruxolitinib phosphate), Jevtana (cabazitaxel), Kadcyla (Ado-trastuzumab emtansine), Keoxifene (raloxifene hydrochloride), Kepivance (palifermin), Kyprolis (carfilzomib), lapatinib ditosylate, lenalidomide, letrozole, folinic acid, chlorambucil (chlorambucil), leuprolide acetate, levan (aminolevulinic acid),Linfolizin (chlorambucil), LipoDox (liposomal doxorubicin hydrochloride), liposomal cytarabine, lomustine, Lupron (leuprolide acetate), Lupron Depot (leuprolide acetate), Lupron Depot-Ped (leuprolide acetate), Lupron Depot-3 Month (leuprolide acetate), Lupron Depot-4 Month (leuprorelin acetate), Marqibo (liposomal vincristine sulfate), Matulane (procarbazine hydrochloride), nitrogen mustard hydrochloride, Megace (megestrol acetate), megestrol acetate, Mekinist (trametinib), mercaptopurine, mesna, Mesnex (mesna), Methazolastone (temozolomide), methotrexate, methotrexate LPF (methotrexate), Mexate (methotrexate), Mexate-AQ (methotrexate), mitomycin C, Mitozytrex (mitomycin C), MOPP, M ozobil (plerixafor), Mustargen (nitrogen mustard hydrochloride), Mutacin (mitomycin C), Malelan (busulfan), Mylosar (azacitidine), Mylotarg (gemtuzumab ozogamicin), NP-paclitaxel (paclitaxel albumin-stable nanoparticle formulation), Navelbine (vinorelbine tartrate), Nelarabine, Neosar (cyclophosphamide), Eubogen (filgrastim), Nexavar (sorafenib tosylate), Nilotinib, Nolvadex (tamoxifen citrate), Nplate (romiplostim), Obinutuzumab, Ofatumumab Anti-, homoharringtonine, Oncaspar (pegaspargase), Ontak (denileukin), OEPA, OPPA, oxaliplatin, paclitaxel, paclitaxel albumin-stable nanoparticle formulation, palifermin, palonosetron hydrochloride, pamidronate disodium, panitumumab, Paraplat (carboplatin), Paraplatin (carboplatin), pazopanib hydrochloride, pegaspargase, pegylated interferon alfa-2b, PEG-Intron (pegylated interferon alfa-2b), pemetrexed disodium, Perjeta (pertuzumab), pertuzumab, Platinol (cisplatin) , Platinol-AQ (cisplatin), Plerixafor, Pomalidomide, Pomalyst (pomalidomide), Ponatinib hydrochloride, Pralatrexate, Prednisone, Procarbazine hydrochloride, Proleukin (aldesleukin), Prolia (denosumab), Promacta (iperapabolamide), Provenge (Cipruset-T), Purinethol (mercaptopurine), radium 223 dichloride, Raloxifene hydrochloride, Rasburicase, R-CHOP, R-CVP, Recombinant HPV bivalent vaccine, Recombinant HPV quadrivalent vaccine, Recombinant interferon α-2b, Regorafenib,Lenalidomide (lenalidomide), Rheumatrex (methotrexate), Retoxan (rituximab), Rituximab, Romidepsin, Romilastin, Daunorubicin (daunorubicin hydrochloride), Ruxolitinib phosphate, Sclerosant intrapleural aerosol (talc), Siprussin-T, Sorafenib tosylate, Sprycel (dasatinib), Stanford V, Sterile Talc (talc), Steritalc (talc), Stivarga (regorafenib), Sunitinib malate, Sutent (sunitinib malate), Sylatron (pegylated interferon alfa-2b), Synovir (thalidomide), Synribo (homoharringtonine), Tafinlar (dabrafenib), Talc, Tamoxifen citrate, Tarabine PFS (cytarabine), Tarceva (erlotinib hydrochloride), Targretin (bexarotene), Tasina (nilotinib), Taxol (paclitaxel), Taxotere (docetaxel), Temodar (temozolomide), temozolomide, temosirolimus, thalidomide, Thalomid (thalidomide), Toposar (etoposide), topotecan hydrochloride, toremifene, Tasina (temosirolimus), tositumomab, and 1 131-iodine tositumomab, Totect (dexrazoxane hydrochloride), trametinib, trastuzumab, Treanda (bendamustine hydrochloride), Trisenox (arsenic trioxide), Tykerb (lapatinib ditosylate), vandetanib, VAMP, Vectib (panitumumab), VelP, Velban (vinblastine sulfate), Velcade (bortezomib), Velsar (vinblastine sulfate), Vemurafenib, Vanbisen (etoposide), Viadur (leuprorelin acetate), Vidaza (azacitidine), vinblastine sulfate, Vincasar PFS (vincristine sulfate), vincristine sulfate, liposomal vincristine sulfate, vinorelbine tartrate, Vismodegib, Voraxaze (carboxypeptidase), Vorinostat, Votrient (pazopanib hydrochloride), Wellcovorin (leucovorin), Xalkori (crizotinib), Xeloda (capecitabine), Xelox, Xgeva (denosumab), Xofigo (radium 223 chloride), Xtandi (enzalutamide), Yervoy (ipilimumab), Zaltrap (ziv-aflibercept), Zelboraf (vemurafenib), Zevalin (ibritumomab tiuxetan), Zinecard (dexrazoxane hydrochloride), Ziv-aflibercept, Noradrelin (goserelin acetate), Zoledronic acid, Zolinza (vorinostat), Zometa (zoledronic acid), and Zytiga (abiraterone acetate).

[0413] In certain embodiments, D is a PBD dimer, calicheamicin, spermycin, tubulysin B, rhizobactin, dolastatin, tunicain B, camptothecin, CBI, temsirolimus, actinomycin D, epothilone B, paclitaxel, moniliformin, SN38, Velcade, brucei, DAVLBH, DM1, Phyllanthoside, Alimta, T2 toxin, MMC, vantalanib, vinorelbine, brefeldin, sunitinib, daunomycin, semaxanib, Tarceva, Iressa, irinotecan, LY-541503, geldanamycin, gemcitabine, methotrexate, Gleevec, topotecan, bleomycin, doxorubicin, cisplatin, N-mustards, etoposide, or 5-FU.

[0414] In certain embodiments, D is an anthracycline. In certain embodiments, D is a taxane. In certain embodiments, D is gemcitabine. In certain embodiments, D is doxorubicin. In certain embodiments, D is docetaxel. In certain embodiments, D is SN38. In certain embodiments, D is monomethyl auristatin E. In certain embodiments, D is dexamethasone. In certain embodiments, D is celecoxib. In certain embodiments, D is gentamicin.

[0415] In certain embodiments, D is an intracellular penetration enhancer. For example, D can be a functionalized keto acid, 6-oxo-6-phenylhexanoic acid, 8-oxo-8-phenyloctanoic acid, 8-(2,5-dichlorophenyl)-8-oxooctanoic acid, a functionalized keto acid ester or aldehyde, a modified amino acid, a modified amino acid, N-[8-(2-hydroxybenzoyl)aminooctanoic acid, N-[8-(2-hydroxybenzoyl)aminodecanoic acid, N-(5-chlorosalicyloyl)-8-aminooctanoic acid, N-[4-(4-chloro-2-hydroxybenzoyl)amino1 Butyric acid, 2-ethylhexyl 2-hydroxybenzoate, 5-cyclohexyl-5-oxopentanoic acid, 6-cyclohexyl-6-oxohexanoic acid, 7-cyclohexyl-7-oxoheptanoic acid, 8-cyclohexyl-8-oxooctanoic acid, 4-cyclopentyl-4-oxobutyric acid, 5-cyclopentyl-5-oxopentanoic acid, 6-cyclopentyl-6-oxohexanoic acid, 7-cyclopentyl-7-oxoheptanoic acid, 8-cyclopentyl-8-oxooctanoic acid, 4-cyclobutyl-4-oxobutyric acid, 5-cyclobutyl-5-oxopentanoic acid, 6-cyclopentyl-6-oxohexanoic acid, 7-cyclopentyl-7-oxoheptanoic acid, 8-cyclopentyl-8-oxooctanoic acid, Cyclobutyl-6-oxohexanoic acid, 7-cyclobutyl-7-oxoheptanoic acid, 8-cyclobutyl-8-oxooctanoic acid, 4-cyclopropyl-4-oxobutanoic acid, 5-cyclopropyl-5-oxopentanoic acid, 6-cyclopropyl-6-oxohexanoic acid, 7-cyclopropyl-7-oxoheptanoic acid, 8-cyclopropyl-8-oxooctanoic acid, 8-[(3-methylcyclohexyl)oxy]octanoic acid, 7-[(3-methylcyclohexyl)oxy]heptanoic acid, 6-[(3-methylcyclohexyl)oxy]hexanoic acid, 5-[(3-methylcyclohexyl)oxy]hexanoic acid [(3-methylcyclohexyl)oxy]pentanoic acid, 4-[(3-methylcyclohexyl)oxy]butanoic acid, 3-[(3-methylcyclohexyl)oxy]propionic acid, octyl salicylate, diketopiperazine, saponin, acylcarnitine, alkanoylcholine, taurodihydrofusidate, sulfoxide, oxazolidinone, pyrrolidone, alcohol or alkanol, benzoic acid, ethylene glycol, surfactant, terpene, any functionally effective salt of the foregoing, any derivative of the foregoing, or a combination thereof.

[0416] 3. Compounds

[0417] In certain embodiments, the present disclosure provides intermediate compounds that can be used in the processes described herein. Thus, in one embodiment, a compound selected from Table 1 or a salt thereof is provided. In another embodiment, an enantiomerically enriched composition comprising a compound selected from Table 1 or a salt thereof is provided.

[0418] In Table 1, and as used throughout, it will be understood that straight bold or dashed bonds are used to indicate relative stereochemistry, while wedge-shaped bold or dashed bonds are used to indicate absolute stereochemistry. When a composition is identified as enantiomerically enriched, it is intended that the composition contains greater than 50% of a single enantiomer, or at least about 55%, or at least about 60%, or at least about 65%, or at least about 70%, or at least about 75%, or at least about 80%, or at least about 85%, or at least about 90%, or at least about 95%, or at least about 97%, or about 99% ee.

[0419] Table 1

[0420]

[0421]

[0422]

[0423] It should be understood that, for clarity, certain features described herein that are described in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, various features described herein that are described in the context of a single embodiment may also be provided individually or in any suitable subcombination. All combinations of embodiments related to the chemical groups represented by the variables included in Formula I are expressly included herein, as are each combination that is individually and expressly stated, to the extent that such combinations include compounds that produce stable compounds (i.e., compounds that can be isolated, characterized, and tested for biological activity).

[0424] Examples

[0425] The disclosed compounds can be prepared using methods disclosed herein and conventional modifications thereof, which will be apparent to the disclosed content and methods well known in the art. In addition to the teachings herein, conventional and well-known synthetic methods can be used. The synthesis of compounds described herein can be achieved as described in the following examples. If available, reagents can be purchased commercially from, for example, Sigma Aldrich or other chemical suppliers. Unless otherwise indicated, the starting materials for the following reactions can be obtained from commercial sources.

[0426] Example 1: Chiral resolution of (Z)-1-methylcyclooct-4-ene-1-carboxylic acid

[0427] The following procedure can produce single enantiomers of compound IV with ≥97% ee on a kilogram scale from racemic starting materials, while previous reports only formed compound IV as a racemate on a small scale (Rossin, R. et al. Bioconjugate Chem. 2016, 1697-1706).

[0428]

[0429] (Z)-1-methylcyclooct-4-ene-1-carboxylic acid (34.3 kg) was dissolved in EA (1355 kg). A solution of cinchonidine (60 kg) in DCM (460 kg) was added at 35-40 ° C. The mixture was stirred at 45 ° C for 4 h and then cooled to 12 ° C for another 8 hours. The resulting suspension was filtered. The collected cake required further purification by recrystallization.

[0430] 1st recrystallization: The salt (220 g) was completely dissolved in isopropanol (1 L) and acetone (5 L) at 60° C. The mixture was kept at 10-15° C. overnight. Filtration and drying gave 148 g of product, 28.0% ee.

[0431] Second recrystallization: The salt (148 g) was completely dissolved in isopropanol (750 mL) and acetone (3850 mL) at 60°C. The mixture was kept at 10-15°C overnight. Filtered and dried to give 85.2 g of product, 54.0% ee. The mother liquor was concentrated to give 62.8 g of solid.

[0432] 3rd recrystallization: The salt (85.2 g) was completely dissolved in isopropanol (420 mL) and acetone (2100 mL) at 60°C. The mixture was kept at 10-15°C overnight. Filtered and dried to give 66.0 g of product, 74.3% ee. The mother liquor was concentrated to give 19.0 g of solid.

[0433] 4th recrystallization: The salt (66.0 g) was completely dissolved in isopropanol (330 mL) and acetone (1650 mL) at 60°C. The mixture was kept at 10-15°C overnight. Filtered and dried to give 42.7 g of product, 83.4% ee. The mother liquor was concentrated to give 13.3 g of solid.

[0434] 5th recrystallization: The salt (42.7 g) was completely dissolved in isopropanol (210 mL) and acetone (1050 mL) at 60°C. The mixture was kept at 10-15°C overnight. Filtered and dried to give 31.8 g of product, 91.0% ee. The mother liquor was concentrated to give 10.9 g of solid.

[0435] 6th recrystallization: The salt (31.8 g) was completely dissolved in isopropanol (160 mL) and acetone (800 mL) at 60°C. The mixture was kept at 10-15°C overnight. Filtered and dried to give 26.2 g of product, 95.3% ee. The mother liquor was concentrated to give 5.6 g of solid.

[0436] The concentrates of the mother liquors (ee>0%) can be combined and purified by several recrystallizations to give more of a single isomer of 4.

[0437] The solid of the salt (single isomer of (Z)-1-methylcyclooct-4-ene-1-carboxylic acid and cinchonidine, 6 kg) was mixed with water (35.0 kg). To the mixture was added HCl solution (2.0 kg of 12N concentrated hydrochloric acid, diluted with 10.0 kg of water) at 20-30°C to adjust the pH to 1. The resulting mixture was washed with hexane (3 x 15.0 kg). The combined organic phases were washed with brine (10 kg). The organic phases were concentrated at 35-55°C to give a single isomer of the title compound in the form of an oil.

[0438] Chromatographic conditions:

[0439] Chromatographic columns Superchiral S-AD Column size 0.46cm ID×15cm L, 5um injection 5μL Mobile phase Hexane / EtOH / DEA=99 / 1 / 0.03 (v / v / v) Flow rate 1.0mL / min wavelength UV 210nm and advanced laser polarimeter temperature 15℃

[0440] After salt decomposition, %ee improves:

[0441] -Conversion of the salt to the free acid of (Z)-1-methylcyclooct-4-ene-1-carboxylic acid by addition of HCl

[0442] - Dissolve (Z)-1-methylcyclooct-4-ene-1-carboxylic acid and add the desired enantiomer seed to induce crystallization

[0443] - Precipitation was observed, although analysis showed no difference in %ee between the solid and the supernatant

[0444] - Suggests that the racemate may exist in the form of solid solution, making enrichment of the acid form unfeasible. Improving the purity of the starting material:

[0445] - 47% low purity racemate was incorporated into pure enriched (Z)-1-methylcyclooct-4-ene-1-carboxylic acid (33% ee)

[0446] - Incorporation of impurities does not affect the relative solubility of the desired and undesired enantiomers

[0447] - Therefore, for the resolution process, no significant %ee improvement was observed using pure (Z)-1-methylcyclooct-4-ene-1-carboxylic acid compared to pure (Z)-1-methylcyclooct-4-ene-1-carboxylic acid

[0448] Dependence of the relative solubility of the desired / unwanted enantiomer on the %ee:

[0449] - The solubility of the desired and undesired enantiomers was measured in samples of various enantiomeric enrichments, ranging from 0-88% ee

[0450] -For racemic materials, the solubility of the undesired enantiomer exceeds that of the desired enantiomer

[0451] - However, once 46% ee is reached, the solubility trend is reversed, with the solubility of the desired enantiomeric salt exceeding that of the undesired enantiomeric salt

[0452] - This switch complicates the development of a high-yield chiral resolution process starting from the racemate to achieve ≥97% ee. Other solvents were screened and the results are shown below.

[0453]

[0454]

[0455] Final purity: 81.2%

[0456] Total yield: 13.7%

[0457] Example 2: Large-Scale Synthesis of Enantiomerically Enriched cis-(Z)-6-Hydroxy-1-methylcyclooct-4-ene-1-carboxylic Acid

[0458] Process for producing kilogram quantities of a single enantiomer (≥97% ee) of cis-(Z)-6-hydroxy-1-methylcyclooct-4-ene-1-carboxylic acid from racemic starting materials. The literature reports the synthesis of 17.5 g of racemic cis-(Z)-6-hydroxy-1-methylcyclooct-4-ene-1-carboxylic acid. (Rossin, R. et al. Bioconjugate Chem. 2016, 1697-1706).

[0459]

[0460] Compound 4 (34.3 kg) was dissolved in EA (1355 kg). A solution of cinchonidine (60 kg) in DCM (460 kg) was added at 35-40 ° C. The mixture was stirred at 45 ° C for 4 h and then cooled to 12 ° C for another 8 hours. The resulting suspension was filtered. The collected cake required further purification by recrystallization.

[0461] 1st recrystallization: The salt (220 g) was completely dissolved in isopropanol (1 L) and acetone (5 L) at 60° C. The mixture was kept at 10-15° C. overnight. Filtration and drying gave 148 g of product, 28.0% ee.

[0462] Second recrystallization: The salt (148 g) was completely dissolved in isopropanol (750 ml) and acetone (3850 mL) at 60°C. The mixture was kept at 10-15°C overnight. Filtered and dried to give 85.2 g of product, 54.0% ee. The mother liquor was concentrated to give 62.8 g of solid.

[0463] 3rd recrystallization: The salt (85.2 g) was completely dissolved in isopropanol (420 ml) and acetone (2100 mL) at 60°C. The mixture was kept at 10-15°C overnight. Filtered and dried to give 66.0 g of product, 74.3% ee. The mother liquor was concentrated to give 19.0 g of solid.

[0464] 4th recrystallization: The salt (66.0 g) was completely dissolved in isopropanol (330 ml) and acetone (1650 mL) at 60°C. The mixture was kept at 10-15°C overnight. Filtered and dried to give 42.7 g of product, 83.4% ee. The mother liquor was concentrated to give 13.3 g of solid.

[0465] 5th recrystallization: The salt (42.7 g) was completely dissolved in isopropanol (210 mL) and acetone (1050 mL) at 60°C. The mixture was kept at 10-15°C overnight. Filtered and dried to give 31.8 g of product, 91.0% ee. The mother liquor was concentrated to give 10.9 g of solid.

[0466] 6th recrystallization: The salt (31.8 g) was completely dissolved in isopropanol (160 mL) and acetone (800 mL) at 60°C. The mixture was kept at 10-15°C overnight. Filtered and dried to give 26.2 g of product, 95.3% ee. The mother liquor was concentrated to give 5.6 g of solid.

[0467] The concentrates of the mother liquors (ee>0%) can be combined and purified by several rounds of recrystallization to obtain more of the single isomer of (Z)-1-methylcyclooct-4-ene-1-carboxylic acid.

[0468] The solid of the salt (single isomer of (Z)-1-methylcyclooctyl-4-ene-1-carboxylic acid and cinchonidine, 6 kg) was mixed with water (35.0 kg). HCl solution (2.0 kg of 12N concentrated hydrochloric acid, diluted with 10.0 kg of water) was added to the mixture at 20-30°C to adjust the pH to 1. The resulting mixture was washed with hexane (3 x 15.0 kg). The combined organic phases were washed with brine (10 kg). The organic phases were concentrated at 35-55°C to give a single isomer of (Z)-1-methylcyclooct-4-ene-1-carboxylic acid in the form of an oil.

[0469] The product (2.8 kg) was dissolved in a mixture of DCM (22.2 kg) and water (16.7 kg), and NaHCO (4.8 kg) was added. The reaction mixture was cooled to 0-5 ° C, and a mixture of KI (8.2 kg) and iodine (8.4 kg) was added in batches. The reactant was stirred at room temperature for 2 h and then sampled for GC. When the starting material was no more than 2.0a% by GC, the reaction mixture was slowly quenched with sodium bisulfite (3.4 kg sodium bisulfite in 16.7 kg water) until a clear solution appeared. The two layers were separated, and the aqueous layer was extracted with DCM (22.2 kg). The combined organic layers were washed with water (8.4 kg), dried with Na SO (1.7 kg) and rotary evaporated to obtain enantiomerically enriched 5-iodo-1-methyl-7-oxabicyclo [4.2.2] decan-8-one (quantitative), which was used for the next step without further purification.

[0470] The obtained product (4.8 kg) was dissolved in toluene (21.3 kg) and DBU (2.9 kg) was then added. The mixture was allowed to stand at 25-30° C. for 12 hours and then heated under reflux for 4 hours, at which time GC showed complete conversion (5-iodo-1-methyl-7-oxabicyclo[4.2.2]dec-8-one<3a%). After cooling, the reaction mixture was washed with water (6.4 kg). The aqueous phase was extracted with toluene (10.7 kg). The organic phase was dried over Na2SO4 (2.9 kg) and then concentrated to a colorless oil (94% yield).

[0471] (Z)-1-methyl-7-oxabicyclo[4.2.2]deca-4-en-8-one (2.66 kg) was mixed with methanol (12.8 kg), KHCO (16.0 kg) and water (0.03 kg). The mixture was kept at 28° C. for 24 hours and monitored by HPLC. The reaction was considered complete when the starting material was NMT 26.0%. Filtered and the filter cake was washed with methanol. The filtrate was concentrated at 35-55° C. and EA (20 kg) and water (12.58 kg) were added. The pH of the mixture was adjusted to 2-3 by 2N HCl solution. The layers were separated and the aqueous phase was washed with EA (4 x 8.0 kg). The combined organic phases were washed with brine (17.2 kg), dried over Na2SO4 (1.3 kg) and then concentrated at 35-55° C. The residue was purified by chromatography using hexanes and EA (50:1) as eluent to give the pure title product as a colorless oil. (Note: The starting material can be recovered and the combined fractions can also be purified again to provide more product.)

[0472] Example 3: Synthesis of enantiomerically enriched 2,5-dioxapyrrolidin-1-yl cis-(E)-6-((((2,5-dioxapyrrolidin-1-yl)oxy)carbonyl)oxy)-1-methylcyclooct-4-ene-1-carboxylate from cis-(E)-6-hydroxy-1-methylcyclooct-4-ene-1-carboxylic acid

[0473] Process for the robust synthesis of high purity 2,5-dioxopyrrolidin-1-yl cis-(E)-6-((((2,5-dioxopyrrolidin-1-yl)oxy)carbonyl)oxy)-1-methylcyclooctyl-4-ene-1-carboxylate in 75% yield on a 100 gram scale from the TCO starting material cis-(E)-6-hydroxy-1-methylcyclooct-4-ene-1-carboxylic acid without the use of chromatography. The reference reports the synthesis of 400 mg of 2,5-dioxapyrrolidin-1-yl cis-(E)-6-((((2,5-dioxapyrrolidin-1-yl)oxy)carbonyl)oxy)-1-methylcyclooct-4-ene-1-carboxylate with a yield of 46%, which required purification by silica gel chromatography (Rossin, R. et al. Bioconjugate Chem. 2016, 1697-1706).

[0474]

[0475] A mixture of cis-(E)-6-hydroxy-1-methylcyclooct-4-ene-1-carboxylic acid (1 eq.), N,N′-disuccinimidyl carbonate (DSC) (4.3 eq.), DIPEA (7.4 eq.) in anhydrous acetonitrile (20 vol.) was stirred at 22 to 25° C. until the intermediate 2,5-dioxapyrrolidin-1-yl cis-(E)-6-hydroxy-1-methylcyclooctyl-4-ene-1-carboxylic acid was less than 0.5% AUC as determined by UPLC (7-9 h). The resulting suspension was added to DI water (50 vol.) at room temperature and the resulting mixture was stirred for about 15 min. The resulting solid suspension was filtered, washed with DI water (3×2 vol.), and dried on the filter under vacuum for about 1 h.

[0476] The solid crude 2,5-dioxapyrrolidin-1-yl cis-(E)-6-((((2,5-dioxapyrrolidin-1-yl)oxy)carbonyl)oxy)-1-methylcyclooct-4-ene-1-carboxylate was suspended in acetonitrile (about 2 volumes) and mixed at a temperature of 35 to 45° C. for about 1 hour. The mixture was cooled to about 15° C. and filtered. The filter cake was washed with acetonitrile (2×1 volume) and dried on the filter under vacuum for about 60 minutes to afford enantiomerically enriched 2,5-dioxapyrrolidin-1-yl cis-(E)-6-((((2,5-dioxapyrrolidin-1-yl)oxy)carbonyl)oxy)-1-methylcyclooct-4-ene-1-carboxylate as a white to off-white crystalline solid in an isolated yield of about 74-80% with an AUC purity of about 99% as determined by UPLC at 214 nm.

[0477] 106.0 g of 2,5-dioxapyrrolidin-1-yl cis-(E)-6-((((2,5-dioxapyrrolidin-1-yl)oxy)carbonyl)oxy)-1-methylcyclooct-4-ene-1-carboxylate was obtained from 61.8 g of cis-(E)-6-hydroxy-1-methylcyclooct-4-ene-1-carboxylic acid with an isolated yield of 75.4% and an AUC purity of 98.6%.

[0478] Analytical methods:

[0479]

[0480] Example 4: Chiral separation of compound VIIC

[0481]

[0482] A resolution screen was performed on Compound VIIC using each of the following chiral bases: (S)-2-amino-1-propanol (L-alaninol), (R)-(+)-1-phenylethylamine, L-(-)-2-amino-1-butanol, (1R,2S)-(-)-ephedrine, (S)-(+)-2-amino-3-methyl-1-butanol (L-valinol), (S)-(-)-N-benzyl-α-methylbenzylamine, (+)-dehydroabietinamine, (1S,2S)-(+)-2 -amino-1-phenyl-1,3-propanediol, (R)-(+)-3-pyrrolidinol, (S)-(+)-2-pyrrolidinemethanol (L-prolinol), (S)-(-)-1-(1-naphthyl)ethylamine, (R)-1-amino-2-propanol, L-prolineamide, (1R,2R)-(-)-pseudoephedrine, L-phenylalaninol, (1R,2R)-2-amino-1-(4-nitrophenyl)propane-1,3-diol, cinchonine, quinidine, quinine, cinchonidine 、(S)-(+)-L-phenylglycinol、(R)-(-)-2-phenylglycine amide、(R)-(+)-2-phenylpropylamine、L-phenylalanine amide、(S)-diphenyl-2-pyrrolidinemethanol、(1S,2S)-(+)-N-p-toluenesulfonyl-1,2-diphenylethylenediamine、(1S,2R)-(+)-2-amino-1,2-diphenyl-ethanol、N-methyl-D-glucamine、(1S,2S)-(-)1,2-diphenylethylenediamine、 (1R,2R)-diaminocyclohexane, (R)-(2-methoxyphenyl)ethylamine, (S)-p-chlorophenylethylamine, (R)-(+)-1-(4-methylphenyl)ethylamine, (S)-2-amino-1,1-diphenyl-1-propanol, (1S,2S)-(-)1,2-diphenylethylenediamine and (1R,2R)-diaminocyclohexane in each of the following solvents: water, acetonitrile, 1:1 water:ethanol, ethanol, IPA, MEK, iPrOAc and dioxane. Selected results for the resolution of Compound VIIC on a 100 mg scale starting from 18 mL of solvent are shown below.

[0483] Toluene EtOAc i TBME <![CDATA[Et2O]]> acetone MEK R-1-amino-2-propanol 1 equivalent × -2 / 48 -7 / 37 -8 / 52 -3 / 54 -62 / 65 -7 / 48 0.5 equivalent -48 / 58 × 0.5 equivalent + 0.5 equivalent NaOH × 0.5 equivalent + 0.5 equivalent NaOH × R-phenyl glycol 1 equivalent x x 56 / -75 x x x x 0.5 equivalent 61 / -42

[0484] 1. 100 mg of compound VIIC and 44 mg of R-aminopropanol in 18 mL of iPrOAc: S = 109 mg -7% ML = 36 mg 37%.

[0485] 2. 100 mg of compound VIIC and 22 mg of R-aminopropanol in 18 mL of iPrOAc: S = 68 mg -48% ML = 54 mg 58%.

[0486] 3. 100 mg of compound VIIC and 81 mg of R-phenyl glycol in 18 mL of iPrOAc: S = 79 mg 56% ML = 98 mg -75%.

[0487] 4. 100 mg of compound VIIC and 40 mg of R-phenyl glycol in 18 mL of iPrOAc: S = 58 mg 61% ML = 86 mg - 42%.

[0488] 5. 100 mg of compound VIIC and 150 mg of L-diphenylpyrrolidinemethanol in 18 mL of iPrOAc: S = 167 mg 0% ML = 50 mg 0%.

[0489] 6. 100 mg of compound VIIC and 75 in 18 mL iPrOAc: S = 104 mg 0% ML = 63 mg 0%

[0490] mg L-diphenylpyrrolidinemethanol.

[0491] Example 5: Chiral Resolution of (Z)-6-Hydroxy-1-methylcyclooct-4-ene-1-carboxylic acid

[0492] The following example shows the chiral resolution of (Z)-6-hydroxy-1-methylcyclooct-4-ene-1-carboxylic acid using (R)-2-amino-2-phenylethane-1-ol, which provides the enantiomer opposite to that provided in Example 2. To distinguish between the two enantiomers, the (-) sign is assigned to the %ee and does not indicate optical rotation.

[0493] (Z)-6-Hydroxy-1-methylcyclooct-4-ene-1-carboxylic acid (8.00 g, 1 eq., 43.4 mmol) was dissolved in 900 isopropyl acetate in a 2 L three-necked flask at 68° C. (some slight white insoluble material smeared on the glass and remained on the glass throughout the first step). (R)-2-amino-2-phenylethan-1-ol (5.96 g, 1 eq., 43.4 mmol) was added and the mixture was stirred at 135 rpm with an overhead stirrer.

[0494] The clarified mixture was cooled at a rate of 10°C / hour and seeded with -61%ee seed crystals every two degrees of cooling. Crystallization occurred at 48°C. The mixture was further cooled to 35°C at a rate of 10°C / hour and then allowed to reach room temperature overnight.

[0495] Filtration gave 7.2 g of solid with ee = -55% and 6.6 g of material in mother liquor with ee = 83%.

[0496] 6.2 g of the solid obtained in step 1 was suspended in 255 mL of isopropyl acetate in a 1 L three-necked flask and heated to 70°C while stirring at 135 rpm with an overhead stirrer. After 5 minutes, the mixture was allowed to reach room temperature overnight. Filtration gave 5.1 g of solid with ee = -76% and 1.0 g of material in a mother liquor with ee = 72%.

[0497] 3.7 grams of solid obtained in step 2 were suspended in 20 mL of 2-butanone in a 100 mL round bottom flask and heated to reflux while stirring with a magnetic stirrer. 12 mL of methanol were added, after which the suspension became a clear solution. The mixture was allowed to reach room temperature overnight, during which it crystallized. Filtration gave 2.0 grams of solid with ee = -92% and 1.7 grams of material in the mother liquor with ee = -71%.

[0498] 1.0 g of the solid obtained in step 3 was suspended in 10 mL of isopropanol in a 100 mL round bottom flask and heated to 60°C while stirring with a magnetic stirrer. After 5 minutes, the mixture was allowed to reach room temperature overnight. Filtration gave 0.85 g of solid with ee = -100% and 0.15 g of material in a mother liquor with ee = -56%.

[0499] Example 6: Synthesis of trans-cyclooctene (TCO)-doxorubicin conjugate 12

[0500] Procedure for compound 11:

[0501] Process for synthesizing trans-cyclooctene (TCO)-doxorubicin conjugate 11 in 98% yield on a gram scale from TCO starting material 10. Literature reference reports the synthesis of 134 mg of 11 in 68% yield. (Rossin, R. et al. Bioconjugate Chem. 2016, 1697-1706).

[0502]

[0503] To a solution of doxorubicin HCl in DMF (10 mL, 10 V) (1.0 g, 1.72 mmol, 1.0 eq) was added 10 (874.0 mg, 1.2 eq) followed by DIPEA (0.9 mL, 3.0 eq). The mixture was stirred at room temperature for 1.5 h and HPLC showed the formation of product 11 (LCAP: 88.5%) and a small amount of doxorubicin (LCAP: 0.3%). The assay yield of 11 was 102.6%. The reaction mixture was diluted with DCM (40 mL) and washed with water (total: 250 mL, monitor HPLC to ensure that most of the DMF was washed out) and brine (10 mL). The DCM extract was dried over Na2SO4 and concentrated to give a crude residue. 1.7 g of crude 11 (95.1% LCAP) was isolated.

[0504] The residue was purified by silica gel chromatography to afford 11 as a red solid. 1.44 g of purified 11 (94.8% LCAP and 97.1% LCWP) was isolated in an uncorrected yield of 98.0%. Neither 10 nor doxorubicin was detected in the isolated material.

[0505] Since the purity of 11 did not seem to be significantly improved after silica gel purification, this step may not be necessary.

[0506] Procedure for compound 12:

[0507]

[0508] One-pot method for obtaining 10 to 12 in 89% purity without chromatography: Compound 10 (874.0 mg, 1.2 eq) was added to a solution of doxorubicin HCl (1.0 g, 1.72 mmol, 1.0 eq) in DMF (10 mL, 10 V) followed by DIPEA (0.9 mL, 3.0 eq). The mixture was stirred at room temperature for 1.25 h, and HPLC showed the formation of product 11 (LCAP: 87.3%) and a small amount of doxorubicin (LCAP: 0.4%). The yield was determined to be 95.2%. To the reaction mixture was added a suspension of glycine (1.29 g, 10.0 eq), N,O-bis(trimethylsilyl)acetamide (BSA) (2.11 mL, 5.0 eq), DIPEA (6.0 mL, 20.0 eq) in DCM (20 mL). The resulting mixture was stirred at room temperature for 24 h. HPLC showed that the LCAP of 12 was 87.7% and that of 11 was 4.4%.

[0509] The reaction mixture was filtered through a pad of celite and washed with DCM (3 mL). The filtrate was concentrated to give a residue (4.9 g), which was mixed with water (26 mL) and 4 mL of 1N NaOH was added to help dissolve the solid at 0°C. The mixture was then extracted with MTBE, but an emulsion was found to have formed, more water (5 mL) and 1N NaOH (2 mL) and MTBE were added, and two layers were eventually formed, the aqueous layer (total volume: about 120 mL) was extracted with MTBE (3 x 60 mL), and then acidified to pH = 1-2 with 1N HCl.

[0510] The orange precipitate was collected by filtration, washed with water and dried in vacuo to afford 1.25 g of 12 (89.3% LCAP) in an uncorrected yield of 89.2%.

[0511] One-pot method for 10 to 12 using chromatography Compound 10 (218.5 mg, 1.2 eq) was added to a solution of doxorubicin HCl (250 mg, 0.431 mmol, 1.0 eq) in DMF (2.5 mL, 10 V), followed by DIPEA (0.23 mL, 3.0 eq). The mixture was stirred at room temperature for 3 h, and HPLC showed the formation of product 11 (LCAP: 91.8%) and a small amount of doxorubicin (LCAP: 0.3%). The yield was determined to be 92%. A pre-prepared suspension of glycine (323.6 mg, 10 eq), BSA (0.528 mL, 5 eq) and DIPEA (1.5 mL, 20 eq) in 5 mL DCM was added to the reaction mixture, which was stirred at room temperature for 2-3 h. The resulting mixture was stirred at room temperature for 25 h. After filtration and concentration, the residue was subjected to reverse phase ISCO. The collected fractions were lyophilized.

[0512] One-pot method for 10 to 12, alternative method - reverse order of addition: Compound 10 (218.5 mg, 1.2 eq) was added to a solution of doxorubicin HCl (250 mg, 0.431 mmol, 1.0 eq) in DMF (2.5 mL, 10 V), followed by DIPEA (0.23 mL, 3.0 eq). The mixture was stirred at room temperature for 1 h, and HPLC showed the formation of product 11 (LCAP: 88.8%) and a small amount of doxorubicin (LCAP: 0.5%). The yield was determined to be 93.3%. The reaction mixture was used directly as described below.

[0513] i) BSA (0.105 mL, 5 equiv.) and DIPEA (0.30 mL, 20 equiv.) were added to a suspension of glycine (64.7 mg, 0.86 mmol, 10 equiv.) in 1 mL DCM at room temperature under nitrogen. The mixture was refluxed for 2 h and cooled at room temperature. The reaction mixture of 11 (0.5 mL) was added thereto. The resulting mixture was heated at 40 ° C for 5 h and monitored by HPLC. The reaction mixture was then cooled to room temperature, filtered through a diatomaceous earth pad, and washed with DCM. The filtrate was concentrated to give a residue (0.64 g), which was mixed with 0.2 N NaOH (0.5 mL), and the mixture was extracted with MTBE (2x1 mL). The aqueous layer was then acidified to pH = 4-5 with 1 N HCl (0.2 mL). A colloidal precipitate was formed and collected, which showed that the LCAP of 12 was 88.7%. The aqueous phase was mainly a mixture of water and DMF.

[0514] ii) To a suspension of glycine (64.7 mg, 0.862 mmol, 10 equiv) in 1 mL of DCM was added BSA (0.105 mL, 10 equiv) and DIPEA (0.30 mL, 40 equiv) at room temperature under nitrogen. The mixture was stirred at room temperature for 5 min, and then the reaction mixture [24-R174] (0.5 mL) was added. The resulting mixture was stirred at room temperature for 22 h. HPLC showed 90.4% LCAP of 12, with no other peaks > 5% LCAP. The reaction mixture was filtered through a celite pad and washed with DCM. The filtrate was concentrated to give a residue, which was subjected to reverse phase ISCO purification. The collected fractions were lyophilized.

[0515] Comparative synthesis: using TMCSI as a protective agent, 11 is converted into 12: under nitrogen at room temperature to glycine (89.3mg, 1.19mmol, 10 equivalents) in 6mL 60% CHCl / MeCN in a suspension of TMSCl (127.7mg, 10 equivalents). The mixture is refluxed for 2h and cooled at room temperature. DIPEA (0.41mL, 20 equivalents) is added, followed by compound 11 (100.0mg, 1.0 equivalents). The resulting mixture is heated at 65°C for 4.5h and monitored by HPLC. The mixture is then diluted with water (10mL) and extracted with DCM (3x 25mL). The DCM extract is purified by silica gel chromatography to obtain compound 12 (7.3mg, 24-165-2, the LCAP of 12 is: 78.8%). The aqueous layer was purified by reverse phase ISCO (0-50% MeCN / water) to afford 12 (19.6 mg, LCAP of 12: 87.6%). The isolated (uncorrected) yield was 28.3%.

[0516] Formation of compound 12-Na:

[0517] The 2000mL flask is equipped with an overhead stirrer, a temperature probe and a nitrogen inlet / outlet. Acetone (5 volumes, 128mL) is added to the flask, and stirred at 20 to 25 ℃ under nitrogen. With crude compound 12 (25.6g, 0.0315mol, 1 equivalent, this amount is from weight analysis %) and a portion of acetone (7 volumes, 179mL) slurry, and load into the flask. The remaining acetone is used for rinsing the free acid flask of compound 12, and is added to the 2000mL flask. This mixture is purged with nitrogen for a short time, then covered under nitrogen.

[0518] To the stirred suspension was added water (3.3 volumes, 84.5 mL, degassed under vacuum to remove oxygen) portionwise at 20 to 25° C. The mixture was completely dissolved (checked with flush light).

[0519] NaHCO3 (0.592 M, 5% in USP water, 53.3 mL, approximately 1 eq., degassed in vacuo) was added portionwise to the mixture with stirring under nitrogen at 20-25° C. The mixture was filtered and stirred at 20 to 25° C. for at least 30 min.

[0520] A solid suspension was formed by adding acetone (about 35 volumes 896 mL) portionwise to the resulting solution in a 2000 mL flask over about 10 min at 19 to 24° C. The mixture was stirred at 20-24° C. for about 30 min.

[0521] Stirring was stopped to allow solids to solidify on the bottom of the flask (over about 1 min). The mixture was filtered. The flask was rinsed with acetone (3 x 75 mL) and the rinses were used to wash the solid filter cake.

[0522] The wet solid was transferred to a 500 mL round bottom flask and dried under vacuum at room temperature (20 to 25 °C) until a constant weight was achieved.

[0523] Example 7: Synthesis of trans-cyclooctene (TCO)-doxorubicin conjugate 12-Ala

[0524]

[0525] General procedure for compound 11: Compound 10 (240 mg, 0.569 mmol, 1.0 equiv) was dissolved in dry DMF (6 mL), followed by the addition of doxorubicin (395 mg, 0.683 mmol, 1.2 equiv, HCl salt) and N,N-diisopropylethylamine (367 mg, 2.85 mmol, 5.0 equiv). The solution was stirred overnight at room temperature under nitrogen and aluminum foil protection. The reaction mixture was diluted with DCM, washed with 5% aqueous citric acid solution, dH2O and brine. The organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to give compound 11 (200 mg) as a red solid.

[0526] TLC: DCM / methanol / acetic acid (20:1:0.2);

[0527] R f (Compound 11) = 0.35;

[0528] LC-MS: 851.2[M+H] + ;

[0529] General procedure for 12-Ala: L-Alanine (52.6 mg, 0.59 mmol, 10 equiv), BSA (60 mg, 0.294 mmol, 5.0 equiv), DIPEA (152 mg, 1.18 mmol, 20.0 equiv) in DCM (0.5 mL) were stirred for 30 min and added to a solution of compound 11 (50.0 mg, 0.059 mmol, 1.0 equiv) in DMF (0.2 mL). The reaction mixture was heated at 32 °C under nitrogen and aluminum foil overnight. The reaction mixture was concentrated to dryness and the residue was diluted with acetonitrile (2 mL). After lyophilizing the fractions with the desired mass to dryness, the resulting crude reaction mixture was purified by preparative HPLC (5% to 100% CH3CN in 30 minutes, neutral, pH 7) to give 12-Ala (2.3 mg). LC-MS: 846.8 [M+H] + ,822.5[MH]-; 1 H NMR (300MHz, CD3OD): δ7.85(m,1H),7.78(m,1H),7.52(m,1H),5.64(m,1H),5.40(m,2H),5.07(m,2H),4.74(s,1H),4.24(m,2H),3.82(s, 3H),3.82(m,1H),3.61(m,2H),2.98(m,2H),2.90(m,2H),2.38–1.74(m,10H),1.33(d,J=6.3Hz,3H),1.26(d,J=6.3Hz,3H),1.09(s,3H).

[0530] Example 8: Comparative Example

[0531] Alternative synthetic methods for trans-cyclooctene (TCO)-doxorubicin conjugates (Dox-TCO-Gly) were evaluated.

[0532] Direct coupling of glycine to Dox-TCO-NHS (without protecting groups) is inefficient

[0533]

[0534] TMS protection route

[0535]

[0536] TMSCl (638 mg, 5.9 mmol, 10 equiv) was added in one portion to a suspension of glycine (440 mg, 5.9 mmol, 10 equiv) in 50 mL of CHCl3: MeCN (60%: 40%). The resulting mixture was stirred at reflux (80°C) for 2 h and then cooled to room temperature. DIPEA (2 mL, 12 mmol, 20 equiv) and Dox-TCO-NHS (500 mg, 0.58 mmol) were added at room temperature and the mixture was stirred at 65°C for 6 h. Analysis of crude HPLC showed a yield of 78%. After removing the solvent, the residue was dissolved in 5 mL of water and purified by reverse phase chromatography (MeCN: water, 0 → 50%) to give Dox-TCO-Gly (235 mg, 49%) as an orange solid.

[0537] Alternative activating groups: PNPs lead to lower Dox conjugation yields compared to NHS

[0538] PNP – 53%

[0539]

[0540] NHS – 72%

[0541]

[0542] Alternative Protecting Groups: Fmoc Route - 4 steps, 17% total

[0543]

[0544] Alternative protecting groups: tBu route - 4 steps, no desired product obtained

[0545]

[0546] Various acidic deprotection conditions (e.g., TFA, AcOH, HCl, H3PO2) were screened in different solvents - the desired product was never observed. Deprotection was always observed by mass along with the addition of water (likely the Dox glycosidic bond or the TCO olefin).

[0547] Direct alcohol mono-activation-carboxylic acid activation of TCO is favored

[0548]

[0549] Alternative pathway for alcohol activation - less efficient than NHS dual activation pathway

[0550] Alternative Path 1

[0551]

[0552] Alternative Path 2

[0553]

[0554] Alternative Path 3

[0555]

[0556] Example 9: Pecidatinib-TCO-Glycine Conjugate by BSA-Protection Method:

[0557] This example provides methods for conjugating TCO starting materials such as those described herein (eg, compound 10) to pesidatinib. Further modification of compound 15 with an amino acid moiety (eg, glycine) is contemplated.

[0558]

[0559] To a solution of compound 13 (418 mg, 1.0 mmol) in DMF (10 mL) was added NaH (about 60%, 44 mg, 1.1 mmol) at 0°C. The reaction mixture was stirred under N2 for 1 h, and then Bis-NHS-TCO (398 mg, 1.0 mmol) was added. The resulting mixture was stirred at room temperature for 18 h. The reactant was quenched with water (10 mL). The product was extracted with ethyl acetate (2×40 mL). The combined organic layers were washed with water (4×15 mL) and brine (15 mL), dried (MgSO4) and evaporated in vacuo. The product was purified by flash chromatography on silica gel, eluted with EA / Hex (0%-100%) to give intermediate 14 (463 mg, 64%) as a white solid. LCMS: R t =1.281,m / z 725[M+1] +

[0560]

[0561] To a solution of N-hydroxysuccinimide ester 14 (200 mg, 0.28 mmol) and DMF (1.6 mL) was added a solution of glycine (207 mg, 2.8 mmol) in N, O-bis(trimethylsilyl)acetamide (285 mg, 1.4 mmol) and DIPEA (724 mg, 5.6 mmol) in DCM (3.2 mL). The resulting mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS and HPLC to show that most (68%) of the content was the desired product compound 15 (m / z 684.8 (+) and 682.5 (-), MW: 685.1). The reaction mixture was diluted with DCM (10 mL) and then filtered through a celite pad. The filtrate was concentrated to dryness. The residue was purified by preparative HPLC (0.1% formic acid as buffer) to give compound 15 (116.2 mg, 61%) as a white solid. LCMS: R t =1.013,m / z 685[M+1] + and 1368[2M+1] + . 1 H NMR (300MHz, CHCl3) δ8.61 (s, 1H), 8.46 (d, J = 1.8Hz, 1H), 7.94-7.80 (m, 2H), 7.68-7.60(m,2H),7.57(s,1H),7.50(dd,J=8.7 and 1.8Hz,1H),6.50(d,J=9.0H z,1H),6.34-6.16(m,2H),5.66(dd,J=16.2 and 2.4Hz,1H),5.57(s,1H),4.59(s ,2H),3.92-3.80(m,4H),2.38-1.80(m,7H),1.74-1.68(m,1H),1.17(s,3H).

[0562] Example 10: Etoposide-TCO conjugate

[0563] This example provides methods for conjugating TCO starting materials such as those described herein (eg, Compound 10) to etoposide.

[0564]

[0565] To a solution of etoposide (50 mg, 0.08 mmol) in DMF 5 mL, TCO-PNP (30 mg, 0.10 mmol) and DMAP (21 mg, 0.16 mmol) were added. The mixture was stirred for 3 days. It was quenched with 20 mL of water. The mixture was extracted with EtOAc (3×30 mL), and the combined organic phases were dried and concentrated. The residue was purified on a column to give etoposide-TCO compound 16 (30 mg, 54%). 1 H NMR(500MHz, CDCl3)δ6.84(s,1H),6.53(s,1H),6.27(s,2H),6.03–5.85(m, 3H),5.51(dd,J=16.5,1.7Hz,1H),5.33(s,1H),4.92(d,J=3.4Hz,1H),4.74( q,J=5.0Hz,1H),4.60(dd,J=15.7,6.4Hz,2H),4.42(dd,J=26.1,16.7Hz,1H) ,4.29–4.12(m,2H),3.68(s,6H),3.62(dd,J=17.9,9.1Hz,1H),3.59–3.52(m ,1H),3.41–3.36(m,1H),3.37–3.22(m,5H),2.86(tdd,J=10.9,8.0,3.3Hz, 1H),2.49(d,J=10.3Hz,1H),2.19(t,J=11.9Hz,1H),2.08–1.94(m,2H),1.89 (dt,J=15.1,6.4Hz,1H),1.68(ddd,J=19.0,15.3,9.7Hz,2H),1.59–1.46(m, 1H),1.38(d,J=5.0Hz,3H),1.22–1.10(m,1H),0.81(td,J=14.6,5.6Hz,1H).

[0566] Example 11: Aniline mustard-TCO conjugate

[0567]

[0568] To a solution of 8-1 (212 mg, 0.72 mmol) in DMF (3 mL) was added 8-2 (305 mg, 0.72 mmol) and DIEA (279 mg, 2.17 mmol). The solution was stirred at room temperature overnight. After the solvent was removed, DCM (30 mL) was added. The organic matter was washed with saturated NaHCO3 (40 mL), water (30 mL), brine (20 mL), dried over Na2SO4, and concentrated to give a residue. The residue was triturated with ether (2×5 mL) and dried to give compound 8-3 (310 mg, 86%) as a light yellow solid.

[0569] To a solution of 8-3 (310 mg, 0.62 mmol) in THF (4 mL) and H2O (1 mL) was added LiOH·H2O (77 mg, 1.86 mmol) and DIEA (279 mg, 2.17 mmol). The solution was stirred at room temperature overnight. After removing the solvent, the residue was purified by preparative HPLC (water and ACN, 0.1% formic acid) to give compound 8-4 (140 mg, 56%).

[0570] To a solution of 8-4 (140 mg, 0.34 mmol) in DCM (4 mL) was added TEA (172 mg, 1.72 mmol) and MsCl (79 mg, 0.85 mmol). The solution was stirred at room temperature for 4 hours. DCM (10 mL) was added. The organics were washed with water (2×10 mL), brine (10 mL), dried over Na2SO4 and concentrated. The residue was dried under high vacuum to give crude 8-5 (171 mg).

[0571] LiCl (252 mg, 6.0 mmol) was added to a solution of 8-5 (170 mg, 0.3 mmol) in DMF (2 mL). The mixture was stirred at room temperature for 2 hours and then stirred at 60 ° C overnight. After removing the solvent, DCM (10 mL) was added. The organic layer was washed with water (10 mL), brine (10 mL), dried over Na2SO4, and concentrated to give a residue. The residue was purified by preparative HPLC (water and ACN, 0.1% formic acid) to give compound 8-6 (86 mg, 64%, in two steps) as an off-white powder. LCMS: 443 [M + H] + . 1 H NMR(300MHz,CD3OD)δ7.25(d,J=7.8Hz,2H),6.72(d,J=7.8Hz,2H),6.00(m,1H),5.71 (m,1H),5.18(s,1H),3.71-3.61(m,8H),2.27-1.67(m,8H),1.28(s,1H),1.12(s,3H).

[0572] ***

[0573] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0574] The disclosure illustratively described herein may be suitably practiced in the absence of any element or elements, limitation or limitations not specifically disclosed herein. Thus, for example, the terms "comprises," "comprising," "containing," and the like should be interpreted broadly and without limitation. In addition, the terms and expressions employed herein have been used as terms of description rather than limitation, and it is not intended that such terms and expressions be used to exclude any equivalents of the features shown and described, or portions thereof, but it is to be recognized that various modifications may be made within the scope of the disclosure claimed.

[0575] Therefore, it should be understood that although the present disclosure has been specifically disclosed through preferred embodiments and optional features, those skilled in the art may make modifications, improvements and variations to the disclosure embodied herein, and these modifications, improvements and variations are considered to be within the scope of the present disclosure. The materials, methods and examples provided herein represent preferred embodiments, are exemplary and are not intended to limit the scope of the present disclosure.

[0576] The disclosure has been described broadly and generally herein. Each narrower species and subgeneric grouping falling within the generic disclosure also forms a part of the disclosure. This includes the generic description of the disclosure with proviso or negative limitation removing any subject matter from the genus, regardless of whether or not excised material is specifically recited herein.

[0577] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0578] All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety to the same extent as if each were individually incorporated by reference. In the event of a conflict, the present specification, including definitions, will control.

[0579] It should be understood that although the present disclosure has been described in conjunction with the above embodiments, the foregoing description and examples are intended to illustrate rather than limit the scope of the present disclosure. For those skilled in the art to which the present disclosure belongs, other aspects, advantages and modifications within the scope of the present disclosure will be apparent.

Claims

1. A process for preparing a compound of formula I and its salt: in R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety; n is 0, 1, 2, 3 or 4; Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 alkoxy; and D is the effective load part; R 4 is hydrogen or C 1-4 alkyl; This includes making a compound of formula II: Contact HO-R 4 , an optionally substituted heterocyclyl or an amino acid moiety thereof or a salt thereof, and a compound of formula III: in, Each R 3 In the presence of a base in an organic solvent, independently C 1-4 alkyl.

2. The process of claim 1, wherein: The payload moiety is an anthracycline moiety, an auristatin moiety, a glycopeptide antibiotic moiety, or a lipopeptide antibiotic moiety.

3. The process of claim 2, wherein: The payload moiety is a doxorubicin moiety, a daunorubicin moiety, a monomethyl auristatin E moiety, a vancomycin moiety or a daptomycin moiety.

4. The process according to any one of claims 1 to 3, wherein R 1 is an amino acid moiety, an alanine moiety or a glycine moiety.

5. The process according to any one of claims 1 to 4, wherein The compound of formula III is N,O-bis(trimethylsilyl)acetamide.

6. The process according to any one of claims 1 to 5, wherein The compound of formula I is represented by formula IA: in R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety; R 4 is hydrogen or C 1-4 alkyl; R 5 is hydrogen or C 1-4 Alkyl; and D is the effective load part; And the compound of formula II is represented by formula IIA:

7. The process of any one of claims 1 to 5, further comprising preparing a compound of formula II by reacting a compound of formula IV: The payload moiety or a salt thereof is contacted in the presence of a base in an organic solvent.

8. The process of claim 6, wherein: The compound of formula IV is represented by formula IVA:

9. A process for preparing a compound of formula X or a salt thereof: in R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety; n is 0, 1, 2, 3 or 4; Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 alkoxy; and R 4 is hydrogen or C 1-4 alkyl; This includes making a compound of formula IV: Contact HO-R 4 , an optionally substituted heterocyclic group or an amino acid thereof or a salt thereof and a compound of formula III: in, Each R 3 In the presence of a base in an organic solvent, independently C 1-4 alkyl.

10. The process of claim 9, wherein: The compound of formula X is represented by formula XA: And the compound of formula IV is represented by formula IVA: Among them, R 5 is hydrogen or C 1-4 alkyl.

11. The process of claim 9, further comprising preparing a compound of formula IV or a salt thereof, the process comprising treating a compound of formula V or a salt thereof: The N,N'-disuccinimidyl carbonate is contacted in the presence of a base in an organic solvent.

12. The process of claim 11, wherein: The compound of formula V is represented by formula VA: Among them, R 5 is hydrogen or C 1-4 alkyl.

13. A process for preparing a compound of formula I or a salt thereof: in R 1 Selected from the group consisting of: -OR 4 , an optionally substituted heterocyclyl, and an amino acid moiety; n is 0, 1, 2, 3 or 4; Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 Alkoxy; D is the payload portion; and R 4 is hydrogen or C 1-4 alkyl; The method comprises making a compound of formula X or a salt thereof: The payload moiety or a salt thereof is contacted in the presence of a base in an organic solvent.

14. The process of claim 13, wherein: The compound of formula I is represented by formula IA: And the compound of formula X is represented by formula XA: Among them, R 5 is hydrogen or C 1-4 alkyl.

15. A process for preparing a composition comprising a compound of formula IV or a salt thereof: in n is 0, 1, 2, 3 or 4; and Each R 2 are independently hydrogen or C 1-4 alkyl; This includes making one equivalent of a compound of formula V or a salt thereof: 3-5 equivalents of N,N'-disuccinimidyl carbonate are contacted in 15-25 volumes of anhydrous organic solvent in the presence of a base at a temperature of 15-30°C for 5-10 hours.

16. The process of claim 15, further comprising adding the composition comprising the compound of Formula IV to 40-60 volumes of water at room temperature to form an aqueous product mixture.

17. The process of claim 16, further comprising separating solids formed in the aqueous product mixture.

18. The process of claim 17, further comprising grinding the solid in acetonitrile at a temperature of about 30°C to about 50°C to form a ground solid.

19. The process of claim 18, further comprising separating the ground solids.

20. The process of claim 15, wherein: The anhydrous organic solvent is anhydrous acetonitrile.

21. The process of any one of claims 15 to 20, wherein: The base is a trialkylamine base.

22. The process of claim 21, wherein: The base is diisopropylethylamine.

23. The process of any one of claims 15 to 22, wherein: The compound of formula IV is represented by formula IVA: And the compound of formula V is represented by formula VA: Among them, R 4 is hydrogen or C 1-4 alkyl; and R 5 is hydrogen or C 1-4 alkyl.

24. A process for resolving a composition comprising one or more stereoisomers of a compound of Formula VI or a salt thereof: in n is 0, 1, 2, 3 or 4; Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 Alkoxy; The following steps are involved: (a) contacting a composition comprising one or more stereoisomers of Formula VI with a chiral base to provide a chiral salt of a compound of Formula VI; (b) cooling the composition of step (a); (c) isolating a chiral salt of the compound of formula VI; as well as (d) hydrolyzing a chiral salt of a compound of Formula VI to provide an enantiomerically enriched composition comprising a compound of Formula VI or a salt thereof.

25. The process of claim 24, wherein: The chiral base is cinchonidine.

26. The process of claim 24, wherein: The contacting of step (a) is performed in a solvent selected from acetone / water or acetone / isopropanol.

27. The process of claim 24, wherein: The cooling of step (b) is performed at a temperature at or below about 15°C.

28. The process of claim 24, wherein: The cooling of step (b) is at a temperature of about 10°C to about 15°C.

29. The process of claim 24, wherein: The cooling of step (b) is maintained for at least about 8 hours.

30. The process of claim 24, wherein: The separation of step (c) is by filtration.

31. The process of claim 24, further comprising dissolving the chiral salt of the compound of formula VI obtained from step (c) in a suitable solvent, and repeating steps (b) and (c) before step (d).

32. The process of claim 30, wherein: a step of dissolving the chiral salt of the compound of formula VI obtained in step (c) in a suitable solvent, and repeating steps (b) and (c) two, three, four, five or six times before step (d).

33. The process of any one of claims 24 to 32, wherein: The compound of formula VI is represented by formula VIA: Among them, R 5 is hydrogen or C 1-4 alkyl.

34. A process for preparing an enantiomerically enriched composition comprising a compound of formula VII or a salt thereof: in n is 0, 1, 2, 3 or 4; Each R 2 Independently for C 1-4 Alkyl, C 1-4 Haloalkyl and C 1-4 alkoxy; and R 4 are independently hydrogen or C 1-4 alkyl; Compositions comprising enantiomerically enriching a compound of Formula VI or a salt thereof: contacting in sequence: (i) a solution comprising KI and I2, (ii) a base, and (iii) a solution comprising a base and a 4 -OH to provide an enantiomerically enriched composition of a compound of Formula VII or a salt thereof.

35. The process of claim 34, wherein: The compound of formula VII is represented by formula VIIA: And the compound of formula VI is represented by formula VIA: Among them, R 5 is hydrogen or C 1-4 alkyl.

36. A process for preparing an enantiomerically enriched composition comprising a compound of Formula VIIB or a salt thereof: in, R 5 is hydrogen or C 1-4 Alkyl, comprising an enantiomerically enriched composition of a hydrolyzed compound of Formula IXB or a salt thereof: Using a base and a formula R 4 The hydrolysis composition of compounds having -OH is hydrolyzed to provide an enantiomerically enriched composition of a compound of Formula VIIB or a salt thereof.

37. A process for resolving a composition comprising one or more stereoisomers of a compound of Formula VIIA or a salt thereof: in R 4 is hydrogen or C 1-4 Alkyl, comprising the following steps: (a) contacting a composition comprising one or more stereoisomers of Formula VIIA with a chiral base to provide a chiral salt of a compound of Formula VIIA; (b) cooling the composition of step (a); (c) isolating a chiral salt of a compound of formula VIIA; as well as (d) hydrolyzing a chiral salt of a compound of Formula VIIA to provide an enantiomerically enriched composition comprising a compound of Formula VIIA or a salt thereof.

38. The process of claim 37, wherein: The chiral base is (R)-1-amino-2-propanol, L-phenylalaninol, (S)-phenyl glycol, and (S)-diphenyl-2-pyrrolidinemethanol.

39. A process for preparing an enantiomerically enriched composition comprising a compound of Formula IXB or a salt thereof: in, R 5 is hydrogen or C 1-4 Alkyl, including compositions that enantiomerically enrich a compound of Formula VIIIB or a salt thereof: Contacting with a base provides an enantiomerically enriched composition of the compound of Formula IXB or a salt thereof.

40. A process for preparing an enantiomerically enriched composition comprising a compound of Formula VIIIB or a salt thereof: in, R 5 is hydrogen or C 1-4 Alkyl, including compositions that enantiomerically enrich a compound of Formula VIB or a salt thereof: Contacting a solution comprising KI and I2 to provide an enantiomerically enriched composition of the compound of Formula VIIIB or a salt thereof.

41. A process for preparing an enantiomerically enriched composition comprising a compound of Formula VIIB or a salt thereof: in R 4 is hydrogen or C 1-4 Alkyl; and R 5 is hydrogen or C 1-4 alkyl; Includes the following step: (a) A composition which enriches the enantiomer of a compound of formula VIB or a salt thereof: contacting a solution comprising KI and I2 to provide an enantiomerically enriched composition of a compound of Formula VIIIB or a salt thereof: (b) contacting the enantiomerically enriched composition of the compound of Formula VIIIB with a base to provide an enantiomerically enriched composition of the compound of Formula IXB or a salt thereof: (c) using a compound comprising a base and a compound of formula R 4 The hydrolysis composition of compounds having -OH hydrolyzes an enantiomerically enriched composition of a compound of Formula IXB or a salt thereof to provide an enantiomerically enriched composition of a compound of Formula VIIB or a salt thereof.

42. A process for preparing an enantiomerically enriched composition comprising a compound of formula VB or a salt thereof: in R 4 is hydrogen or C 1-4 Alkyl; and R 5 is hydrogen or C 1-4 alkyl; An enantiomerically enriched composition comprising an isomerized compound of formula VIIB or a salt thereof:

43. The process of claim 42, wherein: The isomerization comprises exposing the compound of formula VA to UV light.

44. The process of claim 42 or 43, wherein When R 4 C 1-4 When alkyl, the process further comprises the step of 4 The hydrolysis step converts to hydrogen.

45. A process for preparing an enantiomerically enriched composition comprising a compound of Formula IVB or a salt thereof: in, R 5 is hydrogen or C 1-4 Alkyl, including compositions that enantiomerically enrich a compound of formula VB or a salt thereof: and contacting with N,N'-disuccinimidyl carbonate (DSC) under conditions to provide an enantiomerically enriched composition comprising a compound of Formula IVB.

46. ​​A process as claimed in any preceding claim, wherein R 2 or R 5 It is methyl.

47. An enantiomerically enriched composition comprising a compound selected from Table 1 or a salt thereof: Table 1 48. A process as claimed in any preceding claim, wherein The enantiomerically enriched composition comprises a %ee greater than about 25%, or about 50%, or about 70%, or about 80%, or about 90%, or about 95%, or about 98%, or about 99%, or about 100%.

Images