CN109678913A - A kind of synthetic method of sialylated TF antigen lactone and its fluoro analogs - Google Patents
A kind of synthetic method of sialylated TF antigen lactone and its fluoro analogs Download PDFInfo
- Publication number
- CN109678913A CN109678913A CN201811548160.4A CN201811548160A CN109678913A CN 109678913 A CN109678913 A CN 109678913A CN 201811548160 A CN201811548160 A CN 201811548160A CN 109678913 A CN109678913 A CN 109678913A
- Authority
- CN
- China
- Prior art keywords
- sialylated
- antigen
- hydroxyl
- nitrogen
- lactone
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/02—Acyclic radicals, not substituted by cyclic structures
- C07H15/04—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/445—The saccharide radical is condensed with a heterocyclic radical, e.g. everninomycin, papulacandin
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Saccharide Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to a kind of Chemoenzymatic synthesis methods of sialylated TF antigen lactone and its fluoro analogs, and it includes the following steps: (1) the sialylated TF trisaccharide of enzymatic clarification and its fluoro analogs；(2) tri- sugar lactone of chemical method sialic acid synthetase TF and its fluoro analogs.Present invention incorporates the flexibilities of the stereoselectivity of enzyme process, high efficiency and chemical method, the Chemoenzymatic synthesis for realizing sialylated TF antigen lactone and its fluoro analogs for the first time, solve the synthesis step faced in full chemistry sialic acid synthetase TF lactone and its fluoro analogs is various, overall yield is low, stereoselectivity is not high, using heavy metal catalyst the disadvantages of.Therefore, of the invention with important application prospects in molecular probe, antagonist and anti-tumor vaccine.
Description
Technical field
The invention belongs to Carbohydrate drugs fields, are related to the chemical synthesis process more particularly to sialylated TF of glucide
The application of antigen lactone and its fluoro analogs in Field of Drug Discovery.
Technical background
Thomsen-Friedenreich (Gal β 1,3GalNAc α Ser/Thr, TF) antigen is that the most common tumour is related
One of sugar antigens.In tumor tissues, TF antigen has that part is further sialylated by corresponding enzyme as precursor, forms sialic acid
Change TF antigen.The formation of saliva acid lactone is a common natural phenomena, and sialic acid, which lactonizes, can significantly change molecule
Physico-chemical property and biological characteristics, such as charge density, conformation, immunogenicity etc..Largely synthesize this saliva acid lactone glucosides
It is very valuable for studying its function on a molecular scale, and develops efficient, the convenient and fast synthetic method of one kind for pole
The earth promotes using it as the drug discovery process of lead compound.However, due to the complexity of this kind of natural sialic acid glycosidic structure
Property and its unstability, it is difficult directly to carry out isolating and purifying.Therefore, biological in order to study and evaluate its on a molecular scale
Learn meaning, find it is a kind of quickly, to efficiently synthesize sialylated TF antigen lactone synthetic method be current urgent problem to be solved.
Currently, the experiment by us is found, the acylation of sialic acid glucosides is carried out under condition of ice bath, can be formed in Isosorbide-5-Nitrae
Ester structure retains lactone, the available sialic acid TF glucosides containing Isosorbide-5-Nitrae lactone structure using deacylation base.For authentication
The application of method, We conducted the synthesis of serial fluoro sialic acid TF antigen lactone, it was demonstrated that the feasibility of this method and has
Effect property.Therefore, the lactone synthetic method of our Innovative Developments provides thinking and synthesis for the synthesis of sialic acid glucosides lactone
Approach.
Summary of the invention
The object of the present invention is to provide a kind of synthetic methods of sialylated TF antigen lactone, and this method is with sialylated
Tri- sugar antigens of TF are substrate, and pyridine is solvent, and acetic anhydride is acylating reagent, and under condition of ice bath, reaction, which generates, contains Isosorbide-5-Nitrae lactone
The acylate of structure, then using the sialylated TF sugar under methanol/sodium methoxide alkaline condition, obtained containing Isosorbide-5-Nitrae lactone
Glycosides.And adopting said method, obtain its fluoro analogs.This method has synthesis step few, and stereoselectivity is high, and structure is steady
Calmly, the advantages that substrate applicability is wide.
The purpose of the present invention is to overcome above-mentioned the deficiencies in the prior art, provide a kind of sialylated TF antigen lactone and its
The synthetic method of fluoro analogs.
To achieve the above object, the present invention adopts the following technical solutions:
The structure of a kind of sialylated TF antigen lactone, general structure are as follows:
Wherein:
R1, selected from hydrogen atom, α-or beta configuration serine residue, α-or beta configuration threonine residues, azide substitution alkyl,
Alkynyl substituted alkyl, sulfydryl replace alkyl, α-or beta configuration to replace one of alkyl；
R2, it is selected from one of nitrogen acetylamino, nitrogen propionamido, nitrogen trifluoroacetyl amido, nitrogen nitrine acetylamino；
R3, it is selected from one of fluorine atom, hydroxyl；
R4, it is selected from one of fluorine atom, hydroxyl；
R5, it is selected from one of fluorine atom, hydroxyl；
R6, it is selected from one of fluorine atom, hydroxyl；
R7, it is selected from one of fluorine atom, hydroxyl；
R8, it is selected from one of nitrogen acetylamino, nitrogen propionamido, nitrogen trifluoroacetyl amido, nitrogen nitrine acetylamino；
R9, it is selected from one of fluorine atom, hydroxyl；
R10, it is selected from one of fluorine atom, hydroxyl；
R11, it is selected from one of fluorine atom, hydrogen atom, nitrine, hydroxyl.
The Chemoenzymatic synthesis method of above-mentioned sialylated TF antigen lactone and its fluoro analogs, comprising the following steps:
(1) " one pot of double enzyme " method sialic acid synthetase TF antigen trisaccharide and its fluoro analogs are utilized, it is described sialylated
Using sialic acid and the like, the enzyme that " one pot double enzyme " is used during method is sialylated is that cmp sialic acid synthesizes
Enzyme and sialyltransferase；
Sialylated TF antigen and its fluoro analogs shown in general formula II:
Wherein:
R1, selected from hydrogen atom, α-or beta configuration serine residue, α-or beta configuration threonine residues, azide substitution alkyl,
Alkynyl substituted alkyl, sulfydryl replace alkyl, α-or beta configuration to replace one of alkyl；
R2, it is selected from one of nitrogen acetylamino, nitrogen propionamido, nitrogen trifluoroacetyl amido, nitrogen nitrine acetylamino；
R3, it is selected from one of fluorine atom, hydroxyl；
R4, it is selected from one of fluorine atom, hydroxyl；
R5, it is selected from one of fluorine atom, hydroxyl；
R6, it is selected from one of fluorine atom, hydroxyl；
R7, it is selected from one of fluorine atom, hydroxyl；
R8, it is selected from one of nitrogen acetylamino, nitrogen propionamido, nitrogen trifluoroacetyl amido, nitrogen nitrine acetylamino；
R9, it is selected from one of fluorine atom, hydroxyl；
R10, it is selected from one of fluorine atom, hydroxyl；
R11, it is selected from one of fluorine atom, hydrogen atom, nitrine, hydroxyl；
(2) it is acylated using II compound represented of acylation reaction mutual-through type, synthesis contains Isosorbide-5-Nitrae lactone structure, acyl
The intermediate of base protection, later removes acyl group, retains lactone structure, obtains compound ii；
The synthetic method of sialylated TF antigen in the step (1) and the like: disaccharides TF or fluoro is derivative
Object (1.0-5.0 equivalent), N-acetyl-neuraminate and the like (1.0-20.0 equivalent), CTP (cytidine triphosphate (CTP)) (1.0-
20.0 equivalents), MgCl2(5.0-100mM) and Tris-HCl buffer (10-500 mM, pH 5.0-10.5) obtained aqueous solution,
Be added cmp sialic acid synzyme (H.Yu andX.Chen, Org. Lett., 2006,8,2393；H.A.Chokhawala,
H.Cao,H.Yu,X.Chen,J.Am.Chem.Soc., 2007,129,10630；Yu,H.；Cao,H.；Tiwari,V.K.；Li,
Y.；Chen, X.Bioorg.Med.Chem. Lett., 2011,21,5037) and a kind of sialyltransferase (K.Lau, H.Yu,
V.Thon,Z.Khedri,M.E. Leon,B.K.Tran and X.Chen,Org.Biomol.Chem.,2011,9,2784；
H.Yu,H. Chokhawala,R.Karpel,B.Wu,J.Zhang,Y.Zhang,Q.Jia and X.Chen,J.Am.Chem.
Soc., 2005,127,17618), realize that " one pot of double enzyme " method is sialylated.After the reaction was completed, purifying can be obtained sialylated
TF antigen and its fluoro analogs.
The reaction is completed to use TLC (EA:CH3OH:H2O:HOAc=4:2:1:0.1 it) detects.
The enzyme that " one pot double enzyme " is used during method is sialylated in the step (1) is Neisseria meningitidis
CMP-sialic acid synthetase (NmCSS) and Pasteurella multocida sialyltransferase 1
(PmST1), the reaction time is 5 minutes~30 hours.
Reaction temperature is 0~37 DEG C in " one pot of double enzyme " method synthesis in the step (1), and revolving speed is 0-240 rpm；It is described
The method of shutting down of enzyme reaction is to be added into reaction with the isometric methanol of reaction system solution and cultivate 0~30 point at 4 DEG C
Clock.
The synthetic method of sialylated TF antigen lactone and the like in the step (2):
I.e. with compound II (1.0equiv) for raw material, pyridine solvent, acetic anhydride (20.0-50.0equiv) is to be acylated
Reagent, under condition of ice bath, reaction 5~for 24 hours, obtain acylated intermediate, the i.e. acylate of II；Then, anti-by Zempl é n
It answers, i.e., full acetylated object is dissolved in anhydrous methanol/sodium methoxide system, maintain pH value of solution 7.5~9.0, react at room temperature 2h~6h,
The reaction of TLC tracking and monitoring, after the reaction was completed, is isolated and purified by silica gel rapid column chromatography, obtains white solid lactone I.
The advantages and positive effects of the present invention:
1, the present invention provides a kind of efficient, the simple and direct sialylated TF antigen lactones of Chemoenzymatic synthesis and its fluoro class
Like the synthetic method of object.
2, it present invention incorporates the stereoselectivity of the flexibility of chemical synthesis and enzyme process, high efficiency, realizes for the first time
The Chemoenzymatic synthesis of sialic acid TF antigen lactone solves the substrate reactions that current sialic acid synthetase TF antigen lactone is faced
The deficiencies of activity is low, stereoselectivity is poor, synthesis step is various, overall yield is low.
3, the fluoro analogs of sialic acid TF antigen lactone provided by the invention have better stability, thus, this hair
Bright is with important application prospects in Development of Novel antagonist and anti-tumor vaccine.
Detailed description of the invention
Fig. 1 is nucleus magnetic hydrogen spectrum figure of the compound 5 in deuterated water；
Fig. 2 is nuclear-magnetism carbon spectrogram of the compound 5 in deuterated water；
Fig. 3 is nucleus magnetic hydrogen spectrum figure of the compound 6 in deuterated water；
Fig. 4 is nuclear-magnetism carbon spectrogram of the compound 6 in deuterated water；
Fig. 5 is nucleus magnetic hydrogen spectrum figure of the compound 7 in deuterated water；
Fig. 6 is nuclear-magnetism carbon spectrogram of the compound 7 in deuterated water.
Fig. 7 is nucleus magnetic hydrogen spectrum figure of the compound 8 in deuterated water.
Fig. 8 is nuclear-magnetism carbon spectrogram of the compound 8 in deuterated water.
Specific embodiment
In order to understand the present invention, below with reference to embodiment, the invention will be further described: following embodiments are illustrative
, it is not restrictive, cannot be limited the scope of protection of the present invention with following embodiments.
This reaction is carried out in two steps:
1. acylation reaction operates
It is dissolved in compound II (1.0equiv) in pyridine solvent, under condition of ice bath, stirs 20 minutes, into solution
It is added acetic anhydride (20.0-50.0equiv), nitrogen protection, 0 DEG C is reacted 12 hours.The monitoring reaction of TLC plate, after the reaction was completed, to
Q. s. toluene is added in reaction solution, concentrated by rotary evaporation is repeated 3 times.Later, silica gel is admixed, silica gel rapid column chromatography (stone is passed through
Oily ether: ethyl acetate :=1:50) it is isolated and purified, obtain the full acetylated intermediate of white.
2.Zempl é n operation
Vacuum drying obtains full acetylated intermediate, weighs obtained solid and redissolves in anhydrous methanol, sodium methoxide is added, maintains
Solution system pH7.5~9.0.Thin-layer chromatography detection (ethyl acetate:: methanol: water=4:2:1:0.2) reaction, the R of productf=
0.6~0.75.After the reaction was completed, it is separated by silica gel rapid column chromatography (ethyl acetate:: methanol: water=2:1:0.05)
Purifying obtains compound as white solid I.
It is illustrated below by embodiment.
" 1. one pot of double enzyme " method sialic acid synthetase TF antigen and its fluoro analogs
The general operation method of " one pot of double enzyme " method sialic acid synthetase TF trisaccharide
Into 50mL centrifuge tube, TF antigen or fluoro analogs (50-100mg, 1.0 equivalents), N- acetyl nerve ammonia are added
Acid or its fluoro analogs (1.5 equivalent), cytidine triphosphate (CTP) cytidine triphosphate (CTP) (1.5 equivalent of CTP), Tris-HCl buffer
(100mM, pH 8.5) and magnesium chloride (20mM), adding distilled water to adjust total volume is 10mL, and enzyme NmCSS is added after stirring evenly in vibration
(0.5-0.8mg) and PmST1 (0.4-0.6mg) reacts 30 minutes under the conditions of 37 DEG C, 140rpm.Thin-layer chromatography (EA:
CH3OH:H2O:HOAc=4:2:1:0.1) after the reaction was completed, isometric methanol, 4 DEG C of standing 0.5h are added in detection.Later, will
4 DEG C of reaction system, 12000rpm are centrifuged 10 minutes, collect supernatant.After rotary evaporation concentration, pass through the quick column of silicagel column point
From (EtOAc:MeOH:H2O=6:2:1), sialylated TF antigen and its fluoro analogs (85~95%) are obtained.
The following are " one pot of double enzyme " methods to carry out sialylated II representative compound 1-4 obtained:
The following are yields and structural information that " one pot of double enzyme " method carries out sialylated compound 1-4 obtained:
3- nitrine propyl oxygen-(5- acetylaminohydroxyphenylarsonic acid 3,5- dideoxy-D- glycerol-α-nine carbon onosic acid of pyrans)-(2 → 3)-
β-D- galactopyranosyl glycosyl-(1 → 3) -2- acetylaminohydroxyphenylarsonic acid 2- deoxidation-α-D- galactopyranoside (Neu5Ac α 2,3Gal β 1,
3GaNAcαProN31,95%)
Structural parameters:1H NMR(400MHz,D2O) δ 4.87 (d, J=4.0Hz, 1H), 4.51 (d, J=7.6 Hz, 1H),
4.28 (dd, J=10.8,3.6Hz, 1H), 4.21 (d, J=2.8Hz, 1H), 4.06-3.99 (m, 2H), 3.95 (t, J=
6.0Hz, 1H), 3.89 (d, J=2.8Hz, 1H), 3.87-3.36 (m, 19H), 2.72 (dd, J=12.4,4.8Hz, 1H),
1.99 (s, 6H), 1.92-1.82 (m, 2H), 1.75 (t, J=12.0Hz, 1H)13C NMR(101MHz,D2O)δ175.04,
174.60,173.90,104.47,99.75,97.21,77.41,75.70, 74.82,72.84,71.85,70.66,69.12,
68.58,68.39,68.10,67.43,64.98,62.56,61.25, 61.01,51.70,48.72,48.23,39.77,
27.98,22.08,22.06.HRMS(ESI)m/z calcd for [C28H46N5O19,M-H]-,756.2792；found,
756.2785.
3- nitrine propyl oxygen-(5- acetylaminohydroxyphenylarsonic acid 3,5- dideoxy-D- glycerol-α-nine carbon onosic acid of pyrans)-(2 → 3)-
6- deoxidation -6- fluoro-beta-D- galactopyranosyl glycosyl-(1 → 3) -2- acetylaminohydroxyphenylarsonic acid 2- deoxidation-α-D- galactopyranoside
(Neu5Acα2,3Gal6Fβ1,3GalNAcαProN32,91%)
Structural parameters:1H NMR(400MHz,D2O) δ 4.93 (d, J=3.4Hz, 1H), 4.60 (d, J=7.6 Hz, 1H),
4.34 (dd, J=10.8,3.6Hz, 1H), 4.24 (s, 1H), 4.11 (d, J=9.2Hz, 1H), 4.05 (d, J=11.0Hz,
1H), 4.02-3.44 (m, 22H), 2.77 (dd, J=12.0,4.4Hz, 1H), 2.04 (s, 6H), 1.99-1.87 (m, 2H),
1.80 (t, J=12.0Hz, 1H)13C NMR(101MHz,D2O)δ 175.06,174.61,173.84,104.28,99.78,
(97.22,83.18 d, J=166.65Hz), 77.51,75.44,72.87,71.85,70.70,68.98,68.62,68.38,
(68.17,68.12,67.13 d, J=8.08Hz), 65.05,62.59,61.31,51.71,48.70,48.26,39.71,
27.97,22.09.19F NMR(376MHz,D2O)δ -229.63.HRMS(ESI)m/z calcd for[C28H45FN5O18,M-
H]-,758.2749；found, 758.2712.
3- nitrine propyl oxygen-(5- hydroxyl acetylaminohydroxyphenylarsonic acid 3,5- dideoxy-D- glycerol-α-nine carbon onosic acid of pyrans)-(2 →
3) -6- deoxidation -6- fluoro-beta-D- galactopyranosyl glycosyl-(1 → 3) -2- acetylaminohydroxyphenylarsonic acid 2- deoxidation-α-D- galactopyranoside
(Neu5Gcα2,3Gal6Fβ1,3GalNAcαProN3, 3,85%)
Structural parameters:1H NMR(400MHz,D2O) δ 4.87 (d, J=3.6Hz, 1H), 4.66-4.60 (m, 1H), 4.57-
4.45 (m, 2H), 4.34-4.24 (m, 1H), 4.19 (d, J=2.8Hz, 1H), 4.07-3.70 (m, 13H), 3.64-3.39 (m,
6H), 2.73 (dd, J=12.4,4.6Hz, 1H), 1.99 (s, 3H), 1.89-1.83 (m, 2H), 1.76 (t, J=12.0Hz,
1H).13C NMR(101MHz,D2O) δ 175.79,174.60,173.89,104.28,99.79,97.21,83.17 (d, J=
165.64Hz), 77.49,75.42,72.98 (d, J=20.2Hz), 72.58,71.91,70.69,68.97,68.62,68.08
(d, J=10.1Hz), 67.10 (d, J=8.08 Hz), 65.04,62.53,61.30,61.02,59.36,51.40,48.69,
48.25,39.78,27.97,22.08.19F NMR(376MHz,D2O)δ-229.53.HRMS(ESI)m/z calcd for
[C28H46FN5O19Na, M+Na]+798.2669；found,798.2632.
3- nitrine propyl oxygen-(5- trifluoroacetamido -3,5- dideoxy-D- glycerol-α-nine carbon onosic acid of pyrans)-(2
→ 3) -6- deoxidation -6- fluoro-beta-D- galactopyranosyl glycosyl-(1 → 3) -2- acetylaminohydroxyphenylarsonic acid 2- deoxidation-α-D- galactopyranoside
(Neu5NHTFAα2,3Gal6Fβ1,3GalNAcαProN34,95%)
Structural parameters:1H NMR(400MHz,D2O) δ 4.92 (d, J=3.6Hz, 1H), 4.73-4.64 (m, 1H), 4.62-
4.52 (m, 2H), 4.39-4.30 (m, 1H), 4.24 (d, J=2.4Hz, 1H), 4.12 (dd, J=9.6,2.8Hz, 1H),
4.08-3.70 (m, 14H), 3.69-3.43 (m, 6H), 2.80 (dd, J=12.4,4.4Hz, 1H), 2.05 (s, 3H), 1.96-
1.89 (m, 2H), 1.83 (t, J=12.4Hz, 1H)13C NMR(101MHz, D2O)δ174.60,173.75,159.41(d,J
=37.37Hz), 115.80 (d, J=287.85Hz), 104.29,99.81,97.22,83.17 (d, J=166.65Hz),
(77.47,75.45,72.97 d, J=20.2Hz), 72.04,70.70,68.96,68.64,68.06,68.01,67.07 (d, J
=7.07Hz), 65.05,62.51,61.30,59.37,52.25,48.70,48.26,39.77,27.97,22 .09.19F NMR
(400MHz,D2O)δ-75.50,-229.54. HRMS(ESI)m/z calcd for[C28H43F4N5O18Na,M+Na]+,
836.2437；found,836.2495.
2. sialic acid synthetase TF antigen lactone and its fluoro analogs
Embodiment 5~8
Compound I is synthesized by the following way route and obtains:
Embodiment 5
The synthesis of embodiment 5
3- nitrine propyl oxygen-[(5- acetylaminohydroxyphenylarsonic acid 3,5- dideoxy-D- glycerol-α-nine carbon onosic acid of pyrans)-(2 → 3)-
Oxygen-β-D- galactopyranosyl glycosyl-(1 → 3)]-galactopyranoside-(1 " → 4 ')-oxygen -2- acetylaminohydroxyphenylarsonic acid 2- deoxidation-α-D-
Lactone
Into 50mL round-bottomed flask, sialylated TF trisaccharide 1 (106mg), aceticanhydride (3mL), pyridine (6mL), ice bath is added
Under the conditions of 12h is stirred at room temperature.Thin-layer chromatography detects (petroleum ether: ethyl acetate=1:10), after fully reacting, into reaction solution
2mL toluene is added, concentrated by rotary evaporation is repeated 3 times.Later, silica gel is admixed, silica gel rapid column chromatography (petroleum ether: acetic acid is passed through
Ethyl ester :=1:50) it is isolated and purified, obtain full acetylated intermediate, white solid 142mg, yield 94%.Gained is consolidated
Body is redissolved in 4mL anhydrous methanol, and sodium methoxide is added, and maintains solution system pH7.5.Thin-layer chromatography detection (ethyl acetate:: first
Alcohol: water=4:2:1:0.2) reaction, after the reaction was completed, by silica gel rapid column chromatography (ethyl acetate: methanol: water=2:1:
0.05) it is isolated and purified, obtains compound as white solid 541mg, yield 42%.Structural parameters:1H NMR(600MHz,D2O)
δ 5.27 (s, 1H), 4.83 (s, 1H), 4.54 (d, J=8.0Hz, 1H), 4.33-4.23 (m, 2H), 4.15 (s, 1H), 4.00
(d, J=11.2Hz, 1H), 3.95-3.66 (m, 10H), 3.60-3.39 (m, 9H), 2.56 (dd, J=12.0,6.0Hz, 1H),
2.00 (s, 3H), 1.98 (s, 3H), 1.89-1.84 (m, 2H), 1.76 (t, J=12.2Hz, 1H)13C NMR(151MHz,
D2O)δ175.00,174.51, 165.99,103.61,97.16,95.55,77.43,73.39,72.59,72.21,72.08,
70.62,70.18,68.60, 67.56,67.19,64.88,63.18,62.37,61.08,58.93,51.52,48.51,
48.12,39.26,27.88, 21.95,21.90.HRMS(ESI)m/z calcd for[C28H46N5O18,M+H]+,
740.2838；found, 740.2847.
Embodiment 6
The synthesis of embodiment 6
3- nitrine propyl oxygen-[(5- acetylaminohydroxyphenylarsonic acid 3,5- dideoxy-D- glycerol-α-nine carbon onosic acid of pyrans)-(2 → 3)-
Fluoro- oxygen-β-D- galactopyranosyl the glycosyl-(1 → 3) of -6- deoxidation -6-]-oxygen -2- acetylaminohydroxyphenylarsonic acid 2- deoxidation-α-D- galactopyranose
Glycosides-(1 " → 4 ')-lactone
Into 10mL round-bottomed flask, compound 2 (72mg), aceticanhydride (1mL), pyridine (2mL) is added, is stirred at room temperature under ice bath
12h.Thin-layer chromatography detects (petroleum ether: ethyl acetate=1:10), and after fully reacting, 1mL toluene, revolving are added into reaction solution
Concentration, is repeated 3 times.Later, silica gel is admixed, is carried out by silica gel rapid column chromatography (petroleum ether: ethyl acetate :=1:50)
It isolates and purifies, obtains full acetylated intermediate, white solid 93mg, yield 95%.It is all redissolved later in 3mL anhydrous methanol,
Sodium methoxide is added, maintains solution system pH7.5.Thin-layer chromatography detects (EtOAc:MeOH:H2O:AcOH=8:2:0.5:0.25,
V/v it) reacts, after the reaction was completed, is separated by silica gel rapid column chromatography (ethyl acetate:: methanol: water=2:1:0.05)
Purifying obtains the white solid 29mg of compound 6, yield 44%.
Structural parameters:1H NMR(400MHz,D2O) δ 5.36 (d, J=3.6Hz, 1H), 4.92 (d, J=4.0Hz, 1H),
4.68 (d, J=8.0Hz, 1H), 4.42-4.34 (m, 2H), 4.24 (d, J=2.8Hz, 1H), 4.15-3.97 (m, 3H), 3.91
(dd, J=5.6,2.8Hz, 2H), 3.85-3.75 (m, 4H), 3.72-3.43 (m, 7H), 2.67 (dd, J=12.8,4.4Hz,
1H),2.17(s,3H),2.05(s,6H),1.98–1.88(m, 3H).13C NMR(101MHz,D2O)δ175.09,174.62,
(173.26,169.65,104.49,99.15,97.33,82.32 d, J=168.67Hz), 78.23,73.40,72.99,
71.36 (d, J=20.2Hz), 70.80,70.70,70.32 (d, J=6.06Hz), 69.25,68.71,68.14,67.37,
65.15,63.13,61.28,53.64, 51.62,48.58,48.34,38.30,28.01,22.16,22.14,20.12.19F
NMR(376MHz,D2O)δ -232.09.HRMS(ESI)m/z calcd for[C28H43FN5O17,M-H]-,740.2638；
found, 740.2660.
3- nitrine propyl oxygen-[(5- hydroxyl acetylaminohydroxyphenylarsonic acid 3,5- dideoxy-D- glycerol-α-nine carbon onosic acid of pyrans)-(2 →
3) the fluoro- oxygen-β-D- galactopyranosyl glycosyl-(1 → 3) of -6- deoxidation -6-]-oxygen -2- acetylaminohydroxyphenylarsonic acid 2- deoxidation-α-D- galactopyranosyl
Glucosides-(1 " → 4 ')-lactone
Into 10mL round-bottomed flask, compound 3 (102mg), aceticanhydride (2.5mL), pyridine (5.0 mL), condition of ice bath is added
Lower reaction overnight.Thin-layer chromatography detects (petroleum ether: ethyl acetate=1:10), and after fully reacting, 1mL first is added into reaction solution
Benzene, concentrated by rotary evaporation are repeated 3 times.Later, admix silica gel, by silica gel rapid column chromatography (petroleum ether: ethyl acetate :=1:
50) it is isolated and purified, obtains full acetylated product, white solid 127mg, yield 92%.It all redissolves later anhydrous in 4mL
Sodium methoxide is added in methanol, maintains solution system pH7.5.Thin-layer chromatography detects (EtOAc:MeOH:H2O:AcOH=8:2:0.5:
0.25, v/v) it reacts, after the reaction was completed, is carried out by silica gel rapid column chromatography (ethyl acetate:: methanol: water=2:1:0.05)
It isolates and purifies, obtains the white solid 43mg of compound 7, yield 47%.
Structural parameters:1H NMR(400MHz,D2O) δ 5.36 (d, J=3.2Hz, 1H), 4.92 (d, J=3.6 Hz, 1H),
4.69 (d, J=7.9Hz, 1H), 4.43-4.36 (m, 2H), 4.24 (d, J=2.1Hz, 1H), 4.10-3.91 (m, 6H),
3.86-3.75 (m, 5H), 3.70-3.47 (m, 6H), 2.68 (dd, J=13.2,4.0Hz, 1H), 2.17 (s, 3H), 2.05 (s,
3H),1.99–1.89(m,3H).13C NMR(101MHz,D2O)δ 175.82,174.61,173.26,169.63,104.49,
(99.13,97.32,82.32 d, J=168.67Hz), 78.22,73.12,72.99,71.35 (d, J=20.2Hz), 70.83,
(70.69,70.28 d, J=5.05Hz), 69.25,68.71,68.04,67.14,65.13,63.08,61.27,61.07,
53.64,51.27,48.57,48.33,38.34, 28.00,22.13,20.11.19F NMR(376MHz,D2O)δ-
232.12.HRMS(ESI)m/z calcd for [C28H43FN5O18,M-H]-,756.2587found,756.2563.
3- nitrine propyl oxygen-[(5- trifluoroacetamido -3,5- dideoxy-D- glycerol-α-nine carbon onosic acid of pyrans)-(2
→ 3) the fluoro- oxygen-β-D- galactopyranosyl glycosyl-(1 → 3) of -6- deoxidation -6-]-oxygen -2- acetylaminohydroxyphenylarsonic acid 2- deoxidation-α-D- pyrans half
Lactoside-(1 " → 4 ')-lactone
Into 10mL round-bottomed flask, compound 4 (50mg), aceticanhydride (1.0mL), pyridine (2.0 mL), condition of ice bath is added
Lower reaction overnight.Thin-layer chromatography detects (petroleum ether: ethyl acetate=1:10), and after fully reacting, 1mL first is added into reaction solution
Benzene, concentrated by rotary evaporation are repeated 3 times.Later, admix silica gel, by silica gel rapid column chromatography (petroleum ether: ethyl acetate :=1:
50) it is isolated and purified, obtains full acetylated product, white solid 64mg, yield 95%.It all redissolves later anhydrous in 3mL
Sodium methoxide is added in methanol, maintains solution system pH7.5.Thin-layer chromatography detects (EtOAc:MeOH:H2O:AcOH=8:2:0.5:
0.25, v/v) it reacts, after the reaction was completed, is carried out by silica gel rapid column chromatography (ethyl acetate:: methanol: water=2:1:0.05)
It isolates and purifies, obtains the white solid 22mg of compound 8, yield 46%.
Structural parameters:1H NMR(400MHz,D2O) δ 5.37 (d, J=3.4Hz, 1H), 4.92 (d, J=3.7 Hz, 1H),
4.70 (t, J=8.2Hz, 1H), 4.43-4.33 (m, 2H), 4.24 (d, J=2.7Hz, 1H), 4.16-3.97 (m, 4H),
3.96-3.88 (m, 3H), 3.85-3.75 (m, 3H), 3.69-3.44 (m, 6H), 2.70 (dd, J=13.0,4.6Hz, 1H),
2.16(s,3H),2.07(s,3H),1.98–1.89(m,3H).13C NMR(101 MHz,D2O)δ174.60,173.26,
(169.46,104.46,99.10,97.29,82.27 J=169.18Hz), 78.20,73.02,72.57,71.31 (J=
21.21Hz), 70.78,70.67,70.25 (J=5.05Hz), 69.20,68.68,67.99,66.91,65.10,63.03,
61.24,53.65,52.11,48.54,48.29,38.28,27.97, 22.09,20.08.19F NMR(376MHz,D2O)δ-
75.50,-232.17.HRMS(ESI)m/z calcd for[C28H42F4N5O17,M+H]+,796.2512；found,
796.2533。
Claims (8)
1. a kind of sialylated TF antigen lactone, it is characterised in that: general structure is as follows:
Wherein:
R1, selected from hydrogen atom, α-or beta configuration serine residue, α-or beta configuration threonine residues, azide substitution alkyl, alkynyl
Replace alkyl, sulfydryl that alkyl, α-or beta configuration is replaced to replace one of alkyl；
R2, it is selected from one of nitrogen acetylamino, nitrogen propionamido, nitrogen trifluoroacetyl amido, nitrogen nitrine acetylamino；
R3, it is selected from one of fluorine atom, hydroxyl；
R4, it is selected from one of fluorine atom, hydroxyl；
R5, it is selected from one of fluorine atom, hydroxyl；
R6, it is selected from one of fluorine atom, hydroxyl；
R7, it is selected from one of fluorine atom, hydroxyl；
R8, it is selected from nitrogen acetylamino, nitrogen propionamido, nitrogen trifluoroacetyl amido, nitrogen nitrine acetylamino；
R9, it is selected from one of fluorine atom, hydroxyl；
R10, it is selected from one of fluorine atom, hydroxyl；
R11, it is selected from one of fluorine atom, hydrogen atom, nitrine, hydroxyl.
2. a kind of synthetic method of sialylated TF antigen lactone described in claim 1, it is characterised in that: including following step
It is rapid:
(1) " one pot of double enzyme " method sialic acid synthetase TF antigen and its fluoro analogs, " one pot of double enzyme " the method saliva are utilized
The enzyme used in acidification refers to sialic acid CMP- synzyme and sialyltransferase；
Sialylated TF antigen and its fluoro analogs shown in general formula II:
Wherein:
R1, selected from hydrogen atom, α-or beta configuration serine residue, α-or beta configuration threonine residues, azide substitution alkyl, alkynyl
Replace alkyl, sulfydryl that alkyl, α-or beta configuration is replaced to replace one of alkyl；
R2, it is selected from one of nitrogen acetylamino, nitrogen propionamido, nitrogen trifluoroacetyl amido, nitrogen nitrine acetylamino；
R3, it is selected from one of fluorine atom, hydroxyl；
R4, it is selected from one of fluorine atom, hydroxyl；
R5, it is selected from one of fluorine atom, hydroxyl；
R6, it is selected from one of fluorine atom, hydroxyl；
R7, it is selected from one of fluorine atom, hydroxyl；
R8, it is selected from one of nitrogen acetylamino, nitrogen propionamido, nitrogen trifluoroacetyl amido, nitrogen nitrine acetylamino；
R9, it is selected from one of fluorine atom, hydroxyl；
R10, it is selected from one of fluorine atom, hydroxyl；
R11, it is selected from one of fluorine atom, hydrogen atom, nitrine, hydroxyl；
(2) II compound represented of general formula is acylated using acylation reaction, synthesizes the sialylated TF containing lactone structure
Then removing acyl group is retained lactone structure, obtains chemical compounds I by antigen lactone and its fluoro derivatives；
3. the synthetic method of sialylated TF antigen lactone according to claim 2, it is characterised in that: in step (1)
The synthetic method of sialylated TF antigen and the like: by TF disaccharides or fluoro derivatives (1.0-5.0 equivalent), N- acetyl mind
Through propylhomoserin and the like (1.0-20.0 equivalent), CTP (cytidine triphosphate (CTP)) (1.0-20.0 equivalent), MgCl2(5.0-100mM)
With Tris-HCl buffer (10-500mM, pH5.0-10.5) obtained aqueous solution, sialic acid CMP- synzyme and a kind of saliva is added
It is sialylated to realize that " one pot of double enzyme " method carries out receptor for liquid acid transferase；After the reaction was completed, purifying can directly obtain saliva
It is acidified TF antigen and its fluoro analogs.
4. the synthetic method of sialylated TF antigen lactone according to claim 2, it is characterised in that: described " one pot double
The enzyme used during enzyme " method is sialylated is NmCSS and PmST1, and the reaction time is -30 hours 5 minutes.
5. the synthetic method of sialylated TF antigen lactone according to claim 2, it is characterised in that: the enzyme process closes
It is 0-37 DEG C at middle reaction temperature, revolving speed 0-240rpm；The method of shutting down of the enzyme reaction is added in equal volume into reaction
4 DEG C of methanol and cultivated 0-30 minutes at 4 DEG C.
6. the synthetic method of sialylated TF antigen lactone according to claim 2, it is characterised in that: shown in mutual-through type II
Compound, acylation reaction is carried out under condition of ice bath, wherein the use of pyridine is solvent, the acetic anhydride of 10.0-50.0 equivalent is
Acylating reagent.
7. the synthetic method of sialylated TF antigen lactone according to claim 2, it is characterised in that: Zempl é n reaction
Operation, under conditions of anhydrous methanol and sodium methoxide, pH value 7.5~9.0,0-50 DEG C of temperature progress acyl group removing.
8. sialylated TF antigen lactone described in claim 1 is preparing the preparation that interacts between albumen and small molecule, is dividing
Application in sub- probe preparation, anti-tumor vaccine preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811548160.4A CN109678913A (en) | 2018-12-18 | 2018-12-18 | A kind of synthetic method of sialylated TF antigen lactone and its fluoro analogs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811548160.4A CN109678913A (en) | 2018-12-18 | 2018-12-18 | A kind of synthetic method of sialylated TF antigen lactone and its fluoro analogs |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109678913A true CN109678913A (en) | 2019-04-26 |
Family
ID=66186781
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811548160.4A Pending CN109678913A (en) | 2018-12-18 | 2018-12-18 | A kind of synthetic method of sialylated TF antigen lactone and its fluoro analogs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109678913A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112480188A (en) * | 2020-11-04 | 2021-03-12 | 天津科技大学 | Sialyllactose glucoside and/or analog, synthetic method and application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102796144A (en) * | 2012-08-22 | 2012-11-28 | 山东大学 | Thomsen-Friedenreich (TF) antigen and TF antigen analogue and their chemoenzymatic synthesis method and use |
| WO2013166053A2 (en) * | 2012-04-30 | 2013-11-07 | Sloan-Kettering Institute For Cancer Research | Homogenous and fully glycosylated human erythropoietin |
| CN103555796A (en) * | 2013-10-24 | 2014-02-05 | 山东大学 | Synthetic method of double-sialylated tetrasaccharide |
| CN108164573A (en) * | 2017-12-20 | 2018-06-15 | 天津科技大学 | Synthetic method and its application of sialylated TF antigens and its fluoro derivatives |
-
2018
- 2018-12-18 CN CN201811548160.4A patent/CN109678913A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013166053A2 (en) * | 2012-04-30 | 2013-11-07 | Sloan-Kettering Institute For Cancer Research | Homogenous and fully glycosylated human erythropoietin |
| CN102796144A (en) * | 2012-08-22 | 2012-11-28 | 山东大学 | Thomsen-Friedenreich (TF) antigen and TF antigen analogue and their chemoenzymatic synthesis method and use |
| CN103555796A (en) * | 2013-10-24 | 2014-02-05 | 山东大学 | Synthetic method of double-sialylated tetrasaccharide |
| CN108164573A (en) * | 2017-12-20 | 2018-06-15 | 天津科技大学 | Synthetic method and its application of sialylated TF antigens and its fluoro derivatives |
Non-Patent Citations (1)
| Title |
|---|
| XIN MENG，等: "Regioselective Chemoenzymatic Synthesis of Ganglioside Disialyl Tetrasaccharide Epitopes", 《JOURNAL OF THE AMERICAN CHEMICAL SOCIETY》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112480188A (en) * | 2020-11-04 | 2021-03-12 | 天津科技大学 | Sialyllactose glucoside and/or analog, synthetic method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101657462A (en) | The Preparation Method And Their Intermediate of capecitabine | |
| Lázár et al. | Thio-click approach to the synthesis of stable glycomimetics | |
| Khedri et al. | Chemoenzymatic synthesis of sialosides containing C7-modified sialic acids and their application in sialidase substrate specificity studies | |
| Ogawa et al. | Chemical modification of the β-glucocerebrosidase inhibitor N-octyl-β-valienamine: synthesis and biological evaluation of 4-epimeric and 4-O-(β-D-galactopyranosyl) derivatives | |
| CN108164573A (en) | Synthetic method and its application of sialylated TF antigens and its fluoro derivatives | |
| JP2021512069A (en) | Plesiomonas shigeroides O51 Serotype O-antigen oligosaccharide chemical synthesis method | |
| Yi et al. | Synthesis of 4, 5-disubstituted-3-deoxy-d-manno-octulosonic acid (Kdo) derivatives | |
| JPS6345293A (en) | Sialosylceramide compound and production thereof | |
| HU196818B (en) | Process for producing syalinic acid derivatives | |
| CN109678913A (en) | A kind of synthetic method of sialylated TF antigen lactone and its fluoro analogs | |
| Graziani et al. | 2-Deoxy-disaccharide approach to natural and unnatural glycosphingolipids synthesis | |
| JP4036755B2 (en) | Method for producing hyaluronic acid or hyaluronic acid derivative | |
| CN103525888B (en) | Synthesis method of tetrasaccharide MAG antagonist | |
| Li et al. | Chemoenzymatic synthesis and biological evaluation of ganglioside GM3 and lyso-GM3 as potential agents for cancer therapy | |
| WO1996005211A1 (en) | Ganglioside gm3 analog having sialic acid residue fluorinated at the 9-position and intermediate therefor | |
| Zheng et al. | Synthesis of N-modified ganglioside GM3 derivatives | |
| CN111233949A (en) | Ganglioside GM3 and/or its analogue, synthetic method and application | |
| EP0882733B1 (en) | Acylating agent and a method of producing thereof | |
| Kondo et al. | Efficient synthesis of 2α-glycoside of N-acetylneuraminic acid via phenylsulfenyl chloride adduct of 2-deoxy-2, 3-dehydro-N-acetylneuraminic acid methyl ester tetra-O-acetate | |
| JP2009526031A (en) | Preparation of thiosaccharides and their use | |
| KR101259648B1 (en) | A manufacturing process of 2′,2′-difluoronucloside and intermediate | |
| Agnihotri et al. | Synthesis of a di-and a trisaccharide related to the O-antigen of Escherichia coli O83: K24: H31 | |
| Chen et al. | A concise and practical synthesis of antigenic globotriose, α-d-Gal-(1→ 4)-β-d-Gal-(1→ 4)-β-d-Glc | |
| Štěpánek et al. | A short and efficient synthesis of α-C-(1→ 3)-linked disaccharides containing deoxyhexopyranoses | |
| NO169590B (en) | ANALOGY PROCEDURE FOR THE PREPARATION OF NEW THERAPEUTIC-ACTIVE SIALOSYLYL GYCERIDES |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20190426 |
|
| WD01 | Invention patent application deemed withdrawn after publication |