Synthetic process for manufacture of ecteinin compounds

Abstract

The present invention relates to a process for converting a compound of formula (I) to a compound of formula (II), and to the compound of formula (I).

Description

Technical Field

[0001] This invention relates to a method of synthesis, and more particularly to a method of synthesis for producing seasquirting compounds. Background Technology

[0002] Sea tegrins are a group of naturally occurring marine compounds and their synthetic analogues, which have been well identified and structurally characterized and disclosed to possess antibacterial and cytotoxic properties. See, for example, European Patent 309.477; WO 03 / 66638; WO03 / 08423; WO 01 / 77115; WO 03 / 014127; WO 2018 / 197663; R. Sakai et al., 1992, Proc.Natl. Acad. Sci. USA 89, pp. 11456-11460; R. Menchaca et al., 2003, J.Org.Chem.68(23), pp. 8859-8866; and I. Manzanares et al., 2001, Curr. Med. Chem. Anti-Cancer Agents, 1, pp. 257-276; and references therein. Examples of tunicates are provided by ET-743, ET-729, ET-745, ET-759A, ET-759B, ET-759C, ET-770, ET-815, ET-731, ET-745B, ET-722, ET-736, ET-738, ET-808, ET-752, ET-594, ET-552, ET-637, ET-652, ET-583, ET-597, ET-596, ET-639, ET-641 and their synthetic analogues (such as acetylated, formylated, methylated and oxide forms).

[0003] However, there remains a need to develop more efficient methods (especially semi-synthetic routes) to produce tunicate compounds and related compounds. This invention addresses this need. Summary of the Invention

[0004] According to one aspect of the present invention, a method for synthesizing a sucrose compound is provided, the method comprising the step of oxidizing a reaction mixture containing a compound of formula I to obtain a compound of formula II:

[0005]

[0006] in:

[0007] R1 is OH or CN;

[0008] R2 is hydrogen or Prot NH ;and

[0009] R3 is hydrogen or Prot OH ,

[0010] Among them Prot NH It is a protecting group for amino groups, and Prot OH It is a protecting group for OH.

[0011] This invention has identified that compound HQ-3 is highly insoluble in both organic and aqueous media, resulting in precipitate formation and reduced overall process yield. The amount of precipitate generated during the reaction process can be reduced by decreasing the amount of compound HQ-3 produced as a downstream intermediate, particularly by oxidizing compound I to compound II in the early stages of sucrose compound synthesis. Therefore, this yields a more efficient and higher-yield method, especially on a plant scale.

[0012] In a preferred embodiment, R1 is OH.

[0013] In the preferred aspect, R1 is CN.

[0014] In a preferred embodiment, R2 is hydrogen.

[0015] In the preferred embodiment, R2 is Prot NH .

[0016] In terms of preference, Prot NH It is selected from allyl carbamate (Alloc), 2,2,2-trichloroethyl carbamate (Troc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbamate (Fmoc), CONHPh, CSNHPh and tert-butyl carbamate (Boc), preferably tert-butyl carbamate (Boc).

[0017] In terms of alternatives, Prot NH CONHPh is not included.

[0018] In terms of alternatives, Prot NH It is selected from allyl carbamate (Alloc), 2,2,2-trichloroethyl carbamate (Troc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbamate (Fmoc), CSNHPh and tert-butyl carbamate (Boc), preferably tert-butyl carbamate (Boc).

[0019] In a preferred embodiment, R3 is hydrogen.

[0020] In a preferred embodiment, oxidation is performed using an oxidizing agent, biochemical oxidation, or electrochemical oxidation.

[0021] In a preferred aspect, the oxidant is selected from the group consisting of: metal oxidants (such as Cr(VI) compounds, Mn(IV) compounds, Mn(VII) compounds, Fe(III) compounds, Cu(II) compounds, Ag(I) compounds, Pb(IV) compounds, Ce(IV) compounds), molecular oxygen, hydrogen peroxide, hypochlorites (such as NaOCl and Ca(OCl)2), chlorites, 1,4-benzoquinone and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone or combinations thereof.

[0022] In a preferred embodiment, the oxidant is selected from molecular oxygen, hydrogen peroxide, 1,4-benzoquinone, and metal oxidants or combinations thereof.

[0023] In a preferred embodiment, the metal oxidizing agent is a transition metal oxidizing agent. In a preferred embodiment, the transition metal is Fe, Cu, or Mn.

[0024] In a preferred aspect, the transition metal oxidant is selected from Fe(III) compounds (e.g., FeCl3 or K3[Fe(CN)6]), Cu(II) compounds (e.g., CuSO4), Mn(III) compounds (e.g., Mn(OAc)3), Mn(IV) compounds (e.g., MnO2) and Mn(VII) compounds (e.g., KMnO4).

[0025] In a preferred embodiment, the Fe(III) compound is FeCl3.

[0026] In a preferred embodiment, the oxidant may be a halogen-based oxidant. The halogen may be iodine (e.g., I₂ and HIO₃).

[0027] In a preferred embodiment, the oxidant may be a quinone (e.g., tetrachlorobenzoquinone).

[0028] In a preferred aspect, the oxidant has an equivalent number of about 1.0 equivalents to about 15.0 equivalents relative to the compound of formula I in the reaction mixture, preferably about 1.2 equivalents to about 10.0 equivalents, more preferably about 1.5 equivalents to about 8.0 equivalents, and even more preferably about 2.0 equivalents to about 5.0 equivalents.

[0029] In a preferred embodiment, the oxidant is molecular oxygen.

[0030] In a preferred embodiment, molecular oxygen is provided by exposing the reaction mixture to a reaction atmosphere containing molecular oxygen.

[0031] In a preferred aspect, during the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II, a stream containing molecular oxygen is bubbled through the reaction mixture.

[0032] In a preferred aspect, the reaction is carried out at a pH of about 3.0 to about 9.0, preferably about 3.5 to about 8.5, and more preferably about 4.0 to about 8.0.

[0033] In a preferred embodiment, the biochemical oxidation is catalyzed by hydroquinone oxidase or by cells exhibiting hydroquinone oxidase activity.

[0034] In a preferred aspect, the reaction mixture further comprises a compound according to Formula II; preferably wherein R1 in the compound according to Formula I in the reaction mixture is the same as R1 in the compound according to Formula II in the reaction mixture, and / or wherein R2 in the compound according to Formula I in the reaction mixture is the same as R2 in the compound according to Formula II in the reaction mixture, and / or wherein R3 in the compound according to Formula I in the reaction mixture is the same as R3 in the compound according to Formula II in the reaction mixture; more preferably wherein R1, R2 and R3 in the compound according to Formula I in the reaction mixture are the same as R1, R2 and R3 in the compound according to Formula II in the reaction mixture.

[0035] In a preferred aspect, the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II is carried out at about 0°C to about 100°C, preferably about 0°C to about 80°C, more preferably about 5°C to about 60°C, even more preferably about 5°C to about 40°C, even more preferably about 10°C to about 30°C, and most preferably about 10°C to about 25°C.

[0036] In a preferred aspect, the method further includes a step of cyanidation to convert R1 from OH to CN before or after the step of oxidizing the reaction mixture containing compound I to obtain compound II; preferably, the cyanidation reaction is carried out after the step of oxidizing the reaction mixture containing compound I to obtain compound II.

[0037] In a preferred aspect, the method further includes, before or after the step of oxidizing the reaction mixture containing Formula I to obtain the compound of Formula II, performing an amino protection reaction to convert R2 from hydrogen to Prot. NH The step; preferably, the amino protection reaction is carried out after the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II.

[0038] In a preferred embodiment, R1 in Formula I is OH, and R2 in Formula I is hydrogen, and the method:

[0039] The process further includes a step of immediately following the step of oxidizing the reaction mixture containing compound I to obtain compound II, to perform a cyanidation reaction to convert R1 from OH to CN; and

[0040] This further includes immediately following the cyanation reaction to convert R1 from OH to CN, an amino protection reaction to convert R2 from hydrogen to Prot. NH The steps.

[0041] In a preferred embodiment, R1 in Formula I is CN, and R2 in Formula I is hydrogen, and the method:

[0042] This further includes a step of immediately performing a cyanidation reaction to convert R1 from OH to CN before the step of oxidizing the reaction mixture containing compound I to obtain compound II; and

[0043] The process further includes, immediately after the step of oxidizing the reaction mixture containing compound I to obtain compound II, carrying out an amino protection reaction to convert R2 from hydrogen to Prot. NH The steps.

[0044] In a preferred embodiment, the steps of oxidizing the reaction mixture containing compound I to obtain compound II and performing a cyanidation reaction to convert R1 from OH to CN are carried out as a one-pot reaction.

[0045] According to another aspect of the present invention, a compound according to Formula I is provided:

[0046]

[0047] R1 to R3 are defined as described in this paper, and the condition is that when R2 is CONHPh, then R3 is hydrogen.

[0048] In a preferred embodiment, the compound has the following formula:

[0049] .

[0050] In a preferred embodiment, the compound has the following formula:

[0051] .

[0052] In a preferred embodiment, the compound has the following formula:

[0053] .

[0054] According to another aspect of the present invention, a compound of the following formula is provided:

[0055] Detailed Implementation

[0056] The following applies to all aspects of this invention.

[0057] General chemical definition

[0058] In the compounds of the present invention, the alkyl group may be branched or unbranched, and preferably has 1 to about 12 carbon atoms. A more preferred class of alkyl groups has 1 to about 6 carbon atoms. Even more preferred are alkyl groups having 1, 2, 3 or 4 carbon atoms. Methyl, ethyl, n-propyl, isopropyl and butyl (including n-butyl, isobutyl, sec-butyl and tert-butyl) are particularly preferred alkyl groups in the compounds of the present invention.

[0059] In the compounds of the present invention, the alkenyl group may be branched or unbranched, having one or more double bonds and 2 to about 12 carbon atoms. A more preferred class of alkenyl groups has 2 to about 6 carbon atoms. Even more preferred are alkenyl groups having 2, 3, or 4 carbon atoms. Vinyl, 1-propenyl, 2-propenyl, 1-methylvinyl, 1-butenyl, 2-butenyl, and 3-butenyl are particularly preferred alkenyl groups in the compounds of the present invention.

[0060] In the compounds of the present invention, the alkynyl group may be branched or unbranched, having one or more triple bonds and 2 to about 12 carbon atoms. A more preferred class of alkynyl groups has 2 to about 6 carbon atoms. Even more preferred are alkynyl groups having 2, 3, or 4 carbon atoms.

[0061] Suitable aryl groups in the compounds of the present invention include monocyclic and polycyclic compounds, including polycyclic compounds containing separate and / or fused aryl groups. Typical aryl groups contain 1 to 3 separate and / or fused rings and 6 to about 18 carbon ring atoms. Preferably, the aryl group contains 6 to about 10 carbon ring atoms. Particularly preferred aryl groups include substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted biphenyl, substituted or unsubstituted phenanthryl, and substituted or unsubstituted anthraceneyl.

[0062] Suitable heterocyclic groups include heteroaromatic and heterocyclic groups containing 1 to 3 separate and / or fused rings and 5 to 18 ring atoms. Preferably, the heteroaromatic and heterocyclic groups contain 5 to 10 ring atoms, most preferably 5, 6 or 7 ring atoms. The suitable heteroaromatic groups in the compounds of the present invention contain one, two, or three heteroatoms selected from N, O, or S atoms, and include, for example, coumarin (including 8-coumaryl), quinolinyl (including 8-quinolinyl), isoquinolinyl, pyridyl, pyrazinyl, pyrazolyl, pyrimidinyl, furanyl, pyrroleyl, thiopheneyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, imidazoleyl, indolyl, isoyndolyl, indazoleyl, inazinyl, phthalazinyl, pteridinyl, purineyl, oxadiazolyl, thiadiazolyl, furazonyl, pyridazinyl, triazinyl, cenolinyl, benzimidazolyl, benzofuranyl, benzofuranyl, benzothiopheneyl, benzothiazolyl, benzooxazolyl, quinazolinyl, naphthidyl, and furanylpyridinyl. The compounds of the present invention contain one, two, or three heteroatoms selected from N, O, or S, and include, for example, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydrothioranyl, piperidinyl, morpholinyl, thiomorpholinyl, thiaxyl, piperazine, aziridine, oxacyclobutyl, oxacyclobutyl, thiohexacyclobutyl, high-piperidinyl, oxacycloheptyl, thiohexacycloheptyl, oxacycloheptenyl, diazacycloyl, thioaziridine, 1,2,3 ,6-Tetrahydropyridyl, 2-pyrrolinyl, 3-pyrrolinyl, dihydroindolyl, 2H-pyranyl, 4H-pyranyl, dioxane, 1,3-dioxolanecycloyl, pyrazolinyl, dithiaalkyl, dithiocyclopentyl, dihydropyranyl, dihydrothiophenyl, dihydrofuranyl, pyrazolyl, imidazolinyl, imidazolinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, 3H-indolyl and quinazinyl.

[0063] The above groups can be replaced by one or more suitable groups (such as OR', =O, SR', SOR', SO2R', NO2, NHR', NR'R', =N-R', NHCOR', N(COR')2, NHSO2R', NR'C(=NR')NR'R', CN, halogen, COR', COOR', OCOR', OCONHR', OCONR'R', CONHR', CONR'R', protected OH, protected amino, protected SH, substituted or unsubstituted C1-C) at one or more available positions. 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12The R' group is substituted with an alkynyl group, a substituted or unsubstituted aryl group, and a substituted or unsubstituted heterocyclic group, wherein each of the R' groups is independently selected from the group consisting of: hydrogen, OH, NO2, NH2, SH, CN, halogen, COH, COalkyl, CO2H, substituted or unsubstituted C1-C. 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne groups, substituted or unsubstituted aryl groups, and substituted or unsubstituted heterocyclic groups. When such groups are substituted themselves, the substituents may be selected from the aforementioned list. Furthermore, when there is more than one R' group on the substituent, each R' may be the same or different.

[0064] In the compounds of this invention, halogen substituents include F, Cl, Br, and I.

[0065] The term "pharmaceutically acceptable salt" refers to any pharmaceutically acceptable salt that, when administered to a patient, can (directly or indirectly) provide the compounds described herein. However, it should be understood that non-pharmaceutically acceptable salts also fall within the scope of this invention, as those can be used to prepare pharmaceutically acceptable salts. The preparation of salts can be performed by methods known in the art.

[0066] For example, pharmaceutically acceptable salts of the compounds provided herein are synthesized by conventional chemical methods from parent compounds containing a basic or acidic moiety. Typically, such salts are prepared, for example, by reacting the free acid or base of these compounds with a stoichiometric amount of a suitable base or acid in water or an organic solvent, or in a mixture of both. Non-aqueous media (such as ether, ethyl acetate, ethanol, 2-propanol, or acetonitrile) are generally preferred. Examples of acid addition salts include inorganic acid addition salts (such as, for example, hydrochlorides, hydrobroms, hydroiodates, sulfates, nitrates, and phosphates) and organic acid addition salts (such as, for example, acetates, trifluoroacetates, maleates, fumarates, citrates, oxalates, succinates, tartrates, malates, mandelates, methanesulfonates, and p-toluenesulfonates). Examples of base addition salts include inorganic salts (such as, for example, sodium, potassium, calcium, and ammonium salts) and organic base salts (such as, for example, ethylenediamine salts, ethanolamine salts, N,N-dialkylene ethanolamine salts, triethanolamine salts, and basic amino acid salts).

[0067] The compounds of the present invention may be in crystalline or amorphous form as free compounds or solvates (e.g., hydrates), and all forms are within the scope of the present invention. Solvation methods are generally known in the art.

[0068] Stereoisomerism with respect to asymmetric carbons having unspecified stereochemistry is possible, thus in which case the asymmetric carbon can have (R) or (S) configurations. All diastereomers and mixtures thereof arising from the combination of a particular configuration of such asymmetric carbons with other asymmetric carbons present in the molecule are considered to be within the scope of this invention. Stereoisomerism (geometric isomerism) with respect to double bonds is also possible, thus in some cases the molecule may exist as (E)-isomers or (Z)-isomers. If the molecule contains several double bonds, each double bond will have its own stereoisomer, which may be the same as or different from the stereoisomers of the other double bonds in the molecule. Furthermore, the compounds mentioned herein can exist as transisomers. Individual stereoisomers (including diastereomers, geometric isomers, and transisomers) and mixtures thereof of the compounds mentioned herein fall within the scope of this invention.

[0069] Furthermore, the compounds mentioned herein may exist in isotopically labeled forms. All pharmaceutically acceptable salts, esters, and isotopically labeled forms of the compounds mentioned herein, as well as mixtures thereof, are considered to be within the scope of this invention.

[0070] The protected forms of the compounds disclosed herein are considered to be within the scope of this invention. Suitable protecting groups are well known to those skilled in the art. A review of protecting groups in organic chemistry is provided by Wuts, PGM, and Greene TW in Protecting Groups in Organic Synthesis. The 4th edition of Wiley-Interscience, and Kocienski PJ's description in Protecting Groups, 3rd edition, Georg Thieme Verlag. These references provide sections on protecting groups for OH, amino, and SH groups. All of these references are incorporated in their entirety by reference.

[0071] Within the scope of this invention, an OH protecting group is defined as an O-bonded portion formed by protecting OH through the formation of a suitable protected OH group. Examples of such protected OH groups include ethers, silyl ethers, esters, sulfonates, sulfenates and sulfinates, carbonates, and carbamates. In the case of ethers, the protecting group of OH may be selected from methyl, methoxymethyl, methylthiomethyl, (phenyldimethylsilyl)methoxymethyl, benzyloxymethyl, p-methoxybenzyloxymethyl, [(3,4-dimethoxybenzyl)oxy]methyl, p-nitrobenzyloxymethyl, o-nitrobenzyloxymethyl, [(R)-1-(2-nitrophenyl)ethoxy]methyl, (4-methoxyphenoxy)methyl, guaiacol methyl, [(p-phenylphenyl)oxy]methyl, tert-butoxymethyl, 4-pentenoxymethyl, siloxymethyl, 2-methoxyethoxymethyl, 2-cyanoethoxymethyl, bis(2-chloroethoxy)methyl, 2,2,2-trichloro Ethoxymethyl, 2-(trimethylsilyl)ethoxymethyl, mentholoxymethyl, O-bis(2-acetoxy-ethoxy)methyl, tetrahydropyranyl, fluorotetrahydropyranyl, 3-bromotetrahydropyranyl, tetrahydrothiaranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl, 4-methoxy-tetrahydrothiaranyl, 4-methoxytetrahydrothiaranyl S,S-dioxide, 1-[(2-chloro-4-methyl)-phenyl]-4-methoxypiperidin-4-yl, 1-(2-fluorophenyl)-4-methoxypiperidin-4-yl, 1-(4-chlorophenyl)-4-methoxypiperidin-4-yl, 1,4-dioxane-2-yl, tetrahydrofuranyl, tetrahydrothio Furanyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methylbridged benzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 2-hydroxyethyl, 2-bromoethyl, 1-[2-(trimethylsilyl)ethoxy]ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 1-methyl-1-phenoxyethyl, 2,2,2-trichloroethyl, 1,1-dianisyl-2,2,2-trichloroethyl, 1,1,1,3,3,3-hexafluoro-2-phenylisopropyl, 1-(2-cyanoethoxy) 2-Trimethylsilylethyl, 2-(benzylthio)ethyl, 2-(phenylseleno)ethyl, tert-butyl, cyclohexyl, 1-methyl-1'-cyclopropylmethyl, allyl, isoprene, cinnamyl, 2-phenylallyl, propargyl, p-chlorophenyl, p-methoxyphenyl, p-nitrophenyl, 2,4-dinitrophenyl, 2,3,5,6-tetrafluoro-4-(trifluoromethyl)phenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, 2,6-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, pentadienylnitrobenzyl, pentadienylnitropiperyl, halobenzyl, 2,6-dichlorobenzyl, 2,6-Difluorobenzyl, p-cyanobenzyl, fluorobenzyl, 4-fluoroalkoxybenzyl, trimethylsilylxylyl, p-phenylbenzyl, 2-phenyl-2-propyl, p-amide benzyl, p-azidobenzyl, 4-azido-3-chlorobenzyl, 2-trifluoromethylbenzyl, 4-trifluoromethylbenzyl, p-(methylsulfinyl)benzyl, p-silazylbenzyl, 4-acetoxybenzyl, 4-(2-trimethylsilyl)ethoxymethoxybenzyl, 2-naphthylmethyl, 2-pyridinemethyl, 4-pyridinemethyl Pyridinemethyl, 3-methyl-2-pyridinemethyl N-oxide, 2-quinolinylmethyl, 6-methoxy-2-(4-methylphenyl)-4-quinolinmethyl, 1-pyrenemethyl, diphenylmethyl, 4-methoxydiphenylmethyl, 4-phenyldiphenylmethyl, p,p'-dinitrodiphenylmethyl, 5-dibenzocycloheptanyl, triphenylmethyl, tris(4-tert-butylphenyl)methyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenyl-methyl, tri... (p-methoxyphenyl)methyl, 4-(4'-bromobenzoyloxy)phenyldiphenylmethyl, 4,4',4''-tris(4,5-dichlorophthaliminophenyl)methyl, 4,4',4''-tris(acetylpropionyloxyphenyl)methyl, 4,4',4''-tris(benzoyloxyphenyl)methyl, 4,4'-dimethoxy-3''-[N-(imidazolylmethyl)]triphenylmethyl, 4,4'-dimethoxy-3''-[N-(imidazolylethyl)carbamoyl [3] Triphenylmethyl, bis(4-methoxyphenyl)-1'-pyrenemethyl, 4-(17-tetrabenzo[a,c,g,i]fluorenylmethyl)-4,4''-dimethoxytriphenylmethyl, 9-anthrayl, 9-(9-phenyl)xanthenyl, 9-phenylthioxanthenyl, 9-(9-phenyl-10-oxo)anthrayl, 1,3-benzodithiacyclopentane-2-yl, 4,5-bis(ethoxycarbonyl)-[1,3]-dioxolane-2-yl, benzisothiazolyl S,S-dioxide. In the case of silyl ethers, the protecting group of OH can be selected from trimethylsilyl, triethylsilyl, triisopropylsilyl, dimethylisopropylsilyl, diethylisopropylsilyl, dimethylhexylsilyl, 2-norbornyldimethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl, di-tert-butylmethylsilyl, bis(tert-butyl)-1-pyrene methoxysilyl, tri(trimethylsilyl)silyl, (2-hydroxystyryl)dimethylsilyl, (2-hydroxystyryl)diisopropylsilyl, tert-butylmethoxyphenylsilyl, tert-butoxydiphenylsilyl, 1,1,3,3-Tetraisopropyl-3-[2-(triphenylmethoxy)ethoxy]disiloxane-1-yl and fluorosilyl. In the case of esters, the protecting group of OH forms an ester together with the oxygen atom of the unprotected OH to which it is attached, and the ester can be selected from formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trichloroacetamide, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, phenylacetate, diphenyl acetate, 3-phenylpropionate, difluoropropionyl, 4-pentenoate, 4-oxopentanoate, 4,4-(ethylene dithio)pentanoate, 5-[3-bis(4-methoxyphenyl)hydroxymethylphenoxy]acetylpropionate, neopentanoate, 1-adamantate, barium sulfate, etc. Croton ester, 4-methoxycroton ester, benzoate, phenyl p-benzoate, 2,4,6-trimethylbenzoate, 4-bromobenzoate, 2,5-difluorobenzoate, p-nitrobenzoate, pyridinecarboxylate, nicotinic acid ester, 2-(azidomethyl)benzoate, 4-azido-butyrate, phenyl (2-azidomethyl)acetic acid, 2-{[(triphenylmethylthio)oxy]methyl}benzoate, 2-{[(4-methoxytriphenylmethylthio)oxy]methyl}benzoate, 2-{[methyl(triphenylmethylthio)amino]methyl}benzoate, 2-{{[(4-methoxytriphenylmethyl)thio]methylamino}methyl}benzoic acid Esters, 2-(allyloxy)phenyl acetate, 2-(isopreneoxymethyl)benzoate, 6-(acetylpropionyloxymethyl)-3-methoxy-2-nitrobenzoate, 6-(acetylpropionyloxymethyl)-3-methoxy-4-nitrobenzoate, 4-benzyloxybutyrate, 4-trialkylsilyloxy-butyrate, 4-acetoxy-2,2-dimethylbutyrate, 2,2-dimethyl-4-pentenoate, 2-iodobenzoate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate 2-(chloroacetoxymethyl)benzoate, 2-[(2-chloroacetoxy)ethyl]benzoate, 2-[2-(benzyloxy)ethyl]benzoate, 2-[2-(4-methoxybenzyloxy)ethyl]benzoate, 2,6-dichloro-4-methylphenoxyacetic acid, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetic acid, 2,4-bis(1,1-dimethylpropyl)phenoxyacetic acid, dichlorophenyl-acetic acid, isobutyrate, monosuccinate, (E)-2-methyl-2-butenoate, o-(methoxycarbonyl)benzoate, α-naphthoate, nitrate, alkyl N,N,N',N'-Tetramethylphosphonidamide and 2-chlorobenzoate. In the case of sulfonates, sulfenates, and sulfinates, the protecting group of OH, together with the oxygen atom of the unprotected OH atom to which it is attached, forms a sulfonate, sulfenate, or sulfinate, which may be selected from sulfates, allyl sulfonates, methanesulfonates, benzyl sulfonates, toluenesulfonates, 2-[(4-nitrophenyl)ethyl]sulfonate, 2-trifluoromethylbenzenesulfonate, 4-monomethoxytribenzylsulfenate, alkyl 2,4-dinitrophenylsulfenate, 2,2,5,5-tetramethylpyrrolidone-3-one-1-sulfinate, and dimethyl thiophosphonate. In the case of carbonates, the protecting group of OH forms a carbonate together with the oxygen atom of the unprotected OH atom to which it is attached. This carbonate can be selected from methyl carbonate, methoxymethyl carbonate, 9-fluorenylmethyl carbonate, ethyl carbonate, ethyl bromide carbonate, 2-(methylthiomethoxy)ethyl carbonate, 2,2,2-trichloroethyl carbonate, 1,1-dimethyl-2,2,2-trichloroethyl carbonate, 2-(trimethylsilyl)ethyl carbonate, 2-[dimethyl(2-naphthylmethyl)silyl]ethyl carbonate, 2-(benzenesulfonyl)ethyl carbonate, 2-(triphenylphosphino)ethyl carbonate, cis-[4-[[(methoxytriphenylmethyl)sulfinyl]oxy]tetrahydrofuran-3-yl]oxy carbonate, isobutyl carbonate, tert-butyl carbonate, ethylene carbonate, allyl carbonate, cinnamate carbonate, propargyl carbonate, and p-carbonate. Chlorophenyl ester, p-nitrobenzene carbonate, 4-ethoxy-1-naphthyl carbonate, 6-bromo-7-hydroxycoumarin-4-ylmethyl carbonate, benzyl carbonate, o-nitrobenzyl carbonate, p-methoxybenzyl carbonate, 3,4-dimethoxybenzyl carbonate, anthraquinone-2-ylmethyl carbonate, 2-dansulyl ethyl carbonate, 2-(4-nitrophenyl)ethyl carbonate, 2-(2,4-dinitrophenyl)ethyl carbonate, 2-(2-nitrophenyl)propyl carbonate, 2-(3,4-methylenedioxy-6-nitrophenyl)propyl carbonate, 2-cyano-1-phenylethyl carbonate, 2-(2-pyridyl)amino-1-phenylethyl carbonate, 2-[N-methyl-N-(2-pyridyl)]amino-1-phenylethyl carbonate, benzoyl carbonate, 3',5'-dimethoxybenzoic acid carbonate, methyl dithiocarbonate, and S-benzyl thiocarbonate. Furthermore, in the case of urethane esters, the protecting group of the OH group, together with the oxygen atom of the unprotected OH group to which it is attached, forms an urethane ester, which can be selected from dimethyl thiocarbamate, N-phenylcarbamate, and N-methyl-N-(o-nitrophenyl)carbamate.

[0072] Preferred OH protecting groups include methoxyethoxymethyl ether (MEM) and methoxymethyl ether (MOM).

[0073] Within the scope of this invention, an amino protecting group is defined as an N-bonded portion formed by protecting an amino group through the formation of a suitable protected amino group. Examples of protected amino groups include urethanes, ureas, amides, heterocyclic systems, N-alkylamines, N-alkenylamines, N-alkynylamines, N-arylamines, imines, enamines, N-metal derivatives, NN derivatives, NP derivatives, N-Si derivatives, and NS derivatives. In the case of carbamates, the protecting group of the amino group, together with the amino group to which it is attached, forms a carbamate, which can be selected from methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate, 2,6-di-tert-butyl-9-fluorenylmethyl carbamate, 2,7-bis(trimethylsilyl)fluorenylmethyl carbamate, 9-(2-sulfonyl)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluorenylmethyl carbamate, 17-tetrabenzo[a,c,g,i]fluorenylmethyl carbamate, 2-chloro-3-indenylmethyl carbamate, benzene[f]indenyl- 3-Methylcarbamate, 1,1-dioxobenzo[b]-thiophene-2-ylmethylcarbamate, 2-methanesulfonyl-3-phenyl-1-prop-2-enylcarbamate, 2,7-di-tert-butyl-[9,(10,10-dioxo-10,10,10,10-tetrahydrothiophene)]methylcarbamate, 2,2,2-trichloroethylcarbamate, 2-trimethylsilylethylcarbamate, (2-phenyl-2-trimethylsilyl)ethylcarbamate, 2-phenylethylcarbamate, 2-chloroethylcarbamate, 1,1-dimethyl-2 - Haloethyl carbamates, 1,1-dimethyl-2,2-dibromoethyl carbamate, 1,1-dimethyl-2,2,2-trichloroethyl carbamate, 2-(2'-pyridyl)ethyl carbamate, 2-(4'-pyridyl)ethyl carbamate, 2,2-bis(4'-nitrophenyl)ethyl carbamate, 2-[(2-nitrophenyl)dithio]-1-phenylethyl carbamate, 2-(N,N-dicyclohexylcarbamate)ethyl carbamate, tert-butyl carbamate, fluoroBOC carbamate, 1-adamantyl carbamate, 2-adamantane 1-(1-adamantyl)-1-methylethylcarbamate, 1-methyl-1-(4-biphenyl)ethylcarbamate, 1-(3,5-di-tert-butylphenyl)-1-methylethylcarbamate, triisopropylsilyloxycarbamate, vinylcarbamate, allylcarbamate, isopreneylcarbamate, 1-isopropylallylcarbamate, cinnamylcarbamate, 4-nitrocinnamylcarbamate, 3-(3'-pyridyl)propyl-2-enylcarbamate, hexadienylcarbamate, propargylcarbamate, 1,4-Butyl-2-ynyl biscarbamate, 8-quinolinyl carbamate, N-hydroxypiperidinyl carbamate, alkyl dithiocarbamate, benzyl carbamate, 3,5-di-tert-butylbenzyl carbamate, p-methoxybenzyl carbamate, p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate, 4-trifluoromethylbenzyl carbamate, fluorobenzyl carbamate, 2-naphthylmethyl carbamate, 9-anthraylmethyl carbamate, diphenylmethyl carbamate, 4-phenylacetoxybenzyl carbamate, 4-azidobenzyl carbamate, 4-azido-methoxybenzyl carbamate Ester, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)-benzyl carbamate, 5-benzisoxazolyl methyl carbamate, 2-(trifluoromethyl)-6-tryptomethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonyl ethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, 2-(4-nitrobenzenesulfonyl)ethyl carbamate, 2-(2,4-dinitrobenzenesulfonyl)ethyl carbamate, 2-(4-trifluoromethylbenzenesulfonyl)ethyl carbamate, [2-(1,3-dithiaalkyl)]methyl carbamate, 2-phosphonoethyl carbamate, 2-[phenyl(methyl)sulfonyl]ethyl carbamate, 1-methyl-1-(tri-) 1,1-Dimethyl-2-cyanoethylcarbamate, 2-Danylethylcarbamate, 2-(4-nitrophenyl)ethylcarbamate, 4-methylthiophenylcarbamate, 2,4-dimethylthiophenylcarbamate, m-nitrophenylcarbamate, 3,5-dimethoxybenzylcarbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethylcarbamate, α-methylnitropiperylcarbamate, o-nitrobenzylcarbamate, 3,4-dimethoxy-6-nitrobenzylcarbamate, phenyl(o-nitrophenyl)methylcarbamate, 2-nitrophenylethylcarbamate, 6-nitroveratrolylcarbamate, 4-methoxybenzoylcarbamate 3',5'-Dimethoxybenzoic acid ester, 9-Xanthonyl methyl carbamate, N-methyl-N-(o-nitrophenyl)carbamate, tert-amyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, cyclobutyl carbamate, cyclopentyl carbamate, cyclohexyl carbamate, isobutyl carbamate, isobornyl carbamate, cyclopropyl methyl carbamate, p-decoxybenzyl carbamate, diisopropyl methyl carbamate, 2,2-dimethoxy-carbonyl vinyl carbamate, o-(N,N-dimethylformamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-Dimethylformamidopropylcarbamate, butynediurethane, 1,1-Dimethylpropynylcarbamate, 2-Iodoethylcarbamate, 1-Methyl-1-(4'-pyridyl)ethylcarbamate, 1-Methyl-1-(p-phenylazophenyl)ethylcarbamate, p-(p'-methoxyphenylazo)benzylcarbamate, p-(phenylazo)benzylcarbamate, 2,4,6-Trimethylbenzylcarbamate, isonicocarbamate, 4-(trimethylammonium)benzylcarbamate, p-cyanobenzylcarbamate, di(2-pyridyl)methylcarbamate, 2-furanylmethylcarbamate, phenylcarbamate, 2,4,6-tri-tert-butylphenylcarbamate, 1-methyl-1-phenylethylcarbamate, and S-benzylthiocarbamate. In the case of urea, the protecting group of the amino group can be selected from phenothiazinyl-(10)-carbonyl, N'-p-toluenesulfonylaminocarbonyl, N'-phenylaminocarbonyl, 4-hydroxyphenylaminocarbonyl, 3-hydroxyphenylaminocarbonyl, and N'-phenylaminothiocarbonyl. In the case of amide, the protecting group of the amino group together with the amino group to which it is attached forms an amide, which can be selected from formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropionamide, pent-4-enamide, pyridinecarboxamide, 3-pyridylcarboxamide, N-benzoylphenylpropionamide, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, 2,2-dimethyl-2-(o-nitrophenyl)acetamide, o-nitrophenoxyacetamide, 3-(o-nitrophenyl)propionamide, 2-methyl-2-(o-nitrophenoxy)propionamide, 3-methyl-3-nitrobutamide, o-nitrocinnamamide, o-nitrobenzamide 3-(4-tert-butyl-2,6-dinitrophenyl)-2,2-dimethylpropionamide, o-(benzoyloxymethyl)benzamide, 2-(acetoxymethyl)benzamide, 2-[(tert-butyldiphenylsiloxy)methyl]benzamide, 3-(3',6'-dioxo-2',4',5'-trimethylcyclohexyl-1',4'-diene)-3,3-dimethylpropionamide, o-hydroxy-trans-cinnamonamide, 2-methyl-2-(o-phenylazophenoxy)propionamide, 4-chlorobutyramide, acetylacetamide, 3-(p-hydroxyphenyl)propionamide, (N'-dithiobenzyloxycarbonylamino)acetamide, and N-acetylmethionine amide. In the case of heterocyclic systems, the protecting group of the amino group, together with the amino group it is attached to, forms a heterocyclic system, which can be selected from 4,5-diphenyl-3-oxazoline-2-one, N-phthalimide, N-dichlorophthalimide, N-tetrachlorophthalimide, N-4-nitrophthalimide, N-thiodiethanolamide, N-dithiosuccinimide, N-2,3-diphenylmaleimide, N-2,3-dimethylmaleimide, N-2,5-dimethylpyrrole ...5-Bis(triisopropylsiloxy)pyrrole, N-1,1,4,4-tetramethyldimethsilylazine adduct, N-1,1,3,3-tetramethyl-1,3-disilisodihydroindole, N-diphenylsilyldiethylene, N-5-substituted 1,3-dimethyl-1,3,5-triazacyclohexane-2-one, N-5-substituted 1,3-benzyl-1,3,5-triazacyclohexane-2-one, 1-substituted 3,5-dinitro-4-pyridone, and 1,3,5-dioxazine. In the case of N-alkyl, N-alkenyl, N-ynyl, or N-arylamine, the protecting group of the amino group may be selected from N-methyl, N-tert-butyl, N-allyl, N-isopreneyl, N-cinnamyl, N-phenylallyl, N-propyne, N-methoxymethyl, N-[2-(trimethylsilyl)ethoxy]methyl, N-3-acetoxypropyl, N-cyanomethyl, N-2-azanorbornene, N-benzyl, N-4-methoxybenzyl, N-2,4-dimethoxybenzyl N-2-hydroxybenzyl, N-ferroceneylmethyl, N-2,4-dinitrophenyl, o-methoxyphenyl, p-methoxyphenyl, N-9-phenylfluorenyl, N-fluorenyl, N-2-pyridinemethylamine N'-oxide, N-7-methoxycoumar-4-ylmethyl, N-dibenzylmethyl, N-bis(4-methoxyphenyl)methyl, N-5-dibenzocycloheptyl, N-tribenzylmethyl, N-(4-methylphenyl)dibenzylmethyl and N-(4-methoxyphenyl)diphenylmethyl. In the case of imine, the protecting group of the amino group can be selected from N-1,1-dimethylthiomethylene, N-benzylene, N-p-methoxybenzylene, N-diphenylmethylene, N-[2-pyridyl)trimethylmethyl]methylene, N-(N',N'-dimethylaminomethylene), N-(N',N'-dibenzylaminomethylene), N-(N'-tert-butylaminomethylene), N,N'-isopropylene, N-p-nitrobenzylene, N-salicylene, N-5-chlorosalicylene, N-(5-chloro-2-hydroxyphenyl)phenylmethylene, N-cyclohexylene, and N-tert-butylene. In the case of enamines, the protecting group of the amino group can be selected from N-(5,5-dimethyl-3-oxo-1-cyclohexenyl), N-2,7-dichloro-9-fluorenylmethylene, N-1-(4,4-dimethyl-2,6-dioxocyclohexyl)ethyl, N-(1,3-dimethyl-2,4,6-(1H,3H,5H)-trioxopyrimidine-5-ylidene)-methyl, N-4,4,4-trifluoro-3-oxo-1-butenyl, and N-(1-isopropyl-4-nitro-2-oxo-3-pyrrololin-3-yl). In the case of N-metal derivatives, the protecting group of the amino group can be selected from N-borane, N-diphenylboronic acid ester, N-diethylboronic acid ester, N-9-boronibriocyclononane, N-difluoroboronic acid ester, and 3,5-Bis(trifluoromethyl)phenylboronic acid; and further comprising N-phenyl(pentacarbonylchromium)carbenyl, N-phenyl(pentacarbonyltungsten)carbenyl, N-methyl(pentacarbonylchromium)carbenyl, N-methyl(pentacarbonyltungsten)carbenyl, N-copper chelate, N-zinc chelate and 18-crown-6-derived compounds. In the case of NN derivatives, the protecting group of the amino group together with the amino group to which it is attached forms the NN derivative, which may be selected from N-nitroamino, N-nitrosoamino, amine N-oxide, azide, triazine derivative and N-trimethylsilylmethyl-N-benzylhydrazine. In the case of NP derivatives, the protected group of the amino group together with the amino group to which it is attached forms the NP derivative, which may be selected from diphenylphosphamide, dimethylthiophosphamide, diphenylthiophosphamide, dialkylaminophosphate, dibenzylaminophosphate, diphenylaminophosphate and iminotriphenylphosphine. In the case of N-Si derivatives, the protecting group of NH2 can be selected from tert-butyldiphenylsilyl and triphenylsilyl. In the case of NS derivatives, the protected amino group can be selected from N-sulfinyl or N-sulfonyl derivatives. N-sulfinyl derivatives can be selected from benzenesulfinamide, 2-nitrobenzenesulfinamide, 2,4-dinitrobenzenesulfinamide, pentachlorobenzenesulfinamide, 2-nitro-4-methoxybenzenesulfinamide, triphenylmethylsulfinamide, 1-(2,2,2-trifluoro-1,1-diphenyl)ethylsulfinamide, and N-3-nitro-2-pyridinesulfinamide. N-sulfonyl derivatives can be selected from methanesulfonamide, trifluoromethanesulfonamide, tert-butylsulfonamide, benzylsulfonamide, 2-(trimethylsilyl)ethanesulfonamide, p-toluenesulfonamide, benzenesulfonamide, o-anisinylsulfonamide, 2-nitrobenzenesulfonamide, 4-nitrobenzenesulfonamide, 2,4-dinitrobenzenesulfonamide, 2-naphthalenesulfonamide, 4-(4',8'-dimethoxynaphthyl)benzenesulfonamide, 2-(4-methylphenyl)-6-methoxy-4-methylsulfonamide, 9-anthrasulfonamide, pyridine-2-sulfonamide, and benzothiazole. 2-Sulfanamide, benzoylsulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide, 2,4,6-trimethoxybenzenesulfonamide, 2,6-dimethyl-4-methoxy-benzenesulfonamide, pentamethylbenzenesulfonamide, 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide, 4-methoxybenzenesulfonamide, 2,4,6-trimethylbenzenesulfonamide, 2,6-dimethoxy-4-methylbenzenesulfonamide, 3-methoxy-4-tert-butylbenzenesulfonamide, and 2,2,5,7,8-pentamethylsomn-6-sulfonamide.

[0074] Preferred amino protecting groups include allyl urethane (Alloc), 2,2,2-trichloroethyl urethane (Troc), benzyl urethane (Cbz), 9-fluorenylmethyl urethane (Fmoc), N'-phenylaminocarbonyl (CONHPh), N'-phenylaminothiocarbonyl (CSNHPh), and tert-butyl urethane (Boc). Tert-butyl urethane is the most preferred.

[0075] Within the scope of this invention, the protecting group of SH is defined as the S-bonded portion resulting from protecting the SH group by forming a suitable protected SH group. Examples of such protected SH groups include thioethers, disulfides, silyl thioethers, thioesters, thiocarbonates, and thiocarbamates. In the case of thioethers, the protecting group of SH may be selected from S-alkyl, S-benzyl, S-p-methoxybenzyl, S-o-hydroxybenzyl, S-p-hydroxybenzyl, S-o-acetoxybenzyl, S-p-acetoxybenzyl, S-p-nitrobenzyl, S-o-nitrobenzyl, S-2,4,6-trimethylbenzyl, S-2,4,6-trimethoxybenzyl, S-4-pyridinemethyl, S-2-pyridinemethyl-N-oxide, S-2-quinolinylmethyl, and S-9-anthraylmethyl. S-9-fluorenylmethyl, S-xanolyl, S-ferroceneylmethyl, S-diphenylmethyl, S-bis(4-methoxyphenyl)methyl, S-5-dibenzocycloheptyl, S-triphenylmethyl, 4-methoxytriphenylmethyl, S-diphenyl-4-pyridylmethyl, S-phenyl, S-2,4-dinitrophenyl, S-2-quinolinyl, S-tert-butyl, S-1-adamantyl, S-methoxymethyl, S-isobutoxymethyl, S-benzyloxymethyl, S-1-ethoxy Ethyl, S-2-tetrahydropyranyl, S-benzylthiomethyl, S-phenylthiomethyl, S-acetamidomethyl (Acm), S-trimethylacetamidomethyl, S-benzamidomethyl, S-allyloxycarbonylaminomethyl, SN-[2,3,5,6-tetrafluoro-4-(N'-piperidinyl)-phenyl-N-allyloxycarbonylaminomethyl, S-phthalimidemethyl, S-phenylacetamidomethyl, S-acetylmethyl, S-carboxymethyl, S-cyano S-methyl, S-(2-nitro-1-phenyl)ethyl, S-2-(2,4-dinitrophenyl)ethyl, S-2-(4'-pyridyl)ethyl, S-2-cyanoethyl, S-2-(trimethylsilyl)ethyl, S-2,2-bis(ethoxycarbonyl)ethyl, S-(1-m-nitrophenyl-2-benzoyl)ethyl, S-2-benzenesulfonylethyl, S-1-(4-methylphenylsulfonyl)-2-methylpropyl-2-yl, and S-p-hydroxybenzoylmethyl. In the case of disulfides, the protected SH group can be selected from S-ethyl disulfide, S-tert-butyl disulfide, S-2-nitrophenyl disulfide, S-2,4-dinitrophenyl disulfide, S-2-phenylazophenyl disulfide, S-2-carboxyphenyl disulfide, and S-3-nitro-2-pyridyl disulfide. In the case of silyl sulfides, the protecting group of SH can be selected from the list of groups listed above for protecting OH with silyl sulfides.In the case of thioesters, the protecting group of SH can be selected from S-acetyl, S-benzoyl, S-2-methoxyisobutyryl, S-trifluoroacetyl, SN-[[p-biphenyl)-isopropoxy]carbonyl]-N-methyl-γ-aminothiobutyrate, and SN-(tert-butoxycarbonyl)-N-methyl-γ-aminothiobutyrate. In the case of thiocarbonates, the protecting group of SH can be selected from S-2,2,2-trichloroethoxycarbonyl, S-tert-butoxycarbonyl, S-benzyloxycarbonyl, S-p-methoxybenzyloxycarbonyl, and S-fluorenylmethylcarbonyl. In the case of thiocarbamates, the protected SH group can be selected from S-(N-ethylcarbamate) and S-(N-methoxymethylcarbamate).

[0076] Preferred SH protecting groups include substituted or unsubstituted S-9-fluorenylmethyl. The most preferred is S-9-fluorenylmethyl (Fm).

[0077] The references to these groups should not be construed as limiting the scope of the invention, as they are mentioned only as examples of protecting groups of OH, amino, and SH groups, but other groups having the aforementioned functions may be known to those skilled in the art and should also be understood to be covered by the invention.

[0078] The term "sea succulent compound" can refer to compound of formula E:

[0079]

[0080] in:

[0081] R 101 and R 104 Independently selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 alkynyl group, C(=O)R 10c C(=O)OR 10b C(=O)NR 10d R 10e and OH protecting groups;

[0082] R 102 Selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 alkynyl group, C(=O)R 10c C(=O)OR 10b C(=O)NR 10d R 10eand the protecting group of amino groups;

[0083] R 103 CN or OH;

[0084] R 105 and R 106 Together with the carbon atoms it is attached to, it forms a functional group:

[0085] (a)C(=O);

[0086] (b)CH(OR 107 ) or CH(NR 108 R 109 ), where R 107 Selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne group, substituted or unsubstituted aryl group, substituted or unsubstituted heterocyclic group and OH protecting group; and R 108 and R 109 Independently selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne group, substituted or unsubstituted aryl group, substituted or unsubstituted heterocyclic group and protecting group of amino group;

[0087] (c) Groups having the following formula:

[0088]

[0089] in

[0090] X 101 and X 102 Independently selected from hydrogen and substituted or unsubstituted C1-C 12 alkyl;

[0091] R 110 and R 111 Independently selected from hydrogen, C(=O)R 10c C(=O)OR 10b C(=O)NR 10d R 10e Substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 alkynyl group; or

[0092] (d) Groups having the following formula:

[0093]

[0094] in

[0095] X is -NR 112 -or -O-;

[0096] Hydrogen, OR b OC(=O)R c OC(=O)OR b OC(=O)NR d R e SR f SOR c SO2R c C(=O)R c C(=O)OR b C(=O)NR d R e NO2, NR d R e 、N(R d )C(=O)R c 、N(R d )-OR b C(R) c )=NOR b 、N(R d )C(=O)OR b 、N(R d )C(=O)NR d R e CN, halogens, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne, substituted or unsubstituted aryl groups, and substituted or unsubstituted heterocyclic groups;

[0097] Y 101 Selected from hydrogen, OR 10b OC(=O)R 10c OC(=O)OR 10b OC(=O)NR 10d R 10e SR 10f SOR 10c SO2R 10c C(=O)R 10c C(=O)OR 10b C(=O)NR 10d R 10e NO2, NR10d R 10e 、N(R 10d )C(=O)R 10c 、N(R 10d )-OR 10b C(R) 10c )=NOR 10b 、N(R 10d )C(=O)OR 10b 、N(R 10d )C(=O)NR 10d R 10e CN, halogens, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne, substituted or unsubstituted aryl groups, and substituted or unsubstituted heterocyclic groups;

[0098] Y 102 and Y 103 Independently selected from hydrogen and substituted or unsubstituted C1-C 12 alkyl;

[0099] R 112 and R 113 Independently selected from hydrogen, C(=O)R 10c C(=O)OR 10b C(=O)NR 10d R 10e Substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 alkynyl group; and

[0100] Each R 10b Independently selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne group, substituted or unsubstituted aryl group, substituted or unsubstituted heterocyclic group and OH protecting group;

[0101] Each R 10c Independently selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne, substituted or unsubstituted aryl groups, and substituted or unsubstituted heterocyclic groups;

[0102] Each R 10d and R 10e Independently selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne group, substituted or unsubstituted aryl group, substituted or unsubstituted heterocyclic group and protecting group of amino group;

[0103] Each R 10f Independently selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne group, substituted or unsubstituted aryl group, substituted or unsubstituted heterocyclic group and SH protecting group;

[0104] Or its pharmaceutically acceptable salt.

[0105] Preferably, R 101 It can be hydrogen or the same as R4 as described herein.

[0106] Preferably, R 102 It can be methyl.

[0107] Preferably, R 104 It can be the same as R8 as described in this article.

[0108] Preferably, X 101 It can be the same as X1 as described in this article.

[0109] Preferably, X 102 It can be the same as X2 as described in this article.

[0110] Preferably, R 110 It can be hydrogen.

[0111] Preferably, R 111 It can be hydrogen.

[0112] Preferably, Y 101 It can be the same as Y1 as described in this article.

[0113] Preferably, Y 102 It can be the same as Y2 as described in this article.

[0114] Preferably, Y 103 It can be the same as Y3 as described in this article.

[0115] Preferably, R 112 It can be hydrogen.

[0116] Preferably, R 113 It can be hydrogen.

[0117] Oxidation reactions and related synthetic steps of compounds of formula I to II

[0118] According to one aspect of the present invention, a method for synthesizing a sucrose compound is provided, the method comprising the step of oxidizing a reaction mixture containing a compound of formula I to obtain a compound of formula II:

[0119]

[0120] in:

[0121] R1 is OH or CN;

[0122] R2 is hydrogen or Prot NH ;and

[0123] R3 is hydrogen or Prot OH ,

[0124] Among them Prot NH It is a protecting group for amino groups, and Prot OH It is a protecting group for OH.

[0125] In some preferred embodiments, R1 is OH. In other preferred embodiments, R1 is CN.

[0126] In some preferred embodiments, R2 is hydrogen. In other preferred embodiments, R2 is Prot. NH .

[0127] Preferably, Prot NH It is selected from allyl carbamate (Alloc), 2,2,2-trichloroethyl carbamate (Troc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbamate (Fmoc), CONHPh, CSNHPh and tert-butyl carbamate (Boc), more preferably tert-butyl carbamate (Boc).

[0128] Preferably, R3 is hydrogen.

[0129] Preferably, oxidation is performed using an oxidizing agent, biochemical oxidation, or electrochemical oxidation.

[0130] Preferably, the oxidant is selected from the group consisting of: metal oxidants (such as Cr(VI) compounds, Mn(IV) compounds, Mn(VII) compounds, Fe(III) compounds, Cu(II) compounds, Ag(I) compounds, Pb(IV) compounds, Ce(IV) compounds), inorganic catalytic metals (such as ruthenium, cobalt, etc.), molecular oxygen, hydrogen peroxide, hypochlorites (such as NaOCl and Ca(OCl)2), chlorites, 1,4-benzoquinone and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone or combinations thereof. Preferred examples of Cr(VI) compounds include dichromates (e.g., potassium dichromate, pyridinium dichromate), chromium trioxide and chlorochromates (e.g., pyridinium chlorochromate). Preferred examples of Mn(IV) compounds include manganese dioxide. Preferred examples of Mn(VII) compounds include permanganates (e.g., potassium permanganate). Preferred Fe(III) compounds include ferric acetate (III), ferric chloride (III), ferric bromide (III), ferric sulfate (III), and ferric nitrate (III). Preferred Cu(II) compounds include copper acetate (II), copper chloride (II), copper bromide (II), copper sulfate (II), and copper nitrate (II). Preferred Ag(I) compounds include silver oxide (I). Preferred Pb(IV) compounds include lead dioxide, lead acetate (IV), and lead chloride (IV). Preferred Ce(IV) compounds include cerium ammonium nitrate. More preferably, the oxidant is selected from molecular oxygen, hydrogen peroxide, 1,4-benzoquinone, and metal oxidants, or combinations thereof.

[0131] Preferably, the metal oxidant is a transition metal oxidant. More preferably, the transition metal oxidant is selected from Fe(III) compounds (e.g., FeCl3) and Cu(II) compounds (e.g., CuSO4). Even more preferably, the Fe(III) compound is FeCl3.

[0132] Preferably, the oxidant has an equivalent number of about 1.0 equivalents to about 15.0 equivalents relative to the compound of formula I in the reaction mixture, more preferably about 1.2 equivalents to about 10.0 equivalents, more preferably about 1.5 equivalents to about 8.0 equivalents, and even more preferably about 2.0 equivalents to about 5.0 equivalents. When the reaction mixture further comprises a compound according to formula II, the equivalent number of the oxidant can be calculated relative to the combined amount of the compound of formula I and the compound of formula II present in the reaction mixture.

[0133] Preferably, the oxidant is molecular oxygen. More preferably, when the oxidant is molecular oxygen, the reaction is carried out at a pH of about 3.0 to about 9.0, preferably about 5.0 to about 8.5, and more preferably about 6.0 to about 8.0.

[0134] Preferably, molecular oxygen is provided by exposing the reaction mixture to a reaction atmosphere containing molecular oxygen. The reaction atmosphere may contain between about 5% and about 100% molecular oxygen by volume, preferably between about 6% and about 50%, more preferably between about 7% and about 40%, and even more preferably between about 8% and about 30%.

[0135] Preferably, during the step of oxidizing the reaction mixture containing Formula I to obtain Formula II, a stream containing molecular oxygen is bubbled through the reaction mixture. This stream may contain between about 5% and about 100% molecular oxygen by volume, preferably between about 6% and about 50%, more preferably between about 7% and about 40%, and even more preferably between about 8% and about 30%.

[0136] Preferably, the reaction is catalyzed by an inorganic catalytic metal. Preferred examples of inorganic catalytic metals include ruthenium, cobalt, etc.

[0137] Preferably, the reaction is carried out at a pH of about 3.0 to about 9.0, more preferably about 3.5 to about 8.5, and more preferably about 4.0 to about 8.0.

[0138] Preferably, the biochemical oxidation is catalyzed by hydroquinone oxidase or by cells exhibiting hydroquinone oxidase activity.

[0139] Preferably, the reaction mixture further comprises a compound according to Formula II; preferably wherein R1 in the compound according to Formula I in the reaction mixture is the same as R1 in the compound according to Formula II in the reaction mixture, and / or wherein R2 in the compound according to Formula I in the reaction mixture is the same as R2 in the compound according to Formula II in the reaction mixture, and / or wherein R3 in the compound according to Formula I in the reaction mixture is the same as R3 in the compound according to Formula II in the reaction mixture; more preferably wherein R1, R2 and R3 in the compound according to Formula I in the reaction mixture are the same as R1, R2 and R3 in the compound according to Formula II in the reaction mixture.

[0140] Preferably, the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II is carried out at about 0°C to about 100°C, preferably about 0°C to about 80°C, more preferably about 5°C to about 60°C, even more preferably about 5°C to about 40°C, even more preferably about 10°C to about 30°C, and most preferably about 10°C to about 25°C.

[0141] Preferably, the method further includes a cyanidation reaction to convert R1 from OH to CN before or after the step of oxidizing the reaction mixture containing compound I to obtain compound II; preferably, the cyanidation reaction is carried out after the step of oxidizing the reaction mixture containing compound I to obtain compound II.

[0142] Preferably, the step of carrying out the cyanidation reaction to convert R1 from OH to CN is performed using a metal cyanide, preferably an alkali metal cyanide, and most preferably KCN or NaCN.

[0143] Preferably, the method further includes, before or after the step of oxidizing the reaction mixture containing compound I to obtain compound II, performing an amino protection reaction to convert R2 from hydrogen to Prot. NH The step; preferably, the amino protection reaction is carried out after the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II.

[0144] Preferably, R1 in Formula I is OH, and R2 in Formula I is hydrogen, and the method:

[0145] The process further includes a step of immediately following the step of oxidizing the reaction mixture containing compound I to obtain compound II, to perform a cyanidation reaction to convert R1 from OH to CN; and

[0146] This further includes immediately following the cyanation reaction to convert R1 from OH to CN, an amino protection reaction to convert R2 from hydrogen to Prot. NH The steps.

[0147] Preferably, R1 in Formula I is CN, and R2 in Formula I is hydrogen, and the method:

[0148] This further includes a step of immediately performing a cyanidation reaction to convert R1 from OH to CN before the step of oxidizing the reaction mixture containing compound I to obtain compound II; and

[0149] The process further includes, immediately after the step of oxidizing the reaction mixture containing compound I to obtain compound II, carrying out an amino protection reaction to convert R2 from hydrogen to Prot. NH The steps.

[0150] Preferably, the steps of oxidizing the reaction mixture containing compound I to obtain compound II and performing a cyanidation reaction to convert R1 from OH to CN are carried out as a one-pot reaction.

[0151] In a preferred alternative embodiment, the compound according to Formula I does not include compound HQ-3.

[0152] In a preferred embodiment, the reaction mixture is a fermentation broth or obtained from a fermentation broth. The fermentation broth may be produced by *Pseudomonas fluorescens*. *Pseudomonas fluorescens* may be a modified strain. *Pseudomonas fluorescens* may be modified to produce compound I. *Pseudomonas fluorescens* may be modified to produce both compound I and compound II. In embodiments, the fermentation broth may be obtained by following the teachings of Munakata et al., *Journal of Antibiotics*, 1983, 36(10), 1279-1283, US4440752, JPS59225189A, and / or Meyers E et al., *Journal of Antibiotics*, 1983, 36(2), 190-193. The fermentation broth may contain a mixture of compound I and compound II.

[0153] In an alternative embodiment, the reaction mixture is an extract from the fermentation broth. The extract from the fermentation broth includes a concentrate. The extract from the fermentation broth may be a methanol concentrate.

[0154] Further synthesis steps

[0155] The oxidation reactions of compounds of formula I to II as described herein can be used in the early stages of jugillin compound synthesis. For example, the methods described herein can be used to generate starting materials and intermediate compounds previously used in jugillin compound synthesis. The starting materials and intermediates can then be used to obtain the target jugillin compound by following known methods.

[0156] This invention relates to a method for synthesizing tunicate compounds, the method comprising the step of oxidizing a reaction mixture containing a compound of formula I to obtain a compound of formula II. Further synthetic steps are required to obtain the final tunicate compound. These further steps will be known to those skilled in the art. Non-limiting examples of the further synthetic steps are summarized below.

[0157] This method may include further synthetic steps as disclosed in Cragg et al., Anticancer Agents from Natural Products (2012). 2nd Edition – Chapter 12: “Ecteinascidin-743 (Yondelis®), Aplidin®, and Irvalec®” (pp. 291–316); WO 01 / 87895 A1 (and the references contained therein); WO 2011 / 147828 A1 (and the references contained therein); and WO 2018 / 197663A1 (and the references contained therein), all of which (and the references contained therein) are incorporated by reference.

[0158] Preferably, R3 in Formula I is hydrogen. Exemplary further synthesis steps are provided in the following schemes:

[0159]

[0160] Option I

[0161] Where R2 is Prot NH Furthermore, R3, R4, and R5 are as defined in Scheme I.

[0162] Preferably, R2 is selected from allyl carbamate (Alloc), 2,2,2-trichloroethyl carbamate (Troc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbamate (Fmoc), CONHPh, CSNHPh and tert-butyl carbamate (Boc), more preferably tert-butyl carbamate (Boc).

[0163] Preferably, R3 and R4 are independently selected from methoxyethoxymethyl ether (MEM) and methoxymethyl ether (MOM).

[0164] Preferably, R5 is different from R3. More preferably, R5 is allyl.

[0165] The preparation of compounds of formula III from phenols of formula IIa is typically carried out by reaction with a suitable reagent for protecting the phenolic group. Preferred reagents for such reactions are alkoxymethyl chlorides, alkoxymethyl bromides, and alkoxyalkoxymethyl chlorides. Alkoxyalkoxymethyl chlorides are particularly preferred reagents. Preferred reagents are methoxyethoxymethyl chloride (MEMCl) or methoxymethyl chloride (MOMCl).

[0166] The conversion of methoxyquinone of formula III to hydroxyquinone of formula IV is typically carried out by reaction with a suitable reagent for deprotecting the methoxy group or by reaction with a hydroxide. Preferred reagents for such reactions are hydroxides or NaI in the presence of a base. More preferably, the reaction is carried out with a basic hydroxide. The most preferred basic hydroxide is NaOH.

[0167] The reduction of hydroxyquinones of formula IV is typically carried out using transition metal-catalyzed hydrogenation or reducing agents (such as Na₂S₂O₄). Transition metal-catalyzed hydrogenation is particularly preferred. The most preferred transition metal catalyst is Pd / C.

[0168] The cyclization of compounds of formula V is typically carried out by reaction with a suitable reagent for protecting 1,2-diol to methylene acetal. Preferred reagents include dihalomethane reagents, such as dibromomethane, bromochloromethane, dichloromethane, and diiodomethane. The most preferred dihalomethane reagent is bromochloromethane.

[0169] The preparation of compounds of formula VII from phenols of formula VI is typically carried out by reaction with a suitable reagent for protecting the phenolic group. Preferred reagents for such reactions are allyl halides, such as allyl chloride, allyl bromide, and allyl iodide. The most preferred reagent is allyl bromide.

[0170] The conversion of compounds of formula VII to compounds of formula VIII typically involves the deprotection of the amino group at R2. Suitable conditions and reagents for deprotecting these protecting groups are known to those skilled in the art (see Wuts, PGM, and Greene TW's description in Protecting Groups in Organic Synthesis). The 4th edition of Wiley-Interscience, and Kocienski PJ's description in Protecting Groups, (Georg Thieme Verlag, 3rd edition). For example, in the case of tert-butylcarbamates, deprotection is typically carried out under acidic conditions. Trifluoroacetic acid is the preferred reagent.

[0171] The conversion of compounds of formula VII to formula VIII typically involves the deprotection of the phenolic group on R3. Suitable conditions and reagents for deprotecting these protecting groups are known to those skilled in the art (see Wuts, PGM, and Greene TW's description in Protecting Groups in Organic Synthesis). The 4th edition of Wiley-Interscience, and Kocienski PJ's description in Protecting Groups, (Georg Thieme Verlag, 3rd edition). For example, in the cases of MEM and MOM, deprotection is typically carried out under acidic conditions. Trifluoroacetic acid is the preferred reagent.

[0172] The conversion of compounds of formula VII to compounds of formula VIII typically involves an Edman degradation reaction. Suitable reagents include reactions with aryl isocyanates or arylisothiocyanates. Particularly preferred reagents for such reactions are aryl isothiocyanates. The most preferred reagent is phenyl isothiocyanate. Degradation can be carried out in a one-pot manner under acidic conditions to degrade the resulting urea or thiourea. A preferred reagent is hydrogen chloride (e.g., HCl / dioxane).

[0173] Compounds of formula VIII are optionally reprotected to compounds of formula VIIIb, typically by reaction with a suitable reagent for protecting the phenolic group. Preferred reagents for such reactions are alkoxymethyl chlorides, alkoxymethyl bromides, and alkoxyalkoxymethyl chlorides. Alkoxyalkoxymethyl chlorides are particularly preferred reagents. Preferred reagents are methoxyethoxymethyl chloride (MEMCl) or methoxymethyl chloride (MOMCl).

[0174] The method may further include the series of steps shown in Scheme II:

[0175]

[0176] Option II

[0177] R4 is Prot OH R5 is Prot OH R6 is Prot NH And R7 is Prot SH Among them, Prot SH It is a protecting group for SH.

[0178] Preferably, R4 is selected from methoxyethoxymethyl ether (MEM) and methoxymethyl ether (MOM).

[0179] Preferably, R5 is different from R4. More preferably, R5 is allyl.

[0180] Preferably, R6 is selected from allyl carbamate (Alloc), 2,2,2-trichloroethyl carbamate (Troc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbamate (Fmoc), CONHPh, CSNHPh and tert-butyl carbamate (Boc), more preferably tert-butyl carbamate (Boc), and even more preferably tert-butyl carbamate (Boc).

[0181] Preferably, R7 is selected from substituted or unsubstituted S-9-fluorenylmethyl. More preferably, R7 is S-9-fluorenylmethyl (Fm).

[0182] Alternatively, compounds of formula XI can be obtained by the sequence of steps shown in scheme III:

[0183]

[0184] Option III

[0185] R4 is Prot OH R5 is Prot OH R6 is Prot NH And R7 is Prot SH .

[0186] Preferably, R4 is different from R5. More preferably, R4 is methoxyethoxymethyl ether (MEM) or methoxymethyl ether (MOM).

[0187] Preferably, R5 is different from R4. More preferably, R5 is allyl.

[0188] Preferably, R6 is selected from allyl carbamate (Alloc), 2,2,2-trichloroethyl carbamate (Troc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbamate (Fmoc), CONHPh, CSNHPh and tert-butyl carbamate (Boc), more preferably tert-butyl carbamate (Boc), and even more preferably tert-butyl carbamate (Boc).

[0189] Preferably, R7 is selected from substituted or unsubstituted S-9-fluorenylmethyl. More preferably, R7 is S-9-fluorenylmethyl (Fm).

[0190] In schemes II and III:

[0191] The conversion of a primary amine of formula VIII or VIIIb to a primary alcohol of formula IX or Xb is typically carried out by reaction with a suitable oxidizing agent, such as an inorganic nitrite, dinitrogen tetroxide, or nitroferricyanide. A more preferred oxidizing agent is an inorganic nitrite. Sodium nitrite is the most preferred oxidizing agent for this step.

[0192] Esterification of compounds of formula IX or Xb to obtain compounds of formula Xa or XI is typically carried out by reaction with an amino- and sulfur-protected cysteine ​​amino acid, wherein the amino acid is activated by a coupling agent such as carbodiimide, phosphonium salt, urea salt, guanidine salt, imidazolium derivatizing agent, or triazolium derivatizing agent. A particularly preferred coupling agent is carbodiimide. The most preferred coupling agent is 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) and its hydrochloride salt (EDC.HCl).

[0193] In Option II:

[0194] The preparation of compounds of formula XI from phenol with protected formula Xa is typically carried out by reaction with a suitable reagent for protecting the phenolic group. Preferred reagents for such reactions are alkoxymethyl chlorides, alkoxymethyl bromides, and alkoxyalkoxymethyl chlorides. Alkoxyalkoxymethyl chlorides are particularly preferred reagents. The most preferred reagent is methoxyethoxymethyl chloride (MEMCl).

[0195] This method may further include the series of steps shown in scheme IV:

[0196]

[0197] Option IV

[0198] R4 and R6 are defined as in Scheme IV, and R5 is Prot. OH R7 is Prot SH And R8 is -C(=O)R a Group, wherein R a Selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 Alkyne group.

[0199] Preferably, R4 is different from R5. More preferably, R4 is methoxyethoxymethyl ether (MEM) or methoxymethyl ether (MOM).

[0200] Preferably, R5 is different from R4. More preferably, R5 is allyl.

[0201] Preferably, R6 is selected from allyl carbamate (Alloc), 2,2,2-trichloroethyl carbamate (Troc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbamate (Fmoc), CONHPh, CSNHPh and tert-butyl carbamate (Boc), more preferably tert-butyl carbamate (Boc), and even more preferably tert-butyl carbamate (Boc).

[0202] Preferably, R7 is selected from substituted or unsubstituted S-9-fluorenylmethyl. More preferably, R7 is S-9-fluorenylmethyl (Fm).

[0203] Preferably, R8 is an acetyl group.

[0204] The deprotection of compound XI to obtain compound XII involves the deprotection of the phenolic group on R5. Suitable conditions and reagents for deprotecting these protecting groups are known to those skilled in the art (see Wuts, PGM, and Greene TW's description in Protecting Groups in Organic Synthesis). The 4th edition of Wiley-Interscience, and Kocienski PJ's description in Protecting Groups, (Georg Thieme Verlag, 3rd edition). For example, in the case of allyl groups, deprotection is typically carried out using metal catalysis. Preferred catalysts include palladium (0) catalysts and palladium (II) catalysts in the presence of a reducing agent (such as a hydrogenated trialkyltin or a secondary amine).

[0205] The oxidation reaction of compound XII to compound XIII is typically achieved by reacting with a suitable oxidizing agent, such as hydrogen peroxide, organic peroxides, perbenzoic acid, periodate, lead tetraacetate, lead oxide, selenium dioxide, high-valent iodine oxidizing agents (such as 2-iodooxybenzoic acid (IBX)), or with organic selenic anhydrides (such as (PhSeO)₂O). More preferred oxidizing agents are organic selenic anhydrides and high-valent iodine oxidizing agents. Organic selenic anhydrides are even more preferred. The most preferred oxidizing agent is (PhSeO)₂O.

[0206] The cyclization reaction of compounds of formula XIV according to formula XIII is typically achieved by forming an exoquinone methylate at the 4-position of ring B, reacting the methylate with the sulfur atom of a cysteine ​​residue, and capturing the resulting phenolate with an acylation agent. Typically, the methylate is formed by reacting the compound of formula XIII with an in-situ generated Swern reagent (such as by reacting Tf₂O with DMSO), followed by treatment with a base (such as tetramethylguanidine). Cyclolysis is typically carried out by removing the protecting group of SH under conditions allowing for the formation of thiolate ions, followed by the nucleophilic addition of sulfur to the quinone methylate to generate a 10-membered lactone bridge, and capturing the resulting phenolate with an acylation agent (such as acetic anhydride, mixed acetic anhydride, or acetyl chloride) to obtain the acylated compound of formula XIV.

[0207] The conversion of compounds of formula XIV or XV-Int-a to compounds of formula XV and the conversion of compounds of formula XIV to compounds of formula XV-Int-b generally involve the deprotection of the amino group on R6. Suitable conditions and reagents for deprotecting these protecting groups are known to those skilled in the art (see Wuts, PGM, and Greene TW's description in Protecting Groups in Organic Synthesis). The 4th edition of Wiley-Interscience, and Kocienski PJ's description in Protecting Groups, (Georg Thieme Verlag, 3rd edition). For example, in the case of tert-butylcarbamates, deprotection is typically carried out under acidic conditions. Preferred reagents are p-toluenesulfonic acid or p-toluenesulfonic acid monohydrate.

[0208] The conversion of compounds of formula XIV or XV-Int-b to compounds of formula XV and the conversion of compounds of formula XIV to compounds of formula XV-Int-a generally involve the deprotection of the phenolic group on R4. Suitable conditions and reagents for deprotecting these protecting groups are known to those skilled in the art (see Wuts, PGM, and Greene TW's description in Protecting Groups in Organic Synthesis). The 4th edition of Wiley-Interscience, and Kocienski PJ's description in Protecting Groups, (Georg Thieme Verlag, 3rd edition). For example, in the cases of MEM and MOM, deprotection is typically carried out under acidic conditions. Specific reagents include p-toluenesulfonic acid or p-toluenesulfonic acid monohydrate or HCl (e.g., in EtOAc).

[0209] Preferably, the deprotection of the amino group on R6 and the deprotection of the phenolic group on R4 are carried out in a one-pot manner.

[0210] The transfer of amino groups from a compound of formula XV to a compound of formula XVI is typically carried out using a suitable carbonyl reagent (such as hindered 1,2-benzoquinone or pyridinecarboxaldehyde or pyridiniumcarboxaldehyde). More preferred carbonyl reagents are methyl iodide of pyridine-4-carboxaldehyde and methylbenzenesulfonate of pyridine-4-carboxaldehyde.

[0211] The method may further include the series of steps shown in scheme V:

[0212]

[0213] Option V

[0214] Where R8 is -C(=O)R a Group, wherein R a Selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 alkynyl group; and

[0215] X1 and X2 are independently selected from hydrogen and unsubstituted or unsubstituted C1-C. 12 alkyl.

[0216] Preferably, R8 is an acetyl group.

[0217] Preferably, X1 is hydrogen.

[0218] Preferably, X2 is hydrogen.

[0219] The conversion of compound XVI to provide compound XVIIa is typically carried out using the Pictet-Spengler reaction. This reaction is usually carried out under acid catalysis. Silica gel is the preferred catalyst.

[0220] Preferably, compound A is:

[0221] .

[0222] The conversion of compound XVIIa to provide compound XVIIIa is typically carried out by reaction with a nitrile-coordinated transition metal salt. More preferred salts are salts of Ag(I) or Cu(I). Most preferred salts are AgNO3 and CuCl.

[0223] Preferably, the compound of formula XVIIIa is trabectedin:

[0224] .

[0225] In another embodiment, the method may further include the series of steps shown in scheme VI:

[0226]

[0227] Solution VI

[0228] Where R8 is -C(=O)R a Group, wherein R a Selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 alkynyl group;

[0229] X is -NH- or -O-;

[0230] Y1 is selected from hydrogen, OR b OC(=O)R c OC(=O)OR b OC(=O)NR d R e SR f SORc SO2R c C(=O)R c C(=O)OR b C(=O)NR d R e NO2, NR d R e 、N(R d )C(=O)R c 、N(R d )-OR b C(R) c )=NOR b 、N(R d )C(=O)OR b 、N(R d )C(=O)NR d R e CN, halogens, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne, substituted or unsubstituted aryl groups, and substituted or unsubstituted heterocyclic groups;

[0231] Y2 and Y3 are independently selected from hydrogen and substituted or unsubstituted C1-C. 12 alkyl;

[0232] Each R b Independently selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne group, substituted or unsubstituted aryl group, substituted or unsubstituted heterocyclic group and OH protecting group;

[0233] Each R c Independently selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne, substituted or unsubstituted aryl groups, and substituted or unsubstituted heterocyclic groups;

[0234] Each R d and R e Independently selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12The protecting groups of alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclic groups, and amino groups; and

[0235] Each R f Independently selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl, substituted or unsubstituted C2-C 12 Alkyne group, substituted or unsubstituted aryl group, substituted or unsubstituted heterocyclic group and SH protecting group.

[0236] Preferably, R8 is as described herein.

[0237] Preferably, Y1 is selected from hydrogen and OR b In some preferred embodiments, Y1 is hydrogen. In other preferred embodiments, Y1 is OR b Preferably, it is methoxylated.

[0238] Preferably, Y2 is hydrogen.

[0239] Preferably, Y3 is selected from hydrogen, -CH2OH, -CH2OC(=O)R g -CH2NH2 and -CH2NHR h , where R g Selected from substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 Alkyne group, and R h For Prot NH .

[0240] The conversion of compound XVI to provide compound XVIIb is typically carried out using the Pictet-Spengler reaction. This reaction is usually carried out under acid catalysis. Silica gel is the preferred catalyst.

[0241] The conversion of compound XVIIb to provide compound XVIIIb is typically carried out by reaction with a nitrile-coordinated transition metal salt. More preferred salts are salts of Ag(I) or Cu(I). Most preferred salts are AgNO3 and CuCl.

[0242] Possible steps in scheme VI are shown in scheme VII:

[0243]

[0244] Option VII

[0245] Where R8 is -C(=O)R aGroup, wherein R a Selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 alkynyl group;

[0246] X is -NH- or -O-;

[0247] Y1 is hydrogen or -OR b Group, wherein R b Selected from substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 alkynyl group;

[0248] Y3 is selected from hydrogen, -CH2OH, and -CH2OC(=O)R. g -CH2NH2 and -CH2NHR h , where R g Selected from substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 Alkyne group, and R h For Prot NH .

[0249] Preferably, R8 and Y1 are as described herein.

[0250] Preferably, Y3 is selected from -CH2NH2 and -CH2NHR h More preferably -CH2NHR h .

[0251] Preferably, R h For Alloc.

[0252] Preferably, Y3 is -CH2NHR h The method further includes, after the step of reacting compound XVI with compound B to provide compound XVIIb, carrying out a deprotection reaction to remove -CH2NHR h The step of conversion to -CH2NH2. Suitable conditions and reagents for deprotecting this protecting group are known to those skilled in the art (see Wuts, PGM, and Greene TW's description in Protecting Groups in Organic Synthesis). The 4th edition of Wiley-Interscience, and Kocienski PJ's description in Protecting Groups, (Georg Thieme Verlag, 3rd edition). For example, in the case of Alloc, deprotection is typically carried out using metal catalysis. Preferred catalysts include palladium (0) catalysts and palladium (II) catalysts in the presence of a reducing agent (such as hydrogenated trialkyltin or a secondary amine).

[0253] The possible steps in scheme VI are shown in scheme VIII:

[0254]

[0255] Scheme VIII

[0256] Where R8 is -C(=O)R a Group, wherein R a Selected from hydrogen, substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 alkynyl group;

[0257] Y1 is hydrogen or -OR b Group, wherein R b Selected from substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 alkynyl group;

[0258] Y3 is selected from hydrogen, -CH2OH, and -CH2OC(=O)R g , where R g Selected from substituted or unsubstituted C1-C 12 Alkyl, substituted or unsubstituted C2-C 12 Alkenyl and substituted or unsubstituted C2-C 12 Alkyne group.

[0259] Preferably, R8 and Y1 are as described herein.

[0260] Preferably, Y3 is selected from hydrogen and -CH2OH. In some more preferred embodiments, Y3 is hydrogen. In other more preferred embodiments, Y3 is -CH2OH.

[0261] Preferably, compound B is:

[0262] .

[0263] Preferably, compound B is:

[0264] .

[0265] Preferably, compound B is:

[0266] .

[0267] Preferably, compound B is:

[0268] .

[0269] Preferably, the compound of formula XVIIIb is lurbinectedin:

[0270] .

[0271] Preferably, the compound of formula XVIIIb is ecubicedin:

[0272] .

[0273] Preferably, the compound of formula XVIIIb is:

[0274] .

[0275] Preferably, the compound of formula XVIIIb is:

[0276] .

[0277] Preferably, in formulas Xa, XI, XII, and XIII, the stereochemical configuration at the asymmetric carbon bonded to the NHR6 group is (R).

[0278] Preferably, in formulas XIV, XV, XV-Int-a, and XV-Int-b, the stereochemical configuration of the asymmetric carbon bonded to the NHR6 or NH2 group is (R).

[0279] intermediate compounds

[0280] According to another aspect of the invention, what is provided herein is a compound according to Formula I:

[0281]

[0282] R1 to R3 are defined as in this paper.

[0283] Preferably, the compound has the following formula:

[0284] .

[0285] Preferably, the compound has the following formula:

[0286] .

[0287] Preferably, the compound has the following formula:

[0288] .

[0289] In alternative preferred embodiments, the compounds according to Formula I do not include compound HQ-3.

[0290] According to another aspect of the invention, a compound of the following formula is provided herein:

[0291] .

[0292] The present invention will be further described in the following non-limiting examples.

[0293] Example

[0294] Separate examples 1 to 3:

[0295] Separation Example 1 – Compound HQ-1:

[0296]

[0297] Since HQ-1 is difficult to separate without oxidation, HQ-1 was prepared by reduction 1 and then compared with the HQ-1 / 1 mixture by HPLC / UV method.

[0298] Procedure: A container containing 0.7 mL of CD3OD with 25 mg of 1 and 9 mg of 10% Pd / C was bubbled with a stream of hydrogen for 5 min, and then the mixture was stirred under a hydrogen atmosphere for 2 h. The yellow solution turned into a colorless solution. Pd / C was filtered off under a nitrogen atmosphere, and the product was immediately analyzed by NMR and HPLC / UV / MS techniques.

[0299] The HPLC / UV and UV data were compared with the HQ-1 / 1 mixture, confirming that the compound was HQ-1.

[0300] HPLC method:

[0301] Mobile phase A: 10 nM ammonium acetate adjusted to pH 3.0 with formic acid, 1% DEA

[0302] Mobile phase B: Acetonitrile

[0303] Column: waters, Symmetry 300 C18 5μm 250 mm x 4.6 mm

[0304] Pre-column: Waters, Symmetry C18 5μm 20 mm x 3.9 mm

[0305] Oven temperature: 40℃

[0306] Injector temperature: 5℃

[0307] Detection wavelength 268 nm; DAD acquisition

[0308] Gradient: 0 min: 5% B

[0309] From 5% to 13% within 17 minutes B

[0310] From 13% to 38% within 9 minutes B

[0311] From 38% to 5% within 2 minutes B

[0312] and maintain at 5% B for 8 minutes

[0313] Injection volume: 50 µL

[0314] Flow rate: 1.2 mL / min

[0315] 1 of r t : 12 min (UV maximum 268 nm); r of HQ-1 t : 10 min (rrt 0.81) (UV maximum value 285 nm)

[0316] 1 H NMR (400 MHz, CD3OD) δ 6.50 (s, 1H), 4.55 (dd, J = 5.5, 2.6 Hz,1H), 4.17 (d, J = 2.6 Hz, 1H), 4.13 – 4.08 (m, 1H), 3.72 (s, 3H), 3.64 (s,3H), 3.49 – 3.39 (m, 2H), 3.34 – 3.32 (m, 1H), 3.26 – 3.17 (m, 1H), 3.08 (dd,J = 13.2, 5.5 Hz, 1H), 3.05 – 2.94 (m, 1H), 2.73 (q, J = 6.9 Hz, 1H), 2.56 –2.48 (m, 1H), 2.26 – 2.22 (m, 6H), 2.10 (s, 3H), 1.82 (dd, J = 15.8, 12.3 Hz, 1H), 0.94 (d, J = 6.9 Hz, 3H).

[0317] 13 C NMR (101 MHz, CD3OD) δ 176.30, 147.58, 143.99, 139.74, 130.84,129.23, 120.63, 120.32, 120.24, 117.63, 93.52, 59.42, 59.36, 56.54, 53.18,53.09, 52.84, 50.14, 42.41, 40.11, 25.86, 24.54, 19.82, 14.56, 8.37.

[0318] ESI m / z 525.5 [M-H2O+H] +

[0319] Maximum UV value: 285 nm

[0320] Separation Example 1 – Compound HQ-2:

[0321]

[0322] Since HQ-2 is difficult to separate without oxidation, HQ-2 was prepared by reduction 2 and then compared with HQ-2 / 2 prepared from a cyanidated HQ-1 / 1 mixture by HPLC / UV method.

[0323] Procedure: A container containing 0.5 mL of CD3OD with 15 mg Pd / C and 5 mg Pd / C was bubbled with a stream of hydrogen for 5 min, and then the mixture was stirred under a hydrogen atmosphere for 2 h. The yellow solution turned into a colorless solution. Pd / C was filtered off under a nitrogen atmosphere, and the product was immediately analyzed by NMR and HPLC / UV / MS techniques.

[0324] HPLC / UV and UV data were compared with cyaninated HQ-1 / 1 mixtures, confirming that the compound was HQ-2.

[0325] HQ-2 was detected by HPLC / UV / MS technology.

[0326] HPLC method:

[0327] Mobile phase A: 10 nM ammonium acetate adjusted to pH 3.0 with formic acid, 1% DEA

[0328] Mobile phase B: Acetonitrile

[0329] Column: waters, Symmetry 300 C18 5μm 250 mm x 4.6 mm

[0330] Pre-column: Waters, Symmetry C18 5μm 20 mm x 3.9 mm

[0331] Oven temperature: 40℃

[0332] Injector temperature: 4℃

[0333] Detection wavelength 268 nm; DAD acquisition

[0334] Gradient: 0 min: 5% B

[0335] From 5% to 50% within 60 minutes B

[0336] From 50% to 5% within 2 minutes B

[0337] and maintain at 5% B for 10 minutes

[0338] Injection volume: 10 µL

[0339] Flow rate: 1.2 mL / min

[0340] 2 of r t : 22 min (UV maximum 268 nm); r of HQ-2 t 11.6 min (maximum UV wavelength 285 nm)

[0341] 1 H NMR (400 MHz, CD3OD) δ 6.48 (s, 1H), 4.32 (d, J = 2.5 Hz, 1H), 4.16(d, J = 3.0 Hz, 2H), 3.72 (d, J = 0.8 Hz, 3H), 3.65 (d, J = 0.8 Hz, 3H), 3.64– 3.57 (m, 2H), 3.43–3.23 (m, 2H), 3.15–2.97 (m, 2H), 2.79–2.69 (m,1H), 2.69–2.59 (m, 1H), 2.30–2.18 (m, 6H), 2.12 (s, 3H), 1.89 (dd, J =16.1, 11.9 Hz, 1H), 0.88 (dd, J = 7.0, 0.8 Hz, 3H).

[0342] 13C NMR (101 MHz, CD3OD) δ 176.20, 147.66, 144.08, 143.45, 139.82,130.94, 129.22, 120.41, 120.09, 118.43, 117.98, 117.79, 59.55, 59.48, 59.37,56.70, 56.58, 56.56, 55.29, 50.01, 40.76, 40.45, 25.79, 25.08, 19.66, 14.53,8.41.

[0343] ESI m / z 525.5 [M-CN+H] + 552.5 [M+H] +

[0344] Maximum UV value: 285 nm

[0345] Separation Example 1 – Compound HQ-3:

[0346]

[0347] Process A: HQ-3 is separated from the crude mixture of HQ-3 / 3 obtained by cyanidation of the HQ-1 / 1 mixture followed by the introduction of a tert-butoxycarbonyl group (see the reaction below) by evaporating the aqueous phase to dryness.

[0348] The NMR, UV, and HPLC / MS data matched those of the compound obtained by reducing compound 3 as follows, which confirmed its structure.

[0349] Procedure B: A container containing 15 mg of 3 and 2 mg of 10% Pd / C in 0.7 mL of CD3OD was bubbled with a stream of hydrogen for 5 min, and then the mixture was stirred under a hydrogen atmosphere for 2 h. The yellow solution turned into a colorless solution. Pd / C was filtered off under a nitrogen atmosphere, and the product was immediately analyzed by NMR and HPLC / UV / MS techniques.

[0350] HPLC method:

[0351] Mobile phase A: 10 nM ammonium acetate adjusted to pH 3.0 with formic acid, 1% DEA

[0352] Mobile phase B: Acetonitrile

[0353] Column: waters, Symmetry 300 C18 5μm 250 mm x 4.6 mm

[0354] Pre-column: Waters, Symmetry C18 5μm 20 mm x 3.9 mm

[0355] Oven temperature: 40℃

[0356] Detection wavelength 268 nm; DAD acquisition

[0357] Gradient: 0 min: 5% B

[0358] From 5% to 35% within 30 minutes B

[0359] From 35% to 60% within 50 minutes B

[0360] From 60% to 5% within 2 minutes B

[0361] and maintain at 5% B for 8 minutes

[0362] Injection volume: 10 µL

[0363] Flow rate: 1.2 mL / min

[0364] 3 of r t : 42.38 min (UV maximum 268 nm); r of HQ-3 t 29.8 min (maximum UV wavelength 285 nm)

[0365] Characterization data:

[0366] 1 H NMR (400 MHz, CD3OD) δ 6.48 (s, 1H), 4.31 (d, J = 2.5 Hz, 1H), 4.19– 4.12 (m, 2H), 3.71 (d, J = 0.7 Hz, 3H), 3.66 (d, J = 0.7 Hz, 3H), 3.43 (d,J = 6.3 Hz, 1H), 3.38 – 3.19 (m, 3H), 3.11 – 2.95 (m, 2H), 2.71 – 2.58 (m,1H), 2.29 – 2.19 (m, 6H), 2.11 (s, 3H), 1.95 (t, J = 14.1 Hz, 1H), 1.35 (s,9H), 0.92 (d, J = 7.1 Hz, 3H).

[0367] 13C NMR (101 MHz, CD3OD) δ 173.46, 155.80, 147.61, 144.07, 143.63,143.57, 139.66, 130.84, 129.17, 120.64, 120.11, 118.59, 118.02, 117.92,117.71, 79.05, 59.89, 59.48, 59.32, 57.08, 56.73, 56.61, 55.38, 49.69, 41.92,40.42, 27.23, 25.75, 25.15, 17.04, 14.64, 8.43.

[0368] ESI m / z 625.6 [M-CN+H] + 652.5 [M+H] +

[0369] Maximum UV value: 285 nm

[0370] Examples 1 to 5 – Oxidation of compound HQ-1 to compound 1: Effects of different oxidizing agents

[0371]

[0372] Starting materials:

[0373] For Examples 1 through 4, fermentation broth from *Pseudomonas fluorescens* (produced according to JPS59225189A) was used. The starting material pH solution was 6.26. The ratio of compound HQ-1 to compound 1 was:

[0374]

[0375] For Example 5, aliquots of the sample were prepared using 50 mL of methanol concentrate with the following composition:

[0376]

[0377] The methanol concentrate was obtained using the following procedure: SepaBeads 825L or 850 resin was added to the fermentation broth to adsorb compounds HQ-1 and 1. The resin was then filtered off, and HQ-1 and 1 were desorbed with methanol or a 0.1% to 1% formic acid-methanol solution. The solvent was evaporated to a (HQ-1 + 1) concentration of 3-5 g / L.

[0378] process:

[0379] Examples 1 to 4:At 22°C and the specified pH, an air stream was bubbled through 100 mL of solution for 10 min. An oxidant was then added to the mixture under an air atmosphere. The oxidant equivalent was calculated based on the amount of compound HQ-1.

[0380] Samples were taken and analyzed at different times to check the progress of the reaction.

[0381] The contents of compounds HQ-1 and 1 were determined by HPLC.

[0382] Example 5: At 22°C and the specified pH, an air stream is bubbled through 50 mL of methanol concentrate to obtain an aliquot of the sample for 10 min or throughout the experiment. The oxidant 1,4-benzoquinone is added to the mixture under an air atmosphere. The oxidant equivalent is calculated based on the amount of compound HQ-1.

[0383] Samples were taken and analyzed at different times to check the progress of the reaction.

[0384] The contents of compounds HQ-1 and 1 were determined by HPLC area. Considering that the relative response factor of compound 1 / compound HQ-1 is 4.5 at 268 nm, the ratio of compound HQ-1 to compound 1 was calculated. A lower ratio of compound HQ-1 to compound 1 indicates a higher conversion rate of the starting material to the oxidation product.

[0385] HPLC method:

[0386] Mobile phase A: 10 nM ammonium acetate adjusted to pH 3.0 with formic acid, 1% DEA

[0387] Mobile phase B: Acetonitrile

[0388] Column: waters, Symmetry 300 C18 5μm 250 mm x 4.6 mm

[0389] Pre-column: Waters, Symmetry C18 5μm 20 mm x 3.9 mm

[0390] Oven temperature: 40℃

[0391] Injector temperature: 5℃

[0392] Detection wavelength 268 nm

[0393] Gradient: 0 min: 5% B

[0394] From 5% to 13% within 17 minutes B

[0395] From 13% to 38% within 9 minutes B

[0396] From 38% to 5% within 2 minutes B

[0397] and maintain at 5% B for 8 minutes

[0398] 1 of r t Approximately 12 minutes, HQ-1's r t Approximately 10 minutes

[0399] Experimental parameters:

[0400]

[0401] result:

[0402]

[0403]

[0404] Examples 1 through 5 show that a variety of oxidants can be used to oxidize compound HQ-1 to compound 1, including air, H2O2, 1,4-benzoquinone, CuSO4, and FeCl3.

[0405] Specifically, in the experiments using CuSO4 as an oxidant, these were conducted under two different pH conditions: 4.15 and 7.22. At pH 7.22, compound HQ-1 was completely oxidized. After 2 h, CuSO4 also proved to be an effective oxidant at low pH, and furthermore, no degradation of compound 1 was observed over time.

[0406] Similar results were obtained with FeCl3, which is also effectively oxidized at acidic pH. In this case, compound 1 was also stable after 5 h.

[0407] Neither FeCl3 nor CuSO4 exhibited reversible behavior over time.

[0408] In general, various oxidizing agents can be used to oxidize compound HQ-1 to compound 1. FeCl3 and CuSO4 are particularly suitable because both are water-soluble and allow oxidation at acidic pH, where the product is more stable.

[0409] Examples 6 to 10 – Oxidation of compound HQ-1 to compound 1: Effects of atmosphere and water dilution

[0410] Starting materials:

[0411] Solution 1

[0412] The methanol concentrate prepared as described in Example 5 was used. The starting material pH solution was 4.48. The ratio of compound HQ-1 to compound 1 was:

[0413]

[0414] Solution 2

[0415] The methanol concentrate prepared as described in Example 5 was used. The starting material pH solution was 4.26. The ratio of compound HQ-1 to compound 1 was:

[0416]

[0417] process:

[0418] At 22°C, an air stream was bubbled through a 100 mL methanol concentrate sample for 10 min. An oxidant was then added to the mixture under air atmosphere. The oxidant equivalent was calculated based on the amount of compound HQ-1 or the amounts of both compound HQ-1 and compound 1.

[0419] To conduct the experiment under a nitrogen atmosphere, a vacuum was applied to the solution to remove oxygen, and then a nitrogen stream was bubbled through the methanol solution for 10 minutes.

[0420] In Example 8, the inerted methanol concentrate was diluted with water (166 mL per 100 mL of methanol concentrate), and then an oxidant was added.

[0421] In Example 10-2, the oxidized methanol concentrate from Example 10-1 was diluted with water (166 mL per 100 mL of methanol concentrate).

[0422] The contents of compounds HQ-1 and 1 were determined by HPLC.

[0423] Experimental parameters:

[0424]

[0425] *Oxidizing agent equivalent is calculated based on the amount of compound HQ-1.

[0426] **Oxidizing agent equivalent is calculated based on the amounts of compound HQ-1 and compound 1.

[0427] result:

[0428]

[0429] *Time represents the product of Example 10-1 after dilution with water.

[0430] Examples 6 and 7 clearly demonstrate the ability of FeCl3 to oxidize compound HQ-1 to compound 1 in an inert or oxidizing atmosphere at an acidic pH. It should be noted that the amount of compound 1 remains the same even after 5 hours.

[0431] Example 8 shows that, despite the observation of a small amount of incomplete oxidation, oxidation still proceeds smoothly under dilution conditions.

[0432] Example 9 shows that CuSO4 oxidizes compound HQ-1 in an inert atmosphere at an acidic pH. A small amount of incomplete oxidation was observed.

[0433] Example 10 shows complete conversion within 30 min, possibly due to the addition of a higher oxidant equivalent compared to Example 6, and possibly due to a higher concentration compared to Example 8.

[0434] Overall, FeCl3 exhibited the highest oxidation effect among the oxidants tested in an inert atmosphere. This allows oxidation to proceed in the absence of oxygen, an advantage related to safety requirements. Only 3 equivalents of FeCl3 were needed to achieve complete oxidation of compound HQ-1 within 30 minutes. Furthermore, CuSO4 was also confirmed to be usable as an oxidant, and oxidation still occurred under diluted conditions.

[0435] Example 11 – Oxidation of compound HQ-1 to compound 1: The effect of temperature

[0436] Starting materials:

[0437] The methanol concentrate prepared as described in Example 5 was used. The starting material pH solution was 4.26. The ratio of compound HQ-1 to compound 1 was:

[0438]

[0439] process:

[0440] Add 83 mL of water to 50 mL of a methanol solution containing compound HQ-1 and compound 1. Adjust the pH to 4.05 with 10% H2SO4 solution and keep the mixture at a constant temperature of 15.5 °C.

[0441] Nitrogen gas was bubbled through the solution for 5 minutes to remove trace amounts of oxygen. Then, 26% FeCl3·6H2O (500 mg in 1.9 mL H2O, equivalent to 3 equivalents of compounds HQ-1 and 1) was added. The solution turned green.

[0442] The contents of compounds HQ-1 and 1 were determined by HPLC.

[0443] result:

[0444]

[0445] Example 11 shows that oxidation at a lower temperature is comparable to the process at 22°C (Example 8).

[0446] Example 12 – Effect of the oxidation of compound HQ-1 to compound 1: FeCl3 equivalent

[0447] Starting materials:

[0448] The methanol concentrate prepared as described in Example 5 was used. The starting material pH solution was 4.26. The ratio of compound HQ-1 to compound 1 was:

[0449]

[0450] process:

[0451] Using the same procedure as in Example 11, the equivalent number of FeCl3 (relative to the amounts of compound HQ-1 and compound 1) was reduced from 3 to 1.5.

[0452] The contents of compounds HQ-1 and 1 were determined by HPLC.

[0453] result:

[0454]

[0455] Example 12 shows that lower equivalents of oxidation are possible, although the conversion is slightly worse compared to the process using 3 equivalents of FeCl3 (Example 11).

[0456] Example 13 – Oxidation of compound HQ-1 to compound 1: Effects of visible light

[0457] process:

[0458] An experiment was conducted on an equal fraction of 50 mL of methanol concentrate, which was prepared as described in Example 5 and had the following composition:

[0459]

[0460] A series of reactions were conducted at room temperature, with the pH adjusted to 3.8, under either a nitrogen or air atmosphere, and with or without visible light. Some reactions were carried out in a concentrated methanol solution, while others were carried out with a diluted methanol solution.

[0461] For diluted samples, dilute the above aliquots with 125 mL of water.

[0462] To evaluate the visible light effect, a glass container was used in the presence of visible light, and an amber glass container was used when visible light was absent.

[0463] The reaction progress was assessed by HPLC analysis at t = 2–5 h and then at t = 20–24 h.

[0464] Given that the relative response factor of compound 1 / compound HQ-1 is 4.5 at 268 nm, the ratio of compound HQ-1 / compound 1 is calculated.

[0465] result:

[0466]

[0467] Example 13 shows that oxidation is carried out under visible light irradiation and / or in an air atmosphere. If oxidation is carried out in the presence of both visible light and an air atmosphere, the reaction is accelerated.

[0468] The oxidation rate is not significantly affected by dilution.

[0469] Example 14 – Oxidation of compound HQ-1 to compound 1: The bubbling effect during the reaction

[0470] process:

[0471] An experiment was conducted on an equal fraction of 50 mL of methanol concentrate, which was prepared as described in Example 5 and had the following composition:

[0472]

[0473] For the diluted sample, the above aliquots were diluted with 125 mL of water, and then the methanol was partially evaporated to a methanol content of 7.9%. The percentages of compound HQ-1 and compound 1 were measured:

[0474]

[0475] The experiment was run at pH 4 or pH 7 by alkalization with 10% NaOH.

[0476] An air atmosphere is provided using an air balloon. In the case of bubbling, this is done by bubbling air through the solution during the experiment.

[0477] result:

[0478]

[0479] Example 14 shows that oxidation takes place in an air atmosphere at either an acidic or neutral pH. Oxidation proceeds more rapidly at pH 7. Furthermore, bubbling air through the reaction solution increases oxidation.

[0480] Examples 15 to 17 – One-pot cyanidation of compound 1 to form compound 2

[0481]

[0482] Starting materials:

[0483] The methanol concentrate prepared as described in Example 5 was used. The starting material pH solution was 4.26. The ratio of compound HQ-1 to compound 1 was:

[0484]

[0485] process:

[0486] Add cooling water to a predetermined volume of methanol concentrate (1.66 mL water per 1 mL concentrate). Adjust the pH to 4–4.2 with 10% H₂SO₄ or 10% NaOH as needed. Maintain the mixture at 15°C and stir under a nitrogen atmosphere. Then add 26% FeCl₃·6H₂O solution and take samples for analysis at 30 min and 60 min.

[0487] One to 1.5 h after adding the oxidant, an aqueous solution of KCN (31 g / L) was added at the same temperature.

[0488] Samples were continuously drawn from the reaction vessel, and compounds 1 and 2 were measured by HPLC.

[0489] The equivalent changes of FeCl3.6H2O and KCN are as follows:

[0490]

[0491] result:

[0492]

[0493] The addition of KCN is accompanied by a color change. Dark green indicates the formation of K4[Fe(CN)6] (blue) and K3[Fe(CN)6] (yellow) complexes.

[0494] In Examples 15 and 16, where cyanide availability was high, Compound 2 was readily obtained within 3 to 4 hours.

[0495] In Example 17, the mixture was allowed to react overnight, and after 20 h, all of Compound 1 was quantitatively converted to Compound 2.

[0496] In general, Examples 15 to 17 show that compound HQ-1 can be oxidized in a one-pot process to form compound 1, and then compound 1 can be cyanided to form compound 2.

[0497] Examples 18 to 20 – Boc protection of compound 2 to form compound 3

[0498]

[0499] process:

[0500] The solution of compound 2 obtained from Examples 16 to 18 was washed twice with MTBE (50 mL MTBE per 134 mL aqueous phase). The pH was adjusted to 8.3 with 10% NaOH at 15 °C. The cooling bath was removed, and MTBE (1 g / 100 mL) containing Boc2O (4 equivalents) and catalytic DMAP were added.

[0501] After the reaction was complete, the phases were decanted and the organic phase was washed twice with water (50 mL water per 134 mL of compound 2 solution). The organic solvent was evaporated to dryness, and the crude product was purified by silica gel rapid chromatography (DCM / dioxane). The fraction containing compound 3 was evaporated to dryness, dissolved in toluene (4 mL / g), and poured into heptane (38 mL / g) at 0 °C. The resulting solid was filtered off to give pure compound 3.

[0502] result:

[0503]

[0504] Examples 18 to 20 showed similar results in terms of chromatographic purity after reacting overnight.

[0505] Example 21 – Oxidation of compound HQ-2 to compound 2: The effect of pH on oxidation with air

[0506]

[0507] process:

[0508] An aqueous solution of compound HQ-2 + compound 2 was obtained from 50 mL of methanol concentrate, the methanol concentrate having the following composition:

[0509]

[0510] It was then acidified to pH 3.8 with 125 mL of water and subjected to cyanation along with 13.2 equivalents of KCN.

[0511] After the reaction is terminated, adjust the pH to 4.0 or 7.0 with 10% H2SO4 or 10% NaOH as needed, and bubble nitrogen gas through the mixture to remove oxygen.

[0512] The aqueous phase was washed twice with MTBE (2 x 120 mL). This aqueous phase was used as the starting material for subsequent experiments.

[0513] The experiment was conducted at room temperature. Samples were taken to analyze the reaction progress.

[0514] result:

[0515]

[0516] Example 21 shows that oxidation takes place in an air atmosphere at an acidic or neutral pH. Oxidation proceeds more rapidly at pH 7. Furthermore, bubbling air through the reaction solution increases oxidation.

[0517] Example 22 – Oxidation of compound HQ-2 to compound 2: The effect of temperature on oxidation with air

[0518] process:

[0519] At pH 7, the same starting materials as in Example 21 were used. At 10°C, air was introduced via an air stream during the reaction process.

[0520] result:

[0521]

[0522] Example 22 shows that oxidation can be carried out at 10°C.

[0523] Example 23 – Oxidation of compound HQ-1 and subsequent reactions to form urea derivatives

[0524] Step A:

[0525]

[0526] Procedure: The methanol extract containing compounds HQ-1 and 1 (containing 41% HQ-1 area) (550 mL, 2.06 g, 3.81 mmol; HQ-1+1 concentration: 3.74 g / L) was diluted with H2O (650 mL) at 15 °C until a concentration of 1.7 g / L was obtained. FeCl3·6H2O aqueous solution (1.60 g; 5.55 mmol in 5 mL H2O) was added and stirred for 45 min. H2O containing KCN (2.30 g; 34.29 mmol) (75 mL) was added and the pH was adjusted to 3.92 with 10% H2SO4 aqueous solution (10 mL). The mixture was stirred at 15 °C for 3 h, cooled to 0 °C, and 3M... NaOH (21 mL) until pH 8.10.

[0527] MTBE (300 mL) and phenyl isocyanate (1.53 mL; 14.1 mmol) were added to the aqueous mixture and the mixture was heated to 22 °C. After 2 h, the solid was filtered through diatomaceous earth and the diatomaceous earth was washed with MTBE (200 mL). The phases were separated, and the aqueous layer was extracted with MTBE (100 mL). The combined organic phases were washed with H2O (400 mL), dried over Na2SO4, filtered, and concentrated. The desired compound (1.012 g, 90%) was purified by column chromatography (SiO2, hexane-ethyl acetate 0→65%) to obtain an orange solid.

[0528] Rf: 0.42 (hexane-ethyl acetate 1:3)

[0529] 1 H NMR (400 MHz, CD3OD) δ 7.22 (d, J = 4.2 Hz, 4H), 6.93 (m, 1H), 6.52(s, 1H), 6.04 (bs, 1H), 5.49 (s, 1H), 4.43 (d, J = 2.6 Hz, 1H), 4.22 – 4.16(m, 1H), 3.92 (s, 3H), 3.84 (bs, 1H), 3.65 (s, 3H), 3.68 – 3.57 (m, 1H), 3.42– 3.33 (m, 3H), 3.29 – 3.22 (m, 1H), 3.17 – 3.00 (m, 3H), 2.67 (d, J = 18.3Hz, 1H), 2.30 (s, 3H), 2.23 (s, 3H), 2.05 – 1.92 (m, 1H), 1.72 (s, 3H), 0.93(d, J = 7.1 Hz, 3H).

[0530] 13 C NMR (101 MHz, CD3OD) δ 185.6, 181.3, 174.3, 174.2, 155.7, 155.2,148.0, 143.8, 142.7, 139.5, 134.9, 130.9, 129.7, 128.4, 127.8, 121.7, 119.7,118.1, 117.5, 117.4, 59.8, 59.1, 58.3, 56.01, 56.0, 55.2, 54.8, 49.3, 40.5,40.0, 39.9, 24.8, 24.4, 17.5, 14.6, 7.3.

[0531] ESI-MS m / z: C 36 H 40Calculated value of N6O7: 668.30. (M+H) + Measured value: 669.30

[0532] Step B:

[0533]

[0534] Procedure: Compound 4 (18.0 g, ) was cooled to -3°C under a nitrogen atmosphere. 26.92 mmol) of MgCl₂ (3.91 mL, 43.07 mmol) and H₂O (0.270 mL) were added to a solution of THF (261 mL). Then, 60% NaH₂O (0.18 g; 1.29 g; 32.30 mL) was added. The 1 mmol (mmol) was added in portions over a 40 min interval, and the mixture was stirred at 0 °C for an additional 30 min. The reaction was quenched with saturated NH4Cl solution (200 mL) and diluted with DCM (100 mL). The phases were separated, and the aqueous phase was extracted with more DCM (2 x 50 mL). The combined organic phases were dried over Na2SO4 and concentrated under vacuum. The residue was purified by column chromatography (SiO2, DCM: ethyl acetate 0→70%) to give compound 5 (16.0 g, 84%) as an orange solid.

[0535] Rf: 0.33 (hexane:ethyl acetate 1:3).

[0536] 1 H NMR (400 MHz, CD3OD) δ 7.21 (m, 5H), 6.98 – 6.88 (m, 1H), 6.80 (s,1H), 5.18 (d, J = 6.1 Hz, 1H), 5.11 (d, J = 6.1 Hz, 1H), 4.44 (d, J = 2.6 Hz,1H), 4.28 – 4.23 (m, 1H), 3.91 (s, 3H), 3.82 (bs, 1H), 3.68 (s, 3H), 3.60 –3.51 (m, 4H), 3.51 – 3.38 (m, 2H), 3.14 – 3.00 (m, 3H), 2.73 (d, J = 18.0 Hz,1H), 2.32 (s, 3H), 2.22 (s, 3H), 1.98 – 1.79 (m, 1H), 1.69 (s, 3H), 0.92 (d,J = 6.0 Hz, 3H).

[0537] 13 C NMR (101 MHz, CD3OD) δ 187.0, 182.8, 175.5, 157.1, 156.4, 150.4,149.7, 144.0, 141.0, 136.3, 132.5, 132.2, 129.8, 129.2, 126.0, 125.1, 123.0,119.4, 118.8, 100.4, 61.2, 60.3, 59.7, 58.1, 57.9, 57.7, 56.5, 56.1, 50.7,41.9, 40.8, 26.0, 25.8, 20.9, 18.8, 16.0, 14.5, 8.7.

[0538] ESI-MS m / z: C 38 H 44 Calculated value of N6O8: 712.32. (M+H) + Measured value: 713.32

[0539] Example 24 – Oxidation of compound HQ-1 and subsequent reactions to form a thiourea derivative.

[0540]

[0541] Procedure: The methanol extract containing compounds HQ-1 and 1 (containing 3% HQ-1) (750 mL, 3.30 g, 6.10 mmol; HQ-1+1 concentration: 4.4 g / L) was diluted with H2O (1190 mL) at 15 °C until a dilution of 1.7 g / L was obtained. FeCl3·6H2O aqueous solution (1.32 g; 6.10 mmol in 4 mL H2O) was added and stirred for 70 min. NaCN (2.69 g; 54.9 mmol) in H2O (11 mL) was added, and the pH was adjusted with 10% H2SO4 aqueous solution (10 mL) until pH 3.91. The mixture was stirred at 15 °C for 3 h, cooled to 0 °C, and 3M... NaOH (21 mL) until pH 8.1.

[0542] MTBE (472 mL) and phenyl isothiocyanate (2.70 mL; 22.6 mL) were added. 1 mmol) was added to the aqueous mixture, heated to 22 °C and stirred for 18 h. The solid was filtered through diatomaceous earth and washed with MTBE (200 mL). The phases were separated and the aqueous layer was extracted with MTBE (200 mL). The combined organic phases were dried over Na2SO4, filtered and evaporated, and the residue was purified by column chromatography (SiO2, DCM: methanol 0→3%) to give compound 6 (3.35 g, 82%) as an orange solid.

[0543] Rf: 0.41 (DCM-methanol 96:4).

[0544] 1 H NMR (400 MHz, CD3OD) δ 7.41 – 7.30 (m, 5H), 7.19 (m, 1H), 6.25 (s,1H), 4.43 (d, J = 2.6 Hz, 2H), 4.22 – 4.12 (m, 2H), 3.93 (s, 3H), 3.86 – 3.77(m, 1H), 3.67 (s, 3H), 3.63 – 3.47 (m, 2H), 3.38 (d, J = 7.7 Hz, 2H), 3.12 –3.03 (m, 2H), 3.02 – 2.91 (m, 1H), 2.68 (d, J = 18.0 Hz, 1H), 2.26 (s, 3H), 2.14 (s, 3H), 1.97 – 1.84 (m, 1H), 1.83 (s, 3H), 0.90 (d, J = 6.0 Hz, 3H).

[0545] 13 C NMR (126 MHz, CD3OD) δ 187.2, 182.7, 181.4, 174.3, 157.1, 149.3,145.0, 144.2, 139.6, 136.6, 132.3, 131.0, 130.2, 129.4, 126.6, 125.0, 121.1,118.9, 118.4, 61.3, 60.9, 59.7, 58.0, 57.3, 56.5, 56.3, 54.8, 54.4, 41.9,40.7, 30.7, 26.1, 26.0, 18.7, 16.0, 8.9.

[0546] ESI-MS m / z: C 36 H 40Calculated value of N6O6S: 684.27. (M+H) + Measured value: 685.30

[0547] Example 25

[0548] Fermentation broth was obtained according to Examples 1-4.

[0549] Procedure: Oxidant was added to 10 mL of fermentation broth under a nitrogen atmosphere, at 22°C and pH 4. Samples were taken at different time points to analyze the reaction progress by HPLC. The HQ-1+1 content was determined to verify the absence of degradation.

[0550]

[0551] All tested reagents showed effectiveness without degrading HQ-1 or 1. The use of potassium permanganate demonstrates that degradation can be avoided by using lower molar equivalents.

[0552] Example 26

[0553] A methanol concentrate was obtained according to Examples 1-4. Procedure: Under a nitrogen atmosphere, 10 mL of the methanol concentrate was diluted with 11.7 mL of water to obtain a final concentration of HQ-1+1 in the range of approximately 1.58–1.8 g / L. An oxidant was added to this solution. The reagent equivalent was calculated based on the total amount of HQ-1+1. The oxidant was added under a nitrogen atmosphere at 22 °C and pH 4. Samples were taken at different time points to analyze the reaction progress by HPLC. The HQ-1+1 content was determined to verify the absence of degradation.

[0554]

[0555] *After using copper sulfate for 2 hours, bubbling oxygen into the solution for 5 minutes. Then, maintain the mixture in an air atmosphere for 1.5 hours.

[0556] In this example, a mixture of oxidants was tested to accelerate oxidation. In all cases, HQ-1 was effectively oxidized without any observed degradation.

[0557] In summary, examples demonstrate that various oxidants (e.g., air (containing molecular oxygen), H₂O₂, 1,4-benzoquinone, CuSO₄, FeCl₃, tetrachlorobenzoquinone, K₃[Fe(CN)₆], MnO₂, I₂, KMnO₄, HIO₃, and Mn(OAc)₃, with FeCl₃ being particularly preferred) can be used to convert compounds of formula I (e.g., compounds HQ-1 and HQ-2) into compounds of formula II. One-pot methods combining oxidation and cyanidation have also been demonstrated.

[0558] Compound HQ-3 has been found to be highly insoluble in both organic and aqueous media, resulting in precipitate formation and reduced overall process yield. These examples demonstrate that the amount of precipitate generated during the reaction process can be reduced by decreasing the amount of compound HQ-3 produced as a downstream intermediate, for example, by oxidizing compounds HQ-1 and / or HQ-2 at an earlier stage, or potentially by directly oxidizing compound HQ-3. This leads to a more efficient and higher-yield method, particularly at plant scale.

Claims

1. A method for synthesizing a sucrose compound, the method comprising the step of oxidizing a reaction mixture containing a compound of formula I to obtain a compound of formula II: in: R1 is OH or CN; R2 is hydrogen or Prot NH ;and R3 is hydrogen or Prot OH , Among them Prot NH It is a protecting group for amino groups, and Prot OH It is a protecting group for OH.

2. The method according to claim 1, wherein R1 is OH.

3. The method according to claim 1, wherein R1 is CN.

4. The method according to any one of claims 1 to 3, wherein R2 is hydrogen.

5. The method according to any one of claims 1 to 3, wherein R2 is Prot NH .

6. The method of claim 5, wherein Prot NH It is selected from allyl carbamate (Alloc), 2,2,2-trichloroethyl carbamate (Troc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbamate (Fmoc), CONHPh, CSNHPh and tert-butyl carbamate (Boc), preferably tert-butyl carbamate (Boc).

7. The method according to any one of claims 1 to 6, wherein R3 is hydrogen.

8. The method according to any one of claims 1 to 7, wherein oxidation is carried out using an oxidizing agent, biochemical oxidation, or electrochemical oxidation.

9. The method according to claim 8, wherein the oxidant is selected from the group consisting of: metal oxidants (such as Cr(VI) compounds, Mn(IV) compounds, Mn(VII) compounds, Fe(III) compounds, Cu(II) compounds, Ag(I) compounds, Pb(IV) compounds, Ce(IV) compounds, Mn(III) compounds, Mn(IV) compounds, Mn(VII) compounds), halogen compounds (such as I2 and HIO3), quinones (such as tetrachlorobenzoquinone), molecular oxygen, hydrogen peroxide, hypochlorites (such as NaOCl and Ca(OCl)2), chlorites, 1,4-benzoquinone and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone or combinations thereof.

10. The method of claim 9, wherein the oxidant is selected from molecular oxygen, hydrogen peroxide, 1,4-benzoquinone, and metal oxidants or combinations thereof.

11. The method according to claim 9 or claim 10, wherein the metal oxidant is a transition metal oxidant.

12. The method according to claim 11, wherein the transition metal is Fe, Cu or Mn.

13. The method according to claim 11, wherein the transition metal oxidant is selected from Fe(III) compounds (e.g., FeCl3 or K3[Fe(CN)6]), Cu(II) compounds (e.g., CuSO4), Mn(III) compounds (e.g., Mn(OAc)3), Mn(IV) compounds (e.g., MnO2) and Mn(VII) compounds (e.g., KMnO4).

14. The method according to claim 13, wherein the Fe(III) compound is FeCl3.

15. The method of claim 9, wherein the oxidant is a halogen compound (such as I2 and HIO3).

16. The method of claim 9, wherein the oxidant is a quinone (such as tetrachlorobenzoquinone).

17. The method according to any one of claims 8 to 16, wherein the oxidant is in equivalent amounts to about 1.0 equivalents to about 15.0 equivalents relative to the compound of formula I in the reaction mixture, preferably about 1.2 equivalents to about 10.0 equivalents, more preferably about 1.5 equivalents to about 8.0 equivalents, and even more preferably about 2.0 equivalents to about 5.0 equivalents.

18. The method according to any one of claims 8 to 10, wherein the oxidant is molecular oxygen.

19. The method of claim 18, wherein the molecular oxygen is provided by exposing the reaction mixture to a reaction atmosphere containing molecular oxygen.

20. The method according to claim 18 or claim 19, wherein during the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II, a stream containing molecular oxygen is bubbled through the reaction mixture.

21. The method according to any one of claims 1 to 20, wherein the reaction is carried out at a pH of about 3.0 to about 9.0, preferably about 3.5 to about 8.5, more preferably about 4.0 to about 8.

0.

22. The method of claim 8, wherein the biochemical oxidation is catalyzed by hydroquinone oxidase or by cells exhibiting hydroquinone oxidase activity.

23. The method according to any one of claims 1 to 22, wherein the reaction mixture further comprises a compound according to formula II; preferably, R1 in the compound according to formula I in the reaction mixture is the same as R1 in the compound according to formula II in the reaction mixture, and / or R2 in the compound according to formula I in the reaction mixture is the same as R2 in the compound according to formula II in the reaction mixture, and / or R3 in the compound according to formula I in the reaction mixture is the same as R3 in the compound according to formula II in the reaction mixture; more preferably, R1, R2 and R3 in the compound according to formula I in the reaction mixture are the same as R1, R2 and R3 in the compound according to formula II in the reaction mixture.

24. The method according to any one of claims 1 to 23, wherein the step of oxidizing the reaction mixture comprising the compound of formula I to obtain the compound of formula II is carried out at about 0°C to about 100°C, preferably about 0°C to about 80°C, more preferably about 5°C to about 60°C, even more preferably about 5°C to about 40°C, even more preferably about 10°C to about 30°C, and most preferably about 10°C to about 25°C.

25. The method according to any one of claims 1 to 24, wherein the reaction mixture is a fermentation broth.

26. The method according to any one of claims 1 to 24, wherein the reaction mixture is an extract from the fermentation broth.

27. The method according to any one of claims 1 to 26, further comprising a step of performing a cyanation reaction to convert R1 from OH to CN before or after the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II; preferably the cyanation reaction is performed after the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II.

28. The method according to any one of claims 1 to 27, further comprising, before or after the step of oxidizing the reaction mixture comprising the compound of formula I to obtain the compound of formula II, performing an amino protection reaction to convert R2 from hydrogen to Prot. NH The step; preferably, the amino protection reaction is carried out after the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II.

29. The method according to any one of claims 1, 2, 4 and 7 to 26, wherein R1 in formula I is OH, and R2 in formula I is hydrogen, and: The method further includes, immediately after the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II, a cyanidation reaction to convert R1 from OH to CN; and The method further includes, immediately after the step of performing the cyanation reaction to convert R1 from OH to CN, an amino protection reaction to convert R2 from hydrogen to Prot. NH The steps.

30. The method according to any one of claims 1, 3, 4 and 7 to 26, wherein R1 in formula I is CN, and R2 in formula I is hydrogen, and: The method further includes a step of immediately performing a cyanidation reaction to convert R1 from OH to CN before the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II; and The method further includes, immediately after the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II, performing an amino protection reaction to convert R2 from hydrogen to Prot. NH The steps.

31. The method according to any one of claims 27, 29 and 30, wherein the step of oxidizing the reaction mixture containing the compound of formula I to obtain the compound of formula II and the step of performing the cyanidation reaction to convert R1 from OH to CN are carried out as a one-pot reaction.

32. The method according to claim 1, wherein the tunicate compound is selected from: , , , and .

33. A compound according to formula I: ； R1 to R3 are defined as in any one of claims 1 to 7, wherein R3 is hydrogen when R2 is CONHPh.

34. The compound according to claim 33, wherein Prot NH It is selected from allyl carbamate (Alloc), 2,2,2-trichloroethyl carbamate (Troc), benzyl carbamate (Cbz), 9-fluorenylmethyl carbamate (Fmoc), CSNHPh and tert-butyl carbamate (Boc), preferably tert-butyl carbamate (Boc).

35. The compound according to claim 33, wherein R2 is H.

36. The compound according to claim 33, having the following formula: 。 37. The compound according to claim 33, having the following formula: 。 38. The compound according to claim 33, having the following formula: 。 39. A compound of the following formula: 。

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