Vitamin E derivatives, carrier compounds and applications thereof

Abstract

This invention relates to a vitamin E derivative, its carrier compound, and its applications. The vitamin E derivative has the structure shown in formula (1), which can be linked and combined with different linking agents to obtain a carrier for the synthesis of polypeptides. The preparation process is simple, and the resulting carrier has good solubility, good stability, and good recyclability, and can be used for the efficient and high-yield synthesis of high-purity polypeptides.

Description

Technical Field

[0001] This invention belongs to the field of polypeptide synthesis technology, specifically relating to a vitamin E derivative for synthesizing polypeptides, its carrier compound, and its applications. Background Technology

[0002] The main methods for the chemical synthesis and production of peptides currently include solid-phase methods and conventional liquid-phase methods.

[0003] Solid-phase synthesis has advantages such as high versatility and short development cycle, and is suitable for the synthesis of all peptides. Its disadvantages are slow reaction rate, high material consumption, few means of monitoring the reaction process and low accuracy, inability to purify intermediates, low purity of crude peptides, and high production cost due to the need for high-performance preparative liquid chromatography (HPLC) to purify the final product.

[0004] Conventional liquid chromatography (LC) has a fast reaction rate and low material consumption; the reaction process can be monitored by various quantitative and qualitative analysis methods such as TLC, HPLC, and MS; each intermediate can be purified, and the crude peptide has high purity; however, it has poor versatility, a long development cycle, and complex steps.

[0005] Liquid-phase carrier method is a new peptide synthesis method developed to improve upon the two methods mentioned above. It replaces traditional resins with soluble compounds or polymers, transforming the solid-liquid two-phase reaction into a liquid-phase reaction. This accelerates the reaction rate, reduces material usage, and is essentially the same as the conventional liquid-phase method. Analytical methods used in conventional liquid-phase methods can be applied, and intermediates can be purified if necessary. A representative example of this technology is JITSUBO's Molecular Hiving method. TM The technologies include Ajiphase technology from Ajinomoto and EMPHASES technology from Tongjun Pharmaceuticals.

[0006] Molecular Hiving TM The technique uses long-chain alkoxybenzyl alcohols, such as 3,5-bis(eicosyloxy)benzyl alcohol, 2,4-bis(eicosyloxy)benzyl alcohol, and 3,4,5-tris(octadecyloxy)benzyl alcohol, as liquid-phase carriers. The carrier serves as a C-terminal protecting agent. The reaction is carried out in a homogeneous phase, and intermediates and byproducts are separated through precipitation-filtration-washing steps. This method has the following problems: ① Low reaction concentration, which is not conducive to scale-up production; ② Multiple precipitation-filtration-washing steps, making the operation complex; ③ The intermediate is an amorphous solid, and filtration-washing is time-consuming, resulting in poor purification.

[0007] The Ajiphase technology uses multi-branched alkyl groups instead of straight-chain alkyl groups, which increases the reaction concentration. The intermediates are purified by extraction and washing, but the reaction time is long, the solvent ratio is high during washing, the removal of impurities is generally not effective, the product loss is large, and the solubility of the intermediates decreases rapidly during the amino acid inoculation process. When the number of amino acids is greater than 4, gelation will occur.

[0008] EMPHASES technology utilizes amphiphilic carriers, further increasing reaction concentration. In homogeneous or heterogeneous solvent systems, especially in heterogeneous systems, it improves reaction rate and reagent utilization, simplifies post-processing, enhances product purity, and increases the versatility and universality of the operational process. However, the carrier preparation process is extremely complex. Carriers with C-terminal carboxyl and amide groups differ, requiring different preparation processes, and recycling is not readily available. Some carrier structures contain ester bonds, leading to carrier degradation during production. The issue of decreased solubility during amino acid incorporation has not been fully resolved; when the number of amino acids exceeds 8, some peptide intermediates exhibit gelation, reducing conversion and yield, making it unsuitable for peptide synthesis with more than 8 amino acids. Furthermore, it does not provide a method for preparing carriers with C-terminal groups of other types.

[0009] Therefore, it is necessary to develop a recyclable carrier that is simple to prepare, has good solubility, and good stability for peptide synthesis, thereby obtaining a more universal peptide synthesis technology. Summary of the Invention

[0010] Based on this, the purpose of this invention is to provide a recyclable carrier that is simple to prepare, has good solubility, and good stability for peptide synthesis.

[0011] The present invention includes the following technical solutions.

[0012] In a first aspect, the present invention provides a vitamin E derivative having the structure shown in formula (1) or a stereoisomer thereof.

[0013]

[0014] Wherein, R is selected from: hydrogen, one or more R5-substituted or unsubstituted C1-C atoms. 22 alkyl,

[0015] R1 is selected from: one or more R5-substituted or unsubstituted C1-C 22 Alkyl group, one or more R6-substituted or unsubstituted C6-C 22 aryl;

[0016] R2, R3, and R4 are each independently selected from: hydrogen and C1-C6 alkyl groups;

[0017] Each R5 is independently selected from: hydrogen, C6-C10 Aryl, halogen;

[0018] Each R6 is independently selected from: hydrogen, C1-C 22 Alkyl, C1-C 22 Alkyl groups, halogens, or two adjacent R6 groups linked together to form one or more R7-substituted or unsubstituted C3-C8 cycloalkyl groups or 3-8-membered heterocyclic groups;

[0019] Each R7 is independently selected from: hydrogen, C1-C 22 Alkyl, C1-C 22 Alkoxy groups, halogens;

[0020] x is selected from: 1, 2, 3, 4, 5;

[0021] n is selected from: 1, 2, 3;

[0022] y is selected from: 1, 2, 3, 4, 5;

[0023] m is selected from: 1, 2, 3.

[0024] Secondly, the present invention provides the use of the aforementioned vitamin E derivative or its stereoisomer in the preparation of a carrier for the synthesis of polypeptides.

[0025] Thirdly, the present invention provides a carrier compound having the structure shown in formula (2) or a stereoisomer thereof for synthesizing polypeptides:

[0026]

[0027] Wherein, R, R2, R3, R4, x and n are as described in the previous formula (1);

[0028] L is a linking group used for linking with amino acids.

[0029] Fourthly, the present invention provides the application of the vitamin E derivative or its stereoisomer described herein, or the carrier compound or its stereoisomer described herein, in the synthesis of polypeptides.

[0030] Fifthly, the present invention provides a method for synthesizing polypeptides, wherein the synthesis method uses the compound described in the present invention as a carrier to carry out the synthesis reaction.

[0031] Furthermore, the method for synthesizing the polypeptide includes the following steps:

[0032] (1) React the N-terminal protected amino acid or peptide with the carrier compound or its stereoisomer as described in this invention to obtain a carrier-amino acid / peptide conjugate.

[0033] (2) Remove the N-terminal protecting group from the carrier-amino acid / peptide conjugate and then react it with the next N-terminal protected amino acid or peptide.

[0034] (3) Repeat step (2) to elongate the peptide chain and obtain a conjugate of the N-terminally protected target polypeptide and the carrier compound.

[0035] (4) Remove the carrier and / or protecting group from the conjugate obtained in step (3) to obtain the target polypeptide.

[0036] This invention utilizes readily available and inexpensive vitamin E (including various chiral and non-chiral natural vitamin E or synthetic vitamin E) as a raw material to prepare a series of novel vitamin E derivatives through simple chemical reactions. The raw materials for preparing these vitamin E derivatives are inexpensive, and the preparation process is simple. Like solid-phase resins, they can be linked and combined with different linking agents to obtain carriers for polypeptide synthesis, making them suitable for all types of polypeptide production. Using them for polypeptide synthesis offers the following advantages:

[0037] (1) The carrier compound has the lipophilic structure of vitamin E and the weakly polar ethylene glycol ether structure. It has high solubility in nonpolar solvents and a certain solubility in polar solvents. It has amphiphilic properties and has a fast reaction rate in both homogeneous and heterogeneous solvent systems. It can be used to synthesize high-purity peptides with high efficiency and high yield.

[0038] (2) The carrier compound of the present invention is linked to the linker by an amide bond, which has good stability during peptide synthesis and can prevent the carrier from falling off during the synthesis process; and it is easy to recover. After the synthesis reaction is completed, the carrier compound can be recovered with high efficiency and high yield through a simple chemical treatment process for the next round of peptide synthesis, thereby greatly saving peptide synthesis costs.

[0039] (3) The peptides synthesized using the carrier compound described in this invention can still maintain good solubility after multiple amino acids are added, which solves the problem of decreased solubility of existing carriers during amino acid addition. This is very beneficial for the synthesis of long peptides. No peptide chain gelation will occur during the synthesis of long peptides, which can effectively improve the synthesis yield and purity of long peptides.

[0040] (4) The carrier compounds of the present invention, when used in combination with different linkers, can provide not only carriers for preparing peptides with C-terminal COOH and amide groups, but also carriers for preparing peptides with C-terminal groups of other types, thereby enabling the preparation of modified polypeptides with different C-terminal groups. The carrier compounds provided by the present invention have better versatility and universality than the first-generation EMPHASES carriers, and are suitable for the large-scale production of polypeptides, especially long peptides. Detailed Implementation

[0041] To facilitate understanding of the present invention, a more complete description will be provided below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0042] Unless otherwise specified, experimental methods in the following examples are generally performed under standard conditions or as recommended by the manufacturer. All commonly used chemical reagents used in the examples are commercially available products.

[0043] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used in this invention includes any and all combinations of one or more of the associated listed items.

[0044] Furthermore, as used herein, the term "or" is an inclusive "or" sign and is equivalent to the term "and / or" unless the context clearly specifies otherwise. The term "based on" is not exclusive and allows for basing on other factors not described unless the context clearly specifies otherwise. Additionally, throughout the specification, the meanings of "an," "a," and "the" include plural indicators. The meaning of "in" includes both "in" and "on."

[0045] In one embodiment of the present invention, a vitamin E derivative having the structure shown in formula (1) or a stereoisomer thereof is provided.

[0046]

[0047] Wherein, R is selected from: hydrogen, one or more R5-substituted or unsubstituted C1-C atoms. 22 alkyl,

[0048] R1 is selected from: one or more R5-substituted or unsubstituted C1-C 22 Alkyl group, one or more R6-substituted or unsubstituted C6-C 22 aryl;

[0049] R2, R3, and R4 are each independently selected from: hydrogen and C1-C6 alkyl groups;

[0050] Each R5 is independently selected from: hydrogen, C6-C 10 Aryl, halogen;

[0051] Each R6 is independently selected from: hydrogen, C1-C22 Alkyl, C1-C 22 Alkyl groups, halogens, or two adjacent R6 groups linked together to form one or more R7-substituted or unsubstituted C3-C8 cycloalkyl groups or 3-8-membered heterocyclic groups;

[0052] Each R7 is independently selected from: hydrogen, C1-C 22 Alkyl, C1-C 22 Alkoxy groups, halogens;

[0053] x is selected from: 1, 2, 3, 4, 5;

[0054] n is selected from: 1, 2, 3;

[0055] y is selected from: 1, 2, 3, 4, 5;

[0056] m is selected from: 1, 2, 3.

[0057] In the compounds of this invention, when any variable (e.g., R5, R6, etc.) appears more than once in any component, the definition of each occurrence is independent of the definitions of other occurrences. Similarly, combinations of substituents and variables are permitted, provided such combinations stabilize the compound. It will be understood that those skilled in the art can select the substituents and substitution patterns of the compounds of this invention to provide chemically stable compounds that can be easily synthesized from readily available starting materials using techniques in the art and the methods described below. If a substituent is itself substituted by more than one group, it should be understood that these groups can be on the same carbon atom or on different carbon atoms, as long as structural stability is achieved.

[0058] As used in this invention, the term "alkyl" refers to both branched and straight-chain saturated aliphatic hydrocarbon groups having a specific number of carbon atoms. For example, the definition of "C1-C6" in "C1-C6 alkyl" includes groups having 1, 2, 3, 4, 5, or 6 carbon atoms arranged in a straight or branched chain. Specifically, "C1-C6 alkyl" includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, and hexyl.

[0059] As used in this invention, the term "cycloalkyl" refers to a monocyclic, bicyclic, or polycyclic cyclic hydrocarbon group whose ring atoms are composed of carbon atoms and are saturated or partially unsaturated. Bicyclic or polycyclic groups include spirocyclic, fused, and bridged rings. For example, "cycloalkyl" includes, but is not limited to, the following groups: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. wait.

[0060] The term "alkoxy" as used in this invention refers to a group having an -O-alkyl structure, such as -OCH3, -OCH2CH3, -OCH2CH2CH3, -O-CH2CH(CH3)2, -OCH2CH2CH2CH3, -O-CH(CH3)2, etc.

[0061] As used in this invention, the term "heterocyclic alkyl" or "heterocyclic group" refers to a saturated or partially unsaturated monocyclic, bicyclic, or polycyclic cyclic substituent (including spirocyclic, bridged, fused, and fused rings, etc.), wherein one or more ring atoms are selected from heteroatoms of N, O, or S(O)m (where m is an integer from 0 to 2), and the remaining ring atoms are carbon. Examples include: morpholinyl, piperidinyl, tetrahydropyrrolyl, pyrrolylalkyl, dihydroimidazolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyrazoleyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothiophene, dihydrotriazolyl, dihydroazacyclobutane, tetrahydrofuranyl, tetrahydrothiophene, etc., and their N-oxides. The connection of heterocyclic substituents can be achieved through carbon atoms or through heteroatoms.

[0062] As will be understood by those skilled in the art, the term "halogen" or "halogen" as used in this invention refers to chlorine, fluorine, bromine, and iodine.

[0063] In some embodiments of the present invention, R is selected from: hydrogen, benzyl, C1-C 20 alkyl,

[0064] In some embodiments of the present invention, R1 is selected from: benzyl, C1-C 22 Alkyl group, one or more R6-substituted or unsubstituted C6-C 10 aryl;

[0065] Each R6 is independently selected from: hydrogen, C1-C 18 Alkyl, C1-C 18 Alkyl groups, halogens, or two adjacent R6 groups linked together to form one or more R7-substituted or unsubstituted C5-C6 cycloalkyl groups or 5-6-membered heterocyclic groups;

[0066] Each R7 is independently selected from: hydrogen, C1-C 18 Alkyl, C1-C 18 Alkoxy groups, halogens.

[0067] In some embodiments of the present invention, R1 is selected from: benzyl, C1-C 12 alkyl,

[0068] In some embodiments of the present invention, R2, R3, and R4 are each independently selected from: hydrogen, methyl, ethyl, n-propyl, and isopropyl.

[0069] In some embodiments of the present invention, R2, R3 and R4 are all methyl groups.

[0070] In some embodiments of the present invention, x is selected from 1, 2, and most preferably 1; n is selected from 1, 2.

[0071] In some embodiments of the present invention, y is selected from 1 and 2, with 1 being the most preferred; m is selected from 1 and 2.

[0072] In some embodiments of the present invention, R is selected from: hydrogen, benzyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptyl, octadecyl, nonadecanyl, eicosyl.

[0073] R1 is selected from: benzyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl.

[0074] y is selected from: 1, 2, m is selected from: 1, 2.

[0075] In some embodiments of the present invention, the vitamin E derivative or its stereoisomer is selected from the following compounds:

[0076]

[0077]

[0078] The vitamin E derivative raw material provided by this invention for peptide synthesis is inexpensive, simple to prepare, and easy to recycle. The vitamin E derivative described in this invention can be linked and combined with different linkers, similar to solid-phase resins, to obtain a carrier for peptide synthesis, suitable for all types of peptide production. The linker is an amide bond, exhibiting good stability during peptide synthesis. This carrier compound possesses both the lipophilic structure of vitamin E and the weakly polar ethylene glycol ether structure, exhibiting high solubility in nonpolar solvents and some solubility in polar solvents, demonstrating amphiphilic properties. It exhibits rapid reaction rates in both homogeneous and heterogeneous solvent systems, and its versatility and universality are superior to first-generation EMPHASES carriers, making it suitable for large-scale peptide production.

[0079] In one embodiment of the present invention, a carrier compound having the structure shown in formula (2) or a stereoisomer thereof for synthesizing polypeptides is also provided.

[0080]

[0081] Wherein, R is selected from: hydrogen, one or more R5-substituted or unsubstituted C1-C atoms. 22 alkyl,

[0082] R1 is selected from: one or more R5-substituted or unsubstituted C1-C 22 Alkyl group, one or more R6-substituted or unsubstituted C6-C 22 aryl;

[0083] R2, R3, and R4 are each independently selected from: hydrogen and C1-C6 alkyl groups;

[0084] Each R5 is independently selected from: hydrogen, C6-C 10 Aryl, halogen;

[0085] Each R6 is independently selected from: hydrogen, C1-C 22 Alkyl, C1-C 22 Alkyl groups, halogens, or two adjacent R6 groups linked together to form one or more R7-substituted or unsubstituted C3-C8 cycloalkyl groups or 3-8-membered heterocyclic groups;

[0086] Each R7 is independently selected from: hydrogen, C1-C 22 Alkyl, C1-C 22 Alkoxy groups, halogens;

[0087] x is selected from: 1, 2, 3, 4, 5;

[0088] n is selected from: 1, 2, 3;

[0089] y is selected from: 1, 2, 3, 4, 5;

[0090] m is selected from: 1, 2, 3;

[0091] L is a linking group used for linking with amino acids.

[0092] In some embodiments of the present invention, R is selected from: hydrogen, benzyl, C1-C 20 alkyl,

[0093] In some embodiments of the present invention, R1 is selected from: benzyl, C1-C 22 Alkyl group, one or more R6-substituted or unsubstituted C6-C 10 aryl;

[0094] Each R6 is independently selected from: hydrogen, C1-C 18 Alkyl, C1-C 18 Alkyl groups, halogens, or two adjacent R6 groups linked together to form one or more R7-substituted or unsubstituted C5-C6 cycloalkyl groups or 5-6-membered heterocyclic groups;

[0095] Each R7 is independently selected from: hydrogen, C1-C 18 Alkyl, C1-C 18 Alkoxy groups, halogens.

[0096] In some embodiments of the present invention, R1 is selected from: benzyl, C1-C 12 alkyl,

[0097] In some embodiments of the present invention, R2, R3, and R4 are each independently selected from: hydrogen, methyl, ethyl, n-propyl, and isopropyl.

[0098] In some embodiments of the present invention, R2, R3 and R4 are all methyl groups.

[0099] In some embodiments of the present invention, x is selected from 1, 2, and most preferably 1; n is selected from 1, 2.

[0100] In some embodiments of the present invention, y is selected from 1 and 2, with 1 being the most preferred; m is selected from 1 and 2.

[0101] In some embodiments of the present invention, R is selected from: hydrogen, benzyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptyl, octadecyl, nonadecanyl, eicosyl.

[0102] R1 is selected from: benzyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl.

[0103] y is selected from: 1, 2, m is selected from: 1, 2.

[0104] In the carrier compound of this invention, L is a linking group obtained by reacting a conventional linker capable of linking amino acids with the amino / amino group in the vitamin E derivative described in this invention. It is used to link amino acids during peptide synthesis and can be removed by conventional deprotecting agents in the art after the synthesis reaction, thereby obtaining the final target peptide chain. Specifically, L can be selected from, but is not limited to, the following groups:

[0105]

[0106] In some embodiments of the present invention, the support compound or its stereoisomer is selected from the following compounds:

[0107]

[0108]

[0109] In some embodiments, the carrier compound is obtained by reacting the vitamin E derivative or its stereoisomer described in this invention with a linker for linking to amino acids, wherein the linker for linking to amino acids is selected from the following compounds:

[0110]

[0111] When the linker used for linking with amino acids contains an amino protecting group (e.g., an Fmoc protecting group), the vitamin E derivative of the present invention reacts with the linker, and then the amino protecting group is removed by deprotection to obtain a carrier compound for synthesizing polypeptides, which is then used for the reaction of linking with amino acids.

[0112] In one embodiment of the present invention, the use of the vitamin E derivative or its stereoisomer described herein, or the carrier compound or its stereoisomer described herein, in the synthesis of polypeptides is provided.

[0113] In one embodiment of the present invention, a method for synthesizing polypeptides is provided, wherein the synthesis method uses the compound described in the present invention as a carrier to carry out the synthesis reaction.

[0114] In some embodiments of the present invention, the method for synthesizing polypeptides includes the following steps:

[0115] (1) React the N-terminal protected amino acid or peptide with the carrier compound or its stereoisomer described in this invention to obtain a carrier-amino acid / peptide conjugate.

[0116] (2) Remove the N-terminal protecting group from the carrier-amino acid / peptide conjugate and then react it with the next N-terminal protected amino acid or peptide.

[0117] (3) Repeat step (2) to elongate the peptide chain and obtain a conjugate of the N-terminally protected target polypeptide and the carrier compound.

[0118] (4) Remove the carrier and / or protecting group from the conjugate obtained in step (3) to obtain the target polypeptide.

[0119] When polypeptides are synthesized using the carrier compound of the present invention, the carrier can be safely deprotected from the N-terminal protecting group and / or the amino acid side-chain protecting group, or it can be deprotected together with the N-terminal protecting group and / or the amino acid side-chain protecting group under certain conditions. Therefore, the method of the present invention can synthesize polypeptides with N-terminal protection or with side-chain protecting groups, polypeptides with both N-terminal protecting groups and side-chain protecting groups, and completely deprotected polypeptides.

[0120] In some embodiments, step (4) includes:

[0121] Remove the carrier from the conjugate obtained in step (3) to obtain the N-terminally protected target polypeptide; or,

[0122] Simultaneously, the N-terminal protecting group and the carrier in the conjugate obtained in step (3) are removed to obtain the target polypeptide; or,

[0123] First, remove the carrier from the conjugate obtained in step (3), and then remove the N-terminal protecting group to obtain the target polypeptide.

[0124] In some embodiments, the N-terminal protected amino acid or peptide described in steps (1) and (2) contains a side-chain protecting group; step (4) includes:

[0125] Remove the carrier from the conjugate obtained in step (3) to obtain the target polypeptide with an N-terminal protected side chain; or,

[0126] Simultaneously, the N-terminal protecting group and the carrier in the conjugate obtained in step (3) are removed to obtain the target polypeptide with a side-chain protecting group; or,

[0127] Simultaneously, the N-terminal protecting group, side chain protecting group, and carrier are removed from the conjugate obtained in step (3) to obtain the target polypeptide; or,

[0128] First, remove the carrier from the conjugate obtained in step (3), then remove the N-terminal protecting group and / or the side chain protecting group to obtain the target polypeptide.

[0129] The carrier compound described herein exhibits excellent solubility in peptide synthesis, overcoming the problem of decreased solubility during amino acid incorporation in existing carriers. This is highly beneficial for the synthesis of long peptides, preventing peptide chain gelation and effectively improving the yield and purity of long peptides. Furthermore, the carrier compound demonstrates good stability and is easily recoverable. After the synthesis reaction, it can be efficiently and readily recovered through a simple chemical process for reuse in the next round of peptide synthesis, significantly reducing peptide synthesis costs and facilitating large-scale production.

[0130] The compounds corresponding to the abbreviations and short names involved in this invention are described below:

[0131] i-PrOAc: Isopropyl acetate;

[0132] MTBE: Methyl tert-butyl ether;

[0133] DMF: N,N-dimethylformamide;

[0134] MeOH: Methanol;

[0135] DMSO: Dimethyl sulfoxide;

[0136] HMBA: 4-hydroxymethylbenzoic acid;

[0137] DMT-MM: 4-(4,6-dimethoxytriazine)-4-methylmorpholine hydrochloride; DMAP: 4-dimethylaminopyridine;

[0138] EDCI: 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride; TFA: Trifluoroacetic acid;

[0139] TIS: Triisopropylsilane;

[0140] TBTU: Benzotriazole tetramethyltetrafluoroboric acid;

[0141] HOBt: 1-hydroxybenzotriazole.

[0142] The present invention will be further described in detail below with reference to specific embodiments.

[0143] Example 1: Synthesis of compounds VEC-001 and VEC-002

[0144]

[0145] Step 1: Dissolve vitamin E (43.1g, 0.1mol) in N,N-dimethylformamide (160ml), add tetrabutylammonium fluoride trihydrate (0.3g, 0.001mol), and heat to 140-150℃; add dropwise a solution of ethylene carbonate (9.7g, 0.11mol) dissolved in N,N-dimethylformamide (20ml), and continue the reaction at 140-150℃ for half an hour after the addition is complete. Recover N,N-dimethylformamide under reduced pressure, dissolve the remainder in toluene, wash twice with water, and reflux to remove residual water to obtain a toluene solution of intermediate 1-01, which can be used directly in the next step.

[0146] Step 2: Under water bath conditions, SOCl2 (14.3g, 0.12mol) was added dropwise to the toluene solution of intermediate 1-01 obtained in the previous step. After the addition was complete, the mixture was heated to 60-70℃ and reacted for 2 hours. The excess SOCl2 was concentrated to remove it, and then dissolved in toluene and concentrated again to obtain 49.3g of crude chloride 1-02 with a yield of 100%, which was directly used in the next step of the reaction.

[0147] Step 3: Dissolve the crude chloride 1-02 obtained in the previous step in N,N-dimethylformamide (400 ml), add benzylamine (5.38 g, 0.05 mol), potassium carbonate (27.6 g, 0.2 mol), and potassium iodide (33.2 g, 0.2 mol), heat to 100 °C and react for 24 hours, filter to remove the solid, concentrate the filtrate to recover N,N-dimethylformamide, add heptane (200 ml) and water (100 ml); separate the aqueous layer, concentrate the heptane, and purify by column chromatography to obtain compound 1-03 (30.6 g, yield 60%) and VEC-001 (11.3 g, yield 20%) (based on VE).

[0148] Step 4: Dissolve compound 1-03 (30.6 g, 0.03 mol) in ethyl acetate (300 ml), add 10% Pd / C (3 g), and react in an autoclave at room temperature under H2 pressure of 1.5 MPa for 3 hours; filter to remove catalyst, concentrate the filtrate to obtain compound VEC-002 25.1 g, yield 90%.

[0149] VEC-001: 1 H-NMR (500MHz, CDCl3): δ7.40-7.30(t, 2H), 7.30-7.20(t, 3H), 3.85-3.83(t, 2H), 3.80-3.75(m, 2H) 3.12-3.10(t, 2H), 2.59-2.55(t, 2H) ), 2.20(s, 3H), 2.16(s, 3H), 2.08(s, 3H), 1.85-1.75(t, 2H), 1.60-1.50(t, 2H), 1.50-1.00(t, 23H), 0.90-0.80(m, 12H); MS: 564.36[M+H] + .

[0150] VEC-002: 1 H-NMR (500MHz, CDCl3): δ3.85-3.83(t, 4H), 3.12-3.10(t, 4H), 2.59-2.55(t, 4H), 2.20(s, 6H), 2.16(s, 6H), 2.08(s, 6H), 1.85-1.75(t, 4H), 1.60-1.50(t, 4H), 1.50-1.00(t, 45H), 0.90-0.80(m, 24H); MS: 930.80[M+H] + .

[0151] Example 2: Synthesis of compounds VEC-003 and VEC-004

[0152]

[0153] Step 1: Dissolve natural vitamin E-α (43.1g, 0.1mol) in N,N-dimethylformamide (500ml), add 1,3-dibromopropane (40.4g, 0.2mol), tetrabutylammonium bromide (0.32g, 0.001mol), and potassium carbonate (27.6g, 0.2mol), and heat to 80-90℃; react for 48 hours, recover N,N-dimethylformamide under reduced pressure and remove excess 1,3-dibromopropane, dissolve the remainder in heptane, wash twice with water and concentrate to obtain crude product 2-01, which can be directly used in the next step of the reaction;

[0154] Step 2: The crude product 2-01 obtained in the previous step was dissolved in N-methylpyrrolidone (200 ml), and benzylamine (5.38 g, 0.05 mol), potassium carbonate (27.6 g, 0.2 mol), and potassium iodide (16.6 g, 0.1 mol) were added. The mixture was heated to 120 °C and reacted for 12 hours. The solid was removed by filtration, and the concentrated filtrate was added to heptane (200 ml) and water (100 ml). The aqueous layer was separated, and the mixture was washed twice with heptane and water. The mixture was concentrated and purified by column chromatography to obtain compound 1-03 (26.2 g, yield 50%) and VEC-003 (14.5 g, yield 25%) (based on VE).

[0155] Step 3: Dissolve compound 2-02 (26.2 g, 0.025 mol) in ethyl acetate (300 ml), add 10% Pd / C (2.6 g), and react in an autoclave at room temperature under H2 pressure of 1.5 MPa for 3 hours; filter to remove catalyst, concentrate the filtrate to obtain compound VEC-004 24.0 g, yield 90%.

[0156] VEC-003: 1 H-NMR (500MHz, CDCl3): δ7.38-7.30(t, 2H), 7.31-7.222(t, 3H), 3.87-3.85(t, 2H), 3.82-3.77(m, 2H), 3.13-3.10(t, 2 H), 2.60-2.57(t, 2H), 2.19(s, 3H), 2.16(s, 3H), 2.08(s, 3H), 1.90-1.00(m, 29H), 0.90-0.80(m, 12H); MS: 578.49[M+H] + .

[0157] VEC-004: 1H-NMR (500MHz, CDCl3): δ3.88-3.84(t, 4H), 3.15-3.10(t, 4H), 2.62-2.58(t, 4H), 2.19(s , 6H), 2.16 (s, 6H), 2.08 (s, 6H), 1.90-1.00 (m, 57H), 0.90-0.80 (m, 24H); MS: 958.66[M+H] + .

[0158] Example 3: Synthesis of compounds VEC-005 and VEC-006

[0159]

[0160] Step 1: Dissolve vitamin E (43.1g, 0.1mol) in N,N-dimethylformamide (500ml), add chlorodiethylene glycol (41.6g, 0.3mol), tetrabutylammonium bromide (0.32g, 0.001mol), potassium iodide (16.6g, 0.1mol), and potassium carbonate (41.4g, 0.3mol), and heat to 80-90℃; react for 24 hours, recover N,N-dimethylformamide under reduced pressure, dissolve the remainder in heptane, wash twice with ethanol:water (1:1), concentrate to obtain crude intermediate 3-01, which is directly used in the next step of the reaction.

[0161] Step 2: Dissolve the intermediate 3-01 obtained in Step 1 in dichloromethane, add thionyl chloride (14.3g, 0.12mol) dropwise under a water bath. After the addition is complete, heat to 60-70℃ and react for 2 hours. Concentrate to remove excess SOCl2, redissolve in toluene, and concentrate again to obtain crude intermediate 3-02, which can be used directly in the next step of the reaction.

[0162] Step 3: Dissolve intermediate 3-02 (53.7 g, 0.1 mol) in N,N-dimethylformamide (400 ml), add benzylamine (4.3 g, 0.04 mol), potassium carbonate (27.6 g, 0.2 mol) and potassium iodide (33.2 g, 0.2 mol), heat to 100 °C and react for 24 hours. Filter to remove solids, concentrate the filtrate to recover N,N-dimethylformamide, add toluene (500 ml) and water (300 ml); separate the aqueous layer, concentrate the toluene layer, and purify by column chromatography to obtain compound 3-03 (31.0 g, 70% yield) and VEC-005 (9.1 g, 15% yield) (based on benzylamine).

[0163] Step 4: Dissolve compound 3-03 (31.0 g, 0.028 mol) in ethyl acetate (300 ml), add 10% Pd / C (3.1 g), and react in an autoclave at room temperature under H2 pressure of 1.5 MPa for 3 hours; filter to remove catalyst, concentrate the filtrate to obtain compound VEC-006 27.1 g, yield 95%.

[0164] VEC-005: 1 H-NMR (500MHz, CDCl3): δ7.40-7.30(t, 2H), 7.30-7.20(t, 3H), 3.85-3.83(t, 2H), 3.80-3.75(m, 2H), 3.76-3.66(t, 2H), 3.60-3.50(t, 2H), 3.10-3.07(t, 2H) ), 2.59-2.55(t, 2H), 2.20(s, 3H), 2.16(s, 3H), 2.08(s, 3H), 1.85-1.75(t, 2H ), 1.60-1.50(t, 2H), 1.50-1.00(t, 23H), 0.90-0.80(t, 12H); MS: 608.50[M+H] + .

[0165] VEC-006: 1 H-NMR (500MHz, CDCl3): δ3.85-3.83(t, 4H), 3.76-3.66(t, 4H), 3.60-3.50(t, 4H), 3.10-3.08(t, 4H), 2.59-2.55(t, 4H), 2.20(s , 6H), 2.16(s, 6H), 2.08(s, 6H), 1.85-1.75(t, 4H), 1.60-1.50(t, 4H), 1.50-1.00(t, 45H), 0.90-0.80(m, 24H); MS: 1018.88[M+H] + .

[0166] Example 4: Synthesis of compound VEC-007

[0167]

[0168] Step 1: Dissolve chloride 1-02 (24.6 g, 0.05 mol) in N,N-dimethylformamide (300 ml), add benzylamine (21.4 g, 0.2 mol), potassium carbonate (8.3 g, 0.06 mol), and potassium iodide (3.3 g, 0.02 mol), heat to 80-90℃ and react for 8 hours, then add heptane (300 ml) and water (200 ml); separate the aqueous layer, wash twice with heptane and water, concentrate, and purify by column chromatography to obtain compound VEC-001 22.5 g, yield 80%.

[0169] Step 2: Dissolve compound VEC-001 (22.5 g, 0.04 mol) in N-methylpyrrolidone (200 ml), add intermediate 2-01 (22.1 g, 0.04 mol), potassium carbonate (8.3 g, 0.06 mol), and potassium iodide (16.6 g, 0.1 mol), heat to 80-90 °C and react for 24 hours, add heptane (400 ml) and water (200 ml); separate the aqueous layer, wash twice with heptane and water, concentrate, and purify by column chromatography to obtain intermediate 4-01 31.0 g, yield 75%.

[0170] Step 3: Dissolve intermediate 4-01 (31.0 g, 0.03 mol) in ethyl acetate (500 ml), add 10% Pd / C (3.1 g), and react in an autoclave at room temperature for 3 hours under H2 pressure of 1.5 MPa; filter to remove catalyst, and concentrate the filtrate to obtain compound VEC-00726.1 g, yield 92%.

[0171] 1 H-NMR (500MHz, CDCl3): δ3.86-3.80(m, 4H), 3.11-3.07(m, 4H), 2.59-2.55(t, 4H), 2.20(s, 6H), 2.16(s, 6H), 2.08(s, 6H), 2.00-1.70(m, 6H), 1.60-1.50(t, 4H), 1.50-1.00(t, 45H), 0.90-0.80(m, 24H); MS:944.80[M+H] + .

[0172] Example 5: Synthesis of compound VEC-008

[0173]

[0174] Step 1: Dissolve intermediate 3-02 (40.3 g, 0.075 mol) and compound VEC-001 (38.9 g, 0.075 mol) in N-methylpyrrolidone (500 ml), add potassium carbonate (12.5 g, 0.09 mol) and potassium iodide (12.5 g, 0.075 mol), heat to 80-90 °C and react for 24 hours, add heptane (600 ml) and water (200 ml); separate the aqueous layer, wash twice with heptane and water, concentrate, and purify by column chromatography to obtain intermediate 5-01 47.9 g, yield 60%.

[0175] Step 2: Dissolve intermediate 5-01 (47.9 g, 0.045 mol) in ethyl acetate (500 ml), add 10% Pd / C (4.8 g), and react in an autoclave at room temperature under H2 pressure of 1.5 MPa for 3 hours; filter to remove catalyst, concentrate the filtrate to obtain compound VEC-008 39.5 g, yield 90%.

[0176] 1 H-NMR (500MHz, CDCl3): δ3.87-3.82(m, 4H), 3.76-3.66(t, 2H), 3.60-3.50(t, 2H), 3.11-3.06(m, 4H), 2.59-2.55(t, 4H), 2.20(s , 6H), 2.16(s, 6H), 2.08(s, 6H), 1.85-1.75(t, 4H), 1.60-1.50(t, 4H), 1.50-1.00(t, 45H), 0.90-0.80(m, 24H); MS: 974.80[M+H] + .

[0177] Example 6 Synthesis of compounds VEC-009 and VEC-010

[0178]

[0179] Compound VEC-001 (25.0 g, 0.044 mol) was dissolved in ethyl acetate (500 ml), and 10% Pd / C (2.5 g) was added. The mixture was then reacted in an autoclave at room temperature for 3 hours under a pressure of 1.5 MPa H2. The catalyst was removed by filtration, and the filtrate was concentrated to obtain compound VEC-009 19.0 g, with a yield of 91%.

[0180] 1H-NMR (500MHz, CDCl3): δ3.85-3.83(t, 2H), 3.12-3.10(t, 2H), 2.59-2.55(t, 2H), 2.25(bs, 2H), 2.20(s, 3H), 2.16( s, 3H), 2.08 (s, 3H), 1.85-1.75 (t, 2H), 1.60-1.50 (t, 2H), 1.50-1.00 (t, 24H), 0.90-0.80 (m, 12H); MS: 474.23[M+H] + .

[0181] Compound VEC-001 (25.0 g, 0.044 mol) was dissolved in tetrahydrofuran (500 ml) and methanol (100 ml). Paraformaldehyde (2 g) and sodium cyanoborohydride (5.5 g, 0.088 mol) were added, and the mixture was reacted at room temperature for 3 hours. The solvent was removed by concentration, and ethyl acetate (200 ml) and water (200 ml) were added. The aqueous layer was separated, and the ethyl acetate layer was washed twice with water and used directly for the hydrogenation reaction. 10% Pd / C (2.5 g) was added to the resulting ethyl acetate solution, and the mixture was reacted in an autoclave at 1.5 MPa H2 pressure at room temperature for 3 hours. The catalyst was removed by filtration, and the filtrate was concentrated to give compound VEC-01018.0 g, with a yield of 85%.

[0182] 1 H-NMR (500MHz, CDCl3): δ3.85-3.83(t, 2H), 3.12-3.10(t, 2H), 2.59-2.55(t, 2H), 2.50(s, 3H), 2.20(s, 3H), 2.16( s, 3H), 2.08 (s, 3H), 1.85-1.75 (t, 2H), 1.60-1.50 (t, 2H), 1.50-1.00 (t, 25H), 0.90-0.80 (m, 12H); MS: 488.40[M+H] + .

[0183] Example 7 Synthesis of compound VEC-011

[0184]

[0185] Chloride 1-02 (24.6 g, 0.05 mol) was dissolved in N,N-dimethylformamide (300 ml), and octadecylamine (26.9 g, 0.1 mol), potassium carbonate (8.3 g, 0.06 mol), and potassium iodide (3.3 g, 0.02 mol) were added. The mixture was heated to 80-90 °C and reacted for 8 hours. Heptane (300 ml) and water (200 ml) were added. The aqueous layer was separated, and the mixture was washed twice with heptane. The solution was concentrated and purified by column chromatography to obtain compound VEC-011 25.4 g, with a yield of 70%.

[0186] 1 H-NMR (500MHz, CDCl3): δ3.85-3.75(t, 2H), 3.05-2.95(t, 2H), 2.75-2.65(t, 2H), 2.59-2.55(t, 2H), 2.18(s, 3H), 2.1 6(s, 3H), 2.08(s, 3H), 1.83-1.75(t, 2H), 1.60-1.50(t, 2H), 1.50-1.00(t, 53H), 0.90-0.80(t, 15H); MS: 726..70[M+H] + .

[0187] Example 8 Synthesis of compound VEC-012

[0188]

[0189] Compound VEC-012 was prepared by replacing octadecylamine with 2-ethylhexylamine according to the preparation method of Example 7, with a yield of 85%.

[0190] 1 H-NMR (500MHz, CDCl3): δ3.85-3.75(t, 2H), 3.00-2.95(t, 2H), 2.60-2.50(t, 4H), 2.18(s, 3H), 2.16(s, 3H ), 2.08(s, 3H), 1.83-1.75(t, 2H), 1.65(bs, 1H), 1.60-1.00(m, 33H), 0.90-0.80(m, 18H); MS: 586.55[M+H] + .

[0191] Example 9 Synthesis of compound VEC-013

[0192]

[0193] Compound VEC-013 was prepared by replacing octadecylamine with 2-ethylhexyloxypropylamine according to the preparation method of Example 7, with a yield of 87%.

[0194] 1 H-NMR (500MHz, CDCl3): δ3.80-3.75(t, 2H), δ3.52-3.48(t, 2H), δ3.30-3.25(t, 2H), 3.00-2.98(t, 2H), 2.80-2.75(t, 2H), 2.59- 2.55(t, 2H), 2.18(s, 3H), 2.16(s, 3H), 2.08(s, 3H), 1.83-1.70(m, 4H), 1.60-1.00(m, 34H), 0.90-0.80(m, 18H); MS: 644.53[M+H] + .

[0195] Example 10 Synthesis of compound VEC-014

[0196]

[0197] Compound VEC-014 was prepared by replacing octadecylamine with methoxyethylamine according to the preparation method of Example 7, with a yield of 95%.

[0198] 1 H-NMR (500MHz, CDCl3): δ3.85-3.83(t,2H), 3.58-3.55(t,2H), 3.48(3,3H), 3.02-3.00(t,2H), 2.92-2.88(t,2H), 2.59-2.55(t,2H), 2.20(s , 3H), 2.16 (s, 3H), 2.08 (s, 3H), 2.00 (bs, 1H), 1.85-1.75 (t, 2H), 1.60 -1.50(t, 2H), 1.50-1.00(t, 24H), 0.90-0.80(m, 12H); MS: 532.45[M+H] + .

[0199] Example 11 Synthesis of compound VEC-015

[0200]

[0201] Compound VEC-015 was prepared by replacing octadecylamine with benzyloxyethylamine according to the preparation method of Example 7, with a yield of 90%.

[0202] 1H-NMR (500MHz, CDCl3): δ7.35-7.20 (m, 5H), 4.56 (s, 2H), 3.85-3.83 (t, 2H), 3.68-3.60 (t, 2H), 3.02-3.00 (t, 2H), 2.92-2.88 (t, 2H), 2.58-2 .55(t,2H),2.20(s,3H),2.16(s,3H),2.08(s,3H),1.95(bs,1H),1.85 -1.75(t, 2H), 1.60-1.00(t, 26H), 0.90-0.80(m, 12H); MS: 608.50[M+H] + .

[0203] Example 12 Solubility Test

[0204] Take 10 ml of solvent and add 10 g, 1 g, 0.5 g and 0.1 g of the test compound respectively. Stir at room temperature (25-30℃) for half an hour and let stand for half an hour. Record the uniform solution as "+", the presence of undissolved droplets or layering as "-", and no test as " / ". The results are shown in Table 1.

[0205] The test results show that the compounds of the present invention have a solubility greater than 10 g / 10 ml in heptane, isopropyl acetate (i-PrOAc), and methyl tert-butyl ether (MTBE), a solubility less than 1 g / 10 ml in N,N-dimethylformamide, and even lower solubility in methanol and dimethyl sulfoxide (DMSO), mostly less than 0.5 g / 10 ml.

[0206] Solubility data show that the compounds of this invention have significant differences in solubility in different solvents, and are polar and nonpolar amphiphilic compounds with special properties, suitable for different types of reactions and separation and purification; binding to peptide chains can increase the solubility of polypeptide intermediates in low to medium polar solvents, which facilitates separation from polar byproducts and impurities, facilitates purification, and improves the purity of polypeptide products; at the same time, it enhances the affinity with polar reagents and improves reaction efficiency.

[0207] Table 1

[0208]

[0209]

[0210]

[0211] Example 13: Compound VEC-001 used for peptide synthesis

[0212] H-Ala-Ser-Ala-Lys-Trp-Thr-His-Asn-Gly-Gly-Glu-Met-Ser-OH

[0213]

[0214] 1) Dissolve compound VEC-001 (11.3 g, 0.02 mol) in methyl tert-butyl ether (100 ml), add 4-hydroxymethylphenoxyacetic acid (L-01, 3.8 g, 0.021 mol) and N-methylmorpholine (2.23 g, 0.022 mol) in DMF (40 ml) and DMT-MM (6.1 g, 0.022 mol) in water (20 ml), react at room temperature for half an hour, separate the aqueous layer, wash the upper layer with sodium bicarbonate solution to obtain HO-L01-VEC-001 methyl tert-butyl ether solution, which can be directly used for the next step of amino acid inoculation reaction.

[0215] 2) Add Fmoc-Ser(t-Bu)-OH (8.5g, 0.021mol) and DMAP (0.12g, 1mmol) to the methyl tert-butyl ether solution of HO-L01-VEC-001 obtained in the previous step, stir and cool to 0-10℃; add EDCI (5.7g, 0.03mol), maintain the reaction at 0-10℃ for 3 hours, add a DMF (40ml) solution of diethylenetriamine (12.4g, 0.12mol) and mercaptopropionic acid (8.51g, 0.08mol), and heat to 40-50℃ for reaction; after 2 hours, add water (20ml), separate the aqueous layer, wash the organic layer with water until neutral, and proceed directly to the next step of the reaction.

[0216] 3) Add a 40ml solution of DMF containing Fmoc-Met-OH (7.8g, 0.021mol) and N-methylmorpholine (2.23g, 0.022mol) and a 20ml solution of water containing DMT-MM (6.1g, 0.022mol) to the solution obtained in step 2) and react for half an hour. Separate the aqueous layer. Add a 40ml solution of DMF containing diethylenetriamine (12.4g, 0.12mol) and mercaptopropionic acid (8.51g, 0.08mol) to the methyl tert-butyl ether layer and heat to 40-50℃ to react. After 2 hours, add water (20ml), separate the aqueous layer, wash the organic layer with water until neutral, and proceed directly to the next step of the reaction.

[0217] 4) Following the method in step 3, sequentially add Fmoc-Glu(OtBu)-OH, Fmoc-Gly-OH, Fmoc-Gly-OH, Fmoc-Asn(Trt)-OH, Fmoc-His(Trt)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Ala-OH, Fmoc-Ser(tBu)-OH, and Boc-Ala-OH. After Boc-Ala-OH incorporation, the addition of diethylenetriamine and mercaptopropionic acid for the Fmoc removal reaction is not required. Methyl tert-butyl ether layer concentration yields Boc-Ala-Ser(tBu)-Ala-Lys(Boc)-Trp(Boc)-Thr(tBu)-His(Trt)-Asn(Trt)-Gly-Gly-Glu(OtBu)-Met-Ser(t-Bu)-O-L01

[0218] -VEC-001.

[0219] 5) Dissolve Boc-Ala-Ser(tBu)-Ala-Lys(Boc)-Trp(Boc)-Thr(tBu)-His(Trt)-Asn(Trt)-Gly-Gly-Glu(OtBu)-Met-Ser(t-Bu)-O-L01-VEC-001 in a mixture of TFA, TIS, and water at a volume ratio of 94:3:3 (110 ml). React at room temperature for 2 hours. Add methyl tert-butyl ether (660 ml), stir at room temperature for half an hour, filter, wash the filter cake with methyl tert-butyl ether until the eluent is neutral, and dry to obtain...

[0220] Crude H-Ala-Ser-Ala-Lys-Trp-Thr-His-Asn-Gly-Gly-Glu-Met-Ser-OH, 28.8 g, yield 90% (based on 2 TFA salts), HPLC purity >98%, MS: 1375.61 [M+H] + .

[0221] 6) Combine and concentrate all the filtrates from step 5. Add butanol (50 ml) and concentrated hydrochloric acid (50 ml) to the remaining product and heat under reflux for 3 hours. Concentrate to dryness, add heptane (100 ml) and water (100 ml), stir well, let stand to separate the aqueous layer, wash the organic layer with 1N sodium hydroxide aqueous solution, wash with water until the washing solution is neutral, concentrate, and purify by column chromatography to obtain compound VEC-00110.0 g, with a recovery rate of 88.5%.

[0222] Example 14: Compound VEC-002 used for the synthesis of peptide fragments

[0223] H2N-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH2

[0224]

[0225] 1) Dissolve compound VEC-002 (9.30, 0.01 mol) in methyl tert-butyl ether (150 ml), add L-02 (5.18 g, 0.0105 mol) and N-methylmorpholine (1.12, 0.011 mol) in DMF (40 ml) and DMT-MM (3.1 g, 0.011 mol) in water (20 ml) for reaction. After half an hour of reaction, separate the aqueous layer. Add diethylenetriamine (6.2 g, 0.06 mol) and mercaptopropionic acid (4.26 g, 0.04 mol) in DMF (40 ml) to the methyl tert-butyl ether layer and heat to 40-50℃ for reaction. After 2 hours, add water (20 ml) and separate the aqueous layer. Wash the organic layer (H2N-L02-VEC-002 solution) with water until neutral and use it directly in the next reaction.

[0226] 2) Following the method in step 1, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, and Fmoc-Gly-OH were sequentially added. The methyl tert-butyl ether layer was then concentrated to obtain...

[0227] H2N-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH-L02-VEC-002.

[0228] 3) Dissolve H2N-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH-L02-VEC-002 in dichloromethane (400 ml). Add 3% TFA-dichloromethane solution dropwise under a water bath at room temperature. After the addition is complete, react for 1 hour. Add methyl cyclopentyl ether (400 ml), and dichloromethane is evaporated under reduced pressure. Filter and collect the solid as H2N-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH2.TFA 9.7 g, yield 85% (based on single TFA salt), HPLC purity >98%, MS ESI: 1020.40 [M+H] +.

[0229] 4) Combine and concentrate all filtrates from step 3. Add butanol (50 ml) and concentrated hydrochloric acid (50 ml) to the remaining product, and heat under reflux for 3 hours. Concentrate to dryness, add heptane (100 ml) and water (100 ml), stir well, allow to stand, separate the aqueous layer, wash the organic layer with 1N sodium hydroxide aqueous solution, wash with water until the wash solution is neutral, concentrate, and purify by column chromatography to obtain compound VEC-002 7.5 g, with a recovery rate of 80.6%.

[0230] Example 15: Compound VEC-003 was used to synthesize peptide fragments.

[0231] H2N-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH2

[0232]

[0233] Following the method of Example 14, compound VEC-003 was used instead of compound VEC-002 in the reaction to prepare H2N-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH2, with a yield of 83%, HPLC purity >98%, and VEC-003 recovery rate of 91.0%.

[0234] Example 16: Compound VEC-004 used for the synthesis of peptide fragments

[0235] H2N-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH2

[0236]

[0237] Following the method of Example 14, compound VEC-004 was used instead of compound VEC-002 in the reaction to prepare H2N-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH2, with a yield of 88%, HPLC purity >98%, and VEC-004 recovery rate of 85.0%.

[0238] Example 17: Compound VEC-005 was used to synthesize the peptide fragment Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH

[0239]

[0240] 1) Dissolve compound VEC-005 (6.1 g, 0.01 mol) in dichloromethane (150 ml), add a DMF (40 ml) solution of L-03 (3.56 g, 0.0105 mol) and N-methylmorpholine (1.12 g, 0.011 mol) and an aqueous solution of DMT-MM (3.1 g, 0.011 mol) (20 ml) to react. After reacting for half an hour, separate the aqueous layer; wash the organic layer with 1 N potassium hydroxide aqueous solution, wash with water until neutral, and dry with anhydrous sodium sulfate to obtain a dichloromethane solution of HO-L03-VEC-005, which can be directly used for the next step of the reaction.

[0241] 2) Cool the dichloromethane solution HO-L03-VEC-005 obtained in the previous step to 5-10℃, add thionyl chloride (1.8g, 0.015mol) dropwise, reflux for 4 hours after the addition is complete, concentrate to remove dichloromethane and excess thionyl chloride, and obtain chloride Cl-L03-VEC-005, which can be used directly in the next step.

[0242] 3) Dissolve the chloride Cl-L-03-VEC-005 obtained in the previous step in methyl tert-butyl ether (150 ml), add Fmoc-Leu-OH (5.3 g, 0.015 mol) and diisopropylethylamine (2.6 g, 0.02 mol), heat under reflux for 8 hours, and then cool to room temperature; add a DMF (40 ml) solution of diethylenetriamine (6.2 g, 0.06 mol) and mercaptopropionic acid (4.26 g, 0.04 mol), and heat to 40-50 °C; after 2 hours, add water (20 ml), separate the aqueous layer; wash the organic layer with water until neutral, and use it directly in the next step of the reaction.

[0243] 4) Add a 40ml solution of Fmoc-Ile-Aib-OH (4.61g, 0.021mol) and N-methylmorpholine (2.23g, 0.022mol) in DMF and a 20ml solution of DMT-MM (6.1g, 0.022mol) in water to the methyl tert-butyl ether solution obtained in the previous step and react. After half an hour, separate the aqueous layer. Add a 40ml solution of diethylenetriamine (12.4g, 0.12mol) and mercaptopropionic acid (8.51g, 0.08mol) in DMF to the methyl tert-butyl ether layer and heat to 40-50℃. After 2 hours, add water (20ml), separate the aqueous layer, wash the organic layer with water until neutral, and proceed directly to the next step of the reaction.

[0244] 5) Following the method in step 4, sequentially add Fmoc-Ser(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-Gly-OH, Fmoc-Glu(OtBu)-OH, and Boc-Tyr(tBu)-Aib-OH. After incorporating Boc-Tyr(tBu)-Aib-OH, there is no need to add diethylenetriamine and mercaptopropionic acid for the Fmoc removal reaction. The methyl tert-butyl ether layer is concentrated to obtain Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OL-03-VEC-005.

[0245] 6) Dissolve Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-O-L03-VEC-005 in dichloromethane (200 ml), add 3% TFA-dichloromethane solution (100 ml) dropwise, react for 1 hour after addition, wash with water to remove TFA, concentrate, add heptane and slurry, filter to obtain Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH 18.0 g, yield 85%, HPLC purity >95%. MS ESI: 1058.60 [M+2H] 2+ / 2.

[0246] 7) Combine and concentrate all the filtrates from step 6. Add butanol (60 ml) and concentrated hydrochloric acid (60 ml) to the remaining product and heat under reflux for 3 hours. Concentrate to dryness, add heptane (150 ml) and water (100 ml), stir well, let stand to separate the aqueous layer, wash the organic layer with 1N sodium hydroxide aqueous solution, wash with water until the washing solution is neutral, concentrate, and purify by column chromatography to obtain compound VEC-0055.2 g, with a recovery rate of 85.5%.

[0247] Example 18: Compound VEC-006 was used to synthesize the peptide fragment Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH

[0248]

[0249] Following the method of Example 17, compound VEC-006 was used instead of compound VEC-005 to prepare Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH, with a yield of 80.5%, HPLC purity >95%, and VEC-006 recovery of 80.0%.

[0250] Example 19: Compound VEC-007 was used to synthesize the peptide fragment Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH

[0251]

[0252] Following the method of Example 17, compound VEC-007 was used instead of compound VEC-005 to prepare Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH, with a yield of 83.5%, HPLC purity >95%, and VEC-007 recovery of 81.0%.

[0253] Example 20: Compound VEC-008 was used to synthesize the peptide fragment Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH

[0254]

[0255] Following the method of Example 17, compound VEC-008 was used instead of compound VEC-005 to prepare Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH, with a yield of 86.5%, HPLC purity >95%, and VEC-008 recovery rate of 82.0%.

[0256] Example 21: Compound VEC-009 was used to synthesize Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2

[0257]

[0258] 1) Dissolve compound VEC-009 (9.5g, 0.02mol) in methyl tert-butyl ether (200ml), add L-04 (11.36g, 0.021mol) and N-methylmorpholine (2.25g, 0.022mol) in DMF (80ml) and DMT-MM (6.2g, 0.022mol) in water (40ml) for reaction. After half an hour of reaction, separate the aqueous layer. Add diethylenetriamine (12.4g, 0.12mol) and mercaptopropionic acid (8.5g, 0.08mol) in DMF (80ml) to the methyl tert-butyl ether layer and heat to 40-50℃ for reaction. After 2 hours, add water (40ml) and separate the aqueous layer. Wash the organic layer (H2N-L04-VEC-009 solution) with water until neutral and use it directly in the next step of reaction.

[0259] 2) Following the method in step 1), sequentially add Fmoc-Arg(Pbf)-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Met-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Met-OH, and Fmoc-Ala-OH to obtain a methyl tert-butyl ether solution of H2N-Ala-Met-Tyr(tBu)-Glu(OtBu)-Glu(OtBu)-Met-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-NH-L04-VEC-009.

[0260] (3) The H2N-Ala-Met-Tyr(tBu)-Glu(OtBu)-Glu(OtBu)-Met-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-NH-L04-VEC-009 obtained in the previous step was dissolved in a mixture (200 ml) of TFA, TIS and water in a volume ratio of 94:3:3. The reaction was carried out at room temperature for 2 hours. Methyl tert-butyl ether (1000 ml) was added, and the mixture was stirred at room temperature for half an hour. The mixture was filtered, and the filter cake was washed with methyl tert-butyl ether until the eluent was neutral. After drying, 20.1 g of Ala-Met-Tyr-Glu-Glu-Met-Gln-Arg-Arg-NH2 was obtained, with a yield of 90% (based on 2TFA salts). The HPLC purity was >98%, and the MS ESI was 1211.53 [M+H]. + .

[0261] 4) Combine and concentrate all the filtrates from step 3). Add butanol (60 ml) and concentrated hydrochloric acid (30 ml) to the remainder and heat under reflux for 3 hours. Concentrate to dryness, add heptane (100 ml) and water (100 ml), stir well, allow to stand to separate the aqueous layer, wash the organic layer with 1N sodium hydroxide aqueous solution, wash with water until the washing solution is neutral, concentrate, and purify by column chromatography to obtain compound VEC-009.

[0262] 8.5g, recovery rate 89.5%.

[0263] Example 22: Compound VEC-009 was used to synthesize Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2

[0264] 1) Dissolve VEC-009 (9.5g, 0.02mol) in methyl tert-butyl ether (200ml), add DMF (40ml) solutions of L-04 (11.36g, 0.021mol), HOBt (2.7g, 0.02mol), and diisopropylethylamine (7.74g, 0.06mol) and TBTU (9.6g, 0.03mol) (40ml) to react; after reacting for 2 hours, add DMF (80ml) solutions of diethylenetriamine (12.4g, 0.12mol) and mercaptopropionic acid (8.5g, 0.08mol) and heat to 40-50℃ to react; after 2 hours, add water (160ml) and separate the aqueous layer; wash the organic layer (H2N-L04-VEC-009 solution) with water until neutral and use it directly in the next step of the reaction.

[0265] 2) Following the method in step 1), sequentially add Fmoc-Arg(Pbf)-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Met-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Met-OH, and Fmoc-Ala-OH to obtain a methyl tert-butyl ether solution of H2N-Ala-Met-Tyr(tBu)-Glu(OtBu)-Glu(OtBu)-Met-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-NH-L04-VEC-009.

[0266] 3) Following step 3) of Example 21, 19.5 g of Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2.2TFA was obtained, with a yield of 87% (based on 2TFA salt), HPLC purity >98%, MS ESI: 1211.53 [M+H]. + .

[0267] Example 23: Compound VEC-010 was used to synthesize Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2

[0268]

[0269] Following the steps of Example 21, compound VEC-010 was used instead of compound VEC-009 to prepare Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2 with a yield of 89% (based on 2TFA salt) and a purity of >98%. The recovery rate of compound VEC-010 was 92%.

[0270] Example 24: Compound VEC-011 was used to synthesize Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2

[0271]

[0272] Following the steps of Example 21, compound VEC-009 was replaced with compound VEC-011 and L-05 was replaced with L-04 to prepare Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2 with a yield of 85.5% (based on 2TFA salt) and a purity >98%. The recovery rate of compound VEC-011 was 87%.

[0273] Example 25: Compound VEC-012 was used to synthesize Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2

[0274]

[0275] Following the steps of Example 21, compound VEC-009 was replaced with compound VEC-012 and L-06 was replaced with L-04 to prepare Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2 with a yield of 84.0% (based on 2TFA salt) and a purity >95%. The recovery rate of compound VEC-012 was 83%.

[0276] Example 26: Compound VEC-013 was used to synthesize Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH

[0277]

[0278] Following the steps of Example 17, compound VEC-005 was replaced by compound VEC-013, and L-07 was replaced by L-03 to prepare Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH, with a yield of 80.0% and a purity >95%. The recovery rate of compound VEC-013 was 81%.

[0279] Example 27: Compound VEC-014 used for polypeptide synthesis

[0280] H-Ala-Ser-Ala-Lys-Trp-Thr-His-Asn-Gly-Gly-Glu-Met-Ser-OH

[0281]

[0282] Following the steps of Example 13, compound VEC-001 was replaced by compound VEC-014, and L-08 was replaced by L-01 to prepare H-Ala-Ser-Ala-Lys-Trp-Thr-His-Asn-Gly-Gly-Glu-Met-Ser-OH, with a yield of 90.0% and a purity >98%. The recovery rate of compound VEC-014 was 85%.

[0283] Example 28: Compound VEC-015 was used to synthesize Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH-iPr

[0284]

[0285] 1) Following the method of Example 21, compound VEC-009 was replaced by compound VEC-015, and L-09 was replaced by L-04 in the reaction. Fmoc-Arg(Pbf)-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Met-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Met-OH, and Boc-Ala-OH were added to prepare Boc-Ala-Met-Tyr(tBu)-Glu(OtBu)-Glu(OtBu)-Met-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-NH-L09-VEC-015.

[0286] 2) Boc-Ala-Met-Tyr(tBu)-Glu(OtBu)-Glu(OtBu)-Met-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-NH-L09-VEC-015 (14.7 g, 5 mmol) was dissolved in dichloromethane (100 ml), and copper acetate (10 mg, 0.5 mmol) and isopropylamine (0.89 g, 15 mmol) were added. The reaction was carried out at room temperature for 2-3 hours, and TLC analysis showed that the reaction was complete. The copper acetate was removed by diatomaceous earth filtration, the mixture was concentrated, and methyl tert-butyl ether was added to form a slurry. After filtration, 8.8 g of solid Boc-Ala-Met-Tyr(tBu)-Glu(OtBu)-Glu(OtBu)-Met-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-NH-iPr was obtained, with a yield of 87%.

[0287] 3) The obtained Boc-Ala-Met-Tyr(tBu)-Glu(OtBu)-Glu(OtBu)-Met-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-NH-iPr was dissolved in a mixture of TFA, TIS and water in a volume ratio of 94:3:3 (88 ml). The reaction was carried out at room temperature for 3 hours. Methyl tert-butyl ether (440 ml) was added, and the mixture was stirred at room temperature for half an hour. The mixture was filtered, and the filter cake was washed with methyl tert-butyl ether until the eluent was neutral. After drying, 5.8 g of Ala-Met-Tyr-Glu-Glu-Met-Gln-Arg-Arg-NH-iPr was obtained, with a yield of 90% (calculated based on 2 TFA salts). The HPLC purity was >98%, and the MS ESI was 1254.58 [M+H]. +

[0288] 4) Combine and concentrate all the filtrates from step 3). Add butanol (60 ml) and concentrated hydrochloric acid (30 ml) to the remaining product and heat under reflux for 3 hours. Concentrate to dryness, add heptane (100 ml) and water (100 ml), stir well, let stand to separate the aqueous layer, wash the organic layer with 1N sodium hydroxide aqueous solution, wash with water until the washing solution is neutral, concentrate, and purify by column chromatography to obtain compound VEC-015 with a recovery rate of 90%.

[0289] Comparative Example 1: Compound CongenT-003 was used to synthesize the peptide fragment Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH

[0290]

[0291] The steps are the same as in Example 17, except that CongenT-003 is used instead of HO-L03-VEC-005. When amino acids are attached to Fmoc-Asp(OtBu)-OH, an upper gel phenomenon occurs, making it impossible to continue the attachment of subsequent amino acids, and the synthesis fails.

[0292] Comparative Example 2: Compound BM-008 was used for peptide synthesis.

[0293] H-Ala-Ser-Ala-Lys-Trp-Thr-His-Asn-Gly-Gly-Glu-Met-Ser-OH

[0294]

[0295] The steps are the same as in Example 13, except that BM-008 is used instead of HO-L01-VEC001. When amino acids are attached to Fmoc-His(Trt)-OH, an upper gel phenomenon occurs, making it impossible to continue the attachment of subsequent amino acids, and the synthesis fails.

[0296] Comparative Example 3: Compound FL-027 was used to synthesize the protective peptide Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH

[0297]

[0298] The steps are the same as in Example 17, except that FL-027 is used instead of HO-L03-VEC-005. When amino acids are attached to Fmoc-Ser(tBu)-OH, an upper gel phenomenon occurs, making it impossible to continue attaching subsequent amino acids, and the synthesis fails.

[0299] Comparative Example 4: Compound DPA-025 was used to synthesize the peptide Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2

[0300]

[0301] The steps are the same as in Example 22, except that DPA-025 is used instead of H2N-L04-VEC-009. When amino acids are attached to Fmoc-Tyr(tBu)-OH, an upper gel phenomenon occurs, making it impossible to continue attaching subsequent amino acids, and the synthesis fails.

[0302] Example 29 Stability Test

[0303] Experiment 1: Linker-linked carrier compounds HO-L01-VEC-001, H2N-L02-VEC-002, H2N-L02-VEC-003, H2N-L-02-VEC-004, HO-L03-VEC-005, HO-L03-VEC-006, HO-L03-VEC-007, HO-L03-VEC-008, H2N-L04-VEC-009, H2N-L04-VEC-010, H 2N-L05-VEC-011, H2N-L06-VEC-012, HO-L07-VEC-013, HO-L08-VEC-014, H2N-L09-VEC-015, and the comparative compounds CongenT-003, BM-008, FL-027, and DPA025 were dissolved in a mixed solvent of DMF:MTBE:water (1:1:0.05) at a concentration of 0.1 mmol / ml, and then 6 equivalents of diethylenetriamine were added and the mixture was kept at 50°C.

[0304] Experiment 2: Same as Experiment 1, except that 6 equivalents of diethylenetriamine and 4 equivalents of mercaptopropionic acid were added, and the temperature was maintained at 50°C.

[0305] The contents of the above compounds were tested at 0 min, 30 min, 60 min, 120 min and 180 min in two experiments, with the initial contents being 100% (0 min).

[0306] The test results are shown in Table 2: The compounds of the present invention have good stability under alkaline conditions and under the conditions of peptide chain synthesis and deprotection, and the carrier does not detach during peptide synthesis.

[0307] Table 2. Stability of the carrier compounds of the present invention in DMF and MTBE solvent systems.

[0308]

[0309]

[0310]

[0311] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A vitamin E derivative having the structure shown in formula (1) or a stereoisomer thereof, in, R is selected from: one or more R5-substituted or unsubstituted C1-C 22 alkyl, R1 is selected from: one or more R5-substituted or unsubstituted C1-C 22 Alkyl group, one or more R6-substituted or unsubstituted C6-C 10 aryl; R2, R3, and R4 are each independently selected from: hydrogen and C1-C6 alkyl groups; Each R5 is independently selected from: hydrogen, C6-C 10 aryl; Each R6 is independently selected from: hydrogen, C1-C 22 Alkyl groups, or two adjacent R6 groups linked together to form one or more R7-substituted or unsubstituted 5-6 membered heterocyclic groups; Each R7 is independently selected from: hydrogen, C1-C 22 alkyl; x is selected from: 1, 2, 3, 4, 5; n is selected from: 1, 2, 3; y is selected from: 1, 2, 3, 4, 5; m is selected from: 1, 2, 3.

2. The vitamin E derivative or its stereoisomer according to claim 1, characterized in that, R is selected from: benzyl, C1-C 20 alkyl, 3. The vitamin E derivative or its stereoisomer according to claim 1, characterized in that, R1 is selected from: benzyl, C1-C 22 Alkyl group, one or more R6-substituted or unsubstituted C6-C 10 aryl; Each R6 is independently selected from: hydrogen, C1-C 18 Alkyl groups, or two adjacent R6 groups linked together to form one or more R7-substituted or unsubstituted 5-6 membered heterocyclic groups; Each R7 is independently selected from: hydrogen, C1-C 18 alkyl.

4. The vitamin E derivative or its stereoisomer according to claim 3, characterized in that, R1 is selected from: benzyl, C1-C 12 alkyl, 5. The vitamin E derivative or its stereoisomer according to any one of claims 1-4, characterized in that, R2, R3, and R4 are each independently selected from: hydrogen, methyl, ethyl, n-propyl, and isopropyl.

6. The vitamin E derivative or its stereoisomer according to claim 5, characterized in that, R2, R3 and R4 are all methyl groups.

7. The vitamin E derivative or its stereoisomer according to any one of claims 1-4, characterized in that, x is selected from: 1, 2; n is selected from: 1, 2.

8. The vitamin E derivative or its stereoisomer according to claim 7, characterized in that, x is 1.

9. The vitamin E derivative or its stereoisomer according to any one of claims 1-4, characterized in that, y is selected from: 1, 2; m is selected from: 1, 2.

10. The vitamin E derivative or its stereoisomer according to claim 9, characterized in that, y is 1.

11. The vitamin E derivative or its stereoisomer according to claim 1, characterized in that, R is selected from: benzyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecanyl, octadecyl, nonadecanyl, eicosyl. R1 is selected from: benzyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl. y is selected from: 1, 2, m is selected from: 1, 2.

12. The vitamin E derivative or its stereoisomer according to claim 1, characterized in that, Selected from the following compounds:

13. The application of vitamin E derivatives having the structure shown in formula (1) or their stereoisomers in the preparation of carriers for the synthesis of polypeptides, in, R is selected from: hydrogen, one or more R5-substituted or unsubstituted C1-C. 22 alkyl, R1 is selected from: one or more R5-substituted or unsubstituted C1-C 22 Alkyl group, one or more R6-substituted or unsubstituted C6-C 10 aryl; R2, R3, and R4 are each independently selected from: hydrogen and C1-C6 alkyl groups; Each R5 is independently selected from: hydrogen, C6-C 10 aryl; Each R6 is independently selected from: hydrogen, C1-C 22 Alkyl groups, or two adjacent R6 groups linked together to form one or more R7-substituted or unsubstituted 5-6 membered heterocyclic groups; Each R7 is independently selected from: hydrogen, C1-C 22 alkyl; x is selected from: 1, 2, 3, 4, 5; n is selected from: 1, 2, 3; y is selected from: 1, 2, 3, 4, 5; m is selected from: 1, 2, 3.

14. The application according to claim 13, characterized in that, R is selected from: hydrogen, benzyl, C1-C 20 alkyl, 15. The application according to claim 13, characterized in that, R1 is selected from: benzyl, C1-C 22 Alkyl group, one or more R6-substituted or unsubstituted C6-C 10 aryl; Each R6 is independently selected from: hydrogen, C1-C 18 Alkyl groups, or two adjacent R6 groups linked together to form one or more R7-substituted or unsubstituted 5-6 membered heterocyclic groups; Each R7 is independently selected from: hydrogen, C1-C 18 alkyl.

16. The application according to claim 13, characterized in that, R1 is selected from: benzyl, C1-C 12 alkyl, 17. The application according to any one of claims 13-16, characterized in that, R2, R3, and R4 are each independently selected from: hydrogen, methyl, ethyl, n-propyl, and isopropyl.

18. The application according to claim 17, characterized in that, R2, R3 and R4 are all methyl groups.

19. The application according to any one of claims 13-16, characterized in that, x is selected from: 1, 2; n is selected from: 1, 2.

20. The application according to claim 19, characterized in that, x is 1.

21. The application according to any one of claims 13-16, characterized in that, y is selected from: 1, 2; m is selected from: 1, 2.

22. The application according to claim 21, characterized in that, y is 1.

23. The application according to claim 13, characterized in that, R is selected from: hydrogen, benzyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecanyl, octadecyl, nonadecanyl, eicosyl. R1 is selected from: benzyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl. y is selected from: 1, 2, m is selected from: 1, 2.

24. The application according to claim 13, characterized in that, The vitamin E derivative is selected from the following compounds:

25. A carrier compound having the structure shown in formula (2) or a stereoisomer thereof for the synthesis of polypeptides, in, R, R2, R3, R4, x, and n are as described in any one of claims 13-24; L is a linking group for linking with an amino acid, and L is linked to the carrier compound via an amide bond.

26. The support compound or its stereoisomer according to claim 25, characterized in that, L is selected from:

27. The support compound or its stereoisomer according to claim 26, characterized in that, Selected from the following compounds:

28. The support compound or its stereoisomer according to claim 25, characterized in that, The vitamin E derivative or its stereoisomer as described in any one of claims 13-24 is obtained by reacting it with a linker for linking to an amino acid, wherein the linker for linking to the amino acid is selected from the following compounds:

29. The use of the vitamin E derivative or its stereoisomer as described in any one of claims 13-24, or the carrier compound or its stereoisomer as described in any one of claims 25-28, in the synthesis of polypeptides.

30. A method for synthesizing polypeptides, characterized in that, The synthetic method uses the compound or its stereoisomer as a carrier according to any one of claims 25-28 to carry out the synthetic reaction.

31. The method for synthesizing polypeptides according to claim 30, characterized in that, Includes the following steps: (1) Reacting an N-terminal protected amino acid or peptide with a carrier compound or its stereoisomer as described in any one of claims 25-28 to obtain a carrier-amino acid / peptide conjugate; (2) Remove the N-terminal protecting group from the carrier-amino acid / peptide conjugate and then react it with the next N-terminal protected amino acid or peptide. (3) Repeat step (2) to elongate the peptide chain and obtain a conjugate of the N-terminally protected target polypeptide and the carrier compound. (4) Remove the carrier and / or protecting group from the conjugate obtained in step (3) to obtain the target polypeptide.

32. The method for synthesizing polypeptides according to claim 31, characterized in that, Step (4) includes: Remove the carrier from the conjugate obtained in step (3) to obtain the N-terminally protected target polypeptide; or, Simultaneously, the N-terminal protecting group and the carrier in the conjugate obtained in step (3) are removed to obtain the target polypeptide; or, First, remove the carrier from the conjugate obtained in step (3), and then remove the N-terminal protecting group to obtain the target polypeptide.

33. The method for synthesizing polypeptides according to claim 31, characterized in that, The N-terminal protected amino acid or peptide described in steps (1) and (2) contains a side-chain protecting group; step (4) includes: Remove the carrier from the conjugate obtained in step (3) to obtain the target polypeptide with an N-terminal protected side chain; or, Simultaneously, the N-terminal protecting group and the carrier in the conjugate obtained in step (3) are removed to obtain the target polypeptide with a side-chain protecting group; or, Simultaneously, the N-terminal protecting group, side chain protecting group, and carrier are removed from the conjugate obtained in step (3) to obtain the target polypeptide; or, First, remove the carrier from the conjugate obtained in step (3), then remove the N-terminal protecting group and / or the side chain protecting group to obtain the target polypeptide.

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