CN113512083B - Method for synthesizing nucleotide or nucleotide analogue - Google Patents
Method for synthesizing nucleotide or nucleotide analogue Download PDFInfo
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- CN113512083B CN113512083B CN202010280613.0A CN202010280613A CN113512083B CN 113512083 B CN113512083 B CN 113512083B CN 202010280613 A CN202010280613 A CN 202010280613A CN 113512083 B CN113512083 B CN 113512083B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
- C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/02—Phosphorylation
- C07H1/04—Introducing polyphosphoric acid radicals
Abstract
The invention provides a method for synthesizing nucleotide or nucleotide analogue. The method comprises nucleophilic substitution reaction of a nucleoside with a phosphate, pyrophosphate or tripolyphosphate under the catalysis of a phase transfer catalyst to obtain the nucleotide or nucleotide analogue, wherein the phosphate, pyrophosphate or tripolyphosphate is non-ammonium phosphate salt, the nucleoside has a structure shown in a formula (I) or a stereoisomer, a tautomer, a pharmaceutically acceptable salt or a prodrug thereof,
Description
Technical Field
The present invention relates to the field of biosynthesis, in particular, the present invention relates to a method of synthesizing a nucleotide or nucleotide analogue.
Background
Nucleotide compounds have an important role in chemistry, biology and chemical biology. The nucleotide is the basic component of all biological cells, and plays a dominant role in the growth, development, reproduction, inheritance and the like of organisms. Therefore, the nucleotide has wide application in national economy construction, for example, the deoxynucleoside triphosphate and the deoxynucleoside hexaphosphate are widely applied to DNA sequencing as DNA sequencing reagents, and the nucleoside monophosphate can be used as an important additive in infant milk powder.
The synthesis of various nucleotides and nucleotide analogues has important economic and scientific significance. Currently, enzyme-catalyzed synthesis and chemical synthesis are the main methods for synthesizing various nucleotides, and chemical methods have been developed for decades. Although various types of nucleotides can be synthesized by chemical synthesis, there are still many problems in chemical synthesis of nucleotides that need to be solved. For example, the synthesis method is complex, the steps are long, the synthesis cost is high, the reaction yield is low, and the like.
In the prior art, nucleophilic substitution reaction is most widely applied to synthesis of various nucleotides, wherein, ammonium salts of various phosphoric acids are used as nucleophilic reagents, which are one of key raw materials for preparing various nucleotides. However, the preparation of various ammonium phosphate salts is subjected to the steps of acid-base neutralization, cation exchange and the like, so that the preparation is complicated, and various ammonium phosphate salts have high price due to the complicated preparation. And various ammonium phosphate salts are easy to absorb moisture in the air and are required to be stored at low temperature. And the decomposition of the reagent is often accompanied during the use. However, various commercial sodium phosphates, potassium phosphates, sodium pyrophosphate and potassium pyrophosphate are available on the market, and although they are inexpensive, they are not suitable for use in the synthesis of nucleotides directly.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems in the related art to some extent.
To this end, in a first aspect of the invention, the invention proposes a method of synthesizing a nucleotide or nucleotide analogue. According to an embodiment of the present invention, the method comprises subjecting a nucleoside having a structure represented by formula (I) or a stereoisomer, tautomer, pharmaceutically acceptable salt or prodrug thereof to nucleophilic substitution reaction with a phosphate, pyrophosphate or tripolyphosphate under the catalysis of a phase transfer catalyst so as to obtain the nucleotide or nucleotide analogue, the phosphate, pyrophosphate or tripolyphosphate being a non-ammonium phosphate salt,
wherein X, Y are each independently selected from H, D, F, cl, br, CN, NO 2 、-C(＝O)R a 、-C(＝O)OR b 、-C(＝O)NR c R d 、-OR b 、-NR c R d 、R b O-C 1-4 Alkylene, R d R c N-C 1-4 Alkylene, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl groups, or X, Y, together with the carbon atoms to which they are attached, form C 3-6 Carbocycles, 5-6 atomsHeterocyclic ring of C 6-10 An aromatic ring or a heteroaromatic ring of 5 to 6 atoms, wherein C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Carbocycle, heterocycle of 5-6 atoms, C 6-10 The aromatic ring and the heteroaromatic ring consisting of 5 to 6 atoms are each independently unsubstituted OR substituted with 1, 2, 3 OR 4 substituents independently selected from D, F, cl, br, CN, -OR b 、-NR c R d 、C 1-6 Alkyl, C 1-6 Haloalkyl, R b O-C 1-4 Alkylene or R d R c N-C 1-4 Alkylene OR a , H、D、F、Cl、Br、CN、NO 2 、-C(＝O)R a 、-C(＝O)OR b 、-C(＝O)NR c R d 、-OR b 、-NR c R d 、R b O-C 1-4 Alkylene, R d R c N-C 1-4 Alkylene, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl;
r is H or
R 1 、R 2 Each independently selected from H, D, F, cl, br, CN, NO 2 、-C(＝O)R a 、-C(＝O)OR b 、-C(＝O)NR c R d 、-OR b 、-NR c R d 、R b O-C 1-4 Alkylene, R d R c N-C 1-4 Alkylene, C 1-6 Alkyl, C 2-6 Alkenyl or C 2-6 Alkynyl;
R a 、R b 、R c and R is d Each independently is H, D, hydroxy, C 1-6 Haloalkyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Carbocyclyl, C 3-6 carbocyclyl-C 1-4 Alkylene, 3-12 atom heterocyclyl, (3-12 atom heterocyclyl) -C 1-4 Alkylene, C 6-10 Aryl, C 6-10 aryl-C 1-4 Alkylene group, 5-10Heteroaryl of individual atomic composition or (heteroaryl of 5 to 10 atomic composition) -C 1-4 Alkylene group, wherein the C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Carbocyclyl, C 3-6 carbocyclyl-C 1-4 Alkylene, 3-12 atom heterocyclyl, (3-12 atom heterocyclyl) -C 1-4 Alkylene, C 6-10 Aryl, C 6-10 aryl-C 1-4 Alkylene, heteroaryl of 5-10 atoms and (heteroaryl of 5-10 atoms) -C 1-4 Alkylene groups are each independently unsubstituted or substituted with 1, 2, 3 or 4 substituents independently selected from D, F, cl, CN, OH, NH 2 、C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy or C 1-6 An alkylamino group;
a is adenine, guanine, cytosine, uracil or thymine.
The inventors of the present application have cleverly utilized the principle of phase transfer catalysis, i.e., the interaction of ion pairs to increase the reactivity and solubility of anions in the reaction. A catalytic amount of a phase transfer catalyst is dissolved in the organic phase and at the same time, the phase transfer catalyst undergoes anion exchange with phosphate which is insoluble in the organic phase to form amine phosphate in situ. The motive force for ion exchange results from the transfer catalyst positive ions having a larger structure that can bind more tightly to the anions to pull the equilibrium toward the production of amine phosphate. The in-situ generated amine phosphate has better solubility and stronger nucleophilicity in an organic solvent, so that the amine phosphate can react with nucleosides to obtain nucleotide products, and the regenerated phase transfer catalyst completes catalytic circulation. The key point of the catalytic cycle is that after the catalyst and phosphate are subjected to ion exchange, the phosphate amine salt is generated, and the catalyst has better solubility and stronger nucleophilicity, so that the completion of the reaction is effectively promoted.
According to an embodiment of the present invention, the above method may further include at least one of the following additional technical features:
According to an embodiment of the invention, the X, Y groups are each independently selected from H OR-OR b 。
According to an embodiment of the invention, the R 1 、R 2 Each independently selected from H, D, F, cl, br, CN, NO 2 、-OR b 。
According to an embodiment of the invention, R is H,
According to an embodiment of the present invention, further comprising reacting the nucleoside with a proton sponge and POCl 3 The contacting is performed so as to obtain a nucleoside electrophilic intermediate. Further improving the success rate of nucleophilic substitution reaction.
According to an embodiment of the invention, the nucleosides are associated with proton sponges and POCl 3 The contact is made in PO (OMe) 3 Is carried out in the following steps.
According to an embodiment of the invention, the molar ratio of nucleoside to phosphate, pyrophosphate or tripolyphosphate is 1:5. further improving the yield of nucleophilic substitution reaction, shortening the reaction time and reducing the waste of raw materials.
According to an embodiment of the invention, the molar ratio of nucleoside to phosphate, pyrophosphate or tripolyphosphate and phase transfer catalyst is 1:5: (0.5-1.5). Further improving the yield of nucleophilic substitution reaction, shortening the reaction time and reducing the waste of raw materials.
According to an embodiment of the present invention, the phase transfer catalyst includes at least one selected from the following. The reaction according to the embodiment of the invention further improves the raw material utilization rate and the target product yield under the catalysis of the following phase transfer catalyst.
According to an embodiment of the invention, the phosphate, pyrophosphate or tripolyphosphate comprises at least one selected from the group consisting of:
according to an embodiment of the present invention, the nucleotide includes at least one selected from the group consisting of nucleoside monophosphates, nucleoside diphosphate, nucleoside triphosphate, and nucleoside tetraphosphate.
According to an embodiment of the present invention, the nucleoside monophosphates have the structure shown below
According to an embodiment of the present invention, the nucleoside diphosphate has a structure shown below,
according to an embodiment of the present invention, the nucleoside triphosphates have the structure shown below,
according to an embodiment of the present invention, the nucleoside tetraphosphate has a structure as shown below,
the method according to the embodiment of the invention utilizes a phase transfer catalyst to promote the solubility and nucleophilicity of various phosphates and pyrophosphates in an organic solvent, thereby realizing the preparation of the nucleotide. The preparation method of the nucleotide provided by the embodiment of the invention has the advantages of lower cost and simpler, more convenient and faster operation.
Detailed Description
Definitions and general terms
Reference will now be made in detail to certain embodiments of the invention, examples of which are illustrated in the accompanying structural and chemical formulas. The invention is intended to cover all alternatives, modifications and equivalents, which may be included within the scope of the invention. Those skilled in the art will recognize that many methods and materials similar or equivalent to those described herein can be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described herein. In the event of one or more of the incorporated references, patents and similar materials differing from or contradictory to the present application (including but not limited to defined terms, term application, described techniques, etc.), the present application controls.
It should further be appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination.
Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, unless otherwise indicated, all patent publications cited throughout the disclosure of this invention are incorporated by reference in their entirety.
The invention will apply to the following definitions unless otherwise indicated. For the purposes of the present invention, chemical elements are defined according to the periodic Table of the elements, CAS version and chemical handbook, 75, thred, 1994. In addition, the general principles of organic chemistry are found in "Organic Chemistry", thomas Sorrell, university Science Books, sausalato 1999,and"March's Advanced Organic Chemistry", by Michael b.smith jerrry March, john Wiley & Sons, new York 2007, and the disclosure of this application is hereby incorporated by reference in its entirety.
The stereochemical definitions and conventions used in the present invention are generally in accordance with S.P. Parker, ed., mcGraw-Hill Dictionary of ChemicalTerms (1984) McGraw-Hill Book Company, new York; and Eliel, e.and Wilen, s., "Stereochemistry of OrganicCompounds", john Wiley & Sons, inc., new York,1994. The compounds of the invention may contain asymmetric or chiral centers and thus exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers and atropisomers (attopiomers) and mixtures thereof, such as racemic mixtures, are also included within the scope of the present invention. Many organic compounds exist in optically active form, i.e., they have the ability to rotate the plane of plane polarized light. When describing optically active compounds, the prefix D and L or R and S are used to denote the absolute configuration of the molecule in terms of chiral center (or chiral centers) in the molecule. The prefixes d and l or (+) and (-) are symbols for specifying the rotation of plane polarized light by a compound, where (-) or l indicates that the compound is left-handed. The compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers may also be referred to as enantiomers, and mixtures of such isomers are generally referred to as mixtures of enantiomers. The 50:50 mixture of enantiomers is referred to as a racemic mixture or racemate, which may occur when there is no stereoselectivity or stereospecificity in the chemical reaction or process.
Depending on the choice of starting materials and methods, the compounds according to the invention may be present in the form of one of the possible isomers or mixtures thereof, for example as pure optical isomers or as isomer mixtures, for example as racemic and non-corresponding isomer mixtures, depending on the number of asymmetric carbon atoms. Optically active (R) -or (S) -isomers can be prepared using chiral synthons or chiral preparations, or resolved using conventional techniques. If the compound contains a double bond, the substituent may be in the E or Z configuration; if the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may be in cis or trans (cis-or trans-) configuration.
The compounds of the invention may contain asymmetric or chiral centers and thus exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers and atropisomers (attospimers) and geometric (or conformational) isomers and mixtures thereof, such as racemic mixtures, are within the scope of the present invention.
Unless otherwise indicated, structures described herein are also meant to include all isomeric (e.g., enantiomer, diastereomeric atropisomer (attiosomer) and geometric (or conformational)) forms of such structures; for example, the R and S configurations of each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Thus, individual stereochemical isomers as well as enantiomeric mixtures, diastereomeric mixtures, and geometric (or conformational) isomer mixtures of the compounds of the invention are all within the scope of the invention.
The term "tautomer" or "tautomeric form" refers to structural isomers having different energies that can be interconverted by a low energy barrier (low energy barrier). If tautomerism is possible (e.g., in solution), chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also known as proton transfer tautomers (prototropic tautomer)) include interconversions by proton transfer, such as keto-enol isomerisation and imine-enamine isomerisation. Valence tautomers (valance tautomers) include interconversions by recombination of some of the bond-forming electrons. Specific examples of keto-enol tautomerism are tautomerism of pentane-2, 4-dione and 4-hydroxypent-3-en-2-one tautomer. Another example of tautomerism is phenol-ketone tautomerism. One specific example of phenol-ketone tautomerism is the interconversion of pyridin-4-ol and pyridin-4 (1H) -one tautomers. Unless otherwise indicated, all tautomeric forms of the compounds of the invention are within the scope of the invention.
As used herein, "pharmaceutically acceptable salts" refers to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as in the literature: S.M. Berge et al describe pharmaceutically acceptable salts in detail in J.pharmaceutical Sciences,1977,66:1-19. Pharmaceutically acceptable non-toxic acid forming salts include, but are not limited to, inorganic acid salts formed by reaction with amino groups such as hydrochloride, hydrobromide, phosphate, sulfate, perchlorate, and organic acid salts such as acetate, oxalate, maleate, tartrate, citrate, succinate, malonate, or by other methods described in the literature such as ion exchange. Other pharmaceutically acceptable salts include adipic acid salts, alginates, ascorbates, aspartic acid salts, benzenesulfonates, benzoic acid salts, bisulfate salts, borates, butyric acid salts, camphoric acid salts, cyclopentylpropionates, digluconate, dodecylsulfate, ethanesulfonate, formate salts, fumaric acid salts, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, caproate, hydroiodic acid salts, 2-hydroxy-ethanesulfonate salts, lactobionic aldehyde salts, lactate salts, laurate salts, lauryl sulfate, malate salts, malonate salts, methanesulfonate salts, 2-naphthalenesulfonate salts, nicotinate salts, nitrate salts, oleate salts, palmitate salts, pamoate salts, pectate salts, persulfate salts, 3-phenylpropionate salts, picrate salts, pivalate salts, propionate salts, stearate salts, thiocyanate salts, p-toluenesulfonate salts, undecanoate salts, valerate salts, and the like. Salts obtained with suitable bases include salts of alkali metals, alkaline earth metals, ammonium and n+ (C1-4 alkyl) 4. The present invention also contemplates quaternary ammonium salts formed from any compound containing a group of N. The water-soluble or oil-soluble or dispersible product may be obtained by quaternization. Alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include suitable, non-toxic ammonium, quaternary ammonium salts and counter-ion forming amine cations such as halides, hydroxides, carboxylates, sulphates, phosphates, nitrates, C1-8 sulphonates and aromatic sulphonates.
The term "prodrug" as used herein means a compound that is converted in vivo to a compound of formula (I). Such conversion is effected by hydrolysis of the prodrug in the blood or enzymatic conversion to the parent structure in the blood or tissue. The prodrug of the invention can be esters, and in the prior invention, the esters can be phenyl esters, aliphatic (C1-24) esters, acyloxymethyl esters, carbonates, carbamates and amino acid esters serving as the prodrugs. For example, one compound of the invention may contain a hydroxyl group, i.e., it may be acylated to provide the compound in a prodrug form. Other prodrug forms include phosphates, such as those obtained by phosphorylation of a hydroxyl group on the parent. For a complete discussion of prodrugs, reference may be made to the following documents: higuchi and V.stilla, pro-drugs as Novel Delivery Systems, vol.14of the A.C.S. symposium Series, edward B.Roche, ed., bioreversible Carriers in Drug Design, american Pharmaceutical Association and Pergamon Press,1987,J.Rautio et al, prodrug: design and Clinical Applications, nature Review Drug Discovery,2008,7,255-270,and S.J.Hecker et al, prodrugs of Phosphates and Phosphonates, journal of Medicinal Chemistry,2008,51,2328-2345.
Any asymmetric atom (e.g., carbon, etc.) of the compounds of the present invention may exist in racemic or enantiomerically enriched form, for example, in the form of the (R) -, (S) -, (R, R) -, (S, S) -, (S, R) -or (R, S) -configuration. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in the (R) -or (S) -configuration. The substituents on the atoms having unsaturated double bonds may be present in cis- (Z) -or trans- (E) -form, if possible.
Thus, as described herein, the compounds of the present invention may exist as one of the possible isomers, rotamers, atropisomers, tautomers or as a mixture thereof, for example as substantially pure geometric (cis or trans) isomers, diastereomers, optical isomers (enantiomers), racemates or as a mixture thereof.
As described herein, the compounds of the invention may be optionally substituted with one or more substituents, such as those of the general formula above, or as exemplified by the specific examples provided herein, the subclasses, and the compounds encompassed by the invention Is a compound of the formula (I). It is to be understood that the term "optionally substituted" may be used interchangeably with the term "substituted or unsubstituted". The terms "optionally," "optional," or "optionally" mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. In general, the term "optionally" whether or not preceding the term "substituted" means that one or more hydrogen atoms in a given structure are unsubstituted or substituted with a particular substituent. An optional substituent group may be substituted at each substitutable position of the group unless otherwise indicated. When more than one position in a given formula can be substituted with one or more substituents selected from a particular group, then the substituents may be the same or different at each position. Wherein the substituents may be, but are not limited to D, F, cl, br, CN, N 3 、OH、NH 2 、NO 2 Oxo (=o), -C (=o) R a 、-C(＝O)OR b 、-C(＝O)NR c R d 、-S(＝O)2NR c R d 、(RbO) 2 P (=O) -C0-2 alkylene, -OR b 、-NR c R d 、RbO-C 1-4 Alkylene, R d R c N-C 1-4 Alkylene, C 1-12 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Alkylamino, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-12 Cycloalkyl, C 3-12 cycloalkyl-C 1-4 Alkylene, 3-12 atom heterocyclyl, (3-12 atom heterocyclyl) -C 1-4 Alkylene, C 6-10 Aryl, C 6-10 aryl-C 1-4 Alkylene, heteroaryl of 5-16 atoms or (heteroaryl of 5-16 atoms) -C 1-4 Alkylene, wherein each R a 、R b 、R c 、R d Having the definition according to the invention.
In addition, unless explicitly indicated otherwise, the descriptions used in this disclosure of the manner in which each … is independently "and" … is independently "and" … is independently "are to be construed broadly as meaning that particular items expressed between the same symbols in different groups do not affect each other, or that particular items expressed between the same symbols in the same groups do not affect each other.
In the various parts of the present specification, substituents of the presently disclosed compounds are disclosed in terms of the type or scope of groups. It is specifically noted that the present invention includes each individual subcombination of the individual members of these group classes and ranges. For example, the term "C1-6 alkyl" particularly refers to independently disclosed methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl and C6 alkyl, and the term "heteroaryl of 5-10 atomic composition" particularly refers to independently disclosed heteroaryl of 5 atomic composition, heteroaryl of 6 atomic composition, heteroaryl of 7 atomic composition, heteroaryl of 8 atomic composition, heteroaryl of 9 atomic composition and heteroaryl of 10 atomic composition.
In the various parts of the invention, linking substituents are described. When the structure clearly requires a linking group, the markush variables recited for that group are understood to be linking groups. For example, if the structure requires a linking group and the markush group definition for that variable enumerates an "alkyl" or "aryl" group, it will be understood that the "alkyl" or "aryl" represents a linked alkylene group or arylene group, respectively.
The term "alkyl" or "alkyl group" as used herein refers to a saturated straight or branched chain monovalent hydrocarbon radical containing 1 to 20 carbon atoms. Unless otherwise specified, alkyl groups contain 1 to 20 carbon atoms; some of these are alkyl groups containing 1 to 12 carbon atoms; some of these are alkyl groups containing 1 to 10 carbon atoms; still other embodiments are alkyl groups containing 1 to 9 carbon atoms; still other embodiments are alkyl groups containing 1 to 8 carbon atoms; still other embodiments are alkyl groups containing 1 to 6 carbon atoms; still other embodiments are alkyl groups containing 1 to 4 carbon atoms, and still other embodiments are alkyl groups containing 1 to 3 carbon atoms.
Examples of alkyl groups include, but are not limited to, methyl (Me, -CH 3), ethyl (Et, -CH2CH 3), n-propyl (n-Pr,
-CH2CH2CH 3), isopropyl (i-Pr, -CH (CH 3) 2), n-butyl (n-Bu, -CH2CH2CH2CH 3), isobutyl (i-Bu, -CH2CH (CH 3) 2), sec-butyl (s-Bu, -CH (CH 3) CH2CH 3), tert-butyl (t-Bu), -C (CH 3) 3), n-pentyl (-CH 2CH2CH2CH 3), 2-pentyl (-CH (CH 3) CH2CH2CH 3), 3-pentyl (-CH (CH 2CH 3) 2), 2-methyl-2-butyl (-C (CH 3) 2CH2CH 3), 3-methyl-2-butyl (-CH (CH 3) CH (CH 3) 2), 3-methyl-1-butyl (-CH 2CH (CH 3) 2), 2-methyl-1-butyl (-CH 2CH (CH 3) CH2CH 3), n-hexyl (-CH 2CH2CH 3), 2-hexyl (-CH 2CH (CH 3) CH2CH 3), 2-methyl-2-CH (CH 3) 2-methyl (-CH 3) 2 (CH 3) 2CH 3), 3-methyl-3-pentyl (-C (CH 3) (CH 2CH 3) 2), 2-methyl-3-pentyl (-CH (CH 2CH 3) CH (CH 3) 2), 2, 3-dimethyl-2-butyl (-C (CH 3) 2CH (CH 3) 2), 3-dimethyl-2-butyl (-CH (CH 3) C (CH 3) 3), n-heptyl, n-octyl, and the like, wherein the alkyl groups may independently be unsubstituted or substituted with one or more substituents described herein.
The term "alkyl" and its prefix "alkane" as used herein, both include straight and branched saturated carbon chains. The term "alkylene" means a saturated divalent hydrocarbon group obtained by removing two hydrogen atoms from a straight or branched saturated hydrocarbon group. Unless otherwise specified, an alkylene group contains from 1 to 10 carbon atoms, and in other embodiments an alkylene group contains from 1 to 6 carbon atoms, and in other embodiments an alkylene group contains from 1 to 4 carbon atoms, and in other embodiments an alkylene group contains from 1 to 2 carbon atoms. Examples include methylene (-CH 2-), ethylene (-CH 2-), isopropylidene (-CH (CH 3) CH 2-) and the like, wherein the alkylene groups may independently be unsubstituted or substituted with one or more substituents described herein.
The term "alkenyl" means a straight or branched monovalent hydrocarbon radical of 2 to 12 carbon atoms, or 2 to 8 carbon atoms, or 2 to 6 carbon atoms, or 2 to 4 carbon atoms, wherein at leastC-C in one position is an sp2 double bond, wherein the alkenyl group may be independently unsubstituted or substituted with one or more substituents described herein, including the positioning of the group with "cis", "trans" or "Z", "E", specific examples of which include, but are not limited to, vinyl (-CH=CH) 2 ) Allyl (-CH) 2 CH＝CH 2 ) Etc.
The term "alkynyl" means a straight or branched monovalent hydrocarbon radical of 2 to 12 carbon atoms, or 2 to 8 carbon atoms, or 2 to 6 carbon atoms, or 2 to 4 carbon atoms, wherein C-C in at least one position is an sp triple bond, wherein the alkynyl group may be independently unsubstituted or substituted with one or more substituents described herein, specific examples include, but are not limited to, ethynyl (-C≡CH), propargyl (-CH) 2 C.ident.CH), 1-propynyl (-C.ident.C-CH) 3 ) Etc.
The terms "carbocycle", "carbocyclyl" or "carbocyclic" are used interchangeably herein to refer to a non-aromatic carbocyclic ring system containing 3 to 14 ring carbon atoms, saturated or containing one or more unsaturated units, and not containing any aromatic rings. In some embodiments, the number of carbon atoms is 3 to 12; in other embodiments, the number of carbon atoms is 3 to 10; in other embodiments, the number of carbon atoms is 3 to 8; in other embodiments, the number of carbon atoms is 3 to 6; in other embodiments, the number of carbon atoms is 5 to 6; in other embodiments, the number of carbon atoms is from 5 to 8. In other embodiments, the number of carbon atoms is from 6 to 8. The term "carbocyclyl" includes monocyclic, bicyclic or polycyclic fused, spiro or bridged carbocyclic ring systems. Bicyclic carbocyclyl includes bridged bicyclic carbocyclyl, fused bicyclic carbocyclyl, and spiro bicyclic carbocyclyl, the "fused" bicyclic ring system comprising two rings sharing 2 contiguous ring atoms. The bridge Lian Shuanghuan group includes two rings sharing 2, 3, 4 or 5 adjacent ring atoms. The spiro ring system shares 1 ring atom. Suitable carbocyclic groups include, but are not limited to, cycloalkyl, cycloalkenyl, and cycloalkynyl. Examples of carbocyclic groups further include, but are in no way limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-alkenyl, 1-cyclopentyl-2-alkenyl, 1-cyclopentyl-3-alkenyl, cyclohexyl, 1-cyclohexyl-1-alkenyl, 1-cyclohexyl-2-alkenyl, 1-cyclohexyl-3-alkenyl, cyclohexanedienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, and the like. Bridged carbocyclyl groups include, but are not limited to, bicyclo [2.2.2] octyl, bicyclo [2.2.1] heptyl, bicyclo [3.3.1] nonyl, bicyclo [3.2.3] nonyl, and the like.
The term "cycloalkyl" refers to a monocyclic, bicyclic or tricyclic ring system containing 3 to 12 ring carbon atoms that is saturated and has one or more points of attachment to the remainder of the molecule. Some of these embodiments, cycloalkyl groups are ring systems containing 3 to 10 ring carbon atoms; in other embodiments, cycloalkyl is a ring system containing 3 to 8 ring carbon atoms; in other embodiments, cycloalkyl is a ring system containing 5 to 8 ring carbon atoms; in other embodiments, cycloalkyl is a ring system containing 3 to 6 ring carbon atoms; in other embodiments, cycloalkyl is a ring system containing 5 to 6 ring carbon atoms; examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like, and the cycloalkyl groups may independently be unsubstituted or substituted with one or more substituents described herein.
The terms "heterocyclyl" and "heterocycle" are used interchangeably herein to refer to a saturated or partially unsaturated, non-aromatic, monocyclic, bicyclic or tricyclic ring system containing from 3 to 12 ring atoms, at least one of which is selected from the group consisting of nitrogen, sulfur and oxygen atoms, wherein said heterocyclyl is non-aromatic and does not contain any aromatic rings, and wherein said ring system has one or more points of attachment to the remainder of the molecule. The term "heterocyclyl" includes monocyclic, bicyclic or polycyclic fused, spiro or bridged heterocyclic ring systems. Bicyclic heterocyclyl groups include bridged bicyclic heterocyclyl groups, fused bicyclic heterocyclyl groups, and spiro bicyclic heterocyclyl groups. Unless otherwise indicated, the heterocyclyl group may be a carbon or nitrogen group, and the-CH 2-group may optionally be replaced by-C (=o) -. The sulfur atom of the ring may optionally be oxidized to an S-oxide. The nitrogen atom of the ring may optionally be oxidized to an N-oxide. In some embodiments, the heterocyclyl is a ring system consisting of 3 to 8 ring atoms; in other embodiments, the heterocyclyl is a ring system of 3 to 6 ring atoms; in other embodiments, the heterocyclyl is a ring system of 5 to 7 ring atoms; in other embodiments, the heterocyclyl is a ring system of 5 to 8 ring atoms; in other embodiments, the heterocyclyl is a ring system of 6 to 8 ring atoms; in other embodiments, the heterocyclyl is a ring system of 5 to 6 ring atoms; in other embodiments, the heterocyclyl is a ring system of 4 ring atoms; in other embodiments, the heterocyclyl is a ring system of 5 ring atoms; in other embodiments, the heterocyclyl is a ring system of 6 ring atoms; in other embodiments, the heterocyclyl is a ring system of 7 ring atoms; in other embodiments, the heterocyclyl is a ring system of 8 ring atoms.
Examples of heterocyclyl groups include, but are not limited to: oxiranyl, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1, 3-dioxacyclopentyl, dithiocyclopentyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, dioxanyl, dithianyl, thiazanyl, homopiperazinyl, homopiperidinyl, oxepinyl, thiepinyl, and the like. Examples of the substitution of the-CH 2-group in the heterocyclic group by-C (=o) -include, but are not limited to, 2-oxopyrrolidinyl, oxo-1, 3-thiazolidinyl, 2-piperidonyl, 3, 5-dioxopiperidyl, pyrimidinedionyl. Examples of sulfur atoms in the heterocyclic group that are oxidized include, but are not limited to, sulfolane groups and 1, 1-dioxothiomorpholinyl groups. Bridged heterocyclyl groups include, but are not limited to, 2-oxabicyclo [2.2.2] octyl, 1-azabicyclo [2.2.2] octyl, 3-azabicyclo [3.2.1] octyl, and the like. The heterocyclyl group may be optionally substituted with one or more substituents described herein.
The term "heteroatom" refers to any oxidation state form of O, S, N, P and Si, including N, S and P; primary, secondary, tertiary and quaternary ammonium salt forms; or a form in which the hydrogen on the nitrogen atom in the heterocycle is substituted, for example, N (like N in 3, 4-dihydro-2H-pyrrolyl), NH (like NH in pyrrolidinyl) or NR (like NR in N-substituted pyrrolidinyl).
The term "halogen" refers to F, cl, br or I.
The term "D" refers to deuteration, i.e. 2H.
The term "aryl" may be used alone or as part of an "aralkyl", "aralkoxy" or "aryloxyalkyl" group, to denote monocyclic, bicyclic, and tricyclic carbocyclic ring systems containing 6 to 14 ring atoms, or 6 to 12 ring atoms, or 6 to 10 ring atoms, wherein at least one ring system is aromatic, wherein each ring system contains a ring of 3 to 7 atoms, and one or more attachment points are attached to the remainder of the molecule. The term "aryl" may be used interchangeably with the term "aromatic ring" or "aromatic ring", e.g., aryl may include phenyl, 2, 3-dihydro-1H-indenyl, naphthyl, and anthracenyl. The aryl groups may independently be unsubstituted or substituted with one or more substituents described herein.
The term "heteroaryl" may be used alone or as a majority of "heteroarylalkyl" or "heteroarylalkoxy" groups, meaning a monocyclic, bicyclic, and tricyclic ring system containing 5 to 16 ring atoms, or containing 5 to 14 ring atoms, or containing 5 to 12 ring atoms, or containing 5 to 10 ring atoms, or containing 5 to 8 ring atoms, or containing 5 to 7 ring atoms, or containing 5 to 6 ring atoms, wherein at least one ring system is aromatic and at least one ring system contains one or more heteroatoms, wherein each ring system contains a ring of 5 to 7 atoms, and one or more attachment points are attached to the remainder of the molecule. When a heteroaryl group is present, the-CH 2-group may optionally be replaced by a-C (=o) -. The term "heteroaryl" may be used interchangeably with the term "heteroaromatic ring" or "heteroaromatic compound". In some embodiments, heteroaryl is a 5-14 atom composition heteroaryl comprising 1,2,3, or 4 heteroatoms independently selected from O, S, and N. In other embodiments, heteroaryl is a 5-12 atom heteroaryl group comprising 1,2,3, or 4 heteroatoms independently selected from O, S, and N. In other embodiments, heteroaryl is a 5-10 atom composition heteroaryl comprising 1,2,3, or 4 heteroatoms independently selected from O, S, and N. In other embodiments, heteroaryl is a 5-8 atom composition heteroaryl comprising 1,2,3, or 4 heteroatoms independently selected from O, S, and N. In other embodiments, heteroaryl is a 5-7 atom composition heteroaryl comprising 1,2,3, or 4 heteroatoms independently selected from O, S, and N. In other embodiments, heteroaryl is a 5-6 atom composition heteroaryl comprising 1,2,3, or 4 heteroatoms independently selected from O, S, and N. In other embodiments, the heteroaryl is a 5-atom composition heteroaryl comprising 1,2,3, or 4 heteroatoms independently selected from O, S, and N. In other embodiments, the heteroaryl is a 6-atom composition heteroaryl comprising 1,2,3, or 4 heteroatoms independently selected from O, S, and N.
Still other embodiments are heteroaryl groups including, but not limited to, the following monocyclic groups: 2-furyl, 3-furyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (e.g., 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g., 5H-tetrazolyl, 2H-tetrazolyl), triazolyl (e.g., 2-triazolyl, 5-triazolyl, 4H-1,2, 4-triazolyl, 1,2, 3-triazolyl), 2-thienyl, 3-thienyl, pyrazolyl (e.g., 2-pyrazolyl and 3-pyrazolyl), isothiazolyl, 1,2, 3-oxadiazolyl, 1,2, 3-triazolyl, 1, 3-oxadiazolyl, 1,2, 4-triazolyl, 1, 3-oxadiazolyl, 1, 3-triazolyl, 1, 4-oxadiazolyl, 1, 3-thiodiazolyl, 1, 4-triazolyl; also included are the following bi-or tricyclic groups, but in no way limited to these groups: indolinyl, 1,2,3, 4-tetrahydroisoquinolinyl, benzimidazolyl, benzofuranyl, benzothienyl, indolyl (e.g., 2-indolyl), purinyl, quinolinyl (e.g., 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), isoquinolinyl (e.g., 1-isoquinolinyl, 3-isoquinolinyl, or 4-isoquinolinyl), phenoxathianyl, dibenzoimidazolyl, dibenzofuranyl, dibenzothienyl. The heteroaryl group is optionally substituted with one or more substituents described herein.
The terms "comprising" or "including" are used in an open-ended fashion, i.e., including the teachings described herein, but not excluding additional aspects.
The invention will be further illustrated with reference to specific examples. The experimental methods used in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1 without addition of a phase transfer catalyst
Substrate (1.0 eq) and proton sponge (2.0 eq) were mixed in a 100mL two-necked flask and magneton was added. The above mixture was dissolved in 12mL of PO (OMe) 3 Pumping for 5 min by using a high vacuum oil pump, three-way exchanging argon until no bubble is generated, and repeating the operation for 3 times. After stirring at-15℃for 10 minutes, POCl was added to the above mixed solution 3 (2.0 eq) then stirred in an ice bath for 2-3 hours, a dry 100mL two-necked flask was taken and added with magnetic stirrer K 4 PPi(5.0eq)，Bu 3 N (5.0 eq) was injected with dry DMF (12 mL) and pumped on a high vacuum oil pump for 5 minutes, three exchanges of argon were used, repeated 3 times. Dropwise adding the first-step reaction solution into K under the protection of ice bath and nitrogen 4 In a PPi flask, the flask was rinsed with 10mL DMF and stirring was continued for 12 hours under an ice bath. HPLC demonstrated no product formation and the starting material was also decomposed.
EXAMPLE 2 addition of trimethylamine hydrochloride (tertiary ammonium salt) as a phase transfer catalyst
Substrate (1.0 eq) and proton sponge (2.0 eq) were mixed in a 100mL two-necked flask and magneton was added. The above mixture was dissolved in 12mL of PO (OMe) 3 Pumping for 5 min by using a high vacuum oil pump, three-way exchanging argon until no bubble is generated, and repeating the operation for 3 times. After stirring at-15℃for 10 minutes, POCl was added to the above mixed solution 3 (2.0 eq) then stirred in an ice bath for 2-3 hours, a dry 100mL two-necked flask was taken and added with magnetic stirrer K 4 PPi (5.0 eq), trimethylamine hydrochloride (0.5 eq) and Bu 3 N (5.0 eq) was injected with dry DMF (12 mL) and pumped on a high vacuum oil pump for 5 minutes, three exchanges of argon were used, repeated 3 times. Dropwise adding the first-step reaction solution into K under the protection of ice bath and nitrogen 4 In a PPi flask, the flask was washed with 10mL DMF and stirring was continued for 24 hours at room temperature. The reaction was quenched with 10mL TEAB (1.0 m, ph=7.7) and the product was isolated by direct prep HPLC (H 2 O/CH 3 Cn=10:90-90:10), to give a triphosphorylated product in 6% yield.
EXAMPLE 3 tetrabutylammonium bromide (0.2 eq) was added as a phase transfer catalyst
Substrate (1.0 eq) and proton sponge (2.0 eq) were mixed in a 100mL two-necked flask and magneton was added. The above mixture was dissolved in 12mL of PO (OMe) 3 Pumping for 5 min by using a high vacuum oil pump, three-way exchanging argon until no bubble is generated, and repeating the operation for 3 times. After stirring at-15℃for 10 minutes, POCl was added to the above mixed solution 3 (2.0 eq) then stirred in an ice bath for 2-3 hours, a dry 100mL two-necked flask was taken and added with magnetic stirrer K 4 PPi (5.0 eq), tetrabutylammonium bromide (0.2 eq) and Bu 3 N (5.0 eq) was injected with dry DMF (12 mL) and pumped on a high vacuum oil pump for 5 minutes, three exchanges of argon were used, repeated 3 times. Dropwise adding the first-step reaction solution into K under the protection of ice bath and nitrogen 4 In a PPi flask, the flask was washed with 10mL DMF and stirring was continued for 24 hours at room temperature. The reaction was quenched with 10mL TEAB (1.0 m, ph=7.7) and the product was isolated by direct prep HPLC (H 2 O/CH 3 Cn=10:90-90:10), to give a triphosphorylated product in 21% yield.
EXAMPLE 4 tetrabutylammonium bromide (0.5 eq) was added as a phase transfer catalyst
Substrate (1.0 eq) and proton sponge (2.0 eq) were mixed in a 100mL two-necked flask and magneton was added. The above mixture was dissolved in 12mL of PO (OMe) 3 Pumping for 5 min by using a high vacuum oil pump, three-way exchanging argon until no bubble is generated, and repeating the operation for 3 times. After stirring at-15℃for 10 minutes, POCl was added to the above mixed solution 3 (2.0 eq) then stirred in an ice bath for 2-3 hours, a dry 100mL two-necked flask was taken and added with magnetic stirrer K 4 PPi (5.0 eq), tetrabutylammonium bromide (0.5 eq) and Bu 3 N (5.0 eq) was injected with dry DMF (12 mL) and pumped on a high vacuum oil pump for 5 minutes, three exchanges of argon were used, repeated 3 times. Dropwise adding the first-step reaction solution into K under the protection of ice bath and nitrogen 4 In a PPi flask, the flask was washed with 10mL DMF and stirring was continued for 12 hours at room temperature. The reaction was quenched with 10mL TEAB (1.0 m, ph=7.7) and the product was isolated by direct prep HPLC (H 2 O/CH 3 Cn=10:90-90:10), to give a triphosphorylated product in 43% yield.
EXAMPLE 5 tetrabutylammonium bromide (1.5 eq) was added as a phase transfer catalyst
Substrate (1.0 eq) and proton sponge (2.0 eq) were mixed in a 100mL two-necked flask and magneton was added. The above mixture was dissolved in 12mL of PO (OMe) 3 Pumping for 5 min by using a high vacuum oil pump, three-way exchanging argon until no bubble is generated, and repeating the operation for 3 times.After stirring at-15℃for 10 minutes, POCl was added to the above mixed solution 3 (2.0 eq) then stirred in an ice bath for 2-3 hours, a dry 100mL two-necked flask was taken and added with magnetic stirrer K 4 PPi (5.0 eq), tetrabutylammonium bromide (1.5 eq) and Bu 3 N (5.0 eq) was injected with dry DMF (12 mL) and pumped on a high vacuum oil pump for 5 minutes, three exchanges of argon were used, repeated 3 times. Dropwise adding the first-step reaction solution into K under the protection of ice bath and nitrogen 4 In a PPi flask, the flask was washed with 10mL DMF and stirring was continued for 12 hours at room temperature. The reaction was quenched with 10mL TEAB (1.0 m, ph=7.7) and the product was isolated by direct prep HPLC (H 2 O/CH 3 Cn=10:90-90:10), to give a triphosphorylated product in 42% yield.
Example 6 addition of 18-crown-6 (0.5 eq) as a phase transfer catalyst
Substrate (1.0 eq) and proton sponge (2.0 eq) were mixed in a 100mL two-necked flask and magneton was added. The above mixture was dissolved in 12mL of PO (OMe) 3 Pumping for 5 min by using a high vacuum oil pump, three-way exchanging argon until no bubble is generated, and repeating the operation for 3 times. After stirring at-15℃for 10 minutes, POCl was added to the above mixed solution 3 (2.0 eq) then stirred in an ice bath for 2-3 hours, a dry 100mL two-necked flask was taken and added with magnetic stirrer K 4 PPi (5.0 eq), 18-crown-6 (0.5 eq) and Bu 3 N (5.0 eq) was injected with dry DMF (12 mL) and pumped on a high vacuum oil pump for 5 minutes, three exchanges of argon were used, repeated 3 times. Dropwise adding the first-step reaction solution into K under the protection of ice bath and nitrogen 4 In a PPi flask, the flask was washed with 10mL DMF and stirring was continued for 12 hours at room temperature. The reaction was quenched with 10mL TEAB (1.0 m, ph=7.7) and the product was isolated by direct prep HPLC (H 2 O/CH 3 Cn=10:90-90:10), to give the triphosphates in 35% yield.
EXAMPLE 7 addition of 18-crown-6 (1.5 eq) as phase transfer catalyst
Substrate (1.0 eq) and proton sponge (2.0 eq) were mixed in a 100mL two-necked flask and magneton was added. The above mixture was dissolved in 12mL of PO (OMe) 3 Pumping for 5 min by using a high vacuum oil pump, three-way exchanging argon until no bubble is generated, and repeating the operation for 3 times. After stirring at-15℃for 10 minutes, POCl was added to the above mixed solution 3 (2.0 eq) then stirred in an ice bath for 2-3 hours, a dry 100mL two-necked flask was taken and added with magnetic stirrer K 4 PPi (5.0 eq), 18-crown-6 (1.5 eq) and Bu 3 N (5.0 eq) was injected with dry DMF (12 mL) and pumped on a high vacuum oil pump for 5 minutes, three exchanges of argon were used, repeated 3 times. Dropwise adding the first-step reaction solution into K under the protection of ice bath and nitrogen 4 In a PPi flask, the flask was washed with 10mL DMF and stirring was continued for 12 hours at room temperature. The reaction was quenched with 10mL TEAB (1.0 m, ph=7.7) and the product was isolated by direct prep HPLC (H 2 O/CH 3 Cn=10:90-90:10), to give a triphosphorylated product in 30% yield.
Example 8 use of Na 4 PPi nucleotide Synthesis
Substrate (1.0 eq) and proton sponge (2.0 eq) were mixed in a 100mL two-necked flask and magneton was added. The above mixture was dissolved in 12mL of PO (OMe) 3 Pumping for 5 min by using a high vacuum oil pump, three-way exchanging argon until no bubble is generated, and repeating the operation for 3 times. After stirring at-15℃for 10 minutes, POCl was added to the above mixed solution 3 (2.0 eq) then stirred in an ice bath for 2-3 hours, a dry 100mL two-necked flask was taken and added with magnetic stirrer Na 4 PPi (5.0 eq), tetrabutylammonium bromide (0.5 eq) and Bu 3 N (5.0 eq) was injected with dry DMF (12 mL) and pumped on a high vacuum oil pump for 5 minutes, three exchanges of argon were used, repeated 3 times. Dropwise adding the first-step reaction solution under the protection of ice bath and nitrogenNa 4 In a PPi flask, the flask was washed with 10mL DMF and stirring was continued for 12 hours at room temperature. The reaction was quenched with 10mL TEAB (1.0 m, ph=7.7) and the product was isolated by direct prep HPLC (H 2 O/CH 3 Cn=10:90-90:10), to give a triphosphorylated product in 43% yield.
Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In the description of the present invention, the meaning of "plurality" means at least two, for example, two, three, etc., unless specifically defined otherwise.
In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, the different embodiments or examples described in this specification and the features of the different embodiments or examples may be combined and combined by those skilled in the art without contradiction.
While embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the invention, and that variations, modifications, alternatives and variations may be made to the above embodiments by one of ordinary skill in the art within the scope of the invention.
Claims (10)
1. A method for synthesizing a nucleotide or nucleotide analogue, characterized in that a nucleotide is subjected to nucleophilic substitution reaction with a phosphate, pyrophosphate or tripolyphosphate which is a non-ammonium phosphate salt under the catalysis of a phase transfer catalyst to obtain the nucleotide or nucleotide analogue,
Wherein the nucleoside has a structure shown in formula (I) or a stereoisomer, tautomer, pharmaceutically acceptable salt or prodrug thereof,
wherein X, Y are each independently selected from H or-OH; r is H or
The method comprises the steps of carrying out a first treatment on the surface of the A is adenine, guanine, cytosine, uracil or thymine;
the phase transfer catalyst comprises at least one selected from trimethylamine hydrochloride, tetrabutylammonium bromide and 18-crown-6;
the molar ratio of the nucleoside to the phosphate, pyrophosphate or tripolyphosphate and the phase transfer catalyst is 1:5: (0.5-1.5).
2. The method of claim 1, further comprising combining the nucleoside with a proton sponge and POCl 3 The contacting is performed so as to obtain a nucleoside electrophilic intermediate.
3. The method of claim 2, wherein the nucleoside is associated with a proton sponge and POCl 3 The contact is made in PO (OMe) 3 Is carried out in the following steps.
4. The method of claim 1, wherein the molar ratio of nucleoside to phosphate, pyrophosphate, or tripolyphosphate is 1:5.
5. the method of claim 1, wherein the phosphate, pyrophosphate, or tripolyphosphate comprises at least one selected from the group consisting of:
6. The method of claim 1, wherein the nucleotide comprises at least one selected from the group consisting of nucleoside monophosphates, nucleoside diphosphate, nucleoside triphosphate, and nucleoside tetraphosphate.
7. The method according to claim 6, wherein the nucleoside monophosphate has a structure shown below,
8. the method according to claim 6, wherein the nucleoside diphosphate has a structure shown below,
9. the method according to claim 6, wherein the nucleoside triphosphate has a structure shown below,
10. the method according to claim 6, wherein the nucleoside tetraphosphate has a structure as shown below,
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