CN113121594A

CN113121594A - Phosphoryloxybenzaldehyde oximes or phosphoryloxy benzophenone oximes compound and preparation method thereof

Info

Publication number: CN113121594A
Application number: CN202110399279.5A
Authority: CN
Inventors: 钦传光; 李海迪; 晁洁; 田广
Original assignee: Northwestern Polytechnical University
Current assignee: Northwestern Polytechnical University
Priority date: 2019-01-09
Filing date: 2019-01-09
Publication date: 2021-07-16
Also published as: CN109503655B; CN109503655A

Abstract

The invention relates to a phosphoryloxybenzaldehyde oxime or phosphoryloxybenzophenone oxime compound and a preparation method thereof. The phosphoryl oxybenzaldehyde or ketone is obtained by using (RO) ₂ POCl or POCl ₃ and hydroxybenzaldehyde or ketone as raw materials Oxybenzaldehyde or ketone compound 1; using compound 1 as raw material, react with hydroxylamine hydrochloride to obtain phosphoryloxybenzyl alcohol compound 2 corresponding to compound 1; using compound 1 as raw material, hydrogenation reduction treatment to obtain derivatized Product phosphoryloxybenzaldehyde or ketoxime compound 3; condensation coupling with compound 2 and Fmoc-protected amino acid (Fmoc-AA-OH) to obtain compound 4; with compound 3 and Boc-protected amino acid (Boc-AA- OH) condensation coupling to give compound 5. This method combines the advantages of liquid phase and solid phase synthesis methods, and can synthesize and prepare protected amino acids or polypeptides and their derivatives more simply, quickly, economically, and efficiently, and the phosphate ester carrier can be recovered and directly reused, reducing raw materials. Waste, reduce waste pollution, save costs, and be beneficial to environmental protection.

Description

Phosphoryloxybenzaldehyde oximes or phosphoryloxy benzophenone oximes compound and preparation method thereof

The application is a divisional application, the application number of the original application is 201910017593.5, the application date is 09.01.2019, and the invention name is 'phosphoryloxybenzaldehyde (or ketone) and a derivative thereof and a preparation method thereof'.

Technical Field

The invention belongs to the field of organic synthesis and polypeptide chemistry, and relates to phosphoryloxy benzaldehyde (or ketone) and a derivative thereof and a preparation method thereof, in particular to compounds with novel structures, such as phosphoryloxy benzyl alcohol serving as a reduction product of phosphoryloxy benzaldehyde (or ketone), oxime derivatives of phosphoryloxy benzaldehyde (or ketone) and the like, and a synthesis preparation method, a separation and a purification method thereof.

Background

Amino and carboxyl groups are important functional groups widely present in various bioactive compounds, and their protection and deprotection are often essential key links in modern synthetic organic chemistry, see literature:

[1]I.W.Hamley.Chem.Rev.2017,117,14015-14041.

[2]Kocienski,P.J.Protecting Groups；Georg Thieme Verlag:Stuttgart New York,2004.

[3]Green,T.W.；Wuts,P.G.M.Protective Groups in Organic Synthesis；John Wiley and Sons:New York,1999.

in particular, protection and deprotection of the amino and carboxyl functions of α -amino acids is one of the most important issues in the chemical synthesis of polypeptides. In peptide synthesis, once an amino acid is activated, it must be prevented from polymerization or other side reactions. On the other hand, peptide synthesis, whether in solution or on a solid phase, is preferentially carried out from the C-terminal to the N-terminal direction, and requires repeated removal of the temporary protecting group on the α -amino group or carboxyl group several times during the synthesis, and therefore, the deprotection reaction must be carried out under mild conditions without affecting the side chain residue protecting group or even the peptide chain, see document [4 ]: Isidro-Llobet, a.; alvarez, m.; albericio, F.chem.Rev.2009,109,2455-2504.

Although hundreds of protecting groups have been generated and removed by various methods, there remains a need to explore and continue to develop new and gentle strategies to introduce and cleave many existing protecting groups.

Polypeptide and peptidomimetic drugs involve almost all disciplines of chemistry and biomedicine, see literature:

[5]J.Clardy and C.Walsh,Nature,2004,432,829-837.

[6]H.H.Szeto and P.W.Schiller,Pharm.Res.,2011,28,2669–2679.

[7]C.J.White and A.K.Yudin,Nat.Chem.,2011,3,509–524.

[8]N.Assem and A.K.Yudin,Nat.Protoc.,2012,7,1327–1334.

[9]V.J.Hruby,G.Li,C.HaskellLuevano and M.Shenderovich,Biopolymers,1997,43,219–266.

the search for environmentally friendly synthetic methods of these products for academic research and drug production has been ongoing for over 50 years, see literature:

[10]J.L.Gustafson,D.Lim and S.J.Miller,Science,2010,328,1251–1255.

[11]V.R.Pattabiraman and J.W.Bode,Nature,2011,480,471–479.

[12]E.Ko,J.Liu,L.M.Perez,G.Lu,A.Schaefer and K.Burgess.J.Am.Chem.Soc.,2011,133,462-477.

[13]W.-K.Chan,C.-M.Ho,M.-K.Wong and C.-M.Che,J.Am.Chem.Soc.,2006,128,14796-14797.

[14]R.Hirschmann,A.B.Smith III,C.M.Taylor,et al.Science,1994,265,234-237.

[15]A.B.Smith III,A.B.Benowitz,P.A.Sprengeler,et al.J.Am.Chem.Soc.,1999,121,9286-9298.

the Solid Phase Peptide Synthesis (SPPS) invented by bruise Merrifield (Bruce Merrifield) of the nobel prize winner in the 80 s has shown a remarkable achievement of peptide synthesis, see literature: [16] r.b. merrifield.j.am.chem.soc.,1963,85, 2149-.

The milestone discovery can overcome the defects of complicated purification steps, consumption of a large amount of raw materials and reagents (including coupling agents, solvents and silica gel) and the like in the traditional liquid phase peptide synthesis, and can reduce the generation of waste; the method also greatly accelerates the synthesis and research of peptides and proteins. SPPS also exerts a tremendous impact on general chemical synthesis for drug discovery and development, particularly for combinatorial chemistry and high throughput screening, see literature: [17] sharma and d.crich, j.org.chem.,2011, 76, 6518-.

Since then, several synthetic schemes have been developed to complement SPPS to minimize the difficulty of scale-up, using excess reagents and expensive resins for coupling reactions, particularly for the synthesis of longer polypeptides and cyclic peptides of more complex structure, see literature:

[18]K.D.Eom,Z.W.Miao,J.L.Yang and J.P.Tam,J.Am.Chem.Soc.,2003,125,73-82.

[19]S.Liu,B.L.Pentelute and S.B.H.Kent,Angew.Chem.,Int.Ed.,2012,51,993-999.

[20]D.G.Mullen,B.Weigel,G.Barany and M.D.Distefano,J.Pept.Sci.,2010,16,219-222.

[21]Y.Okada,H.Suzuki,T.Nakae,et al.J.Org.Chem.,2012,78,320-327.

[22]B.C.Li,D.C.Montgomery,J.W.Puckett and P.B.Dervan,J.Org.Chem.,2013,78,124-133.

[23]K.Jin,I.H.Sam,K.H.L.Po,et al.Nat.Commn.2016,7,12394

in this respect, using soluble polymers as carriers for coupling of amino acid residues, an alternative method was subsequently developed which enables the polypeptide synthesis to be reacted in the liquid phase, but isolated and purified either in the solid phase or by a convenient extraction procedure. However, the latter method requires a high molecular weight soluble polymer, which is inconvenient for producing large amounts of small peptides having much lower molecular weights than the high molecular weight, and also does not comply with the principles of atomic economy. Furthermore, by carefully controlling the solidification/crystallization conditions, it often takes a longer time to produce a solid or crystalline product.

In recent years, a research group led by professor of prunus salicina has made a great progress in the field of purification chemistry, with particular attention to avoiding column chromatography and recrystallization, see literature:

[24]J.Wu,G.An,S.Lin,J.Xie,W.Zhou,H.Sun,Y.Pan G.Li.Chem.Commun.,2014,50,1259-1261.

[25]C.W.Seifert,A.Paniagua,G.A.White,L.Cai,G.Li.Eur.J.Org.Chem.2016,1714-1719.

this study and concept is defined as Group Assisted Purification (GAP) chemistry/technology. An organic synthetic chemistry avoids traditional purification methods such as chromatography and/or recrystallization by purposefully introducing functional groups into the starting materials or newly produced products. First, this study is likely to cover the entire field of synthetic organic chemistry. In the initial phase of research they have focused on the synthesis of chiral amines, N-phosphonoimines and N-phosphonoimides, and have met with great success in this regard. By controlling the solubility, the chiral amine product can be selectively precipitated from the crude mixture, thereby avoiding chromatography and recrystallization. In the second stage of research, they are developing methods to extend this technology to other substrates and functional groups. To this end, they have used the GAP properties of chiral auxiliaries and, by modification, have proposed the concept of GAP protecting groups. Protecting groups are present in almost every complex synthesis where multiple functional groups are present. Good protecting groups need to be stable to a variety of conditions and must be added and removed in high yield. An ideal example of GAP chemistry is that semi-permanent protecting groups result in the necessary solubility characteristics of GAP. The problem is that most conventional protecting groups are non-polar and therefore do not give the GAP solubility required for most substrates. If a protecting group can be developed that gives adequate solubility control, then the GAP chemistry can potentially be extended to all syntheses that require the use of that protecting group. By using GAP technology, organic synthesis and ternary coprecipitation can be carried out efficiently without adopting traditional purification methods such as chromatography, recrystallization and the like. Petroleum solvents or co-solvents. Therefore, the use of raw materials, silica gel, energy, manpower, etc. can be greatly reduced. GAP chemistry strategies can facilitate polypeptide synthesis, which has the advantages of Solid Phase Peptide Synthesis (SPPS) and liquid phase peptide synthesis, avoiding their disadvantages in the above factors.

However, the groups currently used for protecting the amino or carboxyl functional groups of amino acids, including the GAP group under development of the professor of prunus salicina mentioned above, are destroyed or decomposed during the removal process and cannot be recycled at all, which greatly consumes the production cost, and the discharge of waste materials inevitably causes serious environmental pollution problems, which is not considered to be the best solution in terms of economic cost and social benefit. Therefore, with the increasing advocation of the concept of green sustainable development of economic society, the search and discovery of new amino acid carboxyl protecting groups which can be recovered and recycled is still a promising important research topic.

Disclosure of Invention

Technical problem to be solved

In order to avoid the defects of the prior art, the invention provides phosphoryloxy benzaldehyde (or ketone) and a derivative thereof and a preparation method thereof, which solve the problems that in the existing chemical synthesis method of polypeptide, an amino acid protective group and solid-phase resin are easy to damage or decompose in a deprotection or shearing step and cannot be directly recycled, so that reaction byproducts of liquid-phase synthesis are relatively complex, separation steps are multiple, the time consumption period is long, the purification scale is small, the production cost is high, the production scale of solid-phase synthesis is small, the raw materials are expensive, the waste is large, resin wastes are multiple, and the environmental pollution is serious.

Technical scheme

A phosphonooxybenzaldehyde (or ketone) and derivatives thereof, characterized by: the molecular structural general formula of the phosphoryloxy benzaldehyde (or ketone) compound 1 is as follows:

R₂＝CH₃,CH₃O,C₂H₅,C₂H₅O,Ph,PhO,

R₃＝H

(X＝NO₂,F,Cl,Br,I,H,CH₃,OCH₃)

or phosphoryloxy benzyl alcohol compounds 2:

R₃＝H

(X＝NO₂,F,Cl,Br,I,H,CH₃,OCH₃)

or the molecular structure general formula of Fmoc-amino acid ester compound 4 of the phosphoryloxy benzyl alcohol compound 2 is as follows:

R₃＝H

(X＝NO₂，F，Cl，Br，l，H，CH₃，OCH₃)

AA＝amino acid

or the molecular structure general formula of the phosphoryloxybenzaldehyde (or ketone) oxime compound 3 is as follows:

R₃＝H

(X＝NO₂，F，Cl，Br，I，H，CH₃，OCH₃)

or Boc-amino acid oxime ester compounds 5 of phosphoryloxybenzaldehyde (or ketone) oxime compounds 3 have the molecular structure general formula:

R₃＝H

(X＝NO₂，F，Cl，Br，I，H，CH₃，OCH₃)

AA＝amino acid

wherein: AA represents various amino acids.

A preparation method of the phosphoryloxy benzaldehyde (or ketone) and the derivatives thereof is characterized by comprising the following steps:

step 1: adopt (RO)₂POCl or POCl₃Reacting with hydroxybenzaldehyde or ketone as raw materials in solvent A under the catalysis of alkali, recovering solvent by rotary evaporation after the reaction is finished, extracting the residual solution with ethyl acetate, separating from the water phase, combining the organic phases, and washing with deionized water for multiple times;

then separating and purifying to obtain phosphoryloxy benzaldehyde or ketone compound 1;

the (RO)₂POCl or POCl₃The feeding ratio of the hydroxyl benzaldehyde to hydroxybenzaldehyde or hydroxybenzaldehyde ketone is 1: 1-4;

step 2: taking the phosphoryloxy benzaldehyde or ketone compound 1 in the step 1 as a raw material, carrying out hydrogenation reduction treatment reaction in a solvent B by using a reducing agent, after the reaction is finished, carrying out rotary evaporation to recover the solvent, extracting the residual solution by using ethyl acetate and separating the residual solution from a water phase, combining organic phases, and washing the organic phases for multiple times by using deionized water;

then the corresponding derivatization product phosphoryloxy benzyl alcohol compound 2 of the compound 1 is obtained by separation and purification;

and step 3: taking the phosphoryloxy benzaldehyde or ketone compound 1 obtained in the step 1 as a raw material, adding alkali into a solvent A for catalysis reaction with hydroxylamine hydrochloride, after the reaction is finished, carrying out rotary evaporation to recover the solvent, extracting the residual solution with ethyl acetate and separating the residual solution from a water phase, combining organic phases, and washing the organic phases with deionized water for multiple times;

then the corresponding derivatization product of the compound 1, namely phosphoryloxy benzaldehyde or ketoxime compound 3, is obtained through separation and purification;

and 4, step 4: coupling the phosphoryloxy benzyl alcohol compound 2 obtained in the step 2 with Fmoc protected amino acid (Fmoc-AA-OH), and separating and purifying to obtain an Fmoc-amino acid ester derivative product 4 corresponding to the compound 2;

and 5: and (3) carrying out coupling reaction on the phosphoryloxy benzaldehyde or ketoxime compound 3 obtained in the step (3) and Boc protected amino acid (Boc-AA-OH), and separating and purifying to obtain a derivative product 5 of the Boc-amino oxime ester corresponding to the compound 3.

The separation and purification of the steps 1 to 5 are as follows: extracting with ethyl acetate, separating from the water phase, combining the organic phases, and washing with deionized water for 2-3 times; dropping alkane or ether solvent with small polarity into ethyl acetate solution until crystallization or precipitation is separated out, and filtering and washing or recrystallization to complete separation and purification.

The reaction conditions of the alkali-added catalytic reaction or the hydrogenation reduction treatment reaction in the steps 1 to 3 are as follows: stirring for 2-3 hours at room temperature.

The deionized water is used for washing for multiple times in the steps 1 to 3, and the times are 2-3.

The solvent A in the step 1 and the step 3 is: tetrahydrofuran, acetonitrile, benzene, toluene, chloroform CHCl₃One or more of dichloromethane DCM, N-dimethylformamide DMF and N-methylpyrrolidone NMP organic solvent.

The alkali in the step 1 and the step 3 is one or more of organic alkali such as tertiary amine TEA, DIEA, NMM, pyridine and derivative DMAP thereof.

The solvent B in the step 2 is: one or more of alcohols, acetonitrile, tetrahydrofuran, etc.

The reducing agent in the step 2 is: NaBH₄、H₂/Pd/C、B₂H₄Or LiAlH₄。

The alkane or ether solvent is one or more of petroleum ether, n-hexane, cyclohexane, diethyl ether, n-butyl ether, etc.

Advantageous effects

The invention provides a phosphoryloxy benzaldehyde (or ketone) and its derivative and its preparing method, (1) adopting (RO)₂POCl or POCl₃Reacting with hydroxybenzaldehyde (or ketone) as raw material in alkaline solution, separating and purifying to obtain phosphoryloxy benzaldehyde (or ketone) compound 1; (2) reacting the obtained phosphoryloxy benzaldehyde (or ketone) compound 1 serving as a raw material with hydroxylamine hydrochloride, and separating and purifying to obtain a corresponding derivative product phosphoryloxy benzyl alcohol compound 2 of the compound 1; (3) using the obtained phosphoryloxy benzaldehyde (or ketone) compound 1 as a raw material, carrying out hydrogenation reduction treatment, and separating and purifying to obtain a corresponding derivative product phosphoryloxy benzaldehyde (or ketone) oxime compound 3 of the compound 1; (4) condensing and coupling the obtained phosphoryloxy benzyl alcohol compound 2 with Fmoc protected amino acid (Fmoc-AA-OH), and separating and purifying to obtain an Fmoc-amino acid ester derivative product 4 corresponding to the compound 1; (5) condensing and coupling the obtained phosphoryloxybenzaldehyde (or ketone) oxime compound 3 with Boc protected amino acid (Boc-AA-OH), and separating and purifying to obtain a Boc-amino acid ester derivative product 5 corresponding to the compound 1.

Through systematic screening research, the phosphoryloxy benzaldehyde (or ketone) can be coupled with amino of amino acid in a Schiff base mode to protect the amino, and can be easily removed through acidolysis. On the other hand, the small molecules of the reduction product (phosphoryloxy benzyl alcohol derivative) of phosphoryloxy benzaldehyde (or ketone) and the oximation product (phosphoryloxy benzyl alcohol or ketone oxime derivative) of the phosphoryloxy benzaldehyde (or ketone) can be subjected to esterification reaction with the carboxyl of amino acid under the conditions of alkali, normal temperature and normal pressure through the mediation of a coupling agent, so that stable amino acid ester can be generated at high yield for protecting the carboxyl of the amino acid, and the phosphate carrier and the amino acid or polypeptide derivative thereof are found to be easily crystallized and precipitated in a nonpolar solvent, can be removed from the amino acid or polypeptide derivative thereof through simple separation, purification and alkaline hydrolysis, and the by-product is also the original phosphate carrier, can be directly recycled after recovery and purification, and can realize a sustainable large-scale green generation process. From the research and analysis of the existing documents, the strategy for protecting amino acid carboxyl and assisting purification by using the phosphate carrier involved in the invention is still pioneered. Compared with the existing amino acid carboxyl protecting group (including the GAP group which is developed by the professor of the plum-cinnamon root mentioned above) or a solid-phase resin carrier, the phosphoryloxy benzaldehyde (or ketone) and the derivative thereof have the advantages of abundant and easily obtained raw materials, recyclability, simple and convenient operation, mild conditions, low equipment cost, little three wastes, environmental protection and the like.

The phosphate small molecule has obvious practical application value in organic synthesis and polypeptide chemistry, can be used as a protective group of amino acid, is not damaged after deprotection, and is easy to recycle. But also can replace a resin carrier in solid phase polypeptide synthesis, is beneficial to the separation and purification of auxiliary polypeptide, is not damaged after being sheared, and is easy to recycle. The method has the advantages of both liquid phase and solid phase synthesis methods, can synthesize and prepare protected amino acid or polypeptide and derivatives thereof more simply, conveniently, quickly, economically and efficiently, and the phosphate carrier can be recovered and directly reused, thereby reducing the waste of raw materials, reducing the pollution of wastes, saving the cost and being beneficial to environmental protection.

Drawings

FIG. 1: process flow chart of preparation method of phosphoryloxy benzaldehyde (or ketone) and derivative thereof

Detailed Description

The invention will now be further described with reference to the following examples and drawings:

the examples mainly include the following compounds and their preparation steps: (1) adopt (RO)₂POCl or POCl₃Reacting with hydroxybenzaldehyde (or ketone) as raw material in alkaline solution, separating and purifying to obtain phosphoryloxy benzaldehyde (or ketone) compound 1; (2) reacting the obtained phosphoryloxy benzaldehyde (or ketone) compound 1 serving as a raw material with hydroxylamine hydrochloride, and separating and purifying to obtain a corresponding derivative product phosphoryloxy benzyl alcohol compound 2 of the compound 1; (3) using the obtained phosphoryloxy benzaldehyde (or ketone) compound 1 as raw material, hydrogenation reducing treatment, separating and purifying to obtain the compound1 corresponding derivatization product phosphoryloxybenzaldehyde (or ketone) oxime compound 3; (4) condensing and coupling the obtained phosphoryloxy benzyl alcohol compound 2 with Fmoc protected amino acid (Fmoc-AA-OH), and separating and purifying to obtain an Fmoc-amino acid ester derivative product 4 corresponding to the compound 1; (5) condensing and coupling the obtained phosphoryloxybenzaldehyde (or ketone) oxime compound 3 with Boc protected amino acid (Boc-AA-OH), and separating and purifying to obtain a Boc-amino acid ester derivative product 5 corresponding to the compound 1. The detailed synthetic route of the invention is shown in figure 1. Some of the abbreviations commonly used in the present invention have the following meanings:

boc: tert-butyloxycarbonyl radical

DCM: methylene chloride CH₂Cl₂

DMAP 4-dimethylaminopyridine

DMF N, N-dimethylformamide

EDC-HCl 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

Fmoc: fmoc group

GPS green polypeptide synthetic carrier

GSH glutathione

HATU 2- (7-benzotriazol oxide) -N, N, N ', N' -tetramethyluronium hexafluorophosphate

HOBT 1-hydroxybenzotriazole

HBTU O-benzotriazole-tetramethyluronium hexafluorophosphate

NMM N-methylmorpholine

NMP N-methylpyrrolidone

PyBop benzotriazol-1-yl-oxytripyrrolidinylphosphine hexafluorophosphate

t-butyl tBu

TFA: trifluoroacetic acid

THF: tetrahydrofuran (THF)

Trt trityl

The invention is suitable for preparing phosphoryloxy benzaldehyde (or ketone) and its derivative, the reaction principle and technical route are shown in figure 1.

Actual procedure

Synthesis of 4-diphenylphosphoryloxybenzophenone (DBK,1a) by accurately weighing 4-hydroxyAdding benzophenone (5g, 25mmol, 1equiv) into a 250mL reaction bottle, adding 100mL tetrahydrofuran THF, placing in an ice bath, stirring for 30min, and dropwise adding an acid-binding agent triethylamine Et into the reaction system₃N (4.2mL, 30mmol, 1.2equiv), diphenyl phosphoryl chloride (5.7mL, 30mmol, 1.2equiv) is weighed and added into the reaction system drop by drop, the reaction system is placed in an ice bath for reaction for 30min, then the ice bath TLC detection reaction system is removed for reaction for 1.5h at room temperature, dilute sulfuric acid (0.1mol/L, 10mL) is added for quenching reaction, a rotary evaporator is used for concentration to remove THF solvent, 20mL deionized water is added, ethyl acetate is added for extraction to obtain an organic phase, and the organic phase is dried by anhydrous magnesium sulfate. After rotary evaporation to dryness, 2mL of ethyl acetate was added to fully dissolve the sample, 14mL of n-Hexane (VEA/VN-Hexane ═ 1:7) was added dropwise, a large amount of white precipitate appeared in the system, and the white precipitate was filtered and dried to obtain the objective compound (1a) in about 93% yield. Structural characterization:¹HNMR(400MHz,CDCl₃),δ＝7.95-7.90(m,4H),7.76-7.73(m,4H),7.59-7.55(m,3H),7.52-7.44(m,6H),7.36-7.34(d,2H)ppm；³¹PNMR(162MHz,CDCl₃),δ＝31.51ppm；¹³CNMR(100MHz,CDCl₃),δ＝195.4,154.4,137.5,133.9,132.8,132.4,132.1,131.8,131.7,131.2,129.9,128.8,128.7,128.3,120.5ppm；HRMS(ESI)m/z calcd for C₂₅H₂₀O₃P⁺(M+H)⁺＝399.11446,found 399.11469。

the solvent in the reaction system can be tetrahydrofuran THF, acetonitrile, benzene, toluene, chloroform CHCl₃One or more of dichloromethane DCM, N-dimethylformamide DMF and N-methylpyrrolidone NMP organic solvent.

The alkali is one or more of organic alkali such as tertiary amine TEA, DIEA, NMM, pyridine and its derivative DMAP.

Synthesis of 4-diphenylphosphoryloxybenzhydrol (DBM,2a) by accurately weighing 4-diphenylphosphoryloxybenzophenone (1a) (800mg, 2mmol, 1equiv) in a 100mL reaction flask, adding 20mL of methanol solution, placing in an ice bath, stirring for 30min, and adding sodium borohydride into the reaction system in three batchesNaBH₄(92mg, 2.4mmol, 1.2equiv), adding a balloon, transferring to room temperature, sealing, reacting for 2h, detecting by TLC, adding saturated ammonium chloride to quench the reaction after the raw materials are completely consumed, concentrating to remove a methanol solution, adding 10mL of deionized water, extracting by ethyl acetate to obtain an organic phase, and drying by anhydrous magnesium sulfate. After rotary evaporation to dryness, 0.5mL of ethyl acetate was added to fully dissolve the sample, 5mL of n-Hexane (VEA/VN-Hexane ═ 1:10) was added dropwise, a large amount of white precipitate appeared in the system, and the white precipitate was filtered and dried to obtain the objective compound (2a) in a yield of about 97%. Structural characterization:¹H NMR(400MHz,CDCl₃),δ＝7.89-7.84(m,4H),7.56-7.52(m,2H),7.48-7.43(m,4H),7.31-7.23(m,7H),7.14-7.12(d,2H),5.74(s,1H),2.99(s,1H)ppm；³¹PNMR(162MHz,CDCl₃),δ＝30.58ppm；¹³CNMR(100MHz,CDCl₃),δ＝150.0,143.9,140.6,132.5,131.8,131.7,130.1,128.7,128.6,128.4,128.0,127.4,126.6,120.6,75.4ppm；HRMS(ESI)m/z calcd for C₂₅H₂₂O₃P⁺(M+H)⁺＝401.13011,found:401.12985。

the solvent in the reaction system can be one or more of alcohols, acetonitrile, tetrahydrofuran and the like.

The reducing agent is: NaBH₄、H₂/Pd/C、B₂H₄Or LiAlH₄。

Synthesis of 4-Diphenylphosphoryloxy benzophenone oxime (DBO,3a) 4-Diphenylphosphoryloxy benzophenone (DBK,1a) (800mg, 2mmol, 1equiv) was accurately weighed into a 100mL reaction flask, 50mL of anhydrous ethanol solution was added, 5mL of Pyridine (EtOH: 10:1) was added with stirring, and NH was subsequently added₂OH & HCl (280mg, 4mmol, 2equiv) stirred at room temperature for 10h, after the reaction is finished, concentrated to remove ethanol solvent and part of pyridine, then 50mL of ethyl acetate is added for dissolution, diluted HCl is added for continuous washing twice, residual pyridine and excess hydroxylamine are removed, finally anhydrous magnesium sulfate is used for drying for 2h, after spin-drying, 0.5mL of ethyl acetate is added for full dissolution of the sample, and n-ethyl acetate is added dropwiseHexane 4mL (V)_EA/V_N-Hexane1:8), a large amount of white precipitate appeared in the system, and the white precipitate was filtered and dried to obtain the target compound (3a) in about 92% yield. Structural characterization:¹HNMR(400MHz,CDCl₃),δ＝9.50(s,1H),7.99-7.90(m,4H),7.59-7.28(m,14H),7.22-7.20(m,1H)ppm；³¹PNMR(162MHz,CDCl₃),δ＝31.09ppm；¹³CNMR(100MHz,CDCl₃),δ＝156.5,151.3,136.4,132.7,131.9,131.3,130.0,129.8,129.3,129.0,128.8,128.6,128.3,128.0,120.4ppm；HRMS(ESI)m/z calcd for C₂₅H₂₂O₃P⁺(M+H)⁺401.13011,found:401.29789。

4-Diphenylphosphoryloxy-benzhydryltyrosine ester [ DBM-O-Tyr (OtBu) -Fmoc, 4a]The synthesis of (2): accurately weighing 4-diphenylphosphoryloxybenzhydrol (DBM,2a) (200mg, 0.5mmol, 1equiv) into a 100mL reaction bottle, adding 30mL DCM for dissolution, sequentially adding Fmoc-Tyr (OtBu) -OH (276mg, 0.6mmol, 1.2equiv), 4-dimethylaminopyridine DMAP (7.32mg, 0.06mmol, 0.12equiv), dicyclohexylcarbodiimide DCC (123mg, 0.6mol, 1.2equiv) into the reaction system, reacting for 2h at room temperature, cooling to 0 ℃ after TLC detection reaction is finished, filtering to obtain filtrate, concentrating, adding ethyl acetate 30mL for dissolution, sequentially using saturated NH₄Aqueous Cl solution and saturated Na₂CO₃The solution was washed and dried over anhydrous magnesium sulfate. After rotary evaporation and concentration, a sample is dissolved by 1mL of ethyl acetate, 6mL of n-Hexane (VEA/VN-Hexane ═ 1:6) is added dropwise, a large amount of white precipitate appears in the system, the white precipitate is filtered and dried to obtain a target compound DBM-O-Tyr (tBu) -Fmoc (4a), the yield is about 95 percent, and after one-time precipitation, the target compound DBM-O-Tyr (tBu) -Fmoc (4a) can be obtainedAnd (3) taking the obtained product as a raw material to carry out next feeding, and continuously washing the sample for 2-3 times to carry out NMR characterization. Structural characterization:¹HNMR(400MHz,CDCl₃),δ＝7.93-7.88(m,4H),7.80-7.78(d,2H),7.59-7.54(m,4H),7.50-7.40(m,6H),7.33-7.20(m,11H),6.87(s,1H),6.82-6.79(d,4H),5.32-5.29(d,1H),4.79-4.74(m,1H),4.44-4.32(m,2H),4.23-4.19(m,1H),3.16-3.10(m,2H),1.34(s,9H)ppm；³¹PNMR(162MHz,CDCl3),δ＝30.86ppm；¹³CNMR(100MHz,CDCl₃),δ＝170.6,155.6,154.5,150.5,143.8,141.3,139.1,135.7,132.6,131.8,130.2,129.8,129.1,128.7,128.6,128.1,127.7,127.1,125.1,124.1,120.8,120.0,78.4,67.0,54.8,47.2,37.4,33.9,28.9ppm；HRMS(ESI)m/z calcd for C₅₃H₄₉NO₇P⁺(M+H)⁺＝842.32412,found:842.32422.

synthesis of 4-diphenylphosphoryloxydiphenylmethanone oxime Boc-valine ester (DBO-O-Val-Boc,5a) by accurately weighing 4-diphenylphosphoryloxydiphenylmethanone oxime (DBO,3a) (206mg, 0.5mmol, 1equiv) in a 100mL reaction flask, adding 30mL DCM for dissolution, sequentially adding Boc-Val-OH (130mg, 0.6mmol, 1.2equiv), 4-dimethylaminopyridine DMAP (7.32mg, 0.06mmol, 0.12equiv), dicyclohexylcarbodiimide (123mg, 0.6mol, 1.2equiv) to the reaction system, reacting at room temperature for 2h, detecting by TLC, cooling to 0 deg.C, filtering to obtain a filtrate, concentrating, adding ethyl acetate 30mL for dissolution, sequentially adding saturated NH₄Aqueous Cl solution, saturated NaHCO₃The solution was washed, dried over anhydrous magnesium sulfate, concentrated, and the sample was dissolved in 0.5mL of ethyl acetate, and 6mL (V) of n-hexane was added dropwise_EA/V_N-Hexane1: 12) and (3) a large amount of white precipitates appear in the system, the white precipitates are filtered and dried to obtain a compound GAP-C ═ N-O-Val-Boc (5a), the yield is about 98%, the compound GAP-C ═ N-O-Val-Boc can be used as a raw material for feeding in the next step after one-time precipitation, and the sample can be subjected to NMR characterization after 2-3 times of continuous washing. Structural characterization:¹HNMR(400MHz,CDCl₃),δ＝7.97-7.87(m,4H),7.61-7.21(m,15H),5.04-5.02(m,1H),4.20-4.17(m,1H),1.95-1.90(m,1H),1.45(s,9H),0.88-0.77(m,6H)ppm；³¹PNMR(162MHz,CDCl₃),δ＝30.66ppm；¹³CNMR(100MHz,CDCl₃),δ＝169.9,164.9,155.5,151.9,134.2,132.7,132.1,131.8,131.7,131.2,130.6,129.8,129.1,128.8,128.7,128.4,128.3,120.5,79.9,57.7,31.3,28.3,18.8,17.6ppm；HRMS(ESI)m/z calcd for C₃₅H₃₇N₂O₆P⁺(M+H)⁺613.23892,found:613.23795。

synthesis of 4-diphenylphosphoryloxybenzophenone oxime valine ester (DBO-O-Val-Fmoc,5 a'), which is to accurately weigh 4-diphenylphosphoryloxybenzophenone oxime (DBO,3a) (206mg, 0.5mmol, 1equiv) into a 100mL reaction bottle, add 30mL of DCM for dissolution, sequentially add Fmoc-Val-OH (203mg, 0.6mmol, 1.2equiv), 4-dimethylaminopyridine DMAP (7.32mg, 0.06mmol, 0.12equiv), dicyclohexylcarbodiimide (123mg, 0.6mol, 1.2equiv) into the reaction system, react for 2h at room temperature, cool to 0 ℃ after TLC detection reaction, filter the filtrate, concentrate, add ethyl acetate 30mL for dissolution, sequentially use saturated NH to dissolve₄Aqueous Cl solution, saturated NaHCO₃The solution was washed, dried over anhydrous magnesium sulfate, concentrated, and the sample was dissolved in 0.5mL of ethyl acetate, and 6mL (V) of n-hexane was added dropwise_EA/V_N-Hexane1: 12) and a large amount of white precipitate appears in the system, the white precipitate is filtered and dried to obtain a compound GAP-C ═ N-O-Val-Fmoc (5a '), the yield is about 95%, the compound GAP-C ═ N-O-Val-Fmoc (5 a'), the yield can be used as a raw material for feeding in the next step after one-time precipitation, and the sample can be subjected to NMR characterization after 2-3-time continuous washing. Structural characterization:¹HNMR(400MHz,CDCl₃),δ＝7.95-7.89(m,4H),7.79-7.78(d,2H),7.62-7.25(m,21H),5.36-5.31(m,1H),4.43-4.39(m,2H),4.29-4.23(m,2H),2.01-1.99(m,1H),0.98-0.80(m,6H)ppm；³¹PNMR(162MHz,CDCl₃),δ＝30.56ppm；¹³CNMR(100MHz,CDCl₃),δ＝169.6,165.2,156.1,152.1,143.7,141.3,134.1,132.7,131.8,131.7,131.3,130.7,130.6,129.8,129.2,128.8,128.7,128.5,128.3,127.7,127.1,125.1,120.7,120.0,67.1,58.2,47.2,31.6,22.7,17.6ppm；HRMS(ESI)m/z calcd for C₄₅H₃₉N₂O₆P⁺(M+H)⁺735.25457,found:735.25534。

Claims

1. a phosphoryloxybenzaldehyde oxime or phosphoryloxybenzophenone oxime compound, is characterized in that, the general formula of molecular structure is:

2. the preparation method of phosphoryloxybenzaldehyde oxime class or phosphoryloxybenzophenone oxime class compound described in claim 1, is characterized in that, comprises the following steps:

Step 1. Use (RO) ₂ POCl or POCl ₃ and hydroxybenzaldehyde or hydroxybenzophenone as raw materials, add alkali catalysis in A solvent to carry out the reaction, after the reaction is completed, the solvent is recovered by rotary evaporation, and the residual solution is extracted with ethyl acetate And separate from the water phase, combine the organic phases and wash with deionized water for many times;

Then separate and purify to obtain phosphoryloxybenzaldehydes or hydroxybenzophenones:

The charging ratio of the (RO) ₂ POCl or POCl ₃ to hydroxybenzaldehyde or hydroxybenzaldehyde ketone is 1:1 to 4;

Step 2. Take the phosphoryloxybenzaldehydes or hydroxybenzophenone compounds obtained in step 1 as raw materials, and react with hydroxylamine hydrochloride in solvent A by adding alkali catalysis. After the reaction is completed, the solvent is recovered by rotary evaporation, and the residual solution is acetic acid. Ethyl ester extraction and separation from aqueous phase, combined organic phases and washed several times with deionized water;

Then, the phosphoryloxybenzaldehyde oximes or phosphoryloxybenzophenone oximes are obtained by separation and purification.

3. a Boc-amino acid oxime ester compound, is characterized in that, molecular structural formula is:

Among them: AA represents various amino acids.

4. the preparation method of Boc-amino acid oxime ester compound described in claim 3, is characterized in that, comprises the following steps:

Step 1. Use (RO) ₂ POCl or POCl ₃ and hydroxybenzaldehydes or hydroxybenzophenone as raw materials, add base to catalyze the reaction in A solvent, after the reaction is completed, revolve the solvent to recycle, and use ethyl acetate for the residual solution. Extract and separate from the aqueous phase, combine the organic phases and wash multiple times with deionized water;

Step 2. Take the phosphoryloxybenzaldehyde or hydroxybenzophenone compounds obtained in step 1 as raw materials, and react with hydroxylamine hydrochloride in solvent A by adding alkali catalysis. After the reaction is completed, the solvent is recovered by rotary evaporation, and the residual solution is treated with ethyl acetate. Ester extraction and separation from aqueous phase, combined organic phases and multiple washes with deionized water;

Then separate and purify to obtain phosphoryloxybenzaldehyde oximes or phosphoryloxybenzophenone oximes:

Step 3. Use the phosphoryloxybenzaldehyde oxime or phosphoryloxybenzophenone oxime compound obtained in step 2 to carry out coupling reaction with Boc-protected amino acid (Boc-AA-OH), and obtain Boc- Amino acid oxime ester derivatives.

5. according to the described preparation method of claim 2 or 4, it is characterized in that, the separation and purification of described step 1～step 3 is:

Use ethyl acetate to extract and separate from the water phase, combine the organic phases and wash with deionized water for 2 to 3 times; add a low polar alkane or ether solvent to the ethyl acetate solution dropwise until crystallization or precipitation occurs, and then Filtration and washing or recrystallization operations complete separation and purification.

6 . The preparation method according to claim 5 , wherein the reaction conditions of the alkali-added catalyzed reaction in the steps 1 and 2 are: stirring at room temperature for 2-3 hours. 7 .

7 . The preparation method according to claim 6 , wherein the number of times of washing with deionized water in the steps 1 and 2 is 2 to 3 times. 8 .

8. preparation method according to claim 7 is characterized in that: the A solvent of described step 1 and step 2 is: tetrahydrofuran, acetonitrile, benzene, toluene, chloroform CHCl ₃ , dichloromethane DCM, N,N-dimethylform One or more of the organic solvents of methylformamide DMF and N-methylpyrrolidone NMP.

9 . The preparation method according to claim 8 , wherein the base in the step 1 and the step 2 is one or more of the tertiary amine TEA, DIEA, NMM, pyridine and its derivative DMAP. 10 .

10 . The preparation method according to claim 9 , wherein the alkane or ether solvent is one or more of petroleum ether, n-hexane, cyclohexane, diethyl ether, n-butyl ether, and the like. 11 .