Vitamin E derivative as well as carrier compound and application thereof

Abstract

The invention relates to a vitamin E derivative as well as a carrier compound and application thereof. The vitamin E derivative has a structure as shown in a formula (1) and can be connected and combined with different linkers to obtain a carrier for synthesizing polypeptide, the preparation process is simple, and the obtained carrier is good in solubility, stability and recoverability and can be used for efficiently synthesizing high-purity polypeptide with high yield. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the technical field of polypeptide synthesis, and particularly relates to a vitamin E derivative for synthesizing polypeptides, its carrier compound and its application. Background Art

[0002] The methods for chemical synthesis production of polypeptides mainly include solid-phase method and ordinary liquid-phase method at present.

[0003] Among them, the solid-phase method has the advantages of strong universality and short development cycle, and is suitable for the synthesis of all peptides; its disadvantages are slow reaction speed, large material consumption, few means for monitoring the reaction process and low accuracy, inability to purify intermediates, and low purity of crude peptides; the final product is usually purified by high-performance preparative liquid chromatography, resulting in high production costs.

[0004] The ordinary liquid-phase method has a fast reaction speed and less 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 purity of the crude peptide is relatively high; however, it has poor universality, long development cycle, and complex steps.

[0005] The liquid-phase carrier method is a new polypeptide synthesis production method developed to improve the above two methods. A soluble compound or polymer is used to replace the traditional resin, converting the solid-liquid two-phase reaction into a liquid-phase reaction, so that the reaction speed can be accelerated, the material usage can be reduced, and it is basically the same as the ordinary liquid-phase method, and the analysis methods of the ordinary liquid-phase method can be used, and the intermediate can also be purified if necessary. This technology is represented by the Molecular Hiving TM technology of JITSUBO company, the Ajiphase technology of Ajinomoto, and the EMPHASES technology of Tongjuan Medicine.

[0006] Molecular Hiving TM The technology uses long-chain alkoxybenzyl alcohols, such as: 3,5-bis(didodecyloxy)benzyl alcohol, 2,4-bis(didodecyloxy)benzyl alcohol, 3,4,5-tris(octadecyloxy)benzyl alcohol, etc., as the liquid-phase carrier. The carrier is used as a C-terminal protecting reagent, the reaction is carried out in a homogeneous phase, and the intermediate products and by-products are separated through the steps of precipitation - filtration - washing. This method has the following problems: ① The reaction concentration is low, which is not conducive to large-scale production; ② There are many processes in precipitation - filtration - washing, and the operation is complex; ③ The intermediate is a non-crystalline solid, and the filtration - washing takes a long time and the purification effect is poor.

[0007] The Ajiphase technology uses multi-branched alkyl groups instead of straight-chain alkyl groups, which increases the reaction concentration. The intermediate is purified by extraction and washing. However, the reaction time is long, the solvent ratio is high during washing, the impurity removal effect is average, the product loss is large, and the solubility of the intermediate drops rapidly during the amino acid attachment process. Gelation occurs when the number of amino acids is greater than 4.

[0008] The EMPHASES technology uses amphiphilic carriers, which further increases the reaction concentration. In homogeneous or heterogeneous solvent systems, especially in heterogeneous solvent systems, it can increase the reaction rate and reagent utilization rate, simplify the post-treatment operation, improve the product purity, and enhance the generality and universality of the operation process. However, the carrier preparation process is extremely complex. The carriers for carboxyl and amide at the C-terminus are different, and the preparation processes are also different, making it impossible to recycle and use them well. Some carrier structures contain ester bonds, and carrier degradation occurs during the production process. The problem of decreasing solubility during the amino acid attachment process has not been completely solved. When the number of amino acids is greater than 8, gelation occurs in some peptide chain intermediates, resulting in a decrease in conversion rate and yield, and it is not suitable for the synthesis of polypeptides with more than 8 amino acids. No carriers for other groups at the C-terminus are provided.

[0009] Therefore, it is necessary to develop a recyclable carrier with simple preparation, good solubility, and good stability for polypeptide synthesis, and further obtain a more universal polypeptide synthesis technology. Summary of the Invention

[0010] Based on this, the object of the present invention is to provide a recyclable carrier with simple preparation, good solubility, and good stability for polypeptide synthesis.

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

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

[0013]

[0014] wherein, R is selected from: hydrogen, a C1-C alkyl group substituted or unsubstituted by one or more R5, 22 alkyl group,

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

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

[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 alkoxy, halogen, or two adjacent R6s are linked together to form a C3-C8 cycloalkyl or 3-8 membered heterocyclic group, which may or may not be substituted by one or more R7s;

[0019] Each R7 is independently selected from: hydrogen, C1-C 22 alkyl, C1-C 22 alkoxy, halogen;

[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] In a second aspect, the present invention provides the use of the vitamin E derivative or its stereoisomer as described above in the preparation of a carrier for synthesizing polypeptides.

[0025] In a third aspect, the present invention provides a carrier compound having the structure shown in formula (2) or its stereoisomer 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 for linking with an amino acid.

[0029] In a fourth aspect, the present invention provides the use of the vitamin E derivative or its stereoisomer as described in the present invention, or the carrier compound or its stereoisomer as described in the present invention in the synthesis of polypeptides.

[0030] In a fifth aspect, the present invention provides a method for synthesizing polypeptides, wherein the synthesis reaction is carried out using the compound as described in the present invention as a carrier.

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

[0032] (1) Reacting an N-terminal protected amino acid or peptide with the carrier compound or its stereoisomer as described in the present 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 extend the peptide chain to obtain a conjugate of the N-terminal 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] The present invention uses inexpensive and readily available vitamin E (including various natural vitamin Es, whether chiral or achiral, or synthetic vitamin Es) as raw materials to prepare a series of new vitamin E derivatives through simple chemical reactions. The raw materials for preparing this type of vitamin E derivative are inexpensive and the preparation process is simple. It can, like solid-phase resin, be connected and combined with different linkers to obtain a carrier for polypeptide synthesis, and is suitable for the production of all types of polypeptides. Using it for polypeptide synthesis has the following advantages:

[0037] (1) This carrier compound has the lipophilic structure of vitamin E and also has the weakly polar ethylene glycol ether structure. It has high solubility in non-polar solvents and also has certain solubility in polar solvents. It has amphiphilic properties and has a fast reaction rate in both homogeneous and heterogeneous solvent systems, and can be used to synthesize high-purity polypeptides efficiently and with high yields.

[0038] (2) In the carrier compound of the present invention, the bond connected to the linker is an amide bond, which has good stability during the process of synthesizing peptides, and can avoid the detachment of the carrier during the synthesis process; and it is easy to recover. After the synthesis reaction is completed, this carrier compound can be recovered with high efficiency and high yield through a simple chemical treatment process for the next round of polypeptide synthesis, thereby greatly saving the cost of polypeptide synthesis.

[0039] (3) When using the carrier compound of the present invention for polypeptide synthesis, it can still maintain good solubility after incorporating multiple amino acids, solving the problem of decreased solubility of existing carriers during the process of incorporating amino acids. It is very beneficial for the synthesis of long peptides, and there will be no phenomenon of peptide chain gelation during the synthesis of long peptides, and it can effectively improve the synthesis yield and purity of long peptides.

[0040] (4) By combining and applying the carrier compound of the present invention with different linkers, it can not only provide carriers for preparing peptides with COOH and amide at the C-terminus, but also provide carriers for preparing peptides with other groups at the C-terminus, thereby enabling the preparation of modified polypeptides with different groups at the C-terminus. The versatility and universality of the carrier compound provided by the present invention are superior to the first-generation EMPHASES carrier, and it is suitable for the large-scale production of polypeptides, especially long peptides. Detailed implementation mode

[0041] To facilitate the understanding of the present invention, the present invention will be described more comprehensively below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, these embodiments are provided to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0042] For the experimental methods without specific conditions noted in the following examples, they are generally in accordance with conventional conditions or the conditions recommended by the manufacturers. All common chemical reagents used in the examples are commercially available products.

[0043] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the technical field to which the present invention belongs. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used in the present invention includes any and all combinations of one or more of the related listed items.

[0044] In addition, as used in the present invention, the term "or" is an inclusive "or" symbol and is equivalent to the term "and / or", unless the context clearly dictates otherwise. The term "based on" is not exclusive and allows for other factors not described, unless the context clearly dictates otherwise. In addition, throughout the specification, the meanings of "a", "an", and "the" include plural referents. The meaning of "in..." includes "in..." and "on...".

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

[0046]

[0047] wherein, R is selected from: hydrogen, a C1-C alkyl substituted or unsubstituted with one or more R5, 22 alkyl,

[0048] R1 is selected from: a C1-C alkyl substituted or unsubstituted with one or more R5, a C6-C 22 alkyl, an aryl substituted or unsubstituted with one or more R6; 22 aryl;

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

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

[0051] Each R6 is independently selected from: hydrogen, a C1-C22 alkyl, C1-C 22 alkoxy, halogen, or two adjacent R6 groups are linked to form a C3-C8 cycloalkyl or 3-8 membered heterocyclic group, which is substituted or unsubstituted by one or more R7;

[0052] Each R7 is independently selected from: hydrogen, C1-C 22 alkyl, C1-C 22 alkoxy, halogen;

[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 the present invention, when any variable (such as R5, R6, etc.) appears more than once in any component, its definition at each occurrence is independent of its definition at each other occurrence. Similarly, combinations of substituents and variables are permitted, provided that such combinations render the compounds stable. It is to be understood that one of ordinary skill in the art may select the substituents and substitution patterns of the compounds of the present invention to provide compounds that are chemically stable and readily synthesized from readily available starting materials by the techniques of the art and the methods set forth hereinafter. If a substituent itself is substituted by more than one group, it is to be understood that these groups may be on the same carbon atom or on different carbon atoms, provided that the structure is stable.

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

[0059] As used herein, the term "cycloalkyl" means a saturated or partially unsaturated monocyclic, bicyclic, or polycyclic hydrocarbon group in which the ring atoms are carbon atoms, and bicyclic or polycyclic includes spiro, fused, and bridged rings. For example: "cycloalkyl" includes, but is not limited to, the following groups: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0060] As used herein, the term "alkoxy" refers to a group having the structure -O-alkyl, such as -OCH3, -OCH2CH3, -OCH2CH2CH3, -O-CH2CH(CH3)2, -OCH2CH2CH2CH3, -O-CH(CH3)2, etc.

[0061] As used herein, the term "heterocycloalkyl" or "heterocyclic group" refers to a saturated or partially unsaturated monocyclic, bicyclic or polycyclic ring substituent (including spiro, bridged, fused, annulated, etc.), wherein one or more ring atoms are heteroatoms selected from N, O or S(O)m (where m is an integer from 0 to 2), and the remaining ring atoms are carbon. For example: morpholinyl, piperidinyl, pyrrolidinyl, pyrrolidinyl, dihydroimidazolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl, tetrahydrofuranyl, tetrahydrothienyl, etc., and their N-oxides. The connection of the heterocyclic substituent can be achieved through a carbon atom or through a heteroatom.

[0062] As will be appreciated by those skilled in the art, "halo" or "halogen" as used herein means 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, a C6-C 10 aryl which is substituted or unsubstituted by one or more R6;

[0065] Each R6 is independently selected from: hydrogen, C1-C 18 alkyl, C1-C 18 alkoxy, halogen, or two adjacent R6 are linked to form a C5-C6 cycloalkyl or 5-6 membered heterocyclic group which is substituted or unsubstituted by one or more R7;

[0066] Each R7 is independently selected from: hydrogen, C1-C 18 alkyl, C1-C 18 alkoxy, halogen.

[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 independently selected from: hydrogen, methyl, ethyl, n-propyl, isopropyl.

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

[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, 2, and most preferably 1; m is selected from: 1, 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, heptadecyl, octadecyl, nonadecyl, 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, and 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 the present invention for polypeptide synthesis is cheap, simple to prepare, and easy to recycle. The vitamin E derivative described in the present invention can, like the solid-phase resin, be connected and combined with different linkers to obtain a carrier for polypeptide synthesis, which is suitable for the production of all types of polypeptides; the connection with the linker is an amide bond, and it has good stability during the synthesis of peptides; the carrier compound has the lipophilic structure of vitamin E and also has a weakly polar ethylene glycol ether structure, has high solubility in non-polar solvents, and also has a certain solubility in polar solvents, has amphiphilic properties, and has a fast reaction rate in both homogeneous and heterogeneous solvent systems. Its versatility and universality are superior to those of the first-generation EMPHASES carrier, and it is suitable for the large-scale production of polypeptides.

[0079] In an embodiment of the present invention, a carrier compound having the structure shown in formula (2) or its stereoisomer for polypeptide synthesis is further provided,

[0080]

[0081] wherein, R is selected from: hydrogen, a C1-C alkyl substituted or unsubstituted by one or more R5, 22 alkyl,

[0082] R1 is selected from: one or more C1-C alkyl groups which are substituted or unsubstituted by R5, one or more C6-C 22 aryl groups which are substituted or unsubstituted by R6; 22

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

[0084] Each R5 is independently selected from: hydrogen, C6-C 10 aryl groups, halogens;

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

[0086] Each R7 is independently selected from: hydrogen, C1-C 22 alkyl groups, 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 for linking with an amino acid.

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

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

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

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

[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, 2, and most preferably 1; m is selected from: 1, 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, heptadecyl, octadecyl, nonadecyl, 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, and m is selected from: 1, 2.

[0104] In the carrier compound of the present invention, L is a linking group obtained by reacting a conventional linker capable of linking with an amino acid with the amino group in the vitamin E derivative described in the present invention, and is used to link with an amino acid during the polypeptide synthesis process. After the synthesis reaction is completed, it can be removed by a conventional deprotection reagent in the art, thereby obtaining the final target peptide chain. Specifically, L can be selected from, but not limited to, the following groups:

[0105]

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

[0107]

[0108]

[0109] In some of these embodiments, the carrier compound is obtained by reacting the vitamin E derivative or its stereoisomer according to the present invention with a linker for linking with an amino acid, and the linker for linking with an amino acid is selected from the following compounds:

[0110]

[0111] Wherein, when the linker for linking with an amino acid contains an amino protecting group (such as Fmoc protecting group), after the vitamin E derivative according to the present invention reacts with the linker, the amino protecting group is removed by deprotection to obtain a carrier compound for synthesizing a polypeptide, and then it is used for the reaction of linking an amino acid.

[0112] In one embodiment of the present invention, there is provided the use of the vitamin E derivative or its stereoisomer according to the present invention, or the carrier compound or its stereoisomer in the synthesis of a polypeptide.

[0113] In one embodiment of the present invention, there is provided a method for synthesizing a polypeptide, and the synthesis method uses the compound according to the present invention as a carrier to carry out a synthesis reaction.

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

[0115] (1) React an N-terminal protected amino acid or peptide with the carrier compound or its stereoisomer according to the present 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 extend the peptide chain to obtain a conjugate of the N-terminal 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 synthesizing a polypeptide with the carrier compound of the present invention, the carrier can be removed under the condition that the N-terminal protecting group and / or the amino acid side chain protecting group are safe, or can be removed together with the N-terminal protecting group and / or the amino acid side chain protecting group under certain conditions. Therefore, with the method of the present invention, a polypeptide with an N-terminal protecting group or a side chain protecting group can be synthesized, a polypeptide with both an N-terminal protecting group and a side chain protecting group can also be obtained, and a completely deprotected polypeptide can also be obtained.

[0120] In some of these 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] Remove the N-terminal protecting group and the carrier from the conjugate obtained in step (3) simultaneously 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-terminally protected amino acid or peptide described in steps (1) and (2) contains side-chain protecting groups; step (4) includes:

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

[0126] Remove the N-terminal protecting group and the carrier from the conjugate obtained in step (3) simultaneously to obtain the target polypeptide with side-chain protecting groups; or,

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

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

[0129] Using the carrier compound described in the present invention for polypeptide synthesis, it has good solubility, solves the problem of decreased solubility of the existing carrier during amino acid coupling, is very beneficial to the synthesis of long peptides, does not show the phenomenon of peptide chain gelation during the synthesis of long peptides, and can effectively improve the synthesis yield and purity of long peptides. Moreover, this carrier compound has good stability and 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 polypeptide synthesis, thereby greatly saving the cost of polypeptide synthesis and being conducive to large-scale production.

[0130] The compounds corresponding to the abbreviations and acronyms involved in the present invention are described as follows:

[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-dimethoxytriazinyl)-4-methylmorpholine hydrochloride; DMAP: 4-dimethylaminopyridine;

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

[0139] TIS: triisopropylsilane;

[0140] TBTU: benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate;

[0141] HOBt: 1-hydroxybenzotriazole.

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

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

[0144]

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

[0146] Step 2: Under water bath conditions, dropwise add SOCl2 (14.3 g, 0.12 mol) to the toluene solution of intermediate 1-01 obtained in the previous step. After dropping, heat to 60 - 70 °C and react for 2 hours. Concentrate to remove the excess SOCl2, redissolve with toluene, and concentrate again to obtain 49.3 g of crude product of chloride 1-02 with a yield of 100%, which is directly used in the next 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%) (calculated 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 at room temperature for 3 hours in a high-pressure autoclave under a H2 pressure of 1.5 MPa; filter to remove the catalyst, and concentrate the filtrate to obtain 25.1 g of compound VEC-002, with a yield of 90%.

[0149] VEC-001: 1 1H-NMR (500 MHz, CDCl3): δ 7.40 - 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 1H-NMR (500 MHz, CDCl3): δ 3.85 - 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 (43.1 g, 0.1 mol) in N,N-dimethylformamide (500 ml), add 1,3-dibromopropane (40.4, 0.2 mol), tetrabutylammonium bromide (0.32 g, 0.001 mol) and potassium carbonate (27.6 g, 0.2 mol), and heat to 80 - 90 °C; React for 48 hours, recover N,N-dimethylformamide under reduced pressure and remove the excess 1,3-dibromopropane. Dissolve the residue in heptane, wash twice with water and concentrate to obtain the crude product 2-01, which is directly used for the next reaction;

[0154] Step 2: Dissolve the crude product 2-01 obtained in the previous step in N-methylpyrrolidone (200 ml), add benzylamine (5.38 g, 0.05 mol), potassium carbonate (27.6 g, 0.2 mol) and potassium iodide (16.6 g, 0.1 mol), heat to 120 °C and react for 12 hours. Filter to remove the solid, add heptane (200 ml) and water (100 ml) to the filtrate; Separate the aqueous layer, wash the heptane layer twice with water, concentrate, and purify by column chromatography to obtain compound 1-03 (26.2 g, yield 50%) and VEC-003 (14.5, yield 25%) (calculated 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), in an autoclave, react at room temperature for 3 hours under a H2 pressure of 1.5 MPa; Filter to remove the catalyst, concentrate the filtrate to obtain 24.0 g of compound VEC-004, with a yield of 90%.

[0156] VEC-003: 1 1H-NMR (500 MHz, CDCl3): δ 7.38 - 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, 2H), 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: 11H-NMR (500 MHz, CDCl3): δ 3.88 - 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] Synthesis of Compounds VEC - 005 and VEC - 006 in Example 3

[0159]

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

[0161] Step 2: Dissolve the intermediate 3 - 01 obtained in Step 1 in dichloromethane, dropwise add thionyl chloride (14.3 g, 0.12 mol) under a water bath. After dropping, heat to 60 - 70 °C and react for 2 hours, concentrate to remove the excess SOCl2, redissolve with toluene, and concentrate again to obtain the crude product of intermediate 3 - 02, which is directly used for the next 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 the solid, 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, yield 70%) and VEC - 005 (9.1, yield 15%) (calculated 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 at room temperature for 3 hours in an autoclave under a H2 pressure of 1.5 MPa; filter to remove the catalyst, and concentrate the filtrate to obtain 27.1 g of compound VEC-006, with a yield of 95%.

[0164] VEC-005: 1 1H-NMR (500 MHz, CDCl3): δ 7.40 - 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 1H-NMR (500 MHz, CDCl3): δ 3.85 - 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] Synthesis of Compound VEC-007 in Example 4

[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 °C and react for 8 hours. Add heptane (300 ml) and water (200 ml); separate the aqueous layer, wash with heptane twice, concentrate, and purify by column chromatography to obtain 22.5 g of compound VEC-001 with a yield of 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 with heptane twice, concentrate, and purify by column chromatography to obtain 31.0 g of intermediate 4-01 with a yield of 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), in an autoclave, react at room temperature for 3 hours under a H2 pressure of 1.5 MPa; filter off the catalyst, concentrate the filtrate to obtain 26.1 g of compound VEC-007 with a yield of 92%.

[0171] 1 1H-NMR (500 MHz, CDCl3): δ 3.86 - 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] Synthesis of Compound VEC-008 in Example 5

[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. Then add heptane (600 ml) and water (200 ml); separate the aqueous layer, wash the heptane layer twice with water, concentrate, and purify by column chromatography to obtain 47.9 g of intermediate 5-01 with a yield of 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), in an autoclave, react at room temperature for 3 hours under a H2 pressure of 1.5 MPa; filter to remove the catalyst, and concentrate the filtrate to obtain 39.5 g of compound VEC-008 with a yield of 90%.

[0176] 1 1H-NMR (500 MHz, CDCl3): δ 3.87 - 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] Dissolve compound VEC-001 (25.0 g, 0.044 mol) in ethyl acetate (500 ml), add 10% Pd / C (2.5 g), in an autoclave, react at room temperature for 3 hours under a H2 pressure of 1.5 MPa; filter to remove the catalyst, and concentrate the filtrate to obtain 19.0 g of compound VEC-009 with a yield of 91%.

[0180] 1H-NMR (500 MHz, CDCl3): δ 3.85 - 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] Dissolve compound VEC-001 (25.0 g, 0.044 mol) in tetrahydrofuran (500 ml) and methanol (100 ml), add paraformaldehyde (2 g) and sodium cyanoborohydride (5.5 g, 0.088 mol), react at room temperature for 3 hours, concentrate to remove the solvent, add ethyl acetate (200 ml) and water (200 ml), separate the aqueous layer, and the ethyl acetate layer is washed twice with water and directly used for the hydrogenation reaction. Add 10% Pd / C (2.5 g) to the obtained ethyl acetate solution, in an autoclave, react at room temperature for 3 hours under a H2 pressure of 1.5 MPa; filter off the catalyst, and concentrate the filtrate to obtain 18.0 g of compound VEC-010 with a yield of 85%.

[0182] 1 H-NMR (500 MHz, CDCl3): δ 3.85 - 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] Dissolve chloride 1-02 (24.6 g, 0.05 mol) in N,N-dimethylformamide (300 ml), add octadecylamine (26.9 g, 0.1 mol), potassium carbonate (8.3 g, 0.06 mol) and potassium iodide (3.3 g, 0.02 mol), heat to 80 - 90 °C and react for 8 hours, then add heptane (300 ml) and water (200 ml); separate the aqueous layer, wash with heptane twice, concentrate, and purify by column chromatography to obtain 25.4 g of compound VEC-01 with a yield of 70%.

[0186] 1 H-NMR (500 MHz, CDCl3): δ 3.85 - 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), 2.16 (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] Synthesis of compound VEC-012 in Example 8

[0188]

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

[0190] 1 H-NMR (500 MHz, CDCl3): δ 3.85 - 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] Synthesis of compound VEC-013 in Example 9

[0192]

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

[0194] 1 H-NMR (500 MHz, CDCl3): δ 3.80 - 3.75 (t, 2H), δ 3.52 - 3.48 (t, 2H), δ 3.30 - 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] Synthesis of Compound VEC-014 in Example 10

[0196]

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

[0198] 1 H-NMR (500 MHz, CDCl3): δ 3.85 - 3.83 (t, 2H), 3.58 - 3.55 (t, 2H), 3.48 (s, 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] Synthesis of Compound VEC-015 in Example 11

[0200]

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

[0202] 11H-NMR (500 MHz, CDCl3): δ 7.35 - 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] 10 mL of solvent was added with 10 g, 1 g, 0.5 g, and 0.1 g of the test compound respectively, stirred at room temperature (25 - 30 °C) for half an hour and then allowed to stand for half an hour. A homogeneous solution was denoted as "+", the presence of undissolved droplets or layering was denoted as "-", and no test was denoted as " / ". The results are shown in Table 1.

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

[0206] The solubility data indicate that the compounds of the present invention have significant solubility differences in different solvents, are polar and non-polar amphiphilic compounds with special properties, and are suitable for different types of reactions and separation and purification; binding with peptide chains can increase the solubility of polypeptide intermediates in medium and low polarity solvents, facilitate separation from polar by-products and impurities, and is convenient for purification, improving the purity of polypeptide products; at the same time, it enhances the affinity with polar reagents and improves the reaction efficiency.

[0207] Table 1

[0208]

[0209]

[0210]

[0211] Example 13 Use of Compound VEC-001 in 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 a DMF (40 ml) solution of 4-hydroxymethylphenoxyacetic acid (L-01, 3.8 g, 0.021 mol) and N-methylmorpholine (2.23 g, 0.022 mol), and an aqueous (20 ml) solution of DMT-MM (6.1 g, 0.022 mol). After reacting at room temperature for half an hour, separate the aqueous layer. Wash the upper layer with sodium bicarbonate solution to obtain a methyl tert-butyl ether solution of HO-L01-VEC-001, which is directly used for the next amino acid coupling reaction.

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

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

[0217] 4) Connect 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 in sequence according to the method in step 3. After the addition of Boc-Ala-OH, there is no need to add diethylenetriamine and mercaptopropionic acid for the Fmoc deprotection reaction. The methyl tert-butyl ether layer is concentrated to obtain 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 mixed solution (110 ml) composed of TFA, TIS, and water with a volume ratio of 94:3:3. 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 washings are neutral, and dry to obtain

[0220] 28.8 g of crude product of H-Ala-Ser-Ala-Lys-Trp-Thr-His-Asn-Gly-Gly-Glu-Met-Ser-OH, with a yield of 90% (calculated based on the 2TFA salt), HPLC purity > 98%, MS: 1375.61 [M+H] + .

[0221] 6) Combine and concentrate all the filtrates in step 5. Add butanol (50 ml) and concentrated hydrochloric acid (50 ml) to the residue, and reflux and react for 3 hours; concentrate to dryness, add heptane (100 ml) and water (100 ml), stir evenly, let stand to separate the aqueous layer, wash the organic layer with 1N aqueous sodium hydroxide solution, wash with water until the washings are neutral, concentrate, and purify by column chromatography to obtain 10.0 g of compound VEC-001 with a recovery rate of 88.5%.

[0222] Example 14 Compound VEC-002 is 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 a solution of L-02 (5.18 g, 0.0105 mol) and N-methylmorpholine (1.12, 0.011 mol) in DMF (40 ml) and a solution of DMT-MM (3.1 g, 0.011 mol) in water (20 ml) to react. After reacting for half an hour, separate the aqueous layer; add a solution of 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 °C 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 for the next step of the reaction.

[0226] 2) Sequentially introduce Fmoc-Ser(tBu)-OH, Fmoc-Pro-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 according to the method of step 1, and concentrate the methyl tert-butyl ether layer 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), dropwise add a 3% TFA-dichloromethane solution under normal temperature water bath. After dropping, react for 1 hour, add methyl cyclopentyl ether (400 ml), and distill off dichloromethane under reduced pressure. Filter to collect the solid as 9.7 g of H2N-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-NH2.TFA, with a yield of 85% (calculated based on the single TFA salt), HPLC purity > 98%, MS ESI: 1020.40 [M + H] +。

[0229] 4) Combine and concentrate all the filtrates from step 3. Add butanol (50 ml) and concentrated hydrochloric acid (50 ml) to the residue, and reflux the reaction for 3 hours; concentrate to dryness, add heptane (100 ml) and water (100 ml), stir evenly, let stand to separate the aqueous layer, wash the organic layer with 1N aqueous sodium hydroxide solution, wash with water until the washing liquid is neutral, concentrate, and purify by column chromatography to obtain 2.75 g of compound VEC-002, with a recovery rate of 80.6%.

[0230] Example 15 Compound VEC-003 is used for the synthesis of peptide fragments

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

[0232]

[0233] According to the method of Example 14, use compound VEC-003 instead of compound VEC-002 for 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 the recovery rate of VEC-003 is 91.0%.

[0234] Example 16 Compound VEC-004 is 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] According to the method of Example 14, use compound VEC-004 instead of compound VEC-002 for 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 the recovery rate of VEC-004 is 85.0%.

[0238] Example 17 Compound VEC-005 is used for the synthesis of 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 solution of L-03 (3.56 g, 0.0105 mol) and N-methylmorpholine (1.12, 0.011 mol) in DMF (40 ml) and a solution of DMT-MM (3.1 g, 0.011 mol) in water (20 ml) to carry out the reaction. After reacting for half an hour, separate the aqueous layer; wash the organic layer with 1N aqueous potassium hydroxide solution, wash with water until neutral, and dry with anhydrous sodium sulfate to obtain a dichloromethane solution of HO-L03-VEC-005, which is directly used for the next step of the reaction.

[0241] 2) Cool the dichloromethane solution of HO-L03-VEC-005 obtained in the previous step to 5 - 10 °C, dropwise add thionyl chloride (1.8 g, 0.015 mol). After adding, reflux and react for 4 hours, concentrate to remove dichloromethane and excess thionyl chloride to obtain the chloride Cl-L03-VEC-005, which is directly used for 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, 0.02 mol), heat under reflux and react for 8 hours, then cool to room temperature; add a solution of diethylenetriamine (6.2 g, 0.06 mol) and mercaptopropionic acid (4.26 g, 0.04 mol) in DMF (40 ml) and raise the temperature to 40 - 50 °C for reaction; after 2 hours, add water (20 ml), separate the aqueous layer; wash the organic layer with water until neutral, and directly use it for the next step of the reaction.

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

[0244] 5) Connect 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 in sequence according to the method in step 4. After connecting Boc-Tyr(tBu)-Aib-OH, there is no need to add diethylenetriamine and mercaptopropionic acid for the Fmoc deprotection reaction. Concentrate the methyl tert-butyl ether layer 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-O-L-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 dropwise 3% TFA-dichloromethane solution (100 ml). After the addition is complete, react for 1 hour, wash with water to remove TFA, concentrate, add heptane for slurrying, and filter to obtain 18.0 g of 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 85% and 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 residue, and heat under reflux for 3 hours; concentrate to dryness, add heptane (150 ml) and water (100 ml), stir evenly, let stand to separate the aqueous layer, wash the organic layer with 1N sodium hydroxide aqueous solution, wash with water until the washing liquid is neutral, concentrate, and purify by column chromatography to obtain 5.2 g of compound VEC-005, with a recovery rate of 85.5%.

[0247] Example 18 Compound VEC-006 is used for synthesizing 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] According to the method of Example 17, using Compound VEC-006 to replace Compound VEC-005 for the reaction, Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH was prepared with a yield of 80.5%, HPLC purity > 95%, and the recovery rate of VEC-006 was 80.0%.

[0250] Example 19 Compound VEC-007 is used for synthesizing 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] According to the method of Example 17, using Compound VEC-007 to replace Compound VEC-005 for the reaction, Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH was prepared with a yield of 83.5%, HPLC purity > 95%, and the recovery rate of VEC-007 was 81.0%.

[0253] Example 20 Compound VEC-008 is used for synthesizing 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] According to the method of Example 17, the reaction was carried out with compound VEC-008 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 for the synthesis of Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2

[0257]

[0258] 1) Dissolve compound VEC-009 (9.50, 0.02 mol) in methyl tert-butyl ether (200 ml), add a DMF (80 ml) solution of L-04 (11.36 g, 0.021 mol) and N-methylmorpholine (2.25, 0.022 mol) and a water (40 ml) solution of DMT-MM (6.2 g, 0.022 mol) to carry out the reaction. After reacting for half an hour, the aqueous layer was separated; the methyl tert-butyl ether layer was added with a DMF (80 ml) solution of diethylenetriamine (12.4 g, 0.12 mol) and mercaptopropionic acid (8.5 g, 0.08 mol), and the temperature was raised to 40 - 50 °C for reaction; after 2 hours, water (40 ml) was added, and the aqueous layer was separated; the organic layer (H2N-L04-VEC-009 solution) was washed with water until neutral and directly used for the next step of the reaction.

[0259] 2) According to the method of step 1), 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 were successively introduced 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) Dissolve 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 in a mixed solution (200 ml) composed of TFA, TIS, and water with a volume ratio of 94:3:3, react at room temperature for 2 hours, add methyl tert-butyl ether (1000 ml), stir at room temperature for half an hour, filter, wash the filter cake with methyl tert-butyl ether until the washing liquid is neutral, and dry to obtain 20.1 g of Ala-Met-Tyr-Glu-Glu-Met-Gln-Arg-Arg-NH2, with a yield of 90% (calculated based on the 2TFA salt), HPLC purity > 98%, MS ESI: 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 residue, and heat under reflux for 3 hours; concentrate to dryness, add heptane (100 ml) and water (100 ml), stir evenly, let stand to separate the aqueous layer, wash the organic layer with 1N sodium hydroxide aqueous solution, wash with water until the washing liquid is neutral, concentrate, and purify by column chromatography to obtain 8.5 g of compound VEC-009, with a recovery rate of 89.5%.

[0262] 8.5 g, recovery rate 89.5%.

[0263] Example 22 Use of compound VEC-009 for the synthesis of Ala-Met-Tyr-Glu-Glu-Met-Gln-Arg-Arg-NH2

[0264] 1) Dissolve VEC-009 (9.50, 0.02 mol) in methyl tert-butyl ether (200 ml), add a DMF (40 ml) solution of L-04 (11.36 g, 0.021 mol), HOBt (2.7 g, 0.02 mol), and diisopropylethylamine (7.74 g, 0.06 mol) and a DMF (40 ml) solution of TBTU (9.6 g, 0.03 mol) to carry out the reaction; after reacting for 2 hours, add a DMF (80 ml) solution of diethylenetriamine (12.4 g, 0.12 mol) and mercaptopropionic acid (8.5 g, 0.08 mol), and raise the temperature to 40 - 50 °C for reaction; after 2 hours, add water (160 ml), and separate the aqueous layer; wash the organic layer (H2N-L04-VEC-009 solution) with water until it is neutral, and directly use it for the next step of the reaction.

[0265] 2) Sequentially introduce 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, Fmoc-Ala-OH in the method of step 1) 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) Obtain 19.5 g of Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2·2TFA with a yield of 87% (calculated as the 2TFA salt), HPLC purity > 98%, and MS ESI: 1211.53 [M+H] according to step 3) of Example 21. + 。

[0267] Compound VEC-010 of Example 23 is used for the synthesis of Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2

[0268]

[0269] According to the steps of Example 21, use compound VEC-010 instead of compound VEC-009 for the reaction to prepare Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2 with a yield of 89% (calculated as the 2TFA salt), purity > 98%, and the recovery rate of compound VEC-010 is 92%.

[0270] Compound VEC-011 of Example 24 is used for the synthesis of Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2

[0271]

[0272] According to the steps of Example 21, use compound VEC-011 instead of compound VEC-009 and L-05 instead of L-04 for the reaction to prepare Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2 with a yield of 85.5% (calculated as the 2TFA salt), purity > 98%, and the recovery rate of compound VEC-011 is 87%.

[0273] Example 25 Compound VEC-012 is used for the synthesis of Ala-Met-Tyr-Glu-Glu-Met-Gln-Arg-Arg-NH2

[0274]

[0275] According to the steps of Example 21, using compound VEC-012 instead of compound VEC-009 and L-06 instead of L-04 for the reaction, Ala-Met-Tyr-Glu-Glu-Met-Gln-Arg-Arg-NH2 is prepared with a yield of 84.0% (calculated as the 2TFA salt), a purity of >95%, and a recovery rate of compound VEC-012 of 83%.

[0276] Example 26 Compound VEC-013 is used for the synthesis of 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] According to the steps of Example 17, using compound VEC-013 instead of compound VEC-005 and L-07 instead of L-03 for the reaction, Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Ser(tBu)-Ile-Aib-Leu-OH is prepared with a yield of 80.0%, a purity of >95%, and a recovery rate of compound VEC-013 of 81%.

[0279] Example 27 Compound VEC-014 is used for the synthesis of polypeptide

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

[0281]

[0282] According to the steps of Example 13, using compound VEC-014 instead of compound VEC-001 and L-08 instead of L-01 for the reaction, H-Ala-Ser-Ala-Lys-Trp-Thr-His-Asn-Gly-Gly-Glu-Met-Ser-OH is prepared with a yield of 90.0%, a purity of >98%, and a recovery rate of compound VEC-014 of 85%.

[0283] Example 28 Compound VEC-015 is used for synthesizing Ala-Met-Tyr-Glu-Glu-Met-Gln-Arg-Arg-NH-iPr

[0284]

[0285] 1) According to the method of Example 21, using compound VEC-015 instead of compound VEC-009 and L-09 instead of L-04 for 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 are introduced 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) is dissolved in dichloromethane (100 ml), copper acetate (10 mg, 0.5 mmol) and isopropylamine (0.89 g, 15 mmol) are added, and the reaction is carried out at room temperature for 2 - 3 hours. TLC analysis shows that the reaction is complete; the copper acetate is removed by filtration through diatomaceous earth, concentrated, slurried with methyl tert-butyl ether, and filtered to obtain 8.8 g of solid Boc-Ala-Met-Tyr(tBu)-Glu(OtBu)-Glu(OtBu)-Met-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-NH-iPr, with a yield of 87%.

[0287] 3) Dissolve the Boc-Ala-Met-Tyr(tBu)-Glu(OtBu)-Glu(OtBu)-Met-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-NH-iPr obtained above in a mixed solution (88 ml) composed of TFA, TIS, and water with a volume ratio of 94:3:3, react at room temperature for 3 hours, add methyl tert-butyl ether (440 ml), stir at room temperature for half an hour, filter, wash the filter cake with methyl tert-butyl ether until the washing liquid is neutral, and dry to obtain 5.8 g of Ala-Met-Tyr-Glu-Glu-Met-Gln-Arg-Arg-NH-iPr, with a yield of 90% (calculated based on the 2TFA salt), HPLC purity > 98%, MS ESI: 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 residue, heat under reflux for 3 hours; concentrate to dryness, add heptane (100 ml) and water (100 ml), stir evenly, let stand to separate the aqueous layer, wash the organic layer with 1N aqueous sodium hydroxide solution, wash with water until the washing liquid is neutral, concentrate, and purify by column chromatography to obtain compound VEC-015, with a recovery rate of 90%.

[0289] Compound CongenT-003 in Comparative Example 1 was used for the synthesis of 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 procedure was the same as in Example 17, replacing HO-L03-VEC-005 with CongenT-003. When the amino acid was attached to Fmoc-Asp(OtBu)-OH, a gel phenomenon occurred in the upper layer, and the subsequent connection of amino acids could not continue, resulting in the failure of the synthesis.

[0292] Compound BM-008 in Comparative Example 2 was used for the synthesis of the peptide

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

[0294]

[0295] The procedure was the same as in Example 13. When BM-008 was used to replace HO-L01-VEC001, a phenomenon of upper gel occurred when the amino acid was attached to Fmoc-His(Trt)-OH, and the subsequent connection of amino acids could not be continued, resulting in the failure of synthesis.

[0296] Compound FL-027 in Comparative Example 3 was used for the synthesis of the protected 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 procedure was the same as in Example 17. When FL-027 was used to replace HO-L03-VEC-005, a phenomenon of upper gel occurred when the amino acid was attached to Fmoc-Ser(tBu)-OH, and the subsequent connection of amino acids could not be continued, resulting in the failure of synthesis.

[0299] Compound DPA-025 in Comparative Example 4 was used for the synthesis of the peptide Ala-Met-Tyr-GLu-Glu-Met-Gln-Arg-Arg-NH2

[0300]

[0301] The procedure was the same as in Example 22. When DPA-025 was used to replace H2N-L04-VEC-009, a phenomenon of upper gel occurred when the amino acid was attached to Fmoc-Tyr(tBu)-OH, and the subsequent connection of amino acids could not be continued, resulting in the failure of synthesis.

[0302] Example 29 Stability Test

[0303] Experiment 1: The vector 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, H2N-L05-VEC-011, H2N-L06-VEC-012, HO-L07-VEC-013, HO-L08-VEC-014, H2N-L09-VEC-015 linked with linker and the comparative compounds CongenT-003, BM-008, FL-027 and DPA025 were respectively 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 temperature was kept at 50 °C.

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

[0305] Samples of the above compounds were taken and tested at 0 min, 30 min, 60 min, 120 min, and 180 min in the two experiments respectively, and the initial content was 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 the conditions of peptide chain synthesis and deprotection, and the carrier will not fall off during the polypeptide synthesis process.

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

[0308]

[0309]

[0310]

[0311] The above embodiments only represent several implementation manners of the present invention, and the description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several deformations and improvements can be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the present invention patent shall be subject to the appended claims.

Claims

1. A vitamin E derivative having a structure represented by formula (1) or a stereoisomer thereof, 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, one or more R6 substituted or unsubstituted C6-C 22 Aryl; R2, R3, and R4 are independently selected from: hydrogen, C1-C6 alkyl; Each R5 is independently selected from: hydrogen, C6-C 10 Aryl, halogen; Each R6 is independently selected from: hydrogen, C1-C 22 Alkyl, C1-C 22 Alkoxy, halogen, or two adjacent R6 are linked to form one or more R7 substituted or unsubstituted C3-C8 cycloalkyl or 3-8 membered heterocyclic group; Each R7 is independently selected from: hydrogen, C1-C 22 Alkyl, C1-C 22 Alkoxy, halogen; 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: hydrogen, 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, one or more R6 substituted or unsubstituted C6-C 10 Aryl; Each R6 is independently selected from: hydrogen, C1-C 18 Alkyl, C1-C 18 Alkoxy, halogen, or two adjacent R6 are linked to form one or more R7 substituted or unsubstituted C5-C6 cycloalkyl or 5-6 membered heterocyclic group; Each R7 is independently selected from: hydrogen, C1-C 18 Alkyl, C1-C 18 Alkoxy, halogen.

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 stereoisomer thereof according to any one of claims 1 to 4, characterized in that: R2, R3, and R4 are independently selected from the group consisting of 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 stereoisomer thereof according to any one of claims 1 to 4, characterized in that: x is selected from: 1, 2, most preferably 1; n is selected from: 1, 2.

8. The vitamin E derivative or stereoisomer thereof according to any one of claims 1 to 4, characterized in that: y is selected from: 1, 2, most preferably 1; m is selected from: 1, 2.

9. The vitamin E derivative or its stereoisomer according to claim 1, characterized in that: R is selected from the group consisting of hydrogen, benzyl, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, R1 is selected from the group consisting of 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.

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

11. Use of the vitamin E derivative or its stereoisomer according to any one of claims 1 to 10 in the preparation of a carrier for synthesizing polypeptides.

12. A carrier compound having a structure represented by formula (2) or a stereoisomer thereof for synthesizing a polypeptide, in, R, R2, R3, R4, x and n are as described in any one of claims 1 to 10; L is a linking group for connecting to an amino acid.

13. The carrier compound or its stereoisomer according to claim 12, characterized in that: L is selected from:

14. The carrier compound or its stereoisomer according to claim 12, characterized in that: Selected from the following compounds:

15. The carrier compound or its stereoisomer according to claim 12, characterized in that: Obtained by reacting the vitamin E derivative or its stereoisomer according to any one of claims 1 to 10 with a linker for connecting with an amino acid, wherein the linker for connecting with an amino acid is selected from the following compounds:

16. Use of the vitamin E derivative or stereoisomer thereof according to any one of claims 1 to 10, or the carrier compound or stereoisomer thereof according to any one of claims 12 to 15 in synthesizing polypeptides.

17. A method for synthesizing a polypeptide, characterized in that: The synthesis method uses the compound or stereoisomer thereof according to any one of claims 12 to 15 as a carrier to carry out the synthesis reaction.

18. The method for synthesizing a polypeptide according to claim 17, characterized in that: The steps include: (1) reacting an N-terminally protected amino acid or peptide with the carrier compound or a stereoisomer thereof according to any one of claims 12 to 15 to obtain a carrier-amino acid / peptide conjugate; (2) removing the N-terminal protecting group in the carrier-amino acid / peptide conjugate and then reacting with the next N-terminally protected amino acid or peptide; (3) repeating step (2) to extend the peptide chain to obtain a conjugate of the N-terminally protected target polypeptide and the carrier compound; (4) Removing the carrier and / or protecting group from the conjugate obtained in step (3) to obtain the target polypeptide.

19. The method for synthesizing a polypeptide according to claim 18, characterized in that: Step (4) comprises: Removing the carrier from the conjugate obtained in step (3) to obtain the N-terminally protected target polypeptide; or, Simultaneously remove the N-terminal protecting group and the carrier in the conjugate obtained in step (3) to obtain the target polypeptide; or, First, the carrier in the conjugate obtained in step (3) is removed, and then the N-terminal protecting group is removed to obtain the target polypeptide.

20. The method for synthesizing a polypeptide according to claim 18, characterized in that: The N-terminally protected amino acid or peptide in steps (1) and (2) contains a side chain protecting group; step (4) comprises: Removing the carrier from the conjugate obtained in step (3) to obtain the target polypeptide with N-terminal protection and side chain protecting groups; or, Simultaneously removing the N-terminal protecting group and the carrier in the conjugate obtained in step (3) to obtain the target polypeptide with side chain protecting groups; or, Simultaneously remove the N-terminal protecting group, side chain protecting group and carrier in the conjugate obtained in step (3) to obtain the target polypeptide; or, First, the carrier in the conjugate obtained in step (3) is removed, and then the N-terminal protecting group and / or the side chain protecting group are removed to obtain the target polypeptide.

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