CN115260262A - Preparation method of cytosine azide - Google Patents
Preparation method of cytosine azide Download PDFInfo
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- CN115260262A CN115260262A CN202210948727.7A CN202210948727A CN115260262A CN 115260262 A CN115260262 A CN 115260262A CN 202210948727 A CN202210948727 A CN 202210948727A CN 115260262 A CN115260262 A CN 115260262A
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- Prior art keywords
- compound
- azide
- cytosine
- solvent
- reacting
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- -1 cytosine azide Chemical class 0.000 title claims abstract description 35
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Natural products NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 title claims abstract description 27
- 229940104302 cytosine Drugs 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 150000001875 compounds Chemical class 0.000 claims abstract description 45
- 238000006243 chemical reaction Methods 0.000 claims abstract description 26
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims abstract description 21
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 19
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000002904 solvent Substances 0.000 claims abstract description 18
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 claims abstract description 13
- 229940125782 compound 2 Drugs 0.000 claims abstract description 12
- JKANAVGODYYCQF-UHFFFAOYSA-N prop-2-yn-1-amine Chemical compound NCC#C JKANAVGODYYCQF-UHFFFAOYSA-N 0.000 claims abstract description 12
- 229940126214 compound 3 Drugs 0.000 claims abstract description 11
- 229940125898 compound 5 Drugs 0.000 claims abstract description 10
- 239000012043 crude product Substances 0.000 claims abstract description 8
- 239000012445 acidic reagent Substances 0.000 claims abstract description 6
- 239000003054 catalyst Substances 0.000 claims abstract description 6
- 229910052763 palladium Inorganic materials 0.000 claims abstract description 6
- 238000004537 pulping Methods 0.000 claims abstract description 6
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 5
- KRHYYFGTRYWZRS-UHFFFAOYSA-M Fluoride anion Chemical compound [F-] KRHYYFGTRYWZRS-UHFFFAOYSA-M 0.000 claims abstract description 4
- 230000003301 hydrolyzing effect Effects 0.000 claims abstract description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 42
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 32
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 30
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 16
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 12
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 229940125904 compound 1 Drugs 0.000 claims description 7
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 6
- BCNZYOJHNLTNEZ-UHFFFAOYSA-N tert-butyldimethylsilyl chloride Chemical compound CC(C)(C)[Si](C)(C)Cl BCNZYOJHNLTNEZ-UHFFFAOYSA-N 0.000 claims description 5
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 5
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 3
- 239000012320 chlorinating reagent Substances 0.000 claims description 3
- 230000007062 hydrolysis Effects 0.000 claims description 2
- 238000006460 hydrolysis reaction Methods 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- 239000006227 byproduct Substances 0.000 abstract description 12
- 150000001540 azides Chemical class 0.000 abstract description 6
- OMPJBNCRMGITSC-UHFFFAOYSA-N Benzoylperoxide Chemical group C=1C=CC=CC=1C(=O)OOC(=O)C1=CC=CC=C1 OMPJBNCRMGITSC-UHFFFAOYSA-N 0.000 abstract 1
- 229960003328 benzoyl peroxide Drugs 0.000 abstract 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 12
- 239000007787 solid Substances 0.000 description 9
- 239000012074 organic phase Substances 0.000 description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 6
- QAEDZJGFFMLHHQ-UHFFFAOYSA-N trifluoroacetic anhydride Chemical compound FC(F)(F)C(=O)OC(=O)C(F)(F)F QAEDZJGFFMLHHQ-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 5
- 125000003277 amino group Chemical group 0.000 description 5
- JRNVZBWKYDBUCA-UHFFFAOYSA-N N-chlorosuccinimide Chemical compound ClN1C(=O)CCC1=O JRNVZBWKYDBUCA-UHFFFAOYSA-N 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 4
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 4
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 4
- 125000000714 pyrimidinyl group Chemical group 0.000 description 4
- 239000000741 silica gel Substances 0.000 description 4
- 229910002027 silica gel Inorganic materials 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- FPGGTKZVZWFYPV-UHFFFAOYSA-M tetrabutylammonium fluoride Chemical compound [F-].CCCC[N+](CCCC)(CCCC)CCCC FPGGTKZVZWFYPV-UHFFFAOYSA-M 0.000 description 4
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 4
- NTJBWZHVSJNKAD-UHFFFAOYSA-N triethylazanium;fluoride Chemical compound [F-].CC[NH+](CC)CC NTJBWZHVSJNKAD-UHFFFAOYSA-N 0.000 description 4
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 3
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 3
- 101100054666 Streptomyces halstedii sch3 gene Proteins 0.000 description 3
- 235000011114 ammonium hydroxide Nutrition 0.000 description 3
- 238000005520 cutting process Methods 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 229930024421 Adenine Natural products 0.000 description 2
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- 101150003085 Pdcl gene Proteins 0.000 description 2
- 229960000643 adenine Drugs 0.000 description 2
- IVRMZWNICZWHMI-UHFFFAOYSA-N azide group Chemical group [N-]=[N+]=[N-] IVRMZWNICZWHMI-UHFFFAOYSA-N 0.000 description 2
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 229940113082 thymine Drugs 0.000 description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 2
- WJKHJLXJJJATHN-UHFFFAOYSA-N triflic anhydride Chemical compound FC(F)(F)S(=O)(=O)OS(=O)(=O)C(F)(F)F WJKHJLXJJJATHN-UHFFFAOYSA-N 0.000 description 2
- FIQMHBFVRAXMOP-UHFFFAOYSA-N triphenylphosphane oxide Chemical compound C=1C=CC=CC=1P(C=1C=CC=CC=1)(=O)C1=CC=CC=C1 FIQMHBFVRAXMOP-UHFFFAOYSA-N 0.000 description 2
- NTNKNFHIAFDCSJ-UHFFFAOYSA-N (2-nitrophenyl) thiohypochlorite Chemical compound [O-][N+](=O)C1=CC=CC=C1SCl NTNKNFHIAFDCSJ-UHFFFAOYSA-N 0.000 description 1
- KZPYGQFFRCFCPP-UHFFFAOYSA-N 1,1'-bis(diphenylphosphino)ferrocene Chemical compound [Fe+2].C1=CC=C[C-]1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=C[C-]1P(C=1C=CC=CC=1)C1=CC=CC=C1 KZPYGQFFRCFCPP-UHFFFAOYSA-N 0.000 description 1
- YJTKZCDBKVTVBY-UHFFFAOYSA-N 1,3-Diphenylbenzene Chemical group C1=CC=CC=C1C1=CC=CC(C=2C=CC=CC=2)=C1 YJTKZCDBKVTVBY-UHFFFAOYSA-N 0.000 description 1
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 description 1
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical class [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 125000000066 S-methyl group Chemical group [H]C([H])([H])S* 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical class [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 238000003477 Sonogashira cross-coupling reaction Methods 0.000 description 1
- 125000003668 acetyloxy group Chemical group [H]C([H])([H])C(=O)O[*] 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 150000008064 anhydrides Chemical class 0.000 description 1
- 229910052786 argon Inorganic materials 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 230000009194 climbing Effects 0.000 description 1
- 230000000536 complexating effect Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 229910001431 copper ion Inorganic materials 0.000 description 1
- 229910001873 dinitrogen Inorganic materials 0.000 description 1
- 125000004185 ester group Chemical group 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000007867 post-reaction treatment Methods 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 239000012265 solid product Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- ZVQXQPNJHRNGID-UHFFFAOYSA-N tetramethylsuccinonitrile Chemical compound N#CC(C)(C)C(C)(C)C#N ZVQXQPNJHRNGID-UHFFFAOYSA-N 0.000 description 1
- SEDZOYHHAIAQIW-UHFFFAOYSA-N trimethylsilyl azide Chemical compound C[Si](C)(C)N=[N+]=[N-] SEDZOYHHAIAQIW-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
- C07H19/073—Pyrimidine radicals with 2-deoxyribosyl as the saccharide radical
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Saccharide Compounds (AREA)
- Low-Molecular Organic Synthesis Reactions Using Catalysts (AREA)
- Plural Heterocyclic Compounds (AREA)
Abstract
The invention discloses a preparation method of a cytosine azide, which comprises the following steps: s1, adding a compound 2, an acidic reagent and an anhydride compound for reaction to obtain a compound 3; s2, hydrolyzing the compound 3 to obtain a compound 4; s3, reacting the compound 4, trifluoroacetylated propargylamine, cuI, a palladium catalyst and triethylamine in a first solvent and an inert atmosphere to obtain a crude product, and pulping the crude product with diethyl ether to obtain a compound 5; s4, reacting the compound 5 with acetic anhydride to obtain a compound 6; s5, reacting the compound 6 with a chlorinated reagent; adding azide compounds at 0-5 ℃ for continuous reaction to obtain compounds 7; s6, reacting the compound 7 with an organic fluoride to obtain a compound 8; the preparation method of the invention avoids the generation of acetoxyl (OAc) by-products and ring-closing by-products, and improves the yield of the azide reaction.
Description
Technical Field
The invention relates to the technical field of organic synthesis, in particular to a preparation method of a cytosine azide.
Background
With the development of the second generation sequencing technology, scientists have continuously updated their studies on the 3' -blocking group of nucleotide triphosphate compounds. From the original amino, allyl, ester groups to disulfide, azide functionalities, azide groups were found to be the best blocking groups to date. It is a very small group, and has little influence on the base recognition of polymerase; the cutting condition is mild, the cutting speed is high, and nitrogen released after cutting is free from pollution.
There are four types of nucleotides, adenine a, guanine G, cytosine C and thymine T, which synthesize an azide compound with the lowest yield of cytosine C. From adenine A, guanine G and thymine T, the amino groups on the pyrimidine ring are protected, and then the azide compound is synthesized. However, the cytosine C pyrimidine ring has a naked amino group, and an electron-donating group can influence the synthetic yield of the step of azide. Previous studies have also recognized this, and would be protected with benzoyl. However, the yield of azide reaction after benzoyl protection is only 30%, and there are also OAc by-products that are difficult to separate, which can affect the depth of sequencing.
Therefore, there is a need to provide a novel method for preparing cytosine azide compounds.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art. Therefore, the invention provides a preparation method of the cytosine azide, which can effectively improve the yield of an azide reaction, has no OAc by-product, and prevents the influence of the OAc by-product on the sequencing depth.
According to the embodiment of the first aspect of the invention, a preparation method of cytosine azide is provided, which comprises the following steps:
s1, adding a compound 2, an acidic reagent and an anhydride compound into dimethyl sulfoxide to react to obtain a compound 3;
s2, hydrolyzing the compound 3 to obtain a compound 4;
s3, reacting the compound 4, trifluoroacetylated propargylamine, cuI, a palladium catalyst and triethylamine in a first solvent and an inert atmosphere to obtain a crude product, and pulping and purifying the crude product with diethyl ether to obtain a compound 5;
s4, reacting the compound 5 with acetic anhydride in a second solvent to obtain a compound 6;
s5, reacting the compound 6 with a chlorinated reagent in a third solvent; adding azide compounds at 0-5 ℃ for continuous reaction to obtain compounds 7;
s6, reacting the compound 7 with organic fluoride in a fourth solvent to obtain a compound 8;
wherein the structural formulas of the compounds 2 to 8 are as follows:
the preparation method of the cytosine azide compound provided by the embodiment of the invention has at least the following beneficial effects:
first, the present invention avoids the generation of acetoxy (OAc) by-product by the smart reaction of step S1 and step S2. This is because, in the reaction in step S1, not only the thiomethylation but also the formation of a similar polar OAc by-product are caused, and in step S2 of the present invention, the OAc by-product is hydrolyzed under the condition of ammonia water to obtain a by-product having a large difference in polarity, and it is sufficient to separate it as it is.
Secondly, according to the invention, firstly, acetyl protection and thiomethyl reaction are respectively carried out on amino and hydroxyl in the compound 2, then acetyl is removed, and then Sonogashira reaction is carried out, so that a ring-closing byproduct can be prevented from being generated.
In the step S3, the compound 5 is extracted by adopting an ether pulping mode, and triphenyl oxygen ether which is difficult to separate is removed; this is because it is difficult to remove triphenylphosphine oxide from both the forward silica gel and the reverse silica gel.
Finally, the exposed amino group on the cytosine is subjected to acetyl protection and then subjected to an azide reaction, and the yield of the product reaches 60%.
Wherein, the structures of the OAc byproduct and the closed-loop byproduct are as follows:
according to some embodiments of the invention, compound 2 is prepared by:
reacting the compound 1, pyridine and tert-butyldimethylsilyl chloride to obtain a compound 2;
the structural formula of the compound 1 is as follows:
according to some embodiments of the invention, the trifluoroacetylated propargylamine is prepared by:
under the existence of dichloromethane, propargylamine and trifluoroacetic anhydride react to obtain trifluoroacetylated propargylamine.
According to some embodiments of the invention, the anhydride-based compound comprises at least one of acetic anhydride, triflic anhydride, or trifluoroacetic anhydride.
According to some embodiments of the invention, the acidic reagent comprises at least one of acetic acid or p-toluenesulfonic acid.
According to some embodiments of the invention, the palladium catalyst comprises Pd (PPh) 3 ) 4 、PdCl 2 (dppf)、PdCl 2 (PPh 3 ) 2 At least one of (1).
According to some embodiments of the invention, the chlorinating agent comprises at least one of N-chlorosuccinimide, o-nitrobenzenesulfenyl chloride or sulfone chloride.
According to some embodiments of the invention, the azide-based compound comprises sodium azide or trimethylsilyl azide (TMSN) 3 ) At least one of (1).
According to some embodiments of the invention, the organofluoro compound is selected from at least one of triethylamine hydrofluoric acid salt, tetrabutylammonium fluoride (TBAF).
According to some embodiments of the invention, the compound 1 and tert-butyldimethylsilyl chloride are present in a molar ratio of 1: (1-1.5).
According to some embodiments of the invention, in step S1, the volume ratio of the acidic reagent, the dimethyl sulfoxide and the acid anhydride compound is 1: (1.5-3): (2.5-4).
According to some embodiments of the invention, in step S2, the compound 3 is hydrolyzed with ammonia to obtain a compound 4.
According to some embodiments of the invention, in step S3, the reaction comprises at least the following conditions:
the temperature is 30-70 ℃; the time is 1-5 h.
According to some embodiments of the invention, in step S3, the compound 4, trifluoroacetylated propargylamine, cuI, palladium catalyst, triethylamine are in a molar ratio of 1: (2-3): (0.1-0.2): (0.05-0.1): (2-3).
According to some embodiments of the invention, in step S4, the temperature of the reaction is 20 ℃ to 30 ℃.
According to some embodiments of the invention, in step S5, the molar ratio of the compound 6, the chlorinating reagent and the azide compound is 1: (1.2-1.5): (4-10).
According to some embodiments of the invention, in step S6, the molar ratio of compound 7 to organofluoro compound is 1: (8-15).
According to some embodiments of the invention, the first solvent is selected from at least one of N, N-dimethylformamide or dimethylsulfoxide.
According to some embodiments of the invention, the second solvent is selected from at least one of dichloromethane or pyridine.
According to some embodiments of the invention, the third solvent is selected from N, N-dimethylformamide.
According to some embodiments of the invention, the fourth solvent is selected from at least one of tetrahydrofuran or ethyl acetate.
According to some embodiments of the invention, the CAS number of the hydrofluoric acid triethylamine salt is 73602-61-6.
According to some embodiments of the invention, the inert atmosphere comprises at least one of nitrogen, argon.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.
Detailed Description
The following are specific examples of the present invention, and the technical solutions of the present invention will be further described with reference to the examples, but the present invention is not limited to the examples.
The reagents, methods and equipment adopted by the invention are conventional in the technical field if no special description is given.
The raw materials in the examples of the invention are as follows:
solvents, reagents and compound 1 used in the present invention were all purchased from annagiki chemicals.
Trifluoroacetylated propargylamine: weighing 25g (0.45mol, 1eq) of propargylamine, dissolving the propargylamine in 600mL of dichloromethane, slowly adding 110g of trifluoroacetic anhydride (0.5mol, 1.1eq) at zero temperature, heating to normal temperature after finishing the dropwise addition, stirring for half an hour, monitoring the reaction by TLC, developing by potassium permanganate, and finishing the consumption of raw materials. Adding saturated sodium bicarbonate to adjust the pH to be about =7, extracting the water phase twice, combining the organic phases, drying the organic phases by anhydrous sodium sulfate, and then concentrating the organic phases to obtain a crude product, and distilling the crude product under reduced pressure to obtain 57.7g of colorless oily liquid; the yield thereof was found to be 85%.
Example 1
Example 1 provides a method of preparing compound 2, comprising the steps of:
preparation of compound 2: compound 1 was dissolved in pyridine, followed by addition of 1.2 equivalents of t-butyldimethylchlorosilane (TBSCl) and reaction overnight. TLC monitored the completion of the reaction (EA-crawl), post-treatment rotary evaporated most of the pyridine, which was then poured into a large volume of water to precipitate a white solid using n-hexane (PE)/Ethyl Acetate (EA) in a volume ratio of 10:1, pulping, and filtering to obtain a compound 2;
the nuclear magnetic data for compound 2 is as follows:
1 H NMR(d6 DMSO)δ0.21(s,6H,CH 3 )，0.98(s,9H,t-Bu)，2.15-2.35(m,2H)，3.85(dd,J＝11.2,3.4Hz,1H)，4.15-4.17(m,1H)，4.324.35(m,1H)，5.85(t,1H)，8.25(s,1H)，9.5(s,1H)，10.03(s,1H)。
example 2
Embodiment 2 provides a cytosine azide preparation method, including the following steps:
dissolving S1 and 2 in dimethyl sulfoxide (DMSO), adding acetic acid, and adding acetic anhydride (Ac) under ice bath 2 O), acetic acid/dimethyl sulfoxide/acetic anhydride volume ratio of 1:2:3; the amino group on the pyrimidine ring was simultaneously protected by acetyl, reacted overnight, monitored by TLC for completion (PE/EA =1, 1-plate, rf = 0.4), post-treatment first added large amount of water and ethyl acetate to remove most of the acetic acid, then added saturated sodium bicarbonate to adjust pH to 7, combined the organic phases and concentrated to give compound 3;
the nuclear magnetic data for compound 3 are as follows:
1 H NMR(d 6 DMSO)δ0.21(s,6H,CH 3 )，0.98(s,9H,t-Bu)，2.10-2.33(m,2H)，2.15(s,3H,SCH3)，2.35(s,3H,COCH 3 )，3.80(dd,J＝11.4,3.2Hz,1H)，4.15-4.17(m,1H),4.30(s,2H),4.32-4.35(m,1H)，4.90(dd,J＝14.4,6.2Hz,2H)，6.03(t,1H)，8.25(s,1H)，9.48(s,1H)，10.02(s,1H)。
s2, dissolving the compound 3 with methanol, adding ammonia water for hydrolysis, separating out a solid from the ammonia water, and filtering to obtain a white solid compound 4 with high purity;
the nuclear magnetic data for compound 4 are as follows:
1 H NMR(d 6 DMSO)δ0.21(s,6H,CH 3 )，0.98(s,9H,t-Bu)，2.10-2.33(m,2H)，2.15(s,3H,SCH3)，3.80(dd,J＝11.4,3.2Hz,1H)，4.15-4.17(m,1H)，4.30(s,2H)，4.32-4.35(m,1H)，4.90(dd,J＝14.4,6.2Hz,2H)，6.03(t,1H)，8.25(s,1H)，9.48(s,1H)，10.02(s,1H)。
s3, compound 4 was dissolved in DMF, trifluoroacetylated propargylamine was added, nitrogen was substituted three times, followed by CuI, pd (PPh) 3 ) 4 After the addition, nitrogen gas replacement is carried out, triethylamine is finally added, stirring is carried out for 2-3 hours at 50 ℃, a TLC plate monitors the reaction (EA climbs the plate, rf = 0.3), the raw materials are basically completely consumed, the product is arranged below the TLC plate, after the reaction is cooled, the reaction liquid is poured into the mixed liquid of water and EA, the mixture is fully stirred, after one-time washing, saturated ammonium chloride is added for complexing copper ions, and finally, saturated salt is addedWashing with water, and drying with anhydrous sodium sulfate; then pulping the concentrated solid product with diethyl ether; filtering the product to obtain a white solid, wherein the yield is 68.5%, and the crude product is directly pulped by diethyl ether to remove the triphenylphosphine oxide to obtain a white solid compound 5 with high purity;
the nuclear magnetic data for compound 5 are as follows:
1 H NMR(d 6 DMSO)δ0.21(s,6H,CH 3 )，0.98(s,9H,t-Bu)，2.10-2.33(m,2H)，2.15(s,3H,SCH3)，3.80(dd,J＝11.4,3.2Hz,1H)，4.15-4.17(m,1H)，4.21(s,2H)，4.30(s,2H)，4.32-4.35(m,1H)，4.90(dd,J＝14.4,6.2Hz,2H)，6.03(t,1H)，8.25(s,1H)，9.48(s,1H)，10.02(s,1H)。
s4, compound 5 is dissolved with Dichloromethane (DCM), then the amino group on the pyrimidine ring is protected with 20 equivalents of acetic anhydride and stirred at room temperature for 3h. TLC plate monitoring reaction (DCM/MEOH =20 on 1 plate, rf = 0.3) showed essentially complete consumption of starting material with main product above. And (3) post-reaction treatment: pouring the reaction solution into DCM, neutralizing the reaction solution to be neutral by using saturated sodium bicarbonate, extracting an organic phase, washing the organic phase twice by using saturated salt water, drying and concentrating. The organic phase was passed through a silica gel column (DCM/MEOH =20 1) to give a white foamy solid. The yield was 99% and the purity was 95%.
The nuclear magnetic data for compound 6 are as follows:
1 H NMR(d 6 DMSO)δ0.12(s,6H,CH 3 )，0.78(s,9H,t-Bu)，2.03(s,3H,SCH 3 )，1.96-2.20(m,2H)，2.35(s,3H,COCH 3 )，3.67(dd,1H)，4.06-4.10(m,1H),，4.17(d,2H,J＝5.6Hz)，4.30(m,1H)，4.65(s,2H)，4.63(s,2H)，5.90(t,1Hz,J＝6.3Hz)，8.20(s,1H)，9.34(s,1H)，9.93(s,1H)；MH(+)＝593。
s5 and the compound 6 were dissolved in DMF, and 1.5 equivalents of N-chlorosuccinimide (NBS-Cl) were added thereto and stirred at room temperature for 1 hour. Subsequent TLC (EA, rf = 0.1) plates showed complete consumption of starting material, sodium azide was added under ice bath and stirred at rt for 1-2h after 10 min. The reaction was monitored on TLC plates (PE/EA =1 plate, rf = 0.3) showing complete consumption of intermediate with product above. The reaction solution was poured into a mixed solution of water and EA, and sufficiently stirred. After washing twice, the mixture was finally washed with saturated saline and dried over anhydrous sodium sulfate. The concentrate was purified on normal phase silica gel, PE/EA =2 and 1 the product was washed out. The product was concentrated to give a pale yellow oil. The mixture was dissolved in DCM, n-hexane was added, a pale yellow solid precipitated, the supernatant was decanted, and the solid was drained at 60% yield.
The nuclear magnetic data for compound 7 is as follows:
1 H NMR(d 6 DMSO)δ0.12(s,6H,CH 3 )，0.88(s,9H,t-Bu)，2.15-2.25(m,1H)，2.35(s,3H,COCH 3 )，2.45-2.60(m,1H)，3.80(dd,J＝11.4,3.2Hz,1H)，3.9(dd,J＝11.6,3.0Hz,1H)，4.15-4.17(m,1H)，4.30(s,2H),4.32-4.35(m,1H)，4.90(dd,J＝14.4,6.2Hz,2H)，6.03(t,1H)，8.25(s,1H)，9.48(s,1H)，10.02(s,1H)。
s6 and the compound 7 were dissolved in Tetrahydrofuran (THF), and 10 equivalents of triethylamine hydrofluoric acid (Et) salt were added 3 N-3 HF) to remove TBS protecting groups, reacting overnight, monitoring the completion of the reaction by TLC (EA plate climbing), performing post-treatment and rotary evaporation to remove THF, adding ethyl acetate and saturated sodium bicarbonate to adjust the pH value to 7, combining organic phases and concentrating to obtain a high-purity pale yellow solid compound 8.
The nuclear magnetic data for compound 8 is as follows:
1 H NMR(d 6 DMSO)δ2.01(s,3H,CH 3 CO)，2.15-2.23(m,2H)，3.45-3.6(m,2H)，3.90-4.01(m,1H)，4.75(s,2H)，5.2-4.17(t,1H)，5.96(t,1H)，8.41(s,1H)，9.32(s,1H)，9.96(t,1H)。
wherein the structural formulas of the compounds 1 to 8 are as follows:
while the present invention has been described in detail with reference to the embodiments thereof, the present invention is not limited to the embodiments, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art.
Claims (10)
1. A preparation method of cytosine azide is characterized by comprising the following steps:
s1, adding a compound 2, an acidic reagent and an anhydride compound into dimethyl sulfoxide to react to obtain a compound 3;
s2, hydrolyzing the compound 3 to obtain a compound 4;
s3, reacting the compound 4, trifluoroacetylated propargylamine, cuI, a palladium catalyst and triethylamine in a first solvent and an inert atmosphere to obtain a crude product, and pulping and purifying with diethyl ether to obtain a compound 5;
s4, reacting the compound 5 with acetic anhydride in a second solvent to obtain a compound 6;
s5, reacting the compound 6 with a chlorinated reagent in a third solvent; adding azide compounds at 0-5 ℃ for continuous reaction to obtain compounds 7;
s6, reacting the compound 7 with organic fluoride in a fourth solvent to obtain a compound 8;
wherein the structural formulas of the compounds 2 to 8 are as follows:
2. the method for preparing cytosine azide according to claim 1, wherein the compound 2 is prepared by the following steps:
reacting the compound 1, pyridine and tert-butyldimethylsilyl chloride to obtain a compound 2;
the structural formula of the compound 1 is as follows:
3. the method for preparing a cytosine azide compound according to claim 1, wherein in the step S1, the volume ratio of the acidic reagent to the dimethyl sulfoxide to the acid anhydride compound is 1: (1.5-3): (2.5-4).
4. The method for preparing cytosine azide according to claim 1, wherein in step S2, hydrolysis of compound 3 with aqueous ammonia is carried out to obtain compound 4.
5. The method for producing a cytosine azide compound according to claim 1 wherein, in the step S3, the reaction includes at least the following conditions:
the temperature is 30-70 ℃; the time is 1-5 h.
6. The method for preparing cytosine azide of claim 1 wherein, in step S3, the molar ratio of said compound 4, trifluoroacetylated propargylamine, cuI, palladium catalyst and triethylamine is 1: (2-3): (0.1-0.2): (0.05-0.1): (2-3).
7. The method for producing a cytosine azide compound according to claim 1, wherein the temperature of the reaction in the step S4 is 20 to 30 ℃.
8. The method for preparing cytosine azide according to claim 1, wherein in step S5, the molar ratio of the compound 6, the chlorinating reagent and the azide compound is 1: (1.2-1.5): (4-10).
9. The method for preparing cytosine azide according to claim 1, wherein the molar ratio of the compound 7 to the organic fluoride in step S6 is 1: (8 to 15).
10. The method for preparing a cytosine azide compound according to any one of claims 1 to 9, wherein the first solvent is at least one selected from the group consisting of N, N-dimethylformamide and dimethylsulfoxide;
preferably, the second solvent is selected from at least one of dichloromethane or pyridine;
preferably, the third solvent is selected from N, N-dimethylformamide;
preferably, the fourth solvent is selected from at least one of tetrahydrofuran or ethyl acetate.
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| CN116854749A (en) * | 2023-09-01 | 2023-10-10 | 深圳赛陆医疗科技有限公司 | Method for synthesizing nucleotide intermediate |
| CN117645636A (en) * | 2024-01-30 | 2024-03-05 | 深圳赛陆医疗科技有限公司 | Preparation method of adenine azide intermediate |
| CN117886850A (en) * | 2024-03-14 | 2024-04-16 | 深圳赛陆医疗科技有限公司 | Preparation method of azide compound |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004018497A2 (en) * | 2002-08-23 | 2004-03-04 | Solexa Limited | Modified nucleotides for polynucleotide sequencing |
| CN108779138A (en) * | 2015-09-28 | 2018-11-09 | 哥伦比亚大学董事会 | The design and synthesis of the nucleotide based on new disulfide bond connector of reversible termination object as DNA synthesis order-checkings |
| CN109053839A (en) * | 2018-09-20 | 2018-12-21 | 上海兆维科技发展有限公司 | The novel processing step of 3 '-O-CH2N3-2 '-O-Me- cytidine of nucleosides modifier |
-
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004018497A2 (en) * | 2002-08-23 | 2004-03-04 | Solexa Limited | Modified nucleotides for polynucleotide sequencing |
| CN108779138A (en) * | 2015-09-28 | 2018-11-09 | 哥伦比亚大学董事会 | The design and synthesis of the nucleotide based on new disulfide bond connector of reversible termination object as DNA synthesis order-checkings |
| CN109053839A (en) * | 2018-09-20 | 2018-12-21 | 上海兆维科技发展有限公司 | The novel processing step of 3 '-O-CH2N3-2 '-O-Me- cytidine of nucleosides modifier |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116854749A (en) * | 2023-09-01 | 2023-10-10 | 深圳赛陆医疗科技有限公司 | Method for synthesizing nucleotide intermediate |
| CN116854749B (en) * | 2023-09-01 | 2023-11-21 | 深圳赛陆医疗科技有限公司 | Method for synthesizing nucleotide intermediate |
| CN117645636A (en) * | 2024-01-30 | 2024-03-05 | 深圳赛陆医疗科技有限公司 | Preparation method of adenine azide intermediate |
| CN117645636B (en) * | 2024-01-30 | 2024-04-16 | 深圳赛陆医疗科技有限公司 | Preparation method of adenine azide intermediate |
| CN117886850A (en) * | 2024-03-14 | 2024-04-16 | 深圳赛陆医疗科技有限公司 | Preparation method of azide compound |
| CN117886850B (en) * | 2024-03-14 | 2024-07-05 | 深圳赛陆医疗科技有限公司 | Preparation method of azide compound |
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