CN1814614A - Nucleic acid, peptide nucleicacid derivatives and their use - Google Patents

Nucleic acid, peptide nucleicacid derivatives and their use Download PDF

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CN1814614A
CN1814614A CN 200510007307 CN200510007307A CN1814614A CN 1814614 A CN1814614 A CN 1814614A CN 200510007307 CN200510007307 CN 200510007307 CN 200510007307 A CN200510007307 A CN 200510007307A CN 1814614 A CN1814614 A CN 1814614A
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group
base
replaces
methyl
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刘克良
何军林
梁远军
魏霞
褚征
张迪
徐亮
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Institute of Pharmacology and Toxicology of AMMS
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Abstract

The invention relates to nucleic acid and peptide nucleic acid derivative, their preparation method, product, and use in medicine and biology field.

Description

Nucleic acid, peptide nucleic acid derivatives and their purposes
Invention field
The present invention relates to nucleic acid and peptide nucleic acid derivatives, their preparation method contains their product, and their purposes in medicine, biological field.
Background technology
Utilize the compound of synthetic to simulate structure and the function of biomacromolecule, the biomacromolecule that especially has specific function is to further understanding with illustrate their effect and mechanism in vivo and all have important scientific meaning.Comprising the interaction between macromole-macromole, the macromole-small molecules, or even the interaction between the molecular complex.As the interaction of enzyme-to-substrate, antibody and antigenic interaction, interaction of acceptor and medicine (comprising chemical medicine and biological medicine) or the like.Based on the result of study of molecular mechanism of action, and then utilize specific function to develop its purposes to have crucial meaning at life science.
The structures shape function.For a biomacromolecule, accurate higher structure determines the biological function that it is complicated.With the synthetic biomacromolecule with specific three dimensional structure of chemical process is very difficult, as a rule, almost can't realize in the test.Therefore, with traditional chemical idea and synthetic method, it is very little to simulate the macromolecule interaction feasibility from the level of three-dimensional structure.The generation of intermolecular information transmission and biological function is to produce by the interaction between the chemical group of molecular surface.Wherein containing extremely complicated structure-function relationship, often is again multidigit point, multi-functional problem.Thereby simulate its function with simple small molecules and then too oversimplify, almost do not reach intended purposes.
According to a certain particular organisms macromole three-dimensional structure, make up and keep a specific three-dimensional frame structure, corresponding chemical functional base is installed on specific site, and is kept accurate relative tertiary location and orientation between each functional group, just may produce function corresponding.
Design new three-dimensional frame structure with a kind of chemistry assembling new ideas, specific region in skeleton construction, introduce the chemical functional base (especially residue of protein) of a group or a series of special properties, their positions in skeleton construction, space distance and spatial orientation all are can design and realize by re-set target.People claim this zone for intending the bioactive functions district.Have the higher structure of intending the bioactive functions zone and can be used to simulate the biomacromolecule function, and between research biomacromolecule-biomacromolecule, between biomacromolecule-small molecules, the interaction between biomacromolecule-molecular complex.
These three-dimensional structure frameworks can be different chemical entities, can be individual molecules, also can be that differing molecular is with multi-form bonded mixture.In principle, these three-dimensional framework structures can predict, controlled, then can be used as the skeleton construction of selection.For example, the various higher structures of oligonucleotide can be realized by design base sequence and accurate basepairing rule, as form the double stranded helix structure, the triple helix structure, the parallel double spirane structure, four strands of spirane structures, and based on " Y " font structure of secondary structure, " ten " font structure etc.The principle that forms the accurate three-dimensional skeleton construction is different basepairing rule, as: W-C base pairing, Hoogsteen base pairing etc.
The skeleton that design is in the present invention adopted has two kinds, and a kind of is nucleic acid backbone, and another kind is the peptide nucleic acid(PNA) skeleton.The accurate identification that forms spirane structure all depends on the nucleic acid base pairing, forms the internal structure of skeleton.As for using which kind of spirane structure, then can design the oligonucleotide sequence according to research object, under different condition, form the three-dimensional spiral structure of expection.The chemical functional base of bearing analog functuion is connected on main chain or the base, but does not influence specificity and stability that base pairing forms.Therefore, on a certain construction unit, introduce which kind of chemical functional base, and behind the synthetic oligomer and then the position on spirane structure, can accurately locate according to the character and the position in the oligonucleotide sequence thereof of nucleic acid base on this element, in other words, the position of construction unit in three-dimensional frame structure just determined the position of specific function base.For example: the structural unit that contains A can match with the T on the duplex complementary strand, has the imidazoles residue on the A construction unit if contain, and we just can accurately know the position of imidazolyl in spiral.The rest may be inferred, and we can be incorporated into different aminoacids residue or different chemical group the specific position of spirane structure, again according to character, position, distribution and the spatial orientation of all kinds of groups on the surface, and the interaction between the research biomacromolecule.These effects can be: protein-protein, protein and nucleic acid or other biomacromolecule, various dissimilar intermolecular interactions such as protein and various small molecules.From other angle, the common protein and the effect of other molecule have: the interaction of enzyme-to-substrate (small molecules and macromole), antibody and antigenic interaction, interaction of acceptor and medicine (comprising chemical medicine and biological medicine) or the like.
Summary of the invention
To the effect that of the present invention: as 1, to design new three-dimensional frame structure, and then study interaction and simulation biomacromolecule function between biomacromolecule, molecular complex, the small molecules with a kind of chemistry assembling new ideas.2, design, synthetic a series of nucleic acid or its analogue construction units that have all kinds of difference in functionality bases, as: contain all types of functionality bases of protein side chain residue, in other words, the synthetic construction unit that contains all kinds of natural amino acid side chain residues, then, 3, again these construction units are pressed the synthetic oligomer of implementation sequence, press the three-dimensional framework that basepairing rule forms expection, can predict on the skeleton of spirane structure at these, the relative position of each amino acid side chain functional group and spatial orientation all can accurately be located.In theory, this design concept can be used to simulate the active function group structural pattern of any biomacromolecule, and then shows different biological functions and further study molecular interaction.
The site of introducing amino-acid residue in the nucleotide structure unit has three, lays respectively at ribose, phosphoric acid and base; The position of introducing amino-acid residue in the peptide nucleic acid sequence is respectively main chain and base, studies their character and biological function.
Usually, the interaction between enzyme and part mainly contains hydrogen bond, electrostatic interaction, and hydrophobic interaction.These interactions make proton and charge transfer take place between each functional group.Main functional group has: hydroxyl, amino, sulfydryl, carboxyl, carbonyl, amide group, ester group, urea groups, guanidine radicals, oximido, imidazolyl, indyl isopolarity group and nonpolar alkyl, aromatic base etc.Again according to the kind and relative three-dimensional space position of the active centre functional group of enzyme, on higher structure total arrangement, adjust these functional group kinds, relative tertiary location and orientation just may reach the purpose of the various enzymes of simulation.
The nucleic acid that the discovery that ribozyme and RNA interfere has convincingly demonstrated certain constructional feature has enzyme catalysis really.Further research shows that also the catalytic mechanism of nucleic acid is identical with proteolytic enzyme in itself, and the hydrogen bond that all relates between functional group forms the transfer of proton or electric charge, and hydrophobic interaction etc., therefore, main points of the present invention also are applicable to ribozyme, in other words, ribozyme is an illustration of the present invention.
The objective of the invention is to design amino acid whose side chain functional group is introduced nucleic acid or peptide nucleic acid sequence, synthetic new nucleic acid analog and peptide nucleic acid(PNA) analogue sought the new type functional macromole with protein enzyme catalysis function.
The inventor's further investigation and bibliographical information all show, five and 7 of 8-nitrogen-7-denitrification VITAMIN B4 of pyrimidine, do not participate in the formation of hydrogen bond between base, substituting group in these two positions stretches to the major groove district of nucleic acid spirane structure, and some substituting groups have the effect of the stability that improves spirane structure.Transformation on the ribose-phosphoric acid skeleton of nucleic acid also helps the improvement to properties of nucleic acids, as improving the stability to hydrolysis of nuclease and the saturating film ability of cell membrane, and improves pharmacokinetic property etc.
The inventor finds after deliberation, by synthesis type I, II
Figure A20051000730700121
The nucleotide sequence that contains formula V that obtains can form secondary structure with the complementary dna sequence dna with the peptide nucleic acid sequence that contains formula VI.
Therefore contain the nucleotide sequence of formula V and contain diagnosis and the treatment that the peptide nucleic acid sequence of VI can be used to simulate biomacromolecule, detection reagent, disease.
A first aspect of the present invention relates to the nucleoside analog of formula I and formula II,
Among its Chinese style I and the II, B is natural base (VITAMIN B4, guanine, uracil, thymus pyrimidine, cytosine(Cyt), xanthine, xanthoglobulin or the like), contains the natural base of protecting group, or shown in formula III, IV the non-natural base,
Figure A20051000730700131
In formula III and IV, introduce substituting group-X-R five of uracil and seven of 8-nitrogen-7-denitrification purine 1, wherein, X can be various connecting arm forms, as (CH 2-) n, (CH=CH-) n, (C ≡ C-) n,-O-,-C (O)-,-OC (O)-,-C (O) O-,-NH-, N (R 1) 2-,-C (O) NH-,-NHC (O)-,-CH=N-,-S-,-S (O)-,-S (O) 2-,-S (O) NH-,-S (O) 2NH-, or their combination, n are 0-10,
Wherein when n is 0, promptly there is not connecting arm X, R 1Can be the contained functional group of proteinic all amino-acid residue side chains, comprising: (1) hydrophobic group: methyl (L-Ala), benzyl (phenylalanine), sec.-propyl (Xie Ansuan), 1-methylmercaptoethyl [CH 3S (CH 2) 2-] (methionine(Met)), 3-methyl-propyl [(CH 3) 2CHCH 2-] (leucine), 1-methyl-propyl [CH 3CH 2CH (CH 3)-] (Isoleucine); (2) hydrophilic radical: as methylol (HOCH 2-) (Serine), 1-hydroxyethyl [CH 3CH (OH)-] (Threonine), 4-imidazoles methyl (Histidine), the amino butyl [NH of 4- 2(CH 2) 4-] (Methionin), thiopurine methyltransferase (HSCH 2-) (halfcystine), 3-indole methyl (tryptophane), Pyrrolidine-2-(proline(Pro)), 4-hydroxybenzene methyl (tyrosine), 1-guanidine radicals propyl group (arginine), carboxamide methyl (l-asparagine), 1-carboxamide ethyl (glutamine), carboxymethyl (aspartic acid), 1-propyloic (L-glutamic acid);
When n is not 0, be connecting arm with X: (1) .R wherein 1Can be carbonatoms and be 2 to 10 alkyl and isomer thereof, comprise straight chain and branched-chain alkyl, cycloalkyl, thiazolinyl, and alkynyl, and their isomer, as methyl, ethyl, vinyl, n-propyl, sec.-propyl, allyl group, propargyl, normal-butyl, isobutyl-, the tertiary butyl, 1-(or 2-, 3-) butenyl, 1-(or 2-, 3-) butynyl, amyl group, cyclopentyl, cyclopentadienyl, hexyl, cyclohexyl, etc.; Heteroatomic (straight chain and side chain) alkyl such as nitrogenous, oxygen, sulphur, selenium, cycloalkyl, thiazolinyl, and alkynyl, its heteroatoms show as following functional group, as sulfydryl, ehter bond, thioether bond, amino, amido, guanidine radicals, urea groups, carboxyl, ester group, hydroxyl, amide group, alkylsulfonyl, sulfoamido, sulphonyl ester group, sulfinyl, sulfonamido, sulfinyl ester group, nitro, itrate group, nitroso-group, nitrous acid ester group, halogen, pseudohalogen etc.; The heterocyclic group is as, 2-(or/and 3-replaces) furyl, 2-(or/and 4-, 5-replaces) imidazolyl, 2-(or/and 3-, N-replaces) pyrryl, (or/and 4-, 5-replaces the) oxazolyl to 2-, 2-(or/and 3-replaces) thienyl, 3-(or/and 2-, 4-, 5-, 6, N-replaces) indyl, 2-(or/and 3-replaces) tetrahydrofuran base, 2-(or/and 3-, N-replaces) Pyrrolidine base, 2-(or/and 3-replaces) tetrahydro-thienyl, 2-(or/and 3-, 4-replaces) piperidyl, 2-(or/and 3-, N-replaces) morpholinyl, 2-(or/and 3-, 5-replaces) thiazolyl, 2-(or/and 3-, 5-replaces) thiazolidine base etc.; (2) .R 1The aryl radical and the assorted aryl radical that can be carbonatoms and be 6 to 20 aryl radical and assorted aryl radical and replace, as phenyl, naphthyl, anthryl, fluorenyl etc.; Assorted aryl radical is nitrogenous, oxygen, and sulphur, heteroatomss such as selenium are as 2-(or 3-, 4-) pyridyl, quinolyl, isoquinolyl.In aryl radical that replaces and assorted aryl radical, its substituting group is defined as methyl, ethyl, vinyl, n-propyl, sec.-propyl, allyl group, propargyl, normal-butyl, isobutyl-, the tertiary butyl, 1-(or 2-, 3-) butenyl, 1-(or 2-, 3-) butynyl, amyl group, cyclopentyl, cyclopentadienyl, hexyl, carbonatomss such as cyclohexyl are the alkyl of 1-10; Or contain sulfydryl, ehter bond, thioether bond, amino, amido; guanidine radicals, urea groups, carboxyl, ester group, hydroxyl; amide group, alkylsulfonyl, sulfoamido, sulphonyl ester group; sulfinyl, sulfonamido, sulfinyl ester group, nitro; itrate group, nitroso-group, nitrous acid ester group, halogen; pseudohalogen, imidazolyl, pyrryl, this class alkyl of functional groups such as indyl.
Among the formula IV, P 2Can be hydrogen, ethanoyl, benzoyl, anisoyl; isobutyryl, ortho-nitrophenyl oxygen ethanoyl, N, N dimethylamine base methene base; benzene oxygen ethanoyl, tert.-butylbenzene oxygen ethanoyl, trifluoroacetyl group is to methyl benzoyl; carbonic acid alkyl methyl esters, carbonic acid alkyl allyl ester etc., protecting group amino and imido grpup has benzoyl; isobutyryl, trifluoroacetyl group, N; the N dimethylamine methylene, N, N-dibutyl amino methylene radical.
Among the formula I, P 1Can be ethanoyl, trimethyl silicon based, diethyl is silica-based; tertiary butyl dimethyl is silica-based, and tert-butyl diphenyl is silica-based, THP trtrahydropyranyl; allyl group, uncle's fourth oxygen methyl, (adjacent nitro; to nitro, to halo, to cyano group) benzyl; trityl; the p-methoxyphenyl phenylbenzene, 4,4 '-methoxyl group trityl etc.
Among the formula I, when B is the natural base of natural base or protection, R 2Can be N, N-diisopropylamino, 4-morphine quinoline base, 1-Pyrrolidine base, 2,2,6,6-tetramethyl-piperidyl; R 3Can be-X-R 1,-NH-R 1,-N (R 1) 2(two R wherein 1Can identically also can be R 1Various combination), but do not comprise the 2-cyanoethyl, methoxyl group, oxyethyl group, N, N-diisopropylamino, N, N dimethylamine base, N, N dimethylamine base;
Among the formula I, when B is the non-natural base, R 2Can be N, N-diisopropylamino, morphine quinoline base, Pyrrolidine base, 2,2,6,6-tetramethyl-piperidyl; R 3Can be the 2-cyanoethyl ,-X-R 1,-NH-R 1,-N (R 1) 2(two R wherein 1Can identically also can be R 1Various combination).
Among the formula II, when B is the non-natural base, R 4, R 5, R 6, R 7, R 8Can be H, R 1Or-X-R 1
Among the formula II, when B is the natural base of natural base or protection, R 4, R 5, R 6, R 7, R 8Be defined as H, R 1Or-X-R 1, but be not H simultaneously; Work as R 5, R 6, R 7, R 8During for hydrogen, R 4Be not methylol and 4-imidazoles methyl;
Among the formula II, R 9Can be H or-X-R 1, R 10Can be H, tertbutyloxycarbonyl (Boc), fluorenylmethyloxycarbonyl (Fmoc), carbobenzoxy-(Cbz), benzyl or trityl;
Among the formula II, R 11Can be hydrogen, methyl, ethyl, allyl group, benzyl, 4-nitrobenzyl, 4-methyl-benzyl, 4-methoxy-benzyl;
Among the formula II, R 5With R 6, R 7With R 8, R 5With R 7, R 6With R 8, R 9/ R 10With R 5/ R 6, R 9/ R 10With R 7/ R 8Between form with (CH 2-) nThe ring texture that structure links to each other, n is 1～4; Or with main chain (N-C-C, or N-C, or C-C) form contain heteroatoms (O, S, N etc.) three to seven-membered ring shape structure, heteroatoms forms as ehter bond, thioether bond etc.
A second aspect of the present invention relates to the nucleotide sequence that contains formula V and contains the peptide nucleic acid sequence of formula VI
Among the formula V, when B is the non-natural base, R 12Can be O -, OH, S -, SH, Se -, the R among SeH or the formula I 1, when B is the natural base of natural base or protection, R 12Can be the R among the formula I 3, but be not O -, OH, S -, SH, Se -, SeH; Y can be O, S, atoms such as Se.
Among the formula VI, R 4, R 5, R 6, R 7, R 8, R 9With the R among the formula II 4, R 5, R 6, R 7, R 8, R 9Define identical.
At the nucleotide sequence that contains formula V and/or contain in the peptide nucleic acid sequence of formula VI, the number that contains formula V and/or formula VI can be at 1～50.
A third aspect of the present invention relates to I, II, and III, IV, the arbitrary combination of V and VI one of them or they is used as simulation biomacromolecule, detection reagent, medical diagnosis on disease and medicine.
The abbreviation of Shi Yonging in the present invention has following implication:
The Boc-tertbutyloxycarbonyl
The Bom-benzyloxymethyl
DCC-N, the N-dicyclohexylcarbodiimide
The DCM-methylene dichloride
The DIEA-diisopropylethylamine
DMF-N, dinethylformamide
The DMSO-methyl-sulphoxide
DMT-4,4 '-dimethoxytrityl
DMT-T-5 '-(4,4 '-dimethoxy)-triphen
Methyl-thymus nucleoside
ESI-MS-electro-spray ionization mass spectrum
The His-Histidine
The gel electrophoresis of PAGE-polyacrylamide
TBDPS-tert-butyl diphenyl silylation
The TFA-trifluoracetic acid
Tol-is to methyl benzoyl
Base prepares with the synthetic reaction scheme 1 and 2 of pressing of the construction unit of hydroxyl modified:
Reaction scheme 1:
(i) hexamethyldisilazane, ammonium sulfate, cuprous iodide/CHCl 3(ii) TBDPS-Cl, imidazoles/CHCl 3(iii) sodium methylate/methyl alcohol; (iv) DMT-Cl/ pyridine; (v) β-cyanoethyl-monochloro-N, N-di-isopropyl-phosphoramidite, DIEA/ methylene dichloride
Reaction scheme 2:
(i) hexamethyldisilazane, ammonium sulfate, cuprous iodide/CHCl 3(ii) sodium methylate/methyl alcohol; (iii) DMT-Cl/ pyridine; (iv) β-cyanoethyl-monochloro-N, N-di-isopropyl-phosphoramidite, DIEA/ methylene dichloride
In reaction scheme 1 and the route 2; the hydroxyl protecting group of base should make a distinction with the DMT protecting group of ribose; the former should stablize in follow-up synthesis step and DNA solid phase synthesis (under the acidic conditions), and can be at relatively mild alkalescence (ammoniacal liquor, 50 ℃) conditionally complete deprotection behind the solid phase synthesis.Therefore, in synthetic route of the present invention design,, protect hydroxyl on the base with the tert-butyl diphenyl silylation at 5 ' of ribose also not before the deprotection.Then, will slough 5 ' and 3 ' hydroxyl of methyl benzoyl respectively with the DMT protection and carry out phosphorus acylatedly, it is synthetic that the construction unit that is obtained can be used for solid phase DNA.
Base contains the synthetic of carboxyl function base construction unit and can prepare by reaction scheme 3:
Reaction scheme 3:
(i) hexamethyldisilazane, ammonium sulfate, cuprous iodide/CHCl 3(ii) sodium methylate/methyl alcohol;
The (iii) methanol solution of ammonia; (iv) DMT-Cl/ pyridine; (v) β-cyanoethyl-monochloro-N, N-di-isopropyl-phosphoramidite, DIEA/ methylene dichloride
Reaction scheme 3 is in order to introduce carboxyl and amide group on the DNA spirane structure; the present invention at first introduces ester group at 5 of uracil with suitable connecting arm; at glycosylation reaction, behind the DMT of 5 ' hydroxyl protection and 3 ' hydroxyl phosphorus acylated, be used for the solid phase synthesis of DNA.Then, handle dna sequence dna,, or, ester group is converted into amide group, so just on the DNA spirane structure, introduced the important function base respectively with the methanol solution processing dna sequence dna of ammonia with hydrolysis of ester group position carboxyl with the methanol/water solution of sodium hydroxide.
Base contains the synthetic of imidazoles functional group construction unit and can prepare by reaction scheme 4:
Reaction scheme 4:
(i) 0.1M Li 2PdCl 4(ii) TFA; (iii) DMT-Cl/ pyridine;
(iv) β-cyanoethyl-monochloro-N, N-di-isopropyl-phosphoramidite, DIEA/ methylene dichloride
In the reaction scheme 4, introduced the imidazolyl that participates in many catalytic activity of proteinase center 5 of uracil, it both can be used as hydrogen bond donor, can be used as hydrogen-bond donor again, played the effect of transmitting hydrogen bond at the catalytic center of Chymotrypsin.At first press the 4-vinyl imidazole of the synthetic trityl as protecting group of literature method; carry out the nucleoside analog that condensation reaction obtains expecting with 5-chloromercuri-2 '-deoxidation urea glycosides then; and with the trityl on the imidazole ring after removing under the acidic conditions; follow-up DMT protection and phosphorus acylated program are routinely carried out, and obtain being suitable for the construction unit of DNA solid phase synthesis.
Base contains the synthetic of hydrophobic group construction unit and can prepare by reaction scheme 5:
Reaction scheme 5:
Figure A20051000730700201
(i) hexamethyldisilazane, ammonium sulfate, cuprous iodide/CHCl3; (ii) phenylethylamine, DCC;
(iii) sodium methylate/methyl alcohol; (iv) DMT-Cl/ pyridine; (v) β-cyanoethyl-monochloro-N, N-di-isopropyl-phosphoramidite, DIEA/ methylene dichloride
In reaction scheme 5; the amide group of band phenyl ring is introduced uracil 5; be in order to obtain macromolecular water repellent region; the nucleosides of at first synthetic carboxylic protection; then with phenylethylamine generation amidate action; obtain required functional group, phosphorus acylated succeeded by 5 ' DMT protection and 3 ' obtains being suitable for the construction unit of DNA solid phase synthesis.
The phosphorous acid ester chain contains synthetic can the preparation by reaction scheme 6 and reaction scheme 7 of construction unit of functional group:
Reaction scheme 6:
R is: fluorenes methoxy carbonyl oxygen ethyl 25
3-methyl-butyl 26
Styroyl 27
Fluorenes methoxy carbonyl oxygen-3-oxygen propyl group 28
Fluorenes methoxy carbonyl oxygen-4-oxygen-butyl 29
Methoxy carbonyl propyl group 30
(i) alcohol compound of introducing group, pyridine/DCM ,-78 ℃; (ii) Diisopropylamine ,-40 ℃; (iii) DMT-T, DIEA
Reaction scheme 7:
(i) Diisopropylamine/DCM ,-78 ℃; (ii) DMT-T, DIEA ,-40 ℃;
(iii) tetrazole, 2-[1-(2, the 4-nitrophenyl)-1H-imidazolyl-4-]-ethanol
In reaction scheme 6 and 7, modify phosphorus by forming phosphorous acid ester with containing hydroxyl, carboxyl, hydrophobicity alkyl, phenyl ring and imidazolyl, obtain being suitable for the construction unit of DNA solid phase synthesis.
The base of peptide nucleic acid(PNA) construction unit is introduced the synthetic of hydroxyl and main chain introducing imidazolyl and can be prepared by reaction scheme 8:
Reaction scheme 8:
Figure A20051000730700221
(i) methylsulfonic acid benzyl ester/DMF, 100 ℃; (ii) ethyl bromoacetate, salt of wormwood/DMF;
(iii) DCC, N-2-tertbutyloxycarbonyl-amine ethyl glycine methyl esters or Boc-His (Bom)-OCH 3(iv) 1M lithium hydroxide
In reaction scheme 8, (iii) in, if reaction raw materials is N-2-tertbutyloxycarbonyl-amine ethyl glycine methyl esters, then obtain construction unit 34, if reaction raw materials is Boc-His (Bom)-OCH 3, then obtaining construction unit 35, by this reaction scheme, can on base, modify, also can on main chain, modify, the construction unit that obtains can be directly used in the synthetic of peptide nucleic acid(PNA).
Embodiment
The following examples are used for further specifying the present invention, but this and do not mean that any limitation of the invention.
Embodiment 1
1.1 1-(2-deoxidation-3,5-two-oxygen-to methyl benzoyl-β-D-erythro-ribose)-5-(2-hydroxyethyl)-1H, 3H-pyrimidine-2,4-diketone (1)
(4g 25.64mmol) is suspended in the hexamethyldisilazane (20ml), adds small amount of ammonium sulfate, heats this mixture, and backflow is spent the night with 5-hydroxyethyl uracil.After remaining hexamethyldisilazane is removed in underpressure distillation, add chloroform (400ml), add 1-chloro-2-deoxidation-3 then, two pairs of methyl benzoyl ribose of 5-(12g, 30.89mmol) and cuprous iodide (5.9g, 30.89mmol), stirring is 3 hours under the room temperature.In the sodium hydrogen carbonate solution that this mixture impouring is saturated, stir 1h, tell organic layer, wash one time, use anhydrous sodium sulfate drying, concentrate after filtering and obtain syrupy liq, column chromatography is isolated the colorless solid product, and 11.5g, productive rate are 88.5%.R f(CH 2Cl 2/CH 3OH?20∶1)0.35。 1H?NMR(d6-DMSO)：2.26(t，J＝6.7Hz，5-CH 2CH 2O)，2.39(2s，CH 3O)，2.54(m，2’-H)，3.41(m，5-CH 2CH 2O)，4.47-4.59(m，4’H，5’-H)，5.59(m，3’-H)，6.28(t，J＝7.6Hz，1’-H)，7.47(s，6-H)，7.34，7.89(2m，Ph)，11.33(s，NH)。
1.2 1-(2-deoxidation-3,5-two-oxygen-to methyl benzoyl-β-D-erythro-ribose)-5-(2-phenylbenzene tertiary butyl silica ethyl)-1H, 3H-pyrimidine-2,4-diketone (2)
(9g 17.7mmol) is suspended in 0.2M sodium methylate/methanol solution compound 1, reacts 3h under the room temperature.Reaction solution with in the Glacial acetic acid and after, concentrate, silica gel column chromatography is isolated product, is colourless crystallization, 4.5g, productive rate are 93.4%.Rf(CH 2Cl 2/CH 3OH?9∶1)0.27。 1H?NMR(d6-DMSO)：2.10(m，2’-H)，2.35(m，5-CH 2CH 2OH)，3.47(m，5-CH 2CH 2O)，3.54(m，5’-H)，3.77(m，4’H)，4.23(m，3’-H)，4.50(t，J＝5.6Hz，5-CH 2CH 2OH)，5.19(d，J＝4.2Hz，3’-OH)，4.94(t，J＝5.2Hz，5’-OH)，6.16(t，J?6.9，1’-H)，7.65(s，6-H)，11.20(s，NH)。
1.3 1-(β-D-erythro-2-deoxidation-ribose)-5-(2-phenylbenzene tertiary butyl silica ethyl)-1H, 3H-pyrimidine-2,4-diketone (3)
To compound 1 (12.6g, add in methylene dichloride 24.8mmol) (50ml) solution imidazoles and tert-butyl diphenyl chlorosilane (7.2ml, 27.5mmol), with this reaction mixture in stirring at room half an hour.After this reaction solution usefulness methylene dichloride (100ml) dilution, with 1M hydrochloric acid and water difference washed twice, use anhydrous sodium sulfate drying then, the pressure reducing and steaming solvent obtains the syrupy shape product, is directly used in next step deprotection reaction.
Above product is dissolved in 0.1M sodium methylate/methanol solution, at room temperature stirs 1h.Reaction solution neutralizes with Glacial acetic acid, and concentrates, and product obtains blister colorless solid (11.4g) through column chromatography purification, and productive rate is 88%.R f(CH 2Cl 2/CH 3OH?9∶1)0.54。Ultimate analysis: C 27H 34N 2O 6Si (M 510.65), theoretical value: C 63.50, H 6.71, and N 5.49; Measured value: C 63.06, H 6.56, and N 5.40. 1H?NMR(d6-DMSO)：0.96(s，tert-Bu)，2.00(m，2’-H)，2.48(m，superimposed，5-CH 2CH 2O)，3.55(dd，5-CH 2CH 2O)，3.71-3.78(m，4’-H，5’-H)，4.22(m，3’-H)，5.03(t，J＝5.2Hz，5’-OH)，5.23(d，J＝4.2Hz，3’-OH)，6.17(t，J＝7.0Hz，1’-H)，7.75(s，6-H)，7.38-7.58(m，Ph)，11.28(s，NH)。
1.4 1-[β-D-erythro-2-deoxidation-ribose-5-(4,4 '-dimethoxytrityl)]-5-(2-phenylbenzene tertiary butyl silica ethyl)-1H, 3H-pyrimidine-2,4-diketone (4)
With compound 3 (5.6g, 11mmol) steam altogether 3 times, make solvent with anhydrous pyridine (10ml) then, in this solution, add 4 with anhydrous pyridine, 4 '-dimethoxytrityl chlorine (4.87g, 14.37mmol), behind the 1.5h, add 2ml methyl alcohol, stirred 5 minutes, reaction solution concentrates and is used for column chromatography, obtains colourless blister solid product 8.86g, productive rate 98.6%.R f(CH 2Cl 2/CH 3OH?20∶1)0.44。Ultimate analysis: C 48H 52N 2O 8Si (M 813.02), theoretical value: C 70.91, H 6.45, and N 3.45; Measured value: C 70.83, H 6.40, and N 3.58. 1H?NMR(d6-DMSO)：0.91(s，tert-Bu)，2.09(m，2’-H)，2.27(t，J＝1.6Hz，5-CH 2H 2O)，3.17(d，J＝4.5Hz，5-CH 2CH 2O)，3.65(m，5’-H)，3.70(s，OCH 3)，3.88(m，4’H)，4.21(m，3’-H)，5.34(d，J＝4.5Hz，3’-OH)，6.16(t，J＝6.7Hz，1’-H)，6.85-7.53(s，6-H；m，Ph)，11.34(s，NH)。
1.5 1-[β-D-erythro-2-deoxidation-ribose-5-(4,4 '-dimethoxytrityl)]-5-(2-phenylbenzene tertiary butyl silica ethyl)-1H, 3H-pyrimidine-2,4-diketone 3 '-[(2-cyanoethyl)-N, N-diisopropylphosphoramidite] (5)
Under stirring at room, (1.8g adds diisopropyl ethyl amine (2.5ml) in dichloromethane solution 2.21mmol), drip β-cyanoethyl-monochloro-N then, and N-di-isopropyl-phosphoramidite transforms fully up to reactant to compound 4.Compound of reaction is diluted with methylene dichloride, and respectively wash once with 5% sodium hydrogen carbonate solution and saturated brine, behind anhydrous sodium sulfate drying, concentrate and obtain colourless amorphous solid 1.33g, productive rate is 59%.R f(CH 2Cl 2/CH 3COCH 3?20∶1)0.65，0.75。 31P?NMR(CDCl 3)：149.06，149.66； 1H?NMR(CDCl 3)：0.99(s，tert-Bu)，1.07，1.17[d，J＝6.7Hz，N(CH(CH 3) 2)]，2.11-2.64(m，2’-H，5-CH 2CH 2，NCCH 2CH 2O)，3.30-3.78(m，5-CH 2CH 2，NC?CH 2CH 2O，5’-H)，3.74(2s，OCH 3)，4.17(m，4’H)，4.52(m，3’-H)，6.28(t，J＝6.9Hz，1’-H)，6.79-7.56(s，6-H；m，Ph)。
Embodiment 2
(2.15-3-tert-butyl diphenyl silica propyl group)-uracil (6)
According to embodiment 1.3 methods, in methylene dichloride (20ml), with 5-(3-three hydroxypropyls) uracil (1.5g, 8.81mmol), imidazoles (0.66g, 9.7mmol), tert-butyl diphenyl chlorosilane (2.56ml, 9.8mmol) obtaining product 3-tert-butyl diphenyl silica propyl group uracil, 3.4g, productive rate are 94.4%.R f(CH 2Cl 2/CH 3OH?20∶1)0.15。Ultimate analysis: C 23H 28N 2O 3Si (M 408.57), theoretical value: C 67.61, H 6.91, and N 6.86; Measured value: C 67.56, H 6.78, and N 6.74. 1H?NMR(d6-DMSO)：0.99(s，tBu)，1.69(m，5-CH 2CH 2CH 2)，2.25(t，J＝7.3Hz，5-CH 2CH 2CH 2)，3.62(t，J＝6.3Hz，5-CH 2CH 2CH 2)，7.15(s，6-H)，7.45，7.61(2m，Ph)，11.01，10.63(s，2NH)。
2.2 1-(2-deoxidation-3,5-two-oxygen-to methyl benzoyl-β-D-erythro-ribose)-5-(2-phenylbenzene tertiary butyl silica propyl group)-1H, 3H-pyrimidine-2,4-diketone (7)
Press embodiment 1.1 methods, with 5-(3-tert-butyl diphenyl silica propyl group uracil (2.4g, 5.88mmol) with behind the hexamethyldisilazane generation Silanization reaction, in chloroform (300ml) with 1-chloro-2-deoxidation-3, two pairs of toluyls of 5--D-ribose (3g, 7.06mmol), CuI (2g), reaction.Obtain colorless solid product 2.45g through column chromatography, productive rate is 54.8%.R f(CH 2Cl 2/CH 3OH?20∶1)0.59。Ultimate analysis: C 44H 48N 2O 8Si (M 760.95), theoretical value: C 69.45, H 6.13, and N 3.68; Measured value: C 69.18, H 6.11, and N 3.88. 1H?NMR(d6-DMSO)：0.96(s，tert-Bu)，1.58(m，5-CH 2CH 2CH 2)，2.11(t，J＝7.4Hz，5-CH 2CH 2CH 2)，2.29，2.40(2s，CH 3)，2.57(m，2’-H)，3.50(t，J＝6.3Hz，5-CH 2CH 2CH 2)，4.50-4.63(m，4’-H，5’-H)，5.62(m，3’-H)，6.30(t，J＝7.1Hz，1’-H)，7.24-7.94(m，6-H，Ph)，11.40(s，NH)。
2.3 1-(β-D-erythro-2-deoxidation-ribose)-5-(2-phenylbenzene tertiary butyl silica propyl group)-1H, 3H-pyrimidine-2,4-diketone (8)
With compound 6 (2.38g, 3.13mmol) be dissolved in sodium methylate/methanol solution (0.2M, 50ml) in, at room temperature react 3hr, reaction solution neutralizes with Glacial acetic acid, concentrates then to be used for the normal pressure column chromatography, product is a colorless solid, 1.15g, productive rate are 63%.R f(CH 2Cl 2/CH 3OH9∶1)0.68。Ultimate analysis: C 28H 36N 2O 6Si.0.5H 2O (M 533.68), theoretical value: C62.96, H 6.93, and N 5.25; Measured value: C 62.70, H 6.90, and N 5.09. 1H?NMR(d6-DMSO)：0.99(s，tert-Bu)，1.72(m，5-CH 2CH 2CH 2)，2.04(m，2’-H)，2.31(m，5-CH 2CH 2CH 2)，3.54(m，5-CH 2CH 2CH 2)，3.64(m，5’-H)，3.77(m，4’-H)，4.22(m，3’-H)，5.01(t，J＝5.2Hz，5’-OH)，5.23(d，J＝4.2Hz，3’-OH)，6.16(t，J＝6.9Hz，1’-H)，7.40-7.62(m，Ph)，7.66(s，6-H)，11.35(s，NH)。
2.4 1-[β-D-erythro-2-deoxidation-ribose-5-(4,4 '-dimethoxytrityl)]-5-(2-phenylbenzene tertiary butyl silica propyl group)-1H, 3H-pyrimidine-2,4-diketone (9)
With compound 8 (1g 1.91mmol) with after anhydrous pyridine steams 3 times altogether, is dissolved in the anhydrous pyridine (2ml), to this solution add DMT-Cl (0.77g, 2.3mmol).After at room temperature reacting 1hr, add methyl alcohol (5ml), and continue reaction 5min.This reaction solution concentrated be used for the normal pressure column chromatography, obtain colorless solid, 1.3g, productive rate are 82.5%.R f(CH 2Cl 2/CH 3OH?20∶1)0.49。Ultimate analysis: C 49H 54N 2O 8Si (M 827.05), theoretical value: C 71.16, H 6.58, and N 3.39; Measured value: C 71.07, H 6.58, and N 3.93. 1H?NMR(d6-DMSO)：0.94(s，tert-Bu)，1.52(m，5-CH 2CH 2CH 2)，1.91(m，2’-H)，2.16(t，5-CH 2CH 2CH 2)，3.17(m，5-CH 2CH 2CH 2)，3.39(m，5’-H)，3.67(2s，OCH 3)，3.89(m，4’H)，4.29(m，3’-H)，5.34(d，J＝4.5Hz，5.34，3’-OH)，6.20(t，J＝6.9Hz，1’-H)，6.82-7.57(m，6-H，arom.H)，11.35(s，NH)。
2.5 1-[β-D-erythro-2-deoxidation-ribose-5-(4,4 '-dimethoxytrityl)]-5-(2-phenylbenzene tertiary butyl silica ethyl)-1H, 3H-pyrimidine-2,4-diketone
3 '-[(2-cyanoethyl)-N, N-diisopropylphosphoramidite] (10)
From compound 9 (1g, 1.21mmol) in the solution of the methylene dichloride that contains diisopropyl ethyl amine with β-cyanoethyl-monochloro-N, the reaction of N-di-isopropyl-phosphoramidite.Reaction solution is after diluting with methylene dichloride, with 5% sodium hydrogen carbonate solution and saturated brine washing.Dry back concentrates, and often the pressure column chromatography purifying obtains colorless solid product 0.65g, and productive rate is 52.4%.R f(CH 2Cl 2/CH 3COCH 3?20∶1)0.60，0.63。 31P?NMR(CDCl 3)：149.06，149.53； 1H?NMR(CDCl 3)：0.98(s，tert-Bu)，1.21，1.29[m，N(CH(CH 3) 2)]，1.46-2.73(m，N(CH(CH 3) 2)，5-CH 2CH 2CH 2，2’-H)，2.41(t，J＝6.5Hz，5-CH 2CH 2CH 2)，3.30-3.68(m，5-CH 2CH 2CH 2，NCCH 2CH 2O，5’-H)，3.71(2s，OCH 3)，4.17(m，4’H)，4.60(m，3’-H)，6.35(m，1’-H)，6.77-7.61(m，6-H，Ph)。
Embodiment 3
3.1 1-(β-D-erythro-2-deoxidation-ribose)-1H, 3H-pyrimidine-2,4-diketone-5-methyl propionate (12)
Press the synthetic method preparation of compound 1, obtain compound 11 after, directly carry out deprotection reaction, obtain compound 12,2.6g, yield 45.6%.R f(CH 2Cl 2/CH 3OH?9∶1)0.42。Ultimate analysis: C 13H 18N 2O 7(M 314.29), theoretical value: C 49.68, H 5.77, and N 8.91; Measured value: C 49.67, H 5.52, and N 8.47. 1H?NMR(d6-DMSO)：2.08(m，2’-H)，2.48(m，5-CH 2CH 2)，3.59(s，CH 3)，3.56(m，5’-H)，3.77(m，4’H)，4.24(m，3’-H)，5.03(t，J＝5.2Hz，5’-OH)，5.24(d，J＝4.2Hz，3’-OH)，6.16(t，J＝6.9Hz，1’-H)，7.72(s，6-H)，11.33(s，NH)。
3.2 1-(β-D-erythro-deoxidation-ribose)-1H, 3H-pyrimidine-2,4-diketone 5-propionic acid amide (13)
(0.5g, ammonia/methanol solution 1.59mmol) (70ml) places encloses container, stirs 7 days under the room temperature with compound 12.This reaction solution is concentrated into product begins to separate out, obtain colorless solid 0.45g, productive rate is 94.5%.Ultimate analysis: C12H17N3O6 (M 299.28), theoretical value: C 48.16, H 5.73, and N 14.04; Measured value: C 48.45, H 5.77, N13.66.R f(CH 2Cl 2/CH 3OH?9∶1)0.1。 1H?NMR(d6-DMSO)：2.08(m，2’-H)，2.23，2.40(2m，5-CH 2CH 2)，3.57(m，5’-OH)，3.76(m，4’H)，4.23(m，3’-H)，5.01(t，J＝5.2Hz，5’-OH)，5.23(d，J4.5，3’-OH)，6.16(t，J＝6.7Hz，1’-H)，7.25，6.77(2s，NH 2)，7.65(s，6-H)，Ph)，11.29(s，NH)。
3.3 1-[(β-D-erythro-2-deoxidation-ribose-5-(4,4 '-dimethoxytrityl))-and 1H, 3H-pyrimidine-2,4-diketone-5-methyl propionate (14)
From compound 12 (1.2g, 3.82mmol) and DMT-Cl (1.55g, 4.58mmol) in pyridine (3ml) reaction, stirred 2 hours under the room temperature, the back that reacts completely adds 2ml methyl alcohol stirred 5 minutes, and reaction solution concentrates after the normal pressure column chromatography obtains colorless solid compounds 14,1.7g productive rate is 72%.R f(CH 2Cl 2/CH 3OH?20∶1)0.41。 1H?NMR(d6-DMSO)：2.15-2.55(m，5-CH 2CH 2，2’-H)，3.20(m，5’-H)，3.52(s，CH 3)，3.73(s，OCH 3)，3.89(m，4’H)，4.29(m，3’-H)，5.34(d，J＝3.6Hz，3’-OH)，6.19(t，J＝6.7Hz，1’-H)，6.87-7.27(m，6-H，Ph)，11.40(s，NH)。
3.4 1-[(β-D-erythro-2-deoxidation-ribose-5-(4,4 '-dimethoxytrityl))-and 1H, 3H-pyrimidine-2,4-diketone 5-methyl propionate 3 '-[(2-cyanoethyl)-N, N-diisopropylphosphoramidite] (15)
From compound 14 (1.09g, 1.767mmol) and β-cyanoethyl-monochloro-N, N-di-isopropyl-phosphoramidite, diisopropyl ethyl amine (1ml) in methylene dichloride (10ml) under room temperature reaction obtain product, through with methylene dichloride dilution, 5%NaHCO 3With the salt water washing, dry and concentrated, obtain product by the normal pressure column chromatography, 0.8g, 55.6%.R f(CH 2Cl 2/CH 3COCH 38∶1)0.40，0.52。 31P?NMR(CDCl 3)：149.57，149.20； 1H?NMR(CDCl 3)：1.06-1.18[m，2CH(CH 3) 2]，2.14-2.64[m，5-CH 2CH 2，2’-H，CH(CH 3) 2]，3.31-3.86(m，5’-H，OCH 2CH 2CN)，3.57(s，CH 3)，3.79(s，OCH 3)，4.16(m，4’H)，4.63(m，3’-H)，6.37(m，1’-H)，6.82-7.59(m，6-H，Ph)。
Embodiment 4
4.1 1-(β-D-erythro-2-deoxidation-ribose)-5-[2-(1-trityl-1H-imidazoles-4-)-(E)-vinyl-1H, 3H-pyrimidine-2,4-diketone (16)
(2g, 4.3mmol), 1-trityl-4-vinyl imidazole adds 0.1M PdLi to 5-chloromercuri-2 '-deoxidation urea glycosides 2Cl 4Methanol solution (44ml), react 16h under the room temperature, in reaction mixture, charge into hydrogen sulfide (20min).Remove by filter insoluble solids, filtrate concentrating is used for the normal pressure column chromatography, obtain colorless solid, 1.1g, productive rate are 45.6%.Ultimate analysis: C 33H 30N 4O 5.H 2O (M 580.62), theoretical value: C 68.20, H 5.51, and N 9.64; Measured value: C 68.55, H 5.39, and N 8.94.R f(CH 2Cl 2/CH 3OH?9∶1)0.5。 1H?NMR(d6-DMSO)：2.15(m，2’-H)，3.55-3.66(m，5’-H)，3.79(m，4’-H)，4.26(m，3’-H)，5.13(t，J4.8，5’-OH)，5.24(d，J＝4.5Hz，3’-OH)，6.19(t，J＝6.6Hz，1’-H)，6.95(s，5”-H)，6.84，7.22(2d，J＝16.1Hz，CH＝CH)，7.12，7.40(2m，2”-H?Ph)，8.10(s，6-H)，11.40(s，NH)。
4.2 1-(β-D-erythro-2-deoxidation-ribose)-5-[2-(the 1H-imidazoles-4-)-(E)-vinyl-1H, 3H-pyrimidine-2,4-diketone (17)
With compound 16 (1g, 1.78mmol) be dissolved in the trifluoroacetic acid/dichloromethane solution, after at room temperature stirring 30min, trifluoroacetic acid is removed in underpressure distillation, with methyl alcohol residue is dissolved repeatedly and distills, isolate product with the normal pressure column chromatography method then, be colorless solid, 0.4g productive rate is 70%.R f(CH 2Cl 2/CH 3OH?5∶1)0.12。 1H?NMR(d6-DMSO)：2.15(m，2’-H)，3.63(m，5’-H)，3.80(m，4’-H)，4.28(m，3’-H)，5.18(t，J＝5.0Hz，5’-OH)，5.28(d，J＝4.2Hz，3’-OH)，6.20(t，J＝6.6Hz，1’-H)，6.76，7.31(2d，J＝15.1Hz，CH＝CH)，7.10(s，imH)，7.62(s，im-H)，8.08(s，6-H)，11.44(s，NH)，12.04(br，NH)。
4.3 1-[(β-D-erythro-2-deoxidation-ribose-5-(4,4 '-dimethoxytrityl))-and 5-[2-(the 1H-imidazoles-4-)-(E)-vinyl-1H, 3H-pyrimidine-2,4-diketone (18)
From compound 17 (0.3g, 0.937mmol) and DMT-Cl (often to obtain product be colourless amorphous solid 0.5g to the pressure column chromatography purifying for 0.38g, the 1.12mmol) reaction in pyridine (0.5ml), and productive rate is 86%, R f(CH 2Cl 2/ CH 3OH 9: 1) 0.32.MS：623.6(M+1)。 1H?NMR(d6-DMSO)：2.15-2.38(m，2’-H)，3.67(s，OCH 3)，3.91(m，4’H)，4.24(m，3’-H)，5.36(br，3’-OH)，6.21(t，J＝6.7Hz，1’-H)，6.69-7.72(m，Ar-H，CH＝CH，6-H，2”-H，5”-H)，11.55，12.04(2br，NH)。
4.4 1-[(β-D-erythro-2-deoxidation-ribose-5-(4,4 '-dimethoxytrityl))-and 5-[2-(the 1H-imidazoles-4-)-(E)-vinyl-1H, 3H-pyrimidine-2,4-diketone 3 '-[(2-cyanoethyl)-N, N-diisopropylphosphoramidite] (19)
From compound 18 (0.5g, 0.75mmol) and β-cyanoethyl-monochloro-N, N-di-isopropyl-phosphoramidite, diisopropyl ethyl amine (0.5ml) reaction obtains product, 0.37g, yield 57%.R f(CH 2Cl 2/CH 3OH?9∶1)0.64。 31P?NMR(CDCl 3)：149.25，149.63。 1H?NMR(CDCl 3)：1.16[2d，J＝4.8Hz，2CH(CH 3) 2]，2.35-2.64[m，2’-H，CH(CH 3) 2]，3.50-4.63(m，5’-H，OCH 2CH 2CN)，3.69(s，OCH 3)，4.20(m，4’H)，4.62(m，3’-H)，6.26-8.03(m，1’-H，Ar-H，CH＝CH，6-H，2”-H，5”-H)，10.31，11.33(2?br，NH)。
Embodiment 5
5.1 1-(2-deoxidation-3,5-two-oxygen-to methyl benzoyl-β-D-erythro-ribose)-1H, 3H-pyrimidine-2,4-diketone-5-propionic acid (20)
5-uracil-3-propionic acid (6g, 32.6mmol), CuI (6g) and 1-chloro-2-deoxidation-3, (15g 38.6mmol), obtains colourless powder shape solid (14g) to two pairs of methyl benzoyl ribose of 5-, and productive rate is 80%.R f(CH 2Cl 2/CH 3OH?20∶1)0.34。Ultimate analysis: C 28H 28N 2O 9.0.5H 2O (M 545.53), theoretical value: C 61.59, H 5.32, and N 5.13; Measured value: C 61.78, H 5.13, and N 5.02. 1H?NMR(d6-DMSO)：2.30(m，5-CH 2CH 2COOH)，2.38，2.40(2s，CH 3)，2.57(m，2’-H)，4.47-4.64(m，4’-H，5’-H)，5.61(m，3’-H)，6.31(t，J＝7.1Hz，1’-H)，7.51(s，6-H)，7.31-7.93(m，Ph)，11.44(s，NH)，12.13(s，COOH)。
5.2 1-(2-deoxidation-3,5-two-oxygen-to methyl benzoyl-β-D-erythro-ribose)-1H, 3H-pyrimidine-2,4-diketone-5-[(N-styroyl)-propionic acid amide] (21)
To compound 20 (1.4g, dichloromethane solution 2.61mmol) (30ml) add DCC (1.4g, 6.79mmol) and phenylethylamine (0.75ml, 6mmol), this reaction mixture at room temperature stirs and spends the night, and concentrates the back and obtains product with column chromatography, be colorless solid 0.85g, productive rate is 51%.R f(CH 2Cl 2/CH 3OH?20∶1)0.59。Ultimate analysis: C 36H 37N 3O 8(M639.69), theoretical value: C 67.59, H 5.83, and N 6.57; Measured value: C 67.70, H5.75, N 6.50. 1H?NMR(d6-DMSO)：2.28(t，J＝7.3Hz，5-CH 2CH 2CO)，2.31-2.617m，2’-H，5-NHCH 2CH 2Ph)，2.36，2.40(2s，CH 3)，2.67(t，J＝7.3Hz，5-CH 2CH 2CO)，3.24(m，5-NHCH 2CH 2Ph)，4.45-4.62(m，4’-H，5’-H)，5.58(m，3’-H)，6.29(t，J＝7.1Hz，1’-H)，7.49(s，6-H)，7.15-7.92(m，Ph，NHCO)，11.42(s，NH)。
5.3 1-(2-deoxidation-β-D-erythro-ribose)-1H, 3H-pyrimidine-2,4-diketone 5-[(N-styroyl)-propionic acid amide] (22)
Compound 21 is suspended in 0.1M sodium methylate/methanol solution (200ml), reacts 3hr under the room temperature.With Glacial acetic acid neutralization reaction liquid, use column chromatography after concentrating and obtain the colorless solid product, 1.1g, productive rate are 85.3%.R f(CH 2Cl 2/CH 3OH?9∶1)0.32。Ultimate analysis: C 20H 25N 3O 6.0.5H 2O (M 412.43), theoretical value: C 58.19, H 6.30, and N 10.18; Measured value: C 58.46, H 6.30, and N 10.04. 1H?NMR(d6-DMSO)：2.25-2.04(m，2’-H，5-NHCH 2CH 2Ph)，2.41(t，J＝7.3Hz，5-CH 2CH 2CO)，2.67(t，J＝7.3Hz，5-CH 2CH 2CO)，3.24(m，5-NHCH 2CH 2Ph)，3.57(m，5’-H)，3.76(m，4’-H)，4.23(m，3’-H)，5.03(t，J＝5.3Hz，5’-OH)，5.24(d，J＝4.2Hz，3’-OH)，6.17(t，J＝6.9Hz，1’-H)，7.16-7.29(m，Ph)，7.64(s，6-H)，7.91(t，J＝5.5Hz，NHCO)，11.32(s，NH)。
5.4 1-[(2-deoxidation-β-D-erythro-ribose-5-(4,4 '-dimethoxytrityl))-and 1H, 3H-pyrimidine-2,4-diketone 5-[(N-styroyl)-propionic acid amide] (23)
(0.8g, 2mmol) (0.81g 2.4mmol) reacts compound 22, obtains the colorless solid product, and 1.12g, productive rate are 79.4%R with DMT-Cl in anhydrous pyridine (2ml) f(CH 2Cl 2/ CH 3OH 20: 1) 0.25.Ultimate analysis: C 41H 43N 3O 8(M 705.80), theoretical value: C 69.77, H 6.14, and N 5.95; Measured value: C 69.82, H 6.10, and N 6.14. 1H?NMR(d6-DMSO)：2.12-2.28(m，2’-H，5-NHCH 2CH 2Ph，5-CH 2CH 2CO)，2.65(t，J＝7.3Hz，5-CH 2CH 2CO)，3.16-3.23(m，5’-H，5-NHCH 2CH 2Ph)，3.73(s，CH 3O)，3.86(m，4’-H)，4.22(m，3’-H)，5.32(d，J＝4.8Hz，3’-OH)，6.17(t，J＝6.7Hz，1’-H)，6.87-7.40(m，6-H，Ph)，7.79(t，J＝5.6Hz，NHCO)，11.38(s，NH)。
5.5 1-[(2-deoxidation-β-D-erythro-ribose-5-(4,4 '-dimethoxytrityl))-and 1H, 3H-pyrimidine-2,4-diketone 5-[(N-styroyl)-propionic acid amide] 3 '-[(2-cyanoethyl)-N, N-diisopropylphosphoramidite] (24)
Compound 22 (0.8g, 1.13mmol) in methylene dichloride (10ml) with β-cyanoethyl-monochloro-N, the reaction of N-di-isopropyl-phosphoramidite and diisopropyl ethyl amine (0.75ml) obtains product, is colorless solid, 0.75g, productive rate are 73.3%. 31P?NMR(CDCl 3)：149.22，149.50； 1H?NMR(CDCl 3)：1.18[m，2CH(CH 3) 2]，2.06-2.76[m，2’-H，5-NHCH 2CH 2Ph，5-CH 2CH 2CO，5-CH 2CH 2CO，CH(CH 3) 2]，3.16-3.23(m，5’-H，5-NHCH 2CH 2Ph，OCH 2CH 2CN)，3.78(s，CH 3O)，4.17(m，4’-H)，4.61(m，3’-H)，6.35(t，J＝6.9Hz，1’-H)，6.83-7.54(m，6-H，Ph)。
Embodiment 6
6.1 1-[(2-deoxidation-β-D-erythro-ribose-5-(4,4 '-dimethoxytrityl))-and 5-methyl isophthalic acid H, 3H-pyrimidine-2,4-diketone 3 '-[(2-fluorenes methoxy carbonyl oxygen ethyl)-N, N-diisopropylphosphoramidite] (25)
550mg (4mmol) phosphorus trichloride adds 5mL dry methylene chloride, 320mg (4mmol) dry pyridine respectively in reaction flask, be cooled to-78 ℃.1.14g (4mmol) fluorenes methoxy carboxylic acid-(2-hydroxyl) ethyl ester is dissolved in the 5mL methylene dichloride, slowly drips in reaction solution, stirring at room 3 hours places refrigerator overnight.Take out, be cooled to-40 ℃, drip 810mg (8mmol) Diisopropylamine, behind the low-temp reaction 3h, room temperature is placed 20h.Get 4.0g (7.3mmol) 5 '-(4,4 '-dimethoxy)-trityl-thymidine, 2.10g DIEA and 8mL methylene dichloride, inject reaction solution, stirred 4 hours under the room temperature.The saturated NaHCO of reaction solution 3, saturated NaCl solution washs secondary, anhydrous Na respectively 2SO 4Dry organic phase.Filtering Na 2SO 4, decompression removes methylene dichloride, obtains faint yellow oily thing, through silica gel column chromatography, obtains the colourless blister product of 1.10g, yield: 28.7% (by phosphorus trichloride).R f: 0..55 (DCM: acetone 9: 1); 31P-NMR (CD 3Cl), δ P: 149.12,149.81. 1H-NMR(CD 3Cl)，δ H：1.05～1.20[m，12H，-CH(CH 3) 2]，1.39(s，3H，-CH 3)，2.30～3.40[m，6H，2’-H，-CH(CH 3) 2，5’-H]，3.44～3.90[m，7H，O-CH 2-CH 2-O，O-CH 2-CH-(fluorene)]，3.78(s，6H，-OCH 3)，4.21(m，1H，4’-H)，4.65(m，1H，3’-H)，6.41(m，1H，1’-H)，6.73～7.77(m，22H，Ar-H)，8.16(br，NH)。
6.2 1-[2-deoxidation-β-D-erythro-ribose-5-(4,4 '-dimethoxytrityl)]-5-methyl isophthalic acid H, 3H-pyrimidine-2,4-diketone 3 '-[(3-fluorenes methoxy carbonyl oxygen propyl group)-N, N-di-isopropyl-phosphoramidite] (26)
722mg (5.25mmol) phosphorus trichloride adds 5mL dry methylene chloride, 416mg (5.25mmol) dry pyridine respectively in reaction flask, be cooled to-78 ℃.1.56g (4mmol) fluorenes methoxy carboxylic acid-1-(3-hydroxyl) propyl ester is dissolved in the 6mL methylene dichloride, slowly drips in reaction solution, stirring at room 3 hours places refrigerator overnight.Take out, be cooled to-40 ℃, drip 1060mg (10.4mmol) Diisopropylamine, behind the low-temp reaction 3h, room temperature is placed 20h.Get 2.8g (5.1mmol) 5 '-(4,4 '-dimethoxy)-trityl-thymidine, 2.70g (20.9mmol) DIEA and 10mL methylene dichloride, inject reaction solution, stirred 6 hours under the room temperature.The saturated NaHCO of reaction solution 3, saturated NaCl solution washs secondary, anhydrous Na respectively 2SO 4Dry organic phase.Filtering Na 2SO 4, decompression removes methylene dichloride, obtains faint yellow oily thing, through silica gel column chromatography, obtains the colourless blister product of 2.13g, yield: 41.8% (by phosphorus trichloride).R f: 0.8 (DCM: acetone 12: 1); 31P-NMR (CD 3Cl): δ P: 147.89,148.31. 1H-NMR(CD 3Cl)，δ H：1.04～1.20[m，12H，-CH(CH 3) 2]，1.40(s，3H，-CH 3)，1.66～2.00(m，2H，O-CH 2-CH 2-CH 2-O)，2.30～3.60[m，6H，2’-H，-CH(CH 3) 2，5’-H]，3.60(m，2H，P-O-CH 2-CH 2-CH 2-O)，3.77(s，6H，-OCH 3)，4.20(m，1H，4’-H)，4.20～4.40[m，5H，P-O-CH 2-CH 2-CH 2-O，O-CH 2-CH-(fluorene)]，4.64(m，1H，3’-H)，6.41(m，1H，1’-H)，6.73～7.77(m，22H，Ar-H)，8.38(br，NH)；
6.3 1-[(2-deoxidation-β-D-erythro-ribose-5-(4,4 '-dimethoxytrityl))-and 5-methyl isophthalic acid H, 3H-pyrimidine-2,4-diketone 3 '-[(4-fluorenes methoxy carbonyl oxygen-butyl)-N, N-di-isopropyl-phosphoramidite] (27)
Get 510mg (3.70mmol) phosphorus trichloride in reaction flask, add 5mL dry methylene chloride, 294mg (3.71mmol) dry pyridine respectively, be cooled to-78 ℃.1.15g (3.70mmol) fluorenes methoxy carboxylic acid-1-(4-hydroxyl) butyl ester is dissolved in the 5mL methylene dichloride, slowly drips in reaction solution, stirring at room 3 hours places refrigerator overnight.Take out, be cooled to-40 ℃, drip 750mg (7.41mmol) Diisopropylamine, behind the low-temp reaction 3h, room temperature is placed 20h.Get 2.0g (3.67mmol) 5 '-(4,4 '-dimethoxy)-trityl-thymidine, 1.90g (14.7mmol) DIEA and 10mL methylene dichloride, inject reaction solution, stirred 6 hours under the room temperature.The saturated NaHCO of reaction solution 3, saturated NaCl solution washs secondary, anhydrous Na respectively 2SO 4Drying concentrates and obtains faint yellow oily thing, through silica gel column chromatography, obtains the colourless blister product of 1.07g, yield: 41.8% (by phosphorus trichloride).R f: 0.49 (DCM: acetone 12: 1). 31P-NMR(CD 3Cl)，δ P：147.5，148.0。 1H-NMR(CD 3Cl)，δ H：1.03～1.20[m，12H，-CH(CH 3) 2]，1.39(s，3H，-CH 3)，1.66～2.00(m，4H，O-CH 2-CH 2-CH 2-CH 2-O)，2.30～3.60[m，6H，2’-H，-CH(CH 3) 2，5’-H]，3.55(m，2H，P-O-CH 2-CH 2-CH 2-CH 2-O)，3.77(s，6H，-OCH 3)，4.16(m，1H，4’-H)，4.20～4.40[m，5H，P-O-CH 2-CH 2-CH 2-CH 2-O，O-CH 2-CH-(fluorene)]，4.64(m，1H，3’-H)，6.41(m，1H，1’-H)，6.73～7.77(m，22H，Ar-H)，8.38(br，NH)。
6.4 1-[(2-deoxidation-β-D-erythro-ribose-5-(4,4 '-dimethoxytrityl))-and 5-methyl isophthalic acid H, 3H-pyrimidine-2,4-diketone 3 '-(styroyl-N, N-di-isopropyl-phosphoramidite) (28)
Get 526mg (3.83mmol) phosphorus trichloride in dry and be full of in the reaction flask of argon gas, add 5mL dry methylene chloride, 299mg (3.78mmol) dry pyridine respectively, be cooled to-78 ℃.468mg (3.83mmol) phenylethyl alcohol is dissolved in the 5mL methylene dichloride, slowly drips in reaction solution, and stirring at room 3 hours places refrigerator overnight.Take out, be cooled to-40 ℃, drip 775mg (7.66mmol) Diisopropylamine, behind the low-temp reaction 3h, room temperature is placed 20h.Get 2.0g (3.65mmol) 5 '-(4,4 '-dimethoxy)-trityl-thymidine, 2.0g DIEA and 10mL methylene dichloride, inject reaction solution, stirred 4 hours under the room temperature.The saturated NaHCO of reaction solution 3, saturated NaCl solution washs secondary, anhydrous Na respectively 2SO 4Dry organic phase.Filtering Na 2SO 4, decompression removes methylene dichloride, obtains faint yellow oily thing, through silica gel column chromatography, obtains the colourless blister product of 1.30g, yield: 42.6% (by phosphorus trichloride).R f: 0.55 (DCM: acetone 9: 1). 31P-NMR(CD 3Cl)，δ P：147.18，147.73。 1H-NMR(CD 3Cl)，δ H：1.02～1.20[m，12H，-CH(CH 3) 2]，1.40(m，3H，-CH 3)，2.20～3.90[m，10H，2’-H，O-CH 2-CH 2-Ar，-N-CH(CH 3)2，5’-H]，3.77(s，6H，-OCH 3)，4.15(d，1H，4’-H)，4.62(m，1H，3’-H)，6.42(m，1H，1’-H)，6.81～7.70(m，19H，Ar-H)，8.86(br，NH)。
6.5 1-[(2-deoxidation-β-D-erythro-ribose-5-(4,4 '-dimethoxytrityl))-and 5-methyl isophthalic acid H, 3H-pyrimidine-2,4-diketone 3 '-[(3-methyl)-butyl-N, N-di-isopropyl-phosphoramidite] (29)
Get 529mg (3.85mmol) phosphorus trichloride in reaction flask, add 5mL dry methylene chloride, 305mg (3.85mmol) dry pyridine respectively, be cooled to-78 ℃.339.5mg (3.85mmol) 3-methyl isophthalic acid-butanols is dissolved in the 5mL methylene dichloride, slowly drips in reaction solution, stirring at room 3 hours places refrigerator overnight.Take out, be cooled to-40 ℃, drip 755mg (7.50mmol) Diisopropylamine, behind the low-temp reaction 3h, room temperature is placed 20h.Get 2.0g (3.65mmol) 5 '-(4,4 '-dimethoxy)-trityl-thymidine, 1.93g DIEA and 8mL methylene dichloride, inject reaction solution, stirred 4 hours under the room temperature.The saturated NaHCO of reaction solution 3, saturated NaCl solution washs secondary, anhydrous Na respectively 2SO 4Dry organic phase.Filtering Na 2SO 4, decompression removes methylene dichloride, obtains faint yellow oily thing, through silica gel column chromatography, obtains the colourless blister product of 0.96g, yield: 33.8% (by phosphorus trichloride).R f: 0.57 (DCM: acetone 9: 1); 1H-NMR (CD 3Cl), δ H: 0.83～1.20[m, 18H ,-CH (CH 3) 2], 1.15[m, 2H ,-CH 2-CH (CH 3) 2], 1.39 (s, 3H ,-CH 3), 1.50[m, 2H ,-CH 2-CH (CH 3) 2], 2.30～3.90[m, 7H, 2 '-H, O-CH 2-CH 2,-N-CH (CH 3) 2, 5 '-H], 3.79 (s, 6H ,-OCH 3), 4.15 (d, 1H, 4 '-H), 4.65 (m, 1H, 3 '-H), 6.44 (m, 1H, 1 '-H), 6.82～7.70 (m, 14H, Ar-H). 31P-NMR(CD 3Cl)，δ P：147.08，147.50。
6.6 1-[(2-deoxidation-β-D-erythro-ribose-5-(4,4 '-dimethoxytrityl))-and 5-methyl isophthalic acid H, 3H-pyrimidine-2,4-diketone 3 '-[(3-methoxy carbonyl propyl group)-N, N-di-isopropyl-phosphoramidite] (30)
Get 425mg (3.10mmol) phosphorus trichloride in reaction flask, add 5mL dry methylene chloride, 245mg (3.10mmol) dry pyridine respectively, be cooled to-78 ℃.365mg (3.10mmol) 4-hydroxyl-methyl-butyrate is dissolved in the 5mL methylene dichloride, slowly drips in reaction solution, and stirring at room 3 hours places refrigerator overnight.Take out, be cooled to-40 ℃, drip 626mg (6.20mmol) Diisopropylamine, behind the low-temp reaction 3h, room temperature is placed 20h.Get 1.69g (3.10mmol) 5 '-(4,4 '-dimethoxy)-trityl-thymidine, 1.60g (12.4mmol) DIEA and 10mL methylene dichloride, inject reaction solution, stirred 6 hours under the room temperature.The saturated NaHCO of reaction solution 3, saturated NaCl solution washs secondary, anhydrous Na respectively 2SO 4Dry organic phase.Filtering Na 2SO 4, decompression removes methylene dichloride, obtains faint yellow oily thing, through silica gel column chromatography, obtains the colourless blister product of 300mg, yield: 14.4% (by phosphorus trichloride).Rf:0.54,0.64 (DCM: acetone 9: 1); 1H-NMR (CD 3Cl), δ H: 1.03～1.30[m, 12H ,-CH (CH 3) 2], 1.40 (s, 3H ,-CH 3), 1.59 (m, 2H, O-CH 2-CH 2-CH 2-C=O), 1.72～1.94 (m, 2H, O-CH 2-CH 2-CH 2-C=O), 2.25～2.50[m, 2H, C (2 ')-H 2], 3.25～3.75[m, 9H ,-CH (CH 3) 2, O=C-OCH 3, O-CH 2-CH 2-CH 2-C=O, C (5 ')-H 2], 3.79 (s, 6H ,-OCH 3), 4.18[m, 1H, C (4 ')-H], 4.63[m, 1H, C (3 ')-H], 6.40[m, 1H, C (1 ')-H], 6.81～7.66 (m, 14H, Ar-H], 8.13 (br, NH); 31P-NMR (CD 3Cl), δ P: 147.75,148.13.
Embodiment 7
7.1 1-[(2-deoxidation-β-D-erythro-ribose-5-(4,4 '-dimethoxytrityl))-and 5-methyl isophthalic acid H, 3H-pyrimidine-2,4-diketone 3 '-[(2-[1-(2, the 4-nitrophenyl)-1H-imidazolyl-4-]-ethyl)-N, N-di-isopropyl-phosphoramidite] (31)
Get 297mg (2.16mmol) phosphorus trichloride in reaction flask, add the 10mL dry methylene chloride, be cooled to-78 ℃ and drip 875mg (8.64mmol) Diisopropylamine, stirred overnight at room temperature.Get 0.94g (1.73mmol) 5 '-(4,4 '-dimethoxy)-trityl-thymidine, 557mg (12.4mmol) DIEA and 10mL methylene dichloride, inject reaction solution, stirred 24 hours under the room temperature.Add the 90mg tetrazole, 360mg 2-[1-(2, the 4-nitrophenyl)-1H-imidazolyl-4-]-ethanol, room temperature is placed 20h.The saturated NaHCO of reaction solution 3, saturated NaCl solution washs secondary, anhydrous Na respectively 2SO 4Dry organic phase.Filtering Na 2SO 4, decompression removes methylene dichloride, obtains faint yellow oily thing, through silica gel column chromatography, obtains the yellow blister product of 490mg, yield: 39.8% (press 2-[1-(2, the 4-nitrophenyl)-1H-imidazolyl-4-]-alcohol meter).R f: 0.31 (ethyl acetate: triethylamine 100: 3); 1H-NMR (CD 3Cl), δ H: 1.03～1.30[m, 12H ,-CH (CH 3) 2], 1.38 (s, 3H ,-CH 3), 2.25～2.50[m, 2H, C (2 ')-H 2], 3.30～3.68[m, 6H ,-CH (CH 3) 2, O-CH 2-CH 2-imidazole, C (5 ')-H 2], 3.77 (m, 8H ,-OCH 3, O-CH 2-CH 2-imidazole), and 4.18[m, 1H, C (4 ')-H], 4.65[m, 1H, C (3 ')-H], 6.40[m, 1H, C (1 ')-H], 6.82～7.38 (m, 17H, Ar-H), 8.16 (br, NH), 8.44[m, 1H, 5-H (Dnp)], 8.77[d, 1H, 3-H (Dnp)]; 31P-NMR (CD 3Cl), δ P: 147.55.
Embodiment 8
8.1 5-(2-benzyloxy ethyl)-uridylic (32)
(10.3g 66mmol) adds in the dimethyl formamide (100ml), and (24.6g 132mmol), is heated to 100 ℃ to the methylsulfonic acid benzyl ester of adding new system, reacts 48 hours with 5-(2-hydroxyethyl) uridylic.Solvent evaporated, residuum ether washed twice.Add dehydrated alcohol (100ml), be heated to 50 ℃, filtered while hot is evaporated to 30ml, puts into refrigerator cold-storage.Filter, leach the thing drying, get colorless solid, 8.89g, Mp 144-150 ℃.Ultimate analysis: C 13H 14N 2O 3(M 246.26), theoretical value: C 63.40, H 5.30, and N 11.38; Measured value: C 63.25, H 5.38, and N 11.46. 1HNMR(d6-DMSO)：2.45(t，2H，CH 2-OH)，3.49(t，2H，CH 2-CH 2)，4.51(s，2H，-CH 2-C 6H 5)，7.30(m，6H，-C 6H 5?and?C＝CH)，10.68(d，1H，CH-NH-CO)，11.04(s，1H，CO-NH-CO)。
8.2 5-(2-benzyloxy ethyl)-uridylic-1-acetate (33)
(8.86g 36mmol) is dissolved in the dimethyl formamide (100ml), adds K with 5-(2-benzyloxy ethyl) uridylic 2CO 3(5.79g, 42mmol), after 30 minutes, the dripping bromine methyl acetate (5.47g, 36mmol), stirred overnight at room temperature.Solvent evaporated adds frozen water (20ml), filters water (20ml) washing.Leach thing and add entry (30ml) and (20ml) in the tetrahydrofuran (THF), keep PH=12 with the lithium hydroxide of 1M, complete to the TLC detection reaction.Solution is transferred pH=2.0 with ethyl acetate extraction twice with 1MHCl, collecting precipitation, and drying gets white solid 5.96g, productive rate 54.3%.mp180～181℃。 1HNMR(d6-DMSO)：2.47(t，2H，CH 2-O)，3.49(t，2H，CH 2-CH 2)，4.40(s，2H，N-CH 2-CO)，4.46(s，2H，-CH 2-C 6H 5)，7.33(m，5H，-C 6H 5)，7.49(s，1H，C＝CH)，11.46(s，1H，CO-NH-CO)。
8.3 N-[(5-(2-benzyloxy ethyl)-uridylic-1-ethanoyl)]-N-(2-tertbutyloxycarbonyl-amino-ethyl)-glycine methyl ester (34)
With 5-(2-benzyloxy ethyl)-uridylic-1-acetate (1.00g, 3.29mmol) be dissolved in the dimethyl formamide (10ml), be cooled to 0 ℃, add DCC (0.81g, 3.93mmol), adding (N-2-tertbutyloxycarbonyl-amino-ethyl)-glycine methyl ester after 30 minutes (0.80g, 3.45mmol), continuation was 0 ℃ of reaction 1 hour, and room temperature reaction spends the night.Solvent evaporated, silica gel column chromatography separate 1.25g, productive rate 73.5%.ESI-MS：519[M+H] +。
8.4 N-[5-(2-benzyloxy ethyl)-uridylic-1-ethanoyl]-N-(2-tertbutyloxycarbonyl-amino-ethyl)-glycine (35)
With N-[5-(2-benzyloxy ethyl)-uridylic-1-ethanoyl]-N-(2-tertbutyloxycarbonyl-amino-ethyl)-glycine methyl ester (1.25g; 2.41mmol) in the mixed solution of water-soluble (10ml) and tetrahydrofuran (THF) (10ml); lithium hydroxide with 1M is kept pH12, to reacting completely.Solution is transferred pH2 with ethyl acetate extraction twice with 1MHCl.Ethyl acetate extraction, anhydrous sodium sulfate drying, ethanol/ether recrystallization gets colorless solid 0.99g, productive rate 81.1%.Ultimate analysis: C 24H 32N 4O 8(M 504.53), theoretical value: C 57.13, H 6.39, and N 11.10; Measured value: C 57.34, H 6.58, and N 11.34. 1H NMR (DMSO): 1.37[s, 9H, C (CH 3) 3], 2.50 (t, 2H, CH 2-O), 3.00-3.50 (m, 4H, NHCH 2CH 2NH), 3.51 (dd, 2H ,-CH 2-C 6H 5), 3.98 (s, 1.4H, CH 2COO), 4.20 (s, 0.6H, CH 2COO), 4.45 (s, 2H ,-CH 2-C 6H 5), 4.49 (s, 0.6H, NCH 2CON), 4.66 (s, 1.4H, NCH 2CON), 7.20-7.35 (m, 6H ,-C 6H 5And C=CH), 11.32 (s, 0.6H, COCHCO), 11.34 (s, 1.4H, COCHCO), 12.70 (br s, 1H, COOH).
Embodiment 9
9.1 N α-tertbutyloxycarbonyl-N π-benzyloxymethyl-L-Histidine methyl esters (9a)
The 500mL reactor adds dimethyl formamide 100mL, 11.25gN α-tertbutyloxycarbonyl-N π-benzyloxymethyl-L-Histidine (30mmol), the 4.14g anhydrous K 2CO 3(30mmol), add methyl mesylate 3.31g (30mmol) again, room temperature reaction spends the night, and stops.The pressure reducing and steaming dimethyl formamide adds suitable quantity of water, and ethyl acetate extraction three times is successively with saturated sodium bicarbonate solution, saturated sodium-chloride water solution washing, anhydrous sodium sulfate drying.Evaporated under reduced pressure obtains the 10.13g light yellow oil, yield 86.8%.Ultimate analysis: C 20H 27N 3O 5(349.51), theoretical value: C 61.68, H 6.99, and N 10.79, measured value: C 61.27, and H 7.38, N10.82. 1HNMR(CDCl 3，)：1.41[s，9H，-C(CH 3) 3]，3.09-3.19(m，2H，-CHCH 2C)，3.72(s，3H，-OCH 3)，4.47(s，2H，-O-CH 2-Ar)，4.58(m，1H，NCH(CH 2)CO)，5.28-5.32(m，3H，-NHCH?and?NCH 2O)，6.87(s，1H，-C＝CH-N)，7.28-7.39(m，5H，Ar-H)，7.48(s，1H，-N-CH＝N-)。ESI-MS：390.2[M+H] +。
9.2 N π-benzyloxymethyl-L-Histidine methyl esters (9b)
The 100mL reactor adds N α-tertbutyloxycarbonyl-N π-benzyloxymethyl-L-Histidine methyl esters (9a) 10.13g (26mmol), dioxane 20mL, all dissolving, add 4M HCl/ dioxane 20mL, room temperature reaction 1 hour, raw material disappears, evaporate to dryness, add suitable quantity of water, extracted with diethyl ether twice adds sodium bicarbonate in batches, is stirred well to no bubble and produces, ethyl acetate extraction three times, anhydrous sodium sulfate drying.Evaporated under reduced pressure obtains 6.06g oily matter, yield 80.5%.
9.3 N α-(2-t-butoxycarbonyl amino ethyl)-N π-benzyloxymethyl-L-Histidine methyl esters (9c)
The 250mL reactor, N π-benzyloxymethyl-L-Histidine methyl esters (9b) 6.62g (21mmol) adds among the anhydrous methanol 70mL, and-20 ℃, (3.33g 21mmol), stirred 30 minutes to add N-t-butoxycarbonyl amino acetaldehyde.Add NaBH 3CN (0.925g, 14.7mmol), HAc2.4g (29.4mmol) reacts 30 clocks, and room temperature reaction 1 hour stops.Add ethyl acetate, again with saturated sodium bicarbonate aqueous solution washing three times, saturated sodium-chloride water solution washing, anhydrous sodium sulfate drying.Evaporated under reduced pressure, column chromatography obtains colorless oil 7.70g, yield 85%.Ultimate analysis: C 22H 32N 4O 5(432.67), theoretical value: C 61.09, H 7.46, and N 12.95, measured value: C 59.84, and H 7.70, and N 12.24. 1HNMR (CDCl 3): 1.44[s, 9H ,-C (CH 3) 3], 2.5-3,2 (m, 6H ,-CH 2CH 2With-CH-CH 2-C), 3.53-3.56 (m, 1H ,-CH-COOCH 3), 3.67 (s, 3H ,-COOCH 3), 4.42 (s, 2H ,-OCH 2-Ar), 5.00 (br, 1H ,-NH-CH-COOCH 3), 5.33-5.38 (m, 2H ,-N-CH 2-O-Bzl), 6.88 (s, 1H ,-C=CH-N-), 7.30-7.36 (m, 5H ,-O-CH 2-Ar-H), 7.50 (s, 1H ,-N-CH=N-).ESI-MS：433.4[M+H] +。
9.4 N-[5-(2-benzyloxy ethyl)-uridylic-1-ethanoyl]-N-(2-tertbutyloxycarbonyl-amino-ethyl)-N π-benzyloxymethyl-L-Histidine methyl esters (9d)
(5.30g 17.4mmol) is dissolved among the DMF (50ml), is cooled to 0 ℃, and (4.77g 23.2mmol), adds (6.81g, 15.8mmol) N after 30 minutes to add DCC with 5-(2-benzyloxy ethyl)-uridylic-1-acetate α-(2-t-butoxycarbonyl amino ethyl)-N π-benzyloxymethyl-L-Histidine methyl esters (9c) continues 0 ℃ of reaction 1 hour, and room temperature reaction spends the night.Solvent evaporated, silica gel column chromatography separate 7.65g oily matter, productive rate 67.6%. 1HNMR (CDCl 3): 1.36 (s, 9H ,-C (CH 3) 3), 2.50 (m, 2H, CH 2-O), 2.70-3.40 (m, 6H ,-CH 2CH 2With-CH-CH 2-C), 3.52 (t, 2H ,-CH 2-C 6H 5), 3.57 (s, 3H ,-OCH 3), 4.21 (m, 1H ,-CH-COOCH 3), 4.40-4.70 (m, 6H ,-OCH 2-Ar and-OCH 2-Ar and NCH 2CON), 5.43 (m, 2H, N-CH 2-O), 6.83 (s, 1H ,-C=CH-N-), 7.20-7.40 (m, 10H ,-O-CH 2-Ar-H), 7.43 (s, 1H ,-N-CH=N-), 7.76 (s, 1H, C=CH-N), 11.37 (s, 1H, COCHCO).
9.5 N-[5-(2-benzyloxy ethyl)-uridylic-1-ethanoyl]-N-(2-tertbutyloxycarbonyl-amino-ethyl)-N π-benzyloxymethyl-L-Histidine (36)
With N-[5-(2-benzyloxy ethyl)-uridylic-1-ethanoyl]-N-(2-tertbutyloxycarbonyl-amino-ethyl)-N π(7.65g 2.41mmol) in the mixed solution of water-soluble (100ml) and tetrahydrofuran (THF) (100ml), keeps pH13 with the lithium hydroxide of 2M, to reacting completely to-benzyloxymethyl-L-Histidine methyl esters (9d).Solution is transferred pH2 with ethyl acetate extraction twice with 1MHCl.Evaporated under reduced pressure adds dehydrated alcohol (300ml), filters anhydrous sodium sulfate drying.Evaporate to dryness, ethanol/ether recrystallization gets white solid 6.94g, productive rate 92.5%.Ultimate analysis: C 36H 44N 6O 9(M 704.77), theoretical value: C 61.35, H 6.29, and N 11.92; Measured value: C 61.55, H 6.18, and N 11.98. 1HNMR(CDCl 3)：1.35(s，9H，-C(CH 3) 3)，2.4736(m，2H，CH 2-O)，2.7-3.4(m，6H，-CH 2CH 2?and-CH-CH 2-C)，3.51(m，2H，-CH 2-C 6H 5)，4.22(m，1H，-CH-COOCH 3)，4.40-4.70(m，6H，-OCH 2-Ar，-OCH 2-Ar，NCH 2CONH)，5.41(s，0.7H，NCH 2CON)，4.48(s，0.3H，NCH 2CON)，6.77(s，0.7H，-C＝CH-N-)，6.81(s，0.3H，-C＝CH-N-)，7.20-7.45(m，11H，-C 6H 5，-N-CH＝N-)，7.74(s，0.7H，C＝CH-N)，7.75(s，0.3，C＝CH-N)，11.28(s，0.3H，COCHCO)，11.36(s，0.7H，COCHCO)。

Claims (12)

1. formula I, the nucleoside analog of II
Among its Chinese style I and the II, B is natural base (VITAMIN B4, guanine, uracil, thymus pyrimidine, cytosine(Cyt), xanthine, xanthoglobulin or the like), contains the natural base of protecting group, or shown in formula III, IV the non-natural base,
In formula III and IV, introduce substituting group-X-R five of uracil and seven of 8-nitrogen-7-denitrification purine 1, wherein, X can be various connecting arm forms, as (CH 2-) n, (CH=CH-) n, (C ≡ C-) n,-O-,-C (O)-,-OC (O)-,-C (O) O-,-NH-, N (R 1) 2-,-C (O) NH-,-NHC (O)-,-CH=N-,-S-,-S (O)-,-S (O) 2-,-S (O) NH-,-S (O) 2NH-, or their combination, n are 0-10,
Wherein when n is 0, promptly there is not connecting arm X, R 1Can be the contained functional group of proteinic all amino-acid residue side chains, comprising: (1) hydrophobic group: methyl (L-Ala), benzyl (phenylalanine), sec.-propyl (Xie Ansuan), 1-methylmercaptoethyl [CH 3S (CH 2) 2-] (methionine(Met)), 3-methyl-propyl [(CH 3) 2CHCH 2-] (leucine), 1-methyl-propyl [CH 3CH 2CH (CH 3)-] (Isoleucine); (2) hydrophilic radical: as methylol (HOCH 2-) (Serine), 1-hydroxyethyl [CH 3CH (OH)-] (Threonine), 4-imidazoles methyl (Histidine), the amino butyl [NH of 4- 2(CH 2) 4-] (Methionin), thiopurine methyltransferase (HSCH 2-) (halfcystine), 3-indole methyl (tryptophane), Pyrrolidine-2-(proline(Pro)), 4-hydroxybenzene methyl (tyrosine), 1-guanidine radicals propyl group (arginine), carboxamide methyl (l-asparagine), 1-carboxamide ethyl (glutamine), carboxymethyl (aspartic acid), 1-propyloic (L-glutamic acid)
When n is not 0, be connecting arm with X: (1) .R wherein 1Can be carbonatoms and be 2 to 10 alkyl and isomer thereof, comprise (straight chain and side chain) alkyl, cycloalkyl, thiazolinyl, and alkynyl, and their isomer, as methyl, ethyl, vinyl, n-propyl, sec.-propyl, allyl group, propargyl, normal-butyl, isobutyl-, the tertiary butyl, 1-(or 2-, 3-) butenyl, 1-(or 2-, 3-) butynyl, amyl group, cyclopentyl, cyclopentadienyl, hexyl, cyclohexyl, etc.; Heteroatomic (straight chain and side chain) alkyl such as nitrogenous, oxygen, sulphur, selenium, cycloalkyl, thiazolinyl, and alkynyl, its heteroatoms show as following functional group, as sulfydryl, ehter bond, thioether bond, amino, amido, guanidine radicals, urea groups, carboxyl, ester group, hydroxyl, amide group, alkylsulfonyl, sulfoamido, sulphonyl ester group, sulfinyl, sulfonamido, sulfinyl ester group, nitro, itrate group, nitroso-group, nitrous acid ester group, halogen, pseudohalogen etc.; The heterocyclic group is as, 2-(or/and 3-replaces) furyl, 2-(or/and 4-, 5-replaces) imidazolyl, 2-(or/and 3-, N-replaces) pyrryl, (or/and 4-, 5-replaces the) oxazolyl to 2-, 2-(or/and 3-replaces) thienyl, 3-(or/and 2-, 4-, 5-, 6, N-replaces) indyl, 2-(or/and 3-replaces) tetrahydrofuran base, 2-(or/and 3-, N-replaces) Pyrrolidine base, 2-(or/and 3-replaces) tetrahydro-thienyl, 2-(or/and 3-, 4-replaces) piperidyl, 2-(or/and 3-, N-replaces) morpholinyl, 2-(or/and 3-, 5-replaces) thiazolyl, 2-(or/and 3-, 5-replaces) thiazolidine base etc.; (2) .R 1The aryl radical and the assorted aryl radical that can be carbonatoms and be 6 to 20 aryl radical and assorted aryl radical and replace, as phenyl, naphthyl, anthryl, fluorenyl etc.; Assorted aryl radical is nitrogenous, oxygen, and sulphur, heteroatomss such as selenium are as 2-(or 3-, 4-) pyridyl, quinolyl, isoquinolyl.In aryl radical that replaces and assorted aryl radical, its substituting group is defined as methyl, ethyl, vinyl, n-propyl, sec.-propyl, allyl group, propargyl, normal-butyl, isobutyl-, the tertiary butyl, 1-(or 2-, 3-) butenyl, 1-(or 2-, 3-) butynyl, amyl group, cyclopentyl, cyclopentadienyl, hexyl, carbonatomss such as cyclohexyl are the alkyl of 1-10; Or contain sulfydryl, ehter bond, thioether bond, amino, amido; guanidine radicals, urea groups, carboxyl, ester group, hydroxyl; amide group, alkylsulfonyl, sulfoamido, sulphonyl ester group, sulfinyl; sulfonamido, sulfinyl ester group, nitro, itrate group; nitroso-group, nitrous acid ester group, halogen, pseudohalogen; imidazolyl, pyrryl, this class alkyl of functional groups such as indyl
Among the formula IV, P 2Can be hydrogen, ethanoyl, benzoyl, anisoyl; isobutyryl, ortho-nitrophenyl oxygen ethanoyl, N, N dimethylamine base methene base; benzene oxygen ethanoyl, tert.-butylbenzene oxygen ethanoyl, trifluoroacetyl group is to methyl benzoyl; carbonic acid alkyl methyl esters, carbonic acid alkyl allyl ester etc., protecting group amino and imido grpup has benzoyl, isobutyryl; trifluoroacetyl group, N, N dimethylamine methylene; N, N-dibutyl amino methylene radical
Among the formula I, P 1Can be ethanoyl, trimethyl silicon based, diethyl is silica-based; tertiary butyl dimethyl is silica-based, and tert-butyl diphenyl is silica-based, THP trtrahydropyranyl; allyl group, uncle's fourth oxygen methyl, (adjacent nitro; to nitro, to halo, to cyano group) benzyl; trityl, p-methoxyphenyl phenylbenzene, 4; 4 '-methoxyl group trityl etc.
Among the formula I, when B is the natural base of natural base or protection, R 2Can be N, N-diisopropylamino, 4-morphine quinoline base, 1-Pyrrolidine base, 2,2,6,6-tetramethyl-piperidyl; R 3Can be-X-R 1,-NH-R 1,-N (R 1) 2(two R wherein 1Can identically also can be R 1Various combination), but do not comprise the 2-cyanoethyl, methoxyl group, oxyethyl group, N, N-diisopropylamino, N, N dimethylamine base, N, N dimethylamine base;
Among the formula I, when B is the non-natural base, R 2Can be N, N-diisopropylamino, morphine quinoline base, Pyrrolidine base, 2,2,6,6-tetramethyl-piperidyl; R 3Can be the 2-cyanoethyl ,-X-R 1,-NH-R 1,-N (R 1) 2(two R wherein 1Can identically also can be R 1Various combination).
Among the formula II, when B is the non-natural base, R 4, R 5, R 6, R 7, R 8Can be H, R 1Or-X-R 1
Among the formula II, R 9Can be H or-X-R 1, R 10Can be H, tertbutyloxycarbonyl (Boc), fluorenylmethyloxycarbonyl (Fmoc), carbobenzoxy-(Cbz), benzyl or trityl;
Among the formula II, R 11Can be hydrogen, methyl, ethyl, allyl group, benzyl, 4-nitrobenzyl, 4-methyl-benzyl, 4-methoxy-benzyl;
Among the formula II, R 5With R 6, R 7With R 8, R 5With R 7, R 6With R 8, R 9/ R 10With R 5/ R 6, R 9/ R 10With R 7/ R 8Between form with (CH 2-) nThe ring texture that structure links to each other, n is 1～4; Or with main chain (N-C-C, or N-C, or C-C) form contain heteroatoms (O, S, N etc.) three to seven-membered ring shape structure, heteroatoms forms as ehter bond, thioether bond etc.
2. the nucleotide sequence and the peptide nucleic acid sequence that contains formula VI that contain formula V
Among the formula V, when B is the non-natural base, R 12Can be O -, OH, S -, SH, Se -, SeH or R 1, when B is the natural base of natural base or protection, R 12Can be R 3, but be not O -, OH, S -, SH, Se -, SeH; Y can be O, S, and atoms such as Se,
Among the formula VI, R 4, R 5, R 6, R 7, R 8, R 9It is identical with the corresponding definition in the claim 1,
At the nucleotide sequence that contains formula V and containing in the peptide nucleic acid sequence of formula VI, the number of formula V and formula VI can be at 1～50.
3. the compound of claim 1; wherein B is natural base (VITAMIN B4; guanine, uracil, thymus pyrimidine; cytosine(Cyt); xanthine, xanthoglobulin etc.), the natural base that contains protecting group; or modified base and the modified base that contains protecting group, modified base is to introduce substituting group-X-R five of uracil and seven of 8-nitrogen-7-denitrification purine 1
4. the compound of claim 1, wherein the X in the modified base is various connecting arm forms, as-CH 2-) n, (CH=CH-) n, (C ≡ C-) n,-O-,-C (O)-,-OC (O)-,-C (O) O-,-NH-, N (R 1) 2-,-C (O) NH-,-NHC (O)-,-CH=N-,-S-,-S (O)-,-S (O) 2-,-S (O) NH-,-S (O) 2NH-, or their combination, n are 0-10.
5. the compound of claim 1, wherein in modified base, when n is zero, when promptly not having connecting arm X, R 1Can be the contained functional group of proteinic all amino-acid residue side chains, comprising: (1) hydrophobic group: methyl (L-Ala), benzyl (phenylalanine), sec.-propyl (Xie Ansuan), 1-methylmercaptoethyl [CH 3S (CH 2) 2-] (methionine(Met)), 3-methyl-propyl [(CH 3) 2CHCH 2-] (leucine), 1-methyl-propyl [CH 3CH 2CH (CH 3)-] (Isoleucine); (2) hydrophilic radical: as methylol (HOCH 2-) (Serine), 1-hydroxyethyl [CH 3CH (OH)-] (Threonine), 4-imidazoles methyl (Histidine), the amino butyl [NH of 4- 2(CH 2) 4-] (Methionin), thiopurine methyltransferase (HSCH 2-) (halfcystine), 3-indole methyl (tryptophane), Pyrrolidine-2-(proline(Pro)), 4-hydroxybenzene methyl (tyrosine), 1-guanidine radicals propyl group (arginine), carboxamide methyl (l-asparagine), 1-carboxamide ethyl (glutamine), carboxymethyl (aspartic acid), 1-propyloic (L-glutamic acid).
6. the compound of claim 1 in modified base, when n is non-vanishing, is a connecting arm with X wherein, (1) .R 1Can be carbonatoms and be 2 to 10 alkyl and isomer thereof, comprise (straight chain and side chain) alkyl, cycloalkyl, thiazolinyl, and alkynyl, and their isomer, as methyl, ethyl, vinyl, n-propyl, sec.-propyl, allyl group, propargyl, normal-butyl, isobutyl-, the tertiary butyl, 1-(or 2-, 3-) butenyl, 1-(or 2-, 3-) butynyl, amyl group, cyclopentyl, cyclopentadienyl, hexyl, cyclohexyl, etc.; Heteroatomic (straight chain and side chain) alkyl such as nitrogenous, oxygen, sulphur, selenium, cycloalkyl, thiazolinyl, and alkynyl, its heteroatoms show as following functional group, as sulfydryl, ehter bond, thioether bond, amino, amido, guanidine radicals, urea groups, carboxyl, ester group, hydroxyl, amide group, alkylsulfonyl, sulfoamido, sulphonyl ester group, sulfinyl, sulfonamido, sulfinyl ester group, nitro, itrate group, nitroso-group, nitrous acid ester group, halogen, pseudohalogen etc.; The heterocyclic group is as, 2-(or/and 3-replaces) furyl, 2-(or/and 4-, 5-replaces) imidazolyl, 2-(or/and 3-, N-replaces) pyrryl, (or/and 4-, 5-replaces the) oxazolyl to 2-, 2-(or/and 3-replaces) thienyl, 3-(or/and 2-, 4-, 5-, 6, N-replaces) indyl, 2-(or/and 3-replaces) tetrahydrofuran base, 2-(or/and 3-, N-replaces) Pyrrolidine base, 2-(or/and 3-replaces) tetrahydro-thienyl, 2-(or/and 3-, 4-replaces) piperidyl, 2-(or/and 3-, N-replaces) morpholinyl, 2-(or/and 3-, 5-replaces) thiazolyl, 2-(or/and 3-, 5-replaces) thiazolidine base etc.; (2) .R 1The aryl radical and the assorted aryl radical that can be carbonatoms and be 6 to 20 aryl radical and assorted aryl radical and replace, as phenyl, naphthyl, anthryl, fluorenyl etc.; Assorted aryl radical is nitrogenous, oxygen, and sulphur, heteroatomss such as selenium are as 2-(or 3-, 4-) pyridyl, quinolyl, isoquinolyl.In aryl radical that replaces and assorted aryl radical, its substituting group is defined as methyl, ethyl, vinyl, n-propyl, sec.-propyl, allyl group, propargyl, normal-butyl, isobutyl-, the tertiary butyl, 1-(or 2-, 3-) butenyl, 1-(or 2-, 3-) butynyl, amyl group, cyclopentyl, cyclopentadienyl, hexyl, carbonatomss such as cyclohexyl are the alkyl of 1-10; Or contain sulfydryl, ehter bond, thioether bond, amino, amido; guanidine radicals, urea groups, carboxyl, ester group, hydroxyl; amide group, alkylsulfonyl, sulfoamido, sulphonyl ester group; sulfinyl, sulfonamido, sulfinyl ester group, nitro; itrate group, nitroso-group, nitrous acid ester group, halogen; pseudohalogen, imidazolyl, pyrryl, the alkyl of functional groups such as indyl.
7. the compound of claim 1, wherein in formula III and IV, five of uracil and seven bit substituents of 8-nitrogen-7-denitrification VITAMIN B4 can be methylol, the 2-hydroxyethyl, 3-hydroxypropyl, 3-hydroxypropyn base, imidazole ethyl, the imidazoles vinyl, acetoxyl, propionyloxy, acetamido, propionamido-, styroyl, the side chain functional group of natural amino acid such as phenylacetylene base and alpha-non-natural amino acid.
8. the compound of claim 1, wherein in formula I, when B is the non-natural base, R 2And R 3Can be R 1,-X-R 1,-NH-R 1,-N (R 1) 2, R 1Can identically also can be different.
9. at the compound of claim 1, wherein in formula II, R 5With R 6, R 7With R 8, R 5With R 7, R 6With R 8, R 9/ R 10With R 5/ R 6, R 9/ R 10With R 7/ R 8Between form with (CH 2-) nThe ring texture that structure links to each other, n is 1～4; Or with main chain (N-C-C, or N-C, or C-C) form contain heteroatoms (O, S, N etc.) three to seven-membered ring shape structure, heteroatoms forms as ehter bond, thioether bond etc.
10. the compound of claim 2, wherein in the nucleotide sequence that contains formula V, when B was the non-natural base, five of uracil and seven replacements of 8-nitrogen-7-denitrification purine were selected from following group:
Methylol, 2-hydroxyethyl, 3-hydroxypropyl, 3-hydroxypropyn base, imidazole ethyl, imidazoles vinyl, acetoxyl, propionyloxy, acetamido, propionamido-, styroyl, the side chain functional group of natural amino acid such as phenylacetylene base and alpha-non-natural amino acid;
In the nucleotide sequence that contains formula V, when B is the non-natural base, R 12Can be O -, OH, S -, SH, Se -, SeH or R 1, when B is the natural base of natural base or protection, R 12Can be R 3, but be not O -, OH, S -, SH, Se -, SeH; Y can be O, S, atoms such as Se.
11. the compound of claim 2, wherein in the peptide nucleic acid sequence that contains formula VI, seven bit substituents of five of uracil and 8-nitrogen-7-denitrification purine are with in the claim 10;
R 4, R 5, R 6, R 7, R 8, R 9Definition with in the claim 1.
12. the formula I in claim 1 and/or the claim 2, II, III, IV, V or VI compound or their arbitrary combination are used as simulation biomacromolecule, detection reagent, medical diagnosis on disease and medicine.
CN 200510007307 2005-02-06 2005-02-06 Nucleic acid, peptide nucleicacid derivatives and their use Pending CN1814614A (en)

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JP2010059116A (en) * 2008-09-05 2010-03-18 Institute Of Physical & Chemical Research Pyrimidine nucleoside compound and utilization of the same
EP3067359A1 (en) * 2008-09-23 2016-09-14 Scott G. Petersen Self delivering bio-labile phosphate protected pro-oligos for oligonucleotide based therapeutics and mediating rna interference
CN104011209A (en) * 2011-10-28 2014-08-27 约翰内斯堡威特沃特斯兰德大学 Inhibition Of Viral Gene Expression
CN103145714A (en) * 2013-03-13 2013-06-12 苏州维泰生物技术有限公司 Feather weight PNA (pentose nucleic acid) synthesis method
CN106460058A (en) * 2014-03-30 2017-02-22 赛沛 Modified cytosine polynucleotide oligomers and methods
CN108948147A (en) * 2017-05-22 2018-12-07 清华大学 It is a kind of for drug resistance gram-positive bacteria and the new antibiotic of tuberculosis therapy
CN107236011A (en) * 2017-06-16 2017-10-10 中国人民解放军军事医学科学院毒物药物研究所 Nucleoside compound or its salt, nucleic acid and its application
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