Preparation method and application of vocyclosporine intermediate

Abstract

The invention relates to the field of medicinal chemistry, in particular to a preparation method and application of a vocyclosporine intermediate, and the preparation method is mild in reaction condition, simple in step, convenient in post-treatment, low in cost and suitable for industrial production.

Description

[0001] This invention claims the priority of a Chinese patent application titled "Voclosporin Intermediate and Its Preparation Method and Application", with the application number 202311606177.1 and filed with the Chinese Patent Office on November 29, 2023. The entire disclosure of this application is incorporated herein by reference in its entirety. Technical Field

[0002] This invention relates to the field of medicinal chemistry, and specifically to a preparation method and application of a voclosporin intermediate. Background Art

[0003] Voclosporin is an intracellular calcium / calmodulin-dependent serine / threonine phosphatase inhibitor, abbreviated as a calcineurin inhibitor. It was approved for marketing by the US Food and Drug Administration (FDA) on January 22, 2021, under the trade name Lupkynis. It is the first oral drug for active lupus nephritis.

[0004] The classical synthetic route of voclosporin starts from cyclosporin A, and through hydroxyl protection, oxidation, Wittig reaction, and dehydroxylation protection to obtain voclosporin. One of the key steps is the synthesis of acetyl cyclosporin A aldehyde. The structural formulas of voclosporin and cyclosporin A are as follows:

[0005]

[0006] Among them, the structure of GB is as follows:

[0007]

[0008] The traditional method for synthesizing acetyl cyclosporin A aldehyde is a one-step synthesis from acetyl cyclosporin A, that is, the oxidation of an alkene to an aldehyde. Common oxidants include ozone (Xu Renjie et al., Research Progress on the Reaction of Ozone Oxidizing Organic Small Molecules [J], Chemical Industry Production and Technology, 2021, 27(3): 16 - 20), potassium permanganate (Wang Lei et al., Research on the Reaction Mechanism of Potassium Permanganate Oxidizing Alkenes [J], Journal of Huaibei Coal Teachers College, 1997, 18(1): 37 - 41), osmium tetroxide, ruthenium catalysis (J. Org. Chem., Vol. 46, No. 19, pp 3736 - 3738 (1981)), etc.

[0009] The prior art has disclosed several methods for the one-step synthesis of acetyl cyclosporin A aldehyde from acetyl cyclosporin A:

[0010] 1. RuCl 3 / NaIO 4Methods: US20030171264A1, CN1571795A and CN100334106C dissolve acetyl cyclosporin A in a mixture of acetonitrile and water, and add sodium periodate and ruthenium chloride hydrate to prepare acetyl cyclosporin A aldehyde. However, there are disadvantages such as RuCl 3 being expensive, high production costs, and prone to metal residues.

[0011] 2. Ozone method: WO2006014872A2 treats acetyl cyclosporin A with ozone in dichloromethane at -78 °C, and then reduces it with methyl sulfide to produce the desired acetyl cyclosporin A aldehyde. However, the ozone method has great danger in the oxidation process, harsh process conditions, high technical and equipment requirements, the reaction occurrence and treatment are relatively complex, the reaction device is expensive, and the energy consumption is high, resulting in high production costs.

[0012] 3. NMO / OsO 4 method: WO1999018120A1, US6605593B1 and EP0991660B1 use OsO 4 / NaIO 4 to oxidize acetyl cyclosporin A to acetyl cyclosporin A aldehyde, with a yield reaching 80%, but OsO 4 is highly toxic, has high operation risk, complex post-treatment, and is expensive.

[0013] 4. Potassium permanganate oxidation method: Oxidize alkene to aldehyde under acidic conditions, with low conversion rate, and requires the combined action of other oxidants, increasing production costs; potassium permanganate has poor stability, can decompose and generate heat automatically, and causes combustion when contacting with organic substances, with high operation risk.

[0014] Therefore, it is particularly important to develop a preparation method of acetyl cyclosporin A aldehyde and voclosporin with mild reaction conditions, simple steps, convenient post-treatment, low cost and high purity. Summary of the Invention

[0015] The object of the present invention is to provide a preparation method of a voclosporin intermediate (Formula II);

[0016] The second object of the present invention is to provide a use of a voclosporin intermediate (Formula II);

[0017] The third object of the present invention is to provide a new preparation method of voclosporin.

[0018] To achieve the above object, the technical solution of the present invention is:

[0019] The intermediate of voclosporin shown in Formula II:

[0020]

[0021] Among them,

[0022] R is a hydroxyl protecting group;

[0023] Preferably, R is any one of (C 1-10 alkyl or aryl) 3 silyl, (C 1-10 alkyl or aryl) acyl, (C 1-6 alkoxy or C 6-10 aryloxy) carbonyl, C 1-10 alkyl or substituted alkyl, aryl;

[0024] More preferably, R is any one of trimethylsilyl, triethylsilyl, triisopropylsilyl, isopropyldimethylsilyl, isopropyldiethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, 2,3,6-trimethyl-4-hydroxybenzoyl, p-nitrobenzoyl, formyl, acetyl, benzoyl, tert-butoxycarbonyl, benzyloxycarbonyl, methoxymethyl, allyl, methyl, tert-butyl, 2-tetrahydropyranyl, benzyl, p-methoxybenzyl, trityl;

[0025] More preferably, R is acetyl.

[0026] Specifically, the compound of formula II can be:

[0027]

[0028] A method for preparing a voclosporin intermediate (formula II): obtained by reacting a compound of formula I with an oxidizing agent:

[0029]

[0030] Among them, R is a hydroxyl protecting group;

[0031] Preferably, R is any one of (C 1-10 alkyl or aryl) 3 silyl, (C 1-10 alkyl or aryl) acyl, (C 1-6 alkoxy or C 6-10 aryloxy) carbonyl, C 1-10 alkyl or substituted alkyl, aryl;

[0032] More preferably, R is any one of trimethylsilyl, triethylsilyl, triisopropylsilyl, isopropyldimethylsilyl, isopropyldiethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, 2,3,6-trimethyl-4-hydroxybenzoyl, p-nitrobenzoyl, formyl, acetyl, benzoyl, tert-butoxycarbonyl, benzyloxycarbonyl, methoxymethyl, allyl, methyl, tert-butyl, 2-tetrahydropyranyl, benzyl, p-methoxybenzyl, trityl;

[0033] More preferably, R is an acetyl group.

[0034] Furthermore, the oxidizing agent is any one or a combination of two or more of osmium tetroxide, 4-methylmorpholine-N-oxide hydrate, trimethylamine N-oxide, potassium permanganate, osmium chloride·potassium ferrocyanide, hydrogen peroxide, oxygen, alkyl peroxide, organic peroxyacid, perimido acid, peroxyacetyl nitrate, dimethyldioxirane, trimethylamine N-oxide dihydrate, anhydrous trimethylamine N-oxide, iodine, silver carboxylate, chlorate, iodate, potassium ferrocyanide, potassium osmate dihydrate, and osmium compound and ligand composition;

[0035] Preferably, the oxidizing agent is any one or a combination of two or more of 4-methylmorpholine-N-oxide hydrate, chlorate, iodate, hydrogen peroxide, oxygen, tert-butyl hydroperoxide, potassium ferrocyanide, and potassium osmate dihydrate;

[0036] More preferably, the oxidizing agent is a combination of 4-methylmorpholine-N-oxide hydrate and potassium osmate dihydrate or a combination of trimethylamine N-oxide dihydrate and potassium osmate dihydrate;

[0037] More preferably, the oxidizing agent is a combination of trimethylamine N-oxide dihydrate and potassium osmate dihydrate.

[0038] Furthermore, when the oxidizing agent is a combination of 4-methylmorpholine-N-oxide hydrate and potassium osmate dihydrate, the molar equivalent ratio of potassium osmate dihydrate to formula I is 0.025-0.125:1;

[0039] Specifically, the molar equivalent ratio of potassium osmate dihydrate to the compound of formula I can be 0.025:1, 0.050:1, 0.075:1, 0.100:1, 0.125:1; preferably, the molar equivalent ratio is 0.050-0.100:1.

[0040] The molar equivalent ratio of 4-methylmorpholine-N-oxide hydrate to formula I is 0.7-3:1;

[0041] Specifically, the molar equivalent ratio of 4-methylmorpholine-N-oxide hydrate to the compound of formula I can be 0.7:1, 1.0:1, 2.0:1, 2.5:1, 3.0:1; preferably, the molar equivalent ratio is 2.0-2.5:1.

[0042] When the oxidizing agent is a combination of trimethylamine N-oxide dihydrate and potassium osmate dihydrate, the molar equivalent ratio of potassium osmate dihydrate to formula I is 0.025-0.125:1;

[0043] Specifically, the molar equivalent ratio of potassium osmate dihydrate to the compound of formula I can be 0.025:1, 0.050:1, 0.075:1, 0.100:1, 0.125:1; preferably, the molar equivalent ratio of potassium osmate dihydrate to formula I is 0.025 - 0.075:1.

[0044] The molar equivalent ratio of trimethylamine N-oxide dihydrate to formula I is 0.7 - 3:1;

[0045] Specifically, the molar equivalent ratio of trimethylamine N-oxide dihydrate to formula I can be 0.7:1, 1.0:1, 1.5:1, 2.0:1, 2.5:1, 3.0:1; preferably, the molar equivalent ratio of trimethylamine N-oxide dihydrate to formula I is 1.5 - 2.5:1.

[0046] Furthermore, the reaction solvent for the reaction is selected from any one or more of acetonitrile, tetrahydrofuran, acetone, toluene, dichloromethane, dichloroethane, pyridine, methanol, ethanol, tert-butanol, and water;

[0047] Preferably, the reaction solvent is a combination of acetonitrile and water;

[0048] More preferably, the volume ratio of acetonitrile to water is 2 - 4:1.

[0049] Furthermore, the reaction temperature of the reaction is -45 - 45 °C;

[0050] Specifically, the reaction temperature can be -40 ± 5 °C, -30 ± 5 °C, -20 ± 5 °C, -10 ± 5 °C, 0 ± 5 °C, 10 ± 5 °C, 20 ± 5 °C, 30 ± 5 °C, 40 ± 5 °C; preferably, the reaction temperature is 15 - 25 °C.

[0051] Furthermore, the reaction needs to be carried out in an acidic system, and the acidic system contains organic acid or inorganic acid;

[0052] Preferably, the acidic system contains any one or more of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, acetic anhydride, and nitric acid;

[0053] Preferably, the acidic system contains acetic acid or acetic anhydride;

[0054] More preferably, the acidic system contains acetic anhydride.

[0055] Furthermore, the molar equivalent ratio of acetic acid or acetic anhydride to formula I is 0.1 - 1:1;

[0056] Preferably, the molar equivalent ratio of acetic acid or acetic anhydride to formula I is 0.2:1.

[0057] A preparation method of a voclosporin intermediate (Formula II): obtained by reacting a compound of Formula I with an oxidizing agent:

[0058] Wherein, R is an acetyl group.

[0059] Further, the oxidizing agent is a combination of trimethylamine N-oxide dihydrate and potassium osmate dihydrate, wherein the molar equivalent ratio of trimethylamine N-oxide dihydrate to the compound of Formula I is 1.5 - 2.5:1, and the molar equivalent ratio of potassium osmate dihydrate to the compound of Formula I is 0.025 - 0.075:1.

[0060] Further, the reaction solvent for the reaction is a combination of acetonitrile and water, wherein the volume ratio of acetonitrile to water is 2 - 4:1.

[0061] Further, the reaction temperature of the reaction is 15 - 25 °C.

[0062] Further, the reaction also needs to be carried out in an acidic system, and the acidic system includes an organic acid or an inorganic acid;

[0063] Further, the acidic system includes any one or more of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, acetic anhydride, nitric acid;

[0064] Further, the acidic system includes acetic acid or acetic anhydride;

[0065] Further, the acidic system includes acetic anhydride.

[0066] Further, the molar equivalent ratio of acetic acid or acetic anhydride to Formula I is 0.2:1.

[0067] Use of the compound of Formula II for the preparation of the compound of Formula III.

[0068] Specifically, a preparation method of a compound of Formula III: mainly obtained by reacting a compound of Formula II with an oxidizing reagent to obtain the compound of Formula III:

[0069]

[0070] Wherein, R is a hydroxyl protecting group;

[0071] Preferably, R is (C 1-10 alkyl or aryl) 3 silyl, (C 1-10 alkyl or aryl) acyl, (C 1-6 alkoxy or C 6-10 aryloxy) carbonyl, C 1-10 alkyl or substituted alkyl, aryl, any one of them;

[0072] More preferably, R is any one of trimethylsilyl, triethylsilyl, triisopropylsilyl, isopropyldimethylsilyl, isopropyldiethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, 2,3,6-trimethyl-4-hydroxybenzoyl, p-nitrobenzoyl, formyl, acetyl, benzoyl, tert-butoxycarbonyl, benzyloxycarbonyl, methoxymethyl, allyl, methyl, tert-butyl, 2-tetrahydropyranyl, benzyl, p-methoxybenzyl, trityl;

[0073] More preferably, R is acetyl.

[0074] Furthermore, the oxidation reagent is any one or a combination of more than one of periodic acid, sodium periodate, lead tetraacetate, thallium nitrate, thallium ethoxide, activated manganese dioxide, pyridinium chlorochromate, bis(acetylacetone)vanadium oxide, oxygen, cobalt acetate, silver ion, persulfate;

[0075] Preferably, the oxidation reagent is periodic acid or sodium periodate;

[0076] More preferably, the oxidation reagent is sodium periodate.

[0077] Furthermore, the molar equivalent ratio of the oxidation reagent to the compound of formula II is 1.0 - 2.0:1;

[0078] Specifically, the molar equivalent ratio of the oxidation reagent to the compound of formula II can be 1.0:1, 1.2:1, 1.4:1, 1.6:1, 1.8:1, 2.0:1; preferably, the molar equivalent ratio is 1.4 - 1.8:1.

[0079] Furthermore, the reaction solvent for the reaction is selected from any one or several of acetonitrile, tetrahydrofuran, acetone, toluene, benzene, water;

[0080] Preferably, the reaction solvent is a combination of tetrahydrofuran and water;

[0081] More preferably, the volume ratio of tetrahydrofuran to water is 1 - 9:1.

[0082] Furthermore, the reaction temperature of the reaction is -5 - 45 °C;

[0083] Specifically, the reaction temperature can be 0 ± 5 °C, 10 ± 5 °C, 20 ± 5 °C, 30 ± 5 °C, 40 ± 5 °C; preferably, the reaction temperature is 15 - 25 °C.

[0084] Furthermore, the use of the compound of formula II in the preparation of voriconazole.

[0085] Specifically, a method for preparing voriconazole includes the following steps:

[0086] (1) The hydroxyl group of cyclosporin A (CsA) is protected to form a compound of Formula I, where R is a hydroxyl-protecting group;

[0087] (2) The compound of Formula I is oxidized to form a compound of Formula II;

[0088] (3) The compound of Formula II is oxidized to form a compound of Formula III;

[0089] (4) The compound of Formula III undergoes a Wittig reaction to form a compound of Formula IV;

[0090] (5) The hydroxyl-protecting group of the compound of Formula IV is removed to form a compound of Formula V.

[0091]

[0092] Among them, R is a hydroxyl-protecting group;

[0093] Preferably, R is (C 1-10 alkyl or aryl) 3 silyl, (C 1-10 alkyl or aryl) acyl, (C 1-6 alkoxy or C 6-10 aryloxy) carbonyl, C 1-10 alkyl or substituted alkyl, aryl;

[0094] More preferably, R is any one of trimethylsilyl, triethylsilyl, triisopropylsilyl, isopropyldimethylsilyl, isopropyldiethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, 2,3,6-trimethyl-4-hydroxybenzoyl, p-nitrobenzoyl, formyl, acetyl, benzoyl, tert-butoxycarbonyl, benzyloxycarbonyl, methoxymethyl, allyl, methyl, tert-butyl, 2-tetrahydropyranyl, benzyl, p-methoxybenzyl, trityl;

[0095] More preferably, R is acetyl.

[0096] The beneficial effects of the present invention are as follows:

[0097] (1) In the present invention, an oxidation reagent is used to first perform dihydroxylation of the compound of Formula I to obtain a new intermediate, the compound of Formula II

[0098] compound, and then an oxidizing agent is used to oxidize this new intermediate to obtain the compound of Formula III; the yields of these two steps can be as high as over 90%, significantly improving the yield of the compound of Formula III;

[0099] (2) The oxidizing agent used in the synthesis process has less toxicity and better stability than OsO 4 ;

[0100] (3) The synthesis method used has mild reaction conditions, simple steps, high purity, short required reaction time, improved reaction efficiency and low cost, and is convenient for post-treatment, making it suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0101] Figure 1 For the high performance liquid chromatography of the compound of Formula II at 1 h of reaction in Example 10 a ;

[0102] Figure 2 For the high performance liquid chromatography of the compound of Formula II at 5 h of reaction in Example 11 a ;

[0103] Figure 3 For the 1H NMR spectrum of the compound of Formula II in Example 14 a ;

[0104] Figure 4 For the 13C NMR spectrum of the compound of Formula II in Example 14 a ;

[0105] Figure 5 For the mass spectrum of the compound of Formula II in Example 14 a ;

[0106] Figure 6 For the high performance liquid chromatography of the compound of Formula II in Example 14 a ;

[0107] Figure 7 For the 1H NMR spectrum of the compound of Formula III in Example 27 a ;

[0108] Figure 8 For the 13C NMR spectrum of the compound of Formula III in Example 27 a ;

[0109] Figure 9 For the mass spectrum of the compound of Formula III in Example 27 a ;

[0110] Figure 10 For the high performance liquid chromatography of the compound of Formula III in Example 27 a ;

[0111] Figure 11 For the high performance liquid chromatography of the compound of Formula II at 2 h of reaction in Comparative Example 1 a ;

[0112] Figure 12 For the high performance liquid chromatography of the compound of Formula II at 5 h of reaction in Comparative Example 1 a ;

[0113] Figure 13For the comparison of the high-performance liquid chromatography of the compound of Formula II in Example 10 with a reaction time of 1 h and in Comparative Example 1 with reaction times of 2 h and 5 h a ;

[0114] Figure 14 For the high-performance liquid chromatography of the compound of Formula II in Comparative Example 2 with a reaction time of 5 h a ;

[0115] Figure 15 For the high-performance liquid chromatography of the compound of Formula II in Comparative Example 3 with a reaction time of 2 h a ;

[0116] Figure 16 For the high-performance liquid chromatography of the compound of Formula II in Comparative Example 3 with a reaction time of 5 h a ; Detailed Description of the Invention

[0117] The present invention will be described in detail below. However, the present invention may be embodied in many different forms and should not be limited to the embodiments described herein. The purpose of providing these embodiments is to make the disclosed content more complete and comprehensive. All reagents and raw materials used, except those provided in the preparation methods, are commercially available. Unless otherwise defined, all scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art to which the claimed subject matter pertains.

[0118] Unless otherwise specified, the terms used herein have the following meanings:

[0119] The term "alkyl" refers to a saturated, straight-chain or branched hydrocarbon group containing one to twelve carbon atoms, preferably one to six carbon atoms. C 1-10 Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, and n-hexyl, octyl, decyl, etc.; C 1-6 Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, and n-hexyl.

[0120] The term "aryl" refers to a 6- to 14-membered fully carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent carbon atom pairs) group having a conjugated π-electron system, preferably 6 to 10 members, more preferably 6 to 8 members, such as phenyl and naphthyl. More preferably phenyl.

[0121] The aryl group can be substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate group. Examples of aryl groups include, but are not limited to, tert-butyldiphenyl, 2,3,6-trimethyl-4-hydroxybenzyl, p-nitrobenzyl, benzyl, benzhydryl.

[0122] The term "alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), where alkyl is defined as above, C 1-6 Examples of alkoxy groups include, but are not limited to: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy.

[0123] The term "aryloxy" refers to -O-(aryl) and -O-(substituted aryl), where aryl is defined as above.

[0124] The term "silyl" refers to an alkyl group substituted by a silyl group, where alkyl is defined as above. Examples of silyl groups include, but are not limited to, trimethylsilyl, triethylsilyl, triisopropylsilyl, isopropyldimethylsilyl, isopropyldiethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl.

[0125] The term "alkyl peroxide" refers to an organic compound containing an -O-O- peroxide functional group formed by replacing the hydrogen atom in hydrogen peroxide with an alkyl group, including, but not limited to, tert-butyl hydroperoxide, cumene hydroperoxide.

[0126] The term "organic peroxyacid" refers to an acid containing -O-O- in the molecule, including, but not limited to, performic acid, peracetic acid, trifluoroperacetic acid, perbenzoic acid, m-chloroperbenzoic acid.

[0127] "Silver carboxylate" in the present invention refers to silver acetate, silver benzoate.

[0128] "Osmium compound and ligand composition" in the present invention refers to a combination of osmium tetroxide or potassium osmate dihydrate and a chiral ligand derivative composed of dihydroquinine or dihydroquinidine, including but not limited to AD-mix-α [K 2 OsO 2 (OH) 4 (cat), K 2 CO 3 , K 3 Fe(CN) 6 , (DHQ) 2 PHAL(cat)] and AD-mix-β [K 2OsO 2 (OH) 4 (cat),K 2 CO 3 ,K 3 Fe(CN) 6 ,(DHQD) 2 PHAL(cat)]。

[0129] Example 1. Synthesis of Cyclo[[(6E)-(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6-octenoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula I a )

[0130]

[0131] Under nitrogen protection, CsA (20 g) and dichloromethane (200 mL) were added to a reaction flask and stirred until dissolved. Acetic anhydride (8.5 g) was added, the temperature was controlled at 20 °C, and dimethylaminopyridine (3.0 g) was added dropwise, followed by holding the reaction at this temperature. After the reaction was completed as detected by high performance liquid chromatography, an aqueous sodium bicarbonate solution and dichloromethane were added for extraction. The organic phase was collected, washed successively with a refined industrial brine solution and dried over anhydrous sodium sulfate. The organic phase was concentrated, crystallized, filtered by suction, and dried to obtain the compound of Formula I a (19.64 g, yield 95%, purity 99.6%).

[0132] Example 2. Synthesis of Cyclo[[(6E)-(2S,3R,4R)-3-(triethylsilyloxy)-4-methyl-2-(methylamino)-6-octenoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula I b )

[0133]

[0134] Under nitrogen protection, CsA (10 g), dichloromethane (120 mL), and N,N-diisopropylethylamine (9.2 g) were added to a reaction flask and stirred until clear. Triethylchlorosilane (5 g) was added dropwise, and the mixture was stirred at room temperature. After the reaction was completed as detected by high performance liquid chromatography, it was washed with a saturated sodium bicarbonate solution and a sodium chloride solution. The organic phase was collected, concentrated, crystallized, filtered by suction, and dried to obtain the compound of Formula I bCompound (9.81 g, yield 90%, purity 97.4%).

[0135] Example 3. Synthesis of Cyclo[[(6E)-(2S,3R,4R)-3-(tert-butoxycarbonyloxy)-4-methyl-2-(methylamino)-6-octenoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula I k )

[0136]

[0137] Add CsA (10 g), dichloromethane (80 mL), and N,N-diisopropylethylamine (2 g) to a reaction flask, stir to dissolve clearly; dropwise add di-tert-butyl dicarbonate (2.6 g), and stir at room temperature; after detecting the completion of the reaction by TLC, add saturated sodium bicarbonate solution, stir and separate layers, collect the organic phase, and wash with water; collect the organic phase, wash with saturated brine, and dry with anhydrous sodium sulfate, then concentrate. The crude product is crystallized, filtered, and dried to obtain the compound of Formula I k Compound (10.02 g, yield 92%, purity 98.2%).

[0138] Example 4. Synthesis of Cyclo[[(6E)-(2S,3R,4R)-3-(methoxymethyl)-4-methyl-2-(methylamino)-6-octenoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula I l ) MW: 1202.61

[0139]

[0140] Under nitrogen protection, add CsA (10 g), dichloromethane (100 mL), and N,N-diisopropylethylamine (3.2 g) to a reaction flask, stir to dissolve clearly; dropwise add chloromethyl methyl ether (1.5 g), and stir at room temperature for reaction; after detecting the completion of the reaction by high performance liquid chromatography, wash with ammonium chloride solution, collect the organic phase, dry with anhydrous sodium sulfate, concentrate, filter by suction, and dry to obtain the compound of Formula I l Compound (10.13 g, yield 93%, purity 97.9%).

[0141] Synthesis of Cyclo[[(6E)-(2S,3R,4R)-3-(benzyloxy)-4-methyl-2-(methylamino)-6-octenoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula I n )

[0142]

[0143] Add CsA (10 g), N,N-dimethylformamide (50 mL), and 60% sodium hydride (0.66 g) into a reaction flask, stir until dissolved and clear; add benzyl bromide (3.5 g) dropwise, and stir the reaction at room temperature; after detecting the completion of the reaction by TLC, add the reaction solution dropwise into water (100 mL), cool down to below 0 °C, stir for 30 min, a large amount of solid precipitates, filter by suction, and dry to obtain the compound of Formula I n (9.82 g, yield 90%, purity 98.1%).

[0144] Synthesis of Cyclo[[(6E)-(2S,3R,4R)-3-(trityloxy)-4-methyl-2-(methylamino)-6-octenoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula I r )

[0145]

[0146] Add CsA (10 g) and pyridine (60 mL) into a reaction flask, stir until dissolved and clear; add trityl chloride (7.0 g), and stir the reaction at 50 °C; after detecting the completion of the reaction by TLC, concentrate under reduced pressure until no obvious liquid drips, add ice acetic acid / dichloromethane for crystallization, filter by suction, and dry to obtain the compound of Formula I r (10.00 g, yield 88%, purity 98.1%).

[0147] Synthesis of Cyclo[[(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0148]

[0149] Under nitrogen protection, add acetonitrile (66 mL), water (33 mL), and Compound of Formula I a (16.5 g) and 4-methylmorpholine-N-oxide hydrate (3.86 g) to the reaction flask. Control the temperature at 20 °C and stir until dissolved. Add potassium osmate dihydrate (0.31 g), control the temperature at 25 °C, and stir the reaction. After the reaction is complete, quench the reaction with an aqueous solution of sodium sulfite and sodium bicarbonate, extract with dichloromethane, combine the organic phases, wash with a refined industrial brine solution, collect the organic phase, and concentrate. The crude product is crystallized from methanol / water, filtered by suction, and dried to obtain Compound of Formula II a (16.54 g, yield 98%, purity 98.3%).

[0150] Example 8. Synthesis of Cyclo[[(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0151] Add Compound of Formula I a (10 g) to the reaction flask, add ethanol (100 mL), stir to dissolve, adjust the pH to 10 - 12 with sodium hydroxide solution, dropwise add 1% - 3% potassium permanganate solution (calculated according to the actual situation), control the temperature at -40 °C, and stir the reaction. After detecting the completion of the reaction by TLC, quench the reaction with sodium sulfite, filter, extract the filtrate with chloroform, combine the organic phases, wash with saturated brine, and dry over anhydrous sodium sulfate. Collect the organic phase and concentrate. The crude product is crystallized from methanol / water, filtered by suction, and dried to obtain Compound II a (8.18 g, yield 80%, purity 86.5%).

[0152] Example 9. Synthesis of Cyclo[[(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0153] Under nitrogen protection, add acetonitrile (66 mL), water (33 mL), and Compound of Formula I aCompound (16.5 g), trimethylamine N-oxide dihydrate (3.7 g), the temperature was controlled at 20 ± 5 °C, and stirred until dissolved; potassium osmate dihydrate (0.37 g) was added, the temperature was controlled at 20 ± 5 °C, and stirred for 1 - 2 h. After the reaction was detected by high performance liquid chromatography to be complete, the reaction was quenched with an aqueous solution of sodium sulfite and sodium bicarbonate, extracted with dichloromethane, the organic phases were combined, washed with a refined industrial brine solution, the organic phase was collected and concentrated. The crude product was crystallized from methanol / water, filtered by suction, and dried to obtain Formula II a Compound (14.81 g, yield 88.5%, purity 89.0%).

[0154] Example 10, Synthesis of Cyclo[[(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0155] Under nitrogen protection, acetonitrile (66 mL), water (33 mL), and Formula I were added to the reaction flask a Compound (16.5 g), trimethylamine N-oxide dihydrate (3.7 g), acetic anhydride (0.27 g), the temperature was controlled at 20 ± 5 °C, and stirred until dissolved; potassium osmate dihydrate (0.37 g) was added, the temperature was controlled at 20 ± 5 °C, and stirred for 1 h. After the reaction was detected by high performance liquid chromatography, there was no remaining raw material and the reaction was completed (purity 98.37%, see high performance liquid chromatography in Figure 1 ). The reaction was quenched with an aqueous solution of sodium sulfite and sodium bicarbonate, extracted with dichloromethane, the organic phases were combined, washed with a refined industrial brine solution, the organic phase was collected and concentrated. The crude product was crystallized from methanol / water, filtered by suction, and dried to obtain Formula II a Compound (16.40 g, yield 98%).

[0156] Example 11, Synthesis of Cyclo[[(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0157] Under nitrogen protection, acetonitrile (66 mL), water (33 mL), and Formula I were added to the reaction flask aCompound (16.5 g), trimethylamine N-oxide dihydrate (2.2 g), acetic anhydride (1.35 g), temperature controlled at 20 ± 5 °C, stirred until dissolved clear; potassium osmate dihydrate (0.12 g) was added, temperature controlled at 20 ± 5 °C, stirred for 1 h, the reaction was detected by high performance liquid chromatography, and 30% of the raw materials remained; stirring the reaction continued for 2 h, and 7.9% of the raw materials remained after detection; stirring the reaction continued for 5 h, the reaction was detected by high performance liquid chromatography, and the raw materials had reacted completely, and the reaction was completed (purity 98.01%, high performance liquid chromatography see Figure 2 ). Sodium sulfite and sodium bicarbonate aqueous solution were added to quench the reaction, extracted with dichloromethane, the organic phases were combined, washed with refined industrial brine solution, the organic phase was collected and concentrated. The crude product was crystallized in methanol / water, filtered by suction and dried to obtain Compound of Formula II a (16.11 g, yield 96%).

[0158] Example 12, Synthesis of Cyclo[[(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0159] Under nitrogen protection, tetrahydrofuran (132 mL), water (33 mL), Compound of Formula I a (16.5 g), 4-methylmorpholine-N-oxide hydrate (3.88 g) were added to the reaction flask, temperature controlled at 0 °C, stirred until dissolved clear; potassium osmate dihydrate (0.37 g) was added, temperature controlled at 15 °C, and stirred for reaction. After the reaction was detected to be complete by high performance liquid chromatography, sodium sulfite and sodium bicarbonate aqueous solution were added to quench the reaction, extracted with dichloromethane, the organic phases were combined, washed with refined industrial brine solution, the organic phase was collected and concentrated. The crude product was crystallized in absolute ethanol / water, filtered by suction and dried to obtain Compound of Formula II a (15.00 g, yield 89%, purity 97.2%).

[0160] Example 13, Synthesis of Cyclo[[(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0161] Under nitrogen protection, toluene (99 mL), Compound I a (16.5 g), and 4-methylmorpholine-N-oxide hydrate (1.09 g) were added to the reaction kettle, and the temperature was controlled at 15 °C and stirred until dissolved clearly; potassium osmate dihydrate (0.24 g) was added, and the temperature was controlled at 35 °C and stirred for reaction. After the reaction was detected to be complete by high performance liquid chromatography, the reaction was quenched with an aqueous solution of sodium sulfite and sodium bicarbonate, extracted with dichloromethane, the organic phases were combined, washed with a refined industrial brine solution, the organic phase was collected and concentrated. The crude product was crystallized from methanol / water, filtered by suction and dried to obtain Compound II a (14.35 g, yield 85%, purity 86.8%).

[0162] Example 14, Synthesis of Cyclo[[(2S,3R,4R)-3-(acetyloxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0163] Under nitrogen protection, acetone (99 mL), water (33 mL), Compound I a (16.5 g), and 4-methylmorpholine-N-oxide hydrate (3.11 g) were added to the reaction kettle, and the temperature was controlled at 25 °C and stirred until dissolved clearly; potassium osmate dihydrate (0.49 g) was added, and the temperature was controlled at 25 °C and stirred for reaction. After the reaction was detected to be complete by high performance liquid chromatography, the reaction was quenched with an aqueous solution of sodium sulfite and sodium bicarbonate, extracted with dichloromethane, the organic phases were combined, washed with a refined industrial brine solution, the organic phase was collected and concentrated. The crude product was crystallized from methanol / water, filtered by suction and dried to obtain Compound II of Formula a (15.53 g, yield 92%, purity 98.3%). The hydrogen spectrum, carbon spectrum, mass spectrum and high performance liquid chromatography of Compound II of Formula a are shown in Figures 3 - 6 .

[0164] Example 15, Synthesis of Cyclo[[(2S,3R,4R)-3-(acetyloxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0165] Under nitrogen protection, add acetonitrile (50 mL), pyridine (50 mL), and the compound of formula I a (16.5 g) to a reaction flask, stir to dissolve and clarify, add osmium tetroxide (4.0 g, 15.84 mmol, 1.2 eq), control the temperature at 25 °C, and stir the reaction. After detecting the completion of the reaction by TLC, rotary evaporate until no obvious liquid drips, add water and dichloromethane for extraction, collect the organic phase, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate. The crude product is refined with ethanol / water, filtered by suction, and dried to obtain the compound of formula II a (14.85 g, yield 88%, purity 98.6%).

[0166] Example 16, Synthesis of Cyclo[[(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0167] Add the compound of formula I a (16.5 g), dichloroethane (100 mL), anhydrous sodium sulfate (6.5 g), and anhydrous sodium carbonate (5.0 g) to a reaction flask, and dropwise add a solution prepared from 40 g of hydrogen peroxide (20% - 30%) and 19 g of anhydrous sodium acetate under stirring, control the temperature ≤ 20 °C, and finish dropping in about 30 min. Continue to stir the reaction. After detecting the completion of the reaction by TLC, filter, and wash the filter cake with dichloroethane. Combine the filtrates, wash with sodium bisulfite and sodium bicarbonate solutions, dry over anhydrous sodium sulfate, and distill off dichloroethane under reduced pressure. Add ethanol / water for refinement, filter by suction, and dry to obtain the compound of formula II a (14.35 g, yield 85%, purity 88.4%)

[0168] Example 17, Synthesis of Cyclo[[(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0169] Add K 3 Fe(CN) 6(1.97 g), potassium osmate dihydrate (1.34 g), potassium carbonate (0.83 g), hydroquinidine 1,4-(phthalazine) diether (15.6 g), tert-butanol-water 1:1 (volume ratio) 100 mL, stirred to 0 °C, added Compound of Formula I a (2.49 g), kept warm and stirred for reaction for 12 h. Added 3.0 g of sodium sulfite powder, slowly warmed up to room temperature and stirred for 1 h. After TLC detected that the reaction was complete, extracted with ethyl acetate for 3 times. Combined the organic phases, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to obtain the crude product, and purified by column chromatography to obtain Compound of Formula II a (1.86 g, yield 73%, purity 98.8%)

[0170] Example 18, Synthesis of Cyclo[[(2S,3R,4R)-3-(triethylsilyloxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II b )

[0171] Under nitrogen protection, add tetrahydrofuran (100 mL), Compound of Formula I b (13.2 g), 4-methylmorpholine-N-oxide hydrate (1.35 g) into the reaction flask, control the temperature at 10 °C, stir until dissolved and clear; add potassium osmate dihydrate (0.27 g), control the temperature at 25 °C, stir for reaction. After TLC detected that the reaction was complete, added sodium sulfite and aqueous sodium bicarbonate solution to quench the reaction, extracted with ethyl acetate, combined the organic phases, added saturated brine for washing, and dried over anhydrous sodium sulfate, collected the organic phase, and concentrated. The crude product was refined in ethanol / water, filtered by suction, and dried to obtain Compound of Formula II b (11.89 g, yield 88%, purity 99.6%)

[0172] Example 19, Synthesis of Cyclo[[(2S,3R,4R)-3-(tert-butoxycarbonyloxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II k )

[0173] Under nitrogen protection, add toluene (50 mL) and pyridine (50 mL), Compound of Formula I kCompound (13.0 g) was stirred until dissolved clearly, then osmium tetroxide (3.0 g) was added. The temperature was controlled at 15 °C and the reaction was stirred. After the reaction was detected to be complete by TLC, it was rotary evaporated until no obvious liquid dropped, then water and ethyl acetate were added for extraction. The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. It was purified by methanol / water, filtered by suction, and dried to obtain Formula II k Compound (10.96 g, yield 82%, purity 95.9%)

[0174] Example 20. Synthesis of Cyclo[[(2S,3R,4R)-3-(methoxymethoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II l )

[0175] Compound of Formula I l Compound (12.5 g) was added to a reaction flask, methanol (100 mL) was added, and it was stirred until dissolved. Sodium hydroxide solution was added to adjust the pH to 10 - 12, and 1 - 3% potassium permanganate solution (calculated according to the actual situation) was added dropwise. The temperature was controlled at -20 °C and the reaction was stirred. After the reaction was detected to be complete by TLC, sodium sulfite was added to quench the reaction, and it was filtered. The filtrate was extracted with dichloromethane, the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The organic phase was collected and concentrated. The crude product was purified in methanol / water, filtered by suction, and dried to obtain II l Compound (9.98 g, yield 78%, purity 85.5%).

[0176] Example 21. Synthesis of Cyclo[[(2S,3R,4R)-3-(benzyloxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II n )

[0177] Under nitrogen protection, tetrahydrofuran (100 mL), Compound of Formula I nThe compound (12.9 g) and 4-methylmorpholine-N-oxide hydrate (2.7 g) were added to a reaction flask, and the temperature was controlled at 25 °C and stirred until dissolved clear. Potassium osmate dihydrate (0.33 g) was added, and the reaction was stirred while maintaining the temperature. After the reaction was detected to be complete by TLC, the reaction was quenched by adding an aqueous solution of sodium sulfite and sodium bicarbonate, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, the organic phase was collected and concentrated. The crude product was purified in methanol / water, filtered by suction, and dried to obtain Compound II n Compound (11.93 g, yield 90%, purity 94.3%)

[0178] Example 22, Synthesis of Cyclo[[(2S,3R,4R)-3-(trityloxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II r )

[0179] Under nitrogen protection, acetone (66 mL), water (33 mL), and Formula I were added to a reaction flask r Compound (14.4 g) and 4-methylmorpholine-N-oxide hydrate (2.7 g) were added, and the temperature was controlled at 35 °C and stirred until dissolved clear. Potassium osmate dihydrate (0.33 g) was added, and the reaction was stirred while maintaining the temperature at 35 °C. After the reaction was detected to be complete by high performance liquid chromatography, the reaction was quenched by adding an aqueous solution of sodium sulfite and sodium bicarbonate, extracted with dichloromethane, the organic phases were combined, washed with a refined industrial brine solution, the organic phase was collected and concentrated. The crude product was purified in methanol / water, filtered by suction, and dried to obtain Compound II r Compound (11.83 g, yield 80%, purity 84.7%). Example 23, Synthesis of Cyclo[6-[(3R,4R)-3-(acetoxy)-N,4-dimethyl-6-oxo-L-norleucine]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula III a )

[0180]

[0181] Under nitrogen protection, tetrahydrofuran (111 mL), water (37 mL), and Formula II were added to a reaction flask aCompound (16.1 g) was stirred and dissolved. The temperature was controlled at 25 °C, and sodium periodate (5.17 g) was added, followed by reaction while maintaining the temperature. After the reaction was detected to be complete by high-performance liquid chromatography, the reaction was quenched with an aqueous solution of sodium sulfite and sodium bicarbonate, and then extracted with ethyl acetate. The organic phases were combined, washed successively with a refined industrial salt solution and dried over anhydrous sodium sulfate. The organic phase was collected, concentrated, to obtain Compound of Formula III a Compound (15.36 g, yield 99%, purity 95.4%).

[0182] Example 24. Synthesis of Cyclo[6-[(3R,4R)-3-(acetyloxy)-N,4-dimethyl-6-oxo-L-norleucine]-L-2-aminobutanoyl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula III a )

[0183] Compound of Formula II a (10 g) and benzene (60 mL) were placed in a reaction flask, and lead tetraacetate (3.81 g) was added in portions over about 20 minutes. The temperature was controlled at 30 °C and the mixture was stirred for reaction; after the reaction was detected to be complete by TLC, it was filtered and the filtrate was collected and evaporated to concentrate; the concentrate was extracted with ethyl acetate and water, the organic phase was collected and concentrated, to obtain Compound of Formula III a Compound (8.19 g, yield 85%, purity 91.2%).

[0184] Example 25. Synthesis of Cyclo[6-[(3R,4R)-3-(acetyloxy)-N,4-dimethyl-6-oxo-L-norleucine]-L-2-aminobutanoyl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula III a )

[0185] Under nitrogen protection, acetonitrile (333 mL), water (37 mL), and Compound of Formula II a (16.1 g) were added to a reaction flask. After stirring and dissolving, the temperature was controlled at 40 ± 5 °C, and sodium periodate (4.90 g) was added, followed by reaction while maintaining the temperature; after the reaction was detected to be complete by high-performance liquid chromatography, the reaction was quenched with an aqueous solution of sodium sulfite and sodium bicarbonate, and then extracted with ethyl acetate; the organic phases were combined, washed successively with a refined industrial salt solution and dried over anhydrous sodium sulfate. The organic phase was collected, concentrated, to obtain Compound of Formula III a Compound (13.66 g, yield 88%, purity 93.8%).

[0186] Synthesis of Cyclo[6-[(3R,4R)-3-(acetoxy)-N,4-dimethyl-6-oxo-L-norleucine]-L-2-aminobutanoyl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula III a )

[0187] Under nitrogen protection, toluene (80 mL) and the compound of Formula II a (16.1 g) were added to a reaction flask. After stirring and dissolving, the temperature was controlled at 20 ± 5 °C, and sodium periodate (5.38 g) was added. After the addition was completed, the reaction was carried out under insulation. After the reaction was detected to be complete by high performance liquid chromatography, the reaction was quenched with an aqueous solution of sodium sulfite and sodium bicarbonate, and extracted with ethyl acetate. The organic phases were combined, washed successively with a refined industrial salt solution and dried over anhydrous sodium sulfate. The organic phase was collected and concentrated to obtain the compound of III a (14.12 g, yield 91%, purity 95.6%).

[0188] Synthesis of Cyclo[6-[(3R,4R)-3-(acetoxy)-N,4-dimethyl-6-oxo-L-norleucine]-L-2-aminobutanoyl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula III a )

[0189] Under nitrogen protection, acetone (130 mL) and the compound of Formula II a (16.1 g) were added to a reaction kettle. After stirring and dissolving, the temperature was controlled at 0 ± 5 °C, and sodium periodate (3.77 g) was added. After the addition was completed, the reaction was carried out under insulation. After the reaction was detected to be complete by high performance liquid chromatography, the reaction was quenched with an aqueous solution of sodium sulfite and sodium bicarbonate, and extracted with ethyl acetate. The organic phases were combined, washed successively with a refined industrial salt solution and dried over anhydrous sodium sulfate. The organic phase was collected and concentrated to obtain the compound of Formula III a (14.43 g, yield 93%, purity 98.2%). The 1H NMR, 13C NMR, mass spectrum and high performance liquid chromatography of the compound of Formula III a are shown in Figures 7 - 10 .

[0190] Example 28, Ring [[(6E)-(2S,3R,4R)-3-(Acetyloxy)-4-methyl-2-(methylamino)-6,8-nonadienoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula IV a )

[0191]

[0192] Dissolve Formula III a (16.0 g) in toluene (60 mL), add potassium carbonate (2.7 g) and allyl triphenylphosphonium bromide (7.5 g), stir and reflux for 6 h. After the reaction is completed as detected by high performance liquid chromatography, add ice water (about 60 mL), stir for 30 min, a solid precipitates, filter by suction, and dry to obtain Formula IV a Compound (14.68 g, yield 90%, purity 98.0%). Example 29, Synthesis of Ring [[(6E)-(2S,3R,4R)-3-hydroxy-4-methyl-2-(methylamino)-6,8-nonadienoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula V)

[0193]

[0194] Add DMA (100 mL), Formula IV a (13.1 g), and sodium hydride (1.0 g) to the reaction flask, control the temperature at 25 °C for reaction; after the reaction is completed as detected by TLC, add water, extract with ethyl acetate, and back-extract the aqueous phase. Combine the organic phases, wash with refined industrial salt, dry over anhydrous sodium sulfate, concentrate to obtain a crude product. After crystallization, filtration, and drying, obtain Formula V (10.76 g, yield 85.0%, purity 99.1%).

[0195] Comparative Example 1 Synthesis of Ring [[(2S,3R,4R)-3-(Acetyloxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0196] Under nitrogen protection, add methyl tert-butyl ether (17 mL), n-butanol (3.6 mL), Compound I a (10.3 g), aqueous N-methylmorpholine N-oxide solution (dissolve 1.415 g in 2.52 mL of water, 50% concentration), and potassium osmate dihydrate (0.076 g). Control the temperature at 20 ± 5 °C and stir for 2 h. Detect the reaction by high performance liquid chromatography. There is still raw material remaining (purity 94.4%, see high performance liquid chromatography in Figure 11 ); continue stirring the reaction for 5 h. Detect the reaction by high performance liquid chromatography. There is no raw material remaining and the reaction is completed (purity 97.41%, see high performance liquid chromatography in Figure 12 ). Add sodium sulfite and aqueous sodium bicarbonate solution to quench the reaction. Extract with dichloromethane. Combine the organic phases, wash with refined industrial brine solution, collect the organic phase, and concentrate. The crude product is crystallized in methanol / water, filtered by suction, and dried to obtain Compound II a (9.8 g, yield 92.6%).

[0197] Compare Example 10 with Comparative Example 1. In Example 10, the reaction was completed after 1 h of reaction, while in Comparative Example 1, there was still raw material remaining after 2 h of reaction; it was necessary to react the raw materials completely at 5 h of reaction (see high performance liquid chromatography comparison in Figure 13 ); moreover, after the reaction in Comparative Example 1 was completed, the main impurities generated were significantly higher than those in Example 10 (1.22% vs 0.42%). It can be seen that according to the method provided by the present invention, the reaction time can be greatly shortened, the reaction efficiency can be improved, and the high purity of the product can be ensured at the same time.

[0198] Synthesis of Cyclo[[(2S,3R,4R)-3-(acetoxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Compound II a )

[0199] Under nitrogen protection, add methyl tert-butyl ether (17 mL), n-butanol (3.6 mL), Compound I a (10.3 g), trimethylamine N-oxide dihydrate (1.37 g). Control the temperature at 20 ± 5 °C and stir until dissolved and clear; add potassium osmate dihydrate (0.07 g). Control the temperature at 20 ± 5 °C and stir for 1 h. Detect the reaction by high performance liquid chromatography. 77% of the raw material still remains; continue stirring the reaction for 5 h. Detect the reaction by high performance liquid chromatography. 19.79% of the raw material still remains, and the content of the main impurities has reached as high as 1.90% (see high performance liquid chromatography comparison in Figure 14); Continue the reaction until it is detected that the raw materials have reacted completely. Add aqueous sodium sulfite and sodium bicarbonate solutions to quench the reaction, extract with dichloromethane, combine the organic phases, wash with a refined industrial brine solution, collect the organic phase, and concentrate. The crude product is crystallized from methanol / water, filtered by suction, and dried to obtain Compound of Formula II a (9.5 g, yield 89.8%).

[0200] Synthesis of Cyclo[[(2S,3R,4R)-3-(Acetyloxy)-4-methyl-2-(methylamino)-6,7-dihydroxy-octanoyl]-L-2-aminobutyryl-N-methyl-glycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] (Formula II a )

[0201] Under nitrogen protection, add methyl tert-butyl ether (17 mL), n-butanol (3.6 mL), Compound of Formula I a (10.3 g), aqueous N-methylmorpholine N-oxide solution (dissolve 1.415 g in 2.52 mL of water, 50% concentration), and potassium osmate dihydrate (0.076 g), 0.17 g of acetic anhydride. Control the temperature at 20 ± 5 °C and stir for 2 h. Detect the reaction by high-performance liquid chromatography. 63% of the raw materials still remain (see high-performance liquid chromatography in Figure 15 ); Continue stirring the reaction for 5 h. Detect the reaction by high-performance liquid chromatography. 10.5% of the raw materials still remain (see high-performance liquid chromatography in Figure 16 ); Continue the reaction until it is detected that the raw materials have reacted completely. Add aqueous sodium sulfite and sodium bicarbonate solutions to quench the reaction, extract with dichloromethane, combine the organic phases, wash with a refined industrial brine solution, collect the organic phase, and concentrate. The crude product is crystallized from methanol / water, filtered by suction, and dried to obtain Compound of Formula II a (9.1 g, yield 86.0%).

[0202] In the comparative examples and comparative examples, study the effects of different reaction conditions on the preparation of the Compound of Formula II. The results are shown in Table 1:

[0203] Table 1 Effects of Different Reaction Conditions on the Preparation of the Compound of Formula II

[0204]

[0205] For those skilled in the art, various corresponding changes and deformations can be made according to the technical solutions and concepts described above, and all such changes and deformations should fall within the protection scope of the claims of the present invention.

Claims

1. A method for preparing a cyclosporine intermediate of formula II, characterized in that: The compound of formula Ⅰ is reacted with an oxidizing agent to obtain a compound of formula Ⅱ: Wherein, R is a hydroxyl protecting group; Preferably, R is (C 1-10 alkyl or aryl)3 silyl, (C 1-10 alkyl or aryl) acyl, (C 1-6 Alkoxy or C 6-10 Aryloxy)carbonyl, C 1-10 Any of alkyl or substituted alkyl, and aryl; More preferably, R is any one of trimethylsilyl, triethylsilyl, triisopropylsilyl, isopropyldimethylsilyl, isopropyldiethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, 2,3,6-trimethyl-4-hydroxybenzoyl, p-nitrobenzoyl, formyl, acetyl, benzoyl, tert-butyloxycarbonyl, benzyloxycarbonyl, methoxymethyl, allyl, methyl, tert-butyl, 2-tetrahydropyranyl, benzyl, p-methoxybenzyl, and trityl; More preferably, R is acetyl.

2. The preparation method according to claim 1, characterized in that: The oxidant is osmium tetroxide, 4-methylmorpholine-N-oxide hydrate, trimethylamine oxide, potassium permanganate, osmium chloride potassium ferrocyanide, hydrogen peroxide, oxygen, alkyl peroxide, organic peroxy acid, perimidic acid, peroxyacetyl nitrate, dimethyldioxirane, trimethylamine N-oxide dihydrate, anhydrous trimethylamine N-oxide, elemental iodine, silver carboxylate, chlorate, iodate, potassium ferrocyanide, potassium osmate dihydrate, and any one or more combinations of osmium compounds and ligand compositions; Preferably, the oxidant is any one or more of 4-methylmorpholine-N-oxide hydrate, chlorate, iodate, hydrogen peroxide, trimethylamine N-oxide dihydrate, oxygen, tert-butyl hydroperoxide, potassium ferrocyanide, and potassium osmate dihydrate. More preferably, the oxidant is a combination of 4-methylmorpholine-N-oxide hydrate and potassium osmate dihydrate or a combination of trimethylamine N-oxide dihydrate and potassium osmate dihydrate; More preferably, the oxidant is a combination of trimethylamine N-oxide dihydrate and potassium osmate dihydrate.

3. The preparation method according to claim 2, characterized in that: When the oxidant is a combination of 4-methylmorpholine-N-oxide hydrate and potassium osmate dihydrate, the molar equivalent ratio of potassium osmate dihydrate to formula I is 0.025-0.125:1; preferably, the molar equivalent ratio of potassium osmate dihydrate to formula I is 0.050-0.100:1; the molar equivalent ratio of 4-methylmorpholine-N-oxide hydrate to formula I is 0.7-3:1; preferably, the molar equivalent ratio of 4-methylmorpholine-N-oxide hydrate to formula I is 2.0-2.5:1; When the oxidant is a combination of trimethylamine N-oxide dihydrate and potassium osmate dihydrate, the molar equivalent ratio of potassium osmate dihydrate to formula I is 0.025-0.125:1; preferably, the molar equivalent ratio of potassium osmate dihydrate to formula I is 0.025-0.075:1; the molar equivalent ratio of trimethylamine N-oxide dihydrate to formula I is 0.7-3:1; preferably, the molar equivalent ratio of trimethylamine N-oxide dihydrate to formula I is 1.5-2.5:

1.

4. The preparation method according to claim 1, characterized in that: The reaction solvent of the reaction is selected from any one or more of acetonitrile, tetrahydrofuran, acetone, toluene, dichloromethane, dichloroethane, pyridine, methanol, ethanol, tert-butanol, and water; Preferably, the reaction solvent is a combination of acetonitrile and water; More preferably, the volume ratio of acetonitrile to water is 2 to 4:

1.

5. The preparation method according to claim 1, characterized in that: The reaction temperature of the reaction is -45 to 45°C; preferably, the reaction temperature is 15 to 25°C.

6. The preparation method according to any one of claims 1 to 5, characterized in that: The reaction also needs to be carried out in an acidic system, and the acidic system includes an organic acid or an inorganic acid; Preferably, the acidic system comprises any one or more of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, acetic anhydride, and nitric acid; Preferably, the acidic system comprises acetic acid or acetic anhydride; More preferably, the acidic system comprises acetic anhydride.

7. The preparation method according to claim 6, characterized in that: The molar equivalent ratio of acetic acid or acetic anhydride to Formula I is 0.1 to 1:1; preferably, the molar equivalent ratio of acetic acid or acetic anhydride to Formula I is 0.2:

1.

8. A method for preparing a compound of formula III, characterized in that: Mainly obtained by reacting the compound of formula II with an oxidizing agent: Wherein, R is a hydroxyl protecting group; Preferably, R is (C 1-10 alkyl or aryl)3 silyl, (C 1-10 alkyl or aryl) acyl, (C 1-6 Alkoxy or C 6-10 Aryloxy)carbonyl, C 1-10 Any of alkyl or substituted alkyl, and aryl; More preferably, R is any one of trimethylsilyl, triethylsilyl, triisopropylsilyl, isopropyldimethylsilyl, isopropyldiethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, 2,3,6-trimethyl-4-hydroxybenzoyl, p-nitrobenzoyl, formyl, acetyl, benzoyl, tert-butyloxycarbonyl, benzyloxycarbonyl, methoxymethyl, allyl, methyl, tert-butyl, 2-tetrahydropyranyl, benzyl, p-methoxybenzyl, and trityl; More preferably, R is acetyl; The compound of formula II is prepared by the preparation method described in claim 1.

9. The preparation method according to claim 8, characterized in that: The oxidizing agent is any one or more of periodic acid, sodium periodate, lead tetraacetate, thallium nitrate, thallium ethoxide, active manganese dioxide, pyridinium chlorochromate, bis(acetylacetonate)vanadium oxide, oxygen, cobalt acetate, silver ions, and peroxodisulfate. Preferably, the oxidizing agent is periodic acid or sodium periodate; More preferably, the oxidizing agent is sodium periodate.

10. The preparation method according to claim 8, characterized in that: The molar equivalent of the oxidizing agent to the compound of formula II is 1.0 to 2.0:1; preferably, the molar equivalent is 1.4 to 1.8:

1.

11. The preparation method according to claim 8, characterized in that: The reaction solvent of the reaction is selected from any one or more of acetonitrile, tetrahydrofuran, acetone, toluene, benzene and water; Preferably, the reaction solvent is a combination of tetrahydrofuran and water; More preferably, the volume ratio of tetrahydrofuran to water is 1 to 9:

1.

12. The preparation method according to claim 8, characterized in that: The reaction temperature of the reaction is -5 to 45°C; preferably, the reaction temperature is 15 to 25°C.

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