CN115260262B - Cytosine azide Process for the preparation of compounds - Google Patents
Cytosine azide Process for the preparation of compounds Download PDFInfo
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- CN115260262B CN115260262B CN202210948727.7A CN202210948727A CN115260262B CN 115260262 B CN115260262 B CN 115260262B CN 202210948727 A CN202210948727 A CN 202210948727A CN 115260262 B CN115260262 B CN 115260262B
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- 150000001875 compounds Chemical class 0.000 title claims abstract description 44
- -1 Cytosine azide Chemical class 0.000 title claims abstract description 31
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Natural products NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 title claims abstract description 28
- 229940104302 cytosine Drugs 0.000 title claims abstract description 24
- 238000000034 method Methods 0.000 title claims description 15
- 238000002360 preparation method Methods 0.000 title abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims abstract description 31
- 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 20
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims abstract description 19
- 239000002904 solvent Substances 0.000 claims abstract description 18
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 claims abstract description 14
- 229940125782 compound 2 Drugs 0.000 claims abstract description 13
- 150000001540 azides Chemical class 0.000 claims abstract description 10
- 229940126214 compound 3 Drugs 0.000 claims abstract description 10
- 229940125898 compound 5 Drugs 0.000 claims abstract description 10
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 8
- 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
- 125000001309 chloro group Chemical group Cl* 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
- GAUSMJHDHCSZOX-UHFFFAOYSA-N 2,2,2-trifluoro-n-prop-2-ynylacetamide Chemical compound FC(F)(F)C(=O)NCC#C GAUSMJHDHCSZOX-UHFFFAOYSA-N 0.000 claims abstract description 4
- 150000004812 organic fluorine compounds Chemical class 0.000 claims abstract description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 39
- 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
- 229940125904 compound 1 Drugs 0.000 claims description 8
- JKANAVGODYYCQF-UHFFFAOYSA-N prop-2-yn-1-amine Chemical compound NCC#C JKANAVGODYYCQF-UHFFFAOYSA-N 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
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 4
- KRHYYFGTRYWZRS-UHFFFAOYSA-M Fluoride anion Chemical compound [F-] KRHYYFGTRYWZRS-UHFFFAOYSA-M 0.000 claims description 2
- 238000006460 hydrolysis reaction Methods 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 4
- 239000006227 byproduct Substances 0.000 abstract description 13
- 125000003668 acetyloxy group Chemical group [H]C([H])([H])C(=O)O[*] 0.000 abstract description 2
- 230000003301 hydrolyzing effect Effects 0.000 abstract description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 12
- 239000007787 solid Substances 0.000 description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 8
- 238000005160 1H NMR spectroscopy Methods 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
- 239000012074 organic phase Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- JRNVZBWKYDBUCA-UHFFFAOYSA-N N-chlorosuccinimide Chemical compound ClN1C(=O)CCC1=O JRNVZBWKYDBUCA-UHFFFAOYSA-N 0.000 description 4
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 4
- 125000003277 amino group Chemical group 0.000 description 4
- 230000009194 climbing Effects 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
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 125000000714 pyrimidinyl group Chemical group 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 238000003756 stirring Methods 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
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 3
- 101100054666 Streptomyces halstedii sch3 gene Proteins 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 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
- KRHYYFGTRYWZRS-UHFFFAOYSA-N Fluorane Chemical compound F KRHYYFGTRYWZRS-UHFFFAOYSA-N 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
- 239000012141 concentrate Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 238000003786 synthesis reaction 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
- 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
- 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
- 229910002666 PdCl2 Inorganic materials 0.000 description 1
- YNHIGQDRGKUECZ-UHFFFAOYSA-L PdCl2(PPh3)2 Substances [Cl-].[Cl-].[Pd+2].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 YNHIGQDRGKUECZ-UHFFFAOYSA-L 0.000 description 1
- 125000000066 S-methyl group Chemical group [H]C([H])([H])S* 0.000 description 1
- 238000003477 Sonogashira cross-coupling reaction Methods 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- TUCNEACPLKLKNU-UHFFFAOYSA-N acetyl Chemical compound C[C]=O TUCNEACPLKLKNU-UHFFFAOYSA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 150000008064 anhydrides Chemical class 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 229910052786 argon Inorganic materials 0.000 description 1
- 238000010009 beating Methods 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
- 239000012320 chlorinating reagent Substances 0.000 description 1
- 229910001431 copper ion Inorganic materials 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- PIBWKRNGBLPSSY-UHFFFAOYSA-L palladium(II) chloride Chemical compound Cl[Pd]Cl PIBWKRNGBLPSSY-UHFFFAOYSA-L 0.000 description 1
- 238000007867 post-reaction treatment Methods 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 238000006798 ring closing metathesis reaction Methods 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 239000012265 solid product Substances 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 150000003457 sulfones Chemical class 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000006228 supernatant Substances 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
- ILWRPSCZWQJDMK-UHFFFAOYSA-N triethylazanium;chloride Chemical compound Cl.CCN(CC)CC ILWRPSCZWQJDMK-UHFFFAOYSA-N 0.000 description 1
- NTJBWZHVSJNKAD-UHFFFAOYSA-N triethylazanium;fluoride Chemical compound [F-].CC[NH+](CC)CC NTJBWZHVSJNKAD-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
- 239000001226 triphosphate Substances 0.000 description 1
- 235000011178 triphosphate Nutrition 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 compound, 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, in a first solvent and an inert atmosphere, reacting the compound 4, trifluoroacetyl propargylamine, cuI, a palladium catalyst and triethylamine 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 chloro reagent; adding azide compounds at 0-5 ℃ for continuous reaction to obtain a compound 7; s6, reacting the compound 7 with an organofluoride to obtain a compound 8; the preparation method of the invention avoids the generation of acetoxy (OAc) byproducts and ring-closing byproducts, 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 second generation sequencing technology, scientists have continuously updated the study of the 3' -blocking group of nucleotide triphosphates. From the original amino, allyl, ester to disulfide, azide functions, the azide group was found to be the best blocking group so far. It is a very small group with little influence on polymerase recognition bases; the excision condition is mild, the excision speed is high, and the nitrogen released after excision is pollution-free.
There are four types of nucleotides, adenine a, guanine G, cytosine C and thymine T, respectively, which synthesize azide, with the lowest yield of cytosine C. From the viewpoints of adenine A, guanine G and thymine T, the amino groups on the pyrimidine ring are protected, and then the azide is synthesized. However, the cytosine C pyrimidine ring has an exposed amino group, and an electron donating group can influence the synthesis yield of the azide in one step. Previous studies have also recognized this, and the protection is provided by benzoyl groups. However, the yield of azide reaction after benzoyl protection is only 30% and OAc byproducts which are difficult to separate are also present, which can affect the depth of sequencing.
Thus, there is a need to provide a new process for the preparation of cytosine azides.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems existing in the prior art. Therefore, the invention provides a preparation method of a cytosine azide compound, which can effectively improve the yield of an azide reaction, does not have OAc byproducts and prevents the OAc byproducts from influencing the sequencing depth.
According to an embodiment of the first aspect of the present invention, there is provided a method for preparing a cytosine azide compound, comprising the steps of:
s1, adding a compound 2, an acidic reagent and an anhydride compound into dimethyl sulfoxide for reaction to obtain a compound 3;
S2, hydrolyzing the compound 3 to obtain a compound 4;
s3, in a first solvent and an inert atmosphere, reacting the compound 4, trifluoroacetyl propargylamine, cuI, a palladium catalyst and triethylamine to obtain a crude product, and pulping and purifying the crude product by diethyl ether to obtain a compound 5;
S4, in a second solvent, reacting the compound 5 with acetic anhydride to obtain a compound 6;
s5, in a third solvent, reacting the compound 6 with a chloro reagent; adding azide compounds at 0-5 ℃ for continuous reaction to obtain a compound 7;
s6, in a fourth solvent, reacting the compound 7 with an organic fluoride 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:
Firstly, the invention avoids the generation of acetoxy (OAc) by-products through smart reactions of step S1 and step S2. This is because in the reaction of step S1, not only sulfur methylation but also OAc byproducts having similar polarity are formed, and in step S2 of the present invention, OAc byproducts are hydrolyzed under the condition of aqueous ammonia to obtain byproducts having greatly different polarities, and the byproducts are directly separated.
Secondly, the invention firstly carries out acetyl protection and thiomethyl reaction on amino and hydroxyl in the compound 2 respectively, then removes acetyl, and then carries out Sonogashira reaction, thus avoiding the generation of ring-closing byproducts.
In the step S3, the method adopts a beating mode of diethyl ether to purify the compound 5, so that the triphenyloxy ether which is difficult to separate is removed; this is because it is difficult to remove triphenylphosphine oxide either on forward or reverse phase silica gel.
Finally, the exposed amino group on the cytosine is subjected to an acetyl protection and then subjected to an azide reaction, and the yield of the azide reaction reaches 60%.
Wherein the structures of OAc byproducts and ring-closure byproducts are as follows:
according to some embodiments of the invention, compound 2 is prepared by:
Reacting the compound 1, pyridine and tert-butyl dimethyl chlorosilane 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:
In the presence of dichloromethane, propargylamine and trifluoroacetic anhydride are reacted to obtain trifluoroacetylated propargylamine.
According to some embodiments of the invention, the anhydride-based compound includes at least one of acetic anhydride, trifluoromethanesulfonic anhydride, or trifluoroacetic anhydride.
According to some embodiments of the invention, the acidic reagent comprises at least one of acetic acid or p-toluene sulfonic acid.
According to some embodiments of the invention, the palladium catalyst comprises at least one of Pd (PPh 3)4、PdCl2(dppf)、PdCl2(PPh3)2).
According to some embodiments of the invention, the chlorinating reagent comprises at least one of N-chlorosuccinimide, o-nitrobenzenesulfochloride, or chlorinated sulfone.
According to some embodiments of the invention, the azide compound includes at least one of sodium azide or trimethylsilyl azide (TMSN 3).
According to some embodiments of the invention, the organofluoride is selected from at least one of triethylamine hydrofluoric acid, tetrabutylammonium fluoride (TBAF).
According to some embodiments of the invention, the molar ratio of compound 1 to t-butyldimethylchlorosilane is 1: (1-1.5).
According to some embodiments of the invention, in step S1, the volume ratio of the acidic reagent, dimethyl sulfoxide and 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 h-5 h.
According to some embodiments of the invention, in step S3, the molar ratio of compound 4, trifluoroacetylated propargylamine, cuI, palladium catalyst, triethylamine is 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 compound 6, chloro reagent and azide 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 triethylamine salt of hydrofluoric acid has a CAS number of 73602-61-6.
According to some embodiments of the invention, the inert atmosphere comprises at least one of nitrogen and 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 embodiments of the present invention, and the technical solutions of the present invention will be further described with reference to the embodiments, but the present invention is not limited to these embodiments.
The reagents, methods and apparatus employed in the present invention, unless otherwise specified, are all conventional in the art.
The raw materials in the examples of the present invention are as follows:
solvents, reagents and compound 1 used in the present invention were purchased from Yu Annai g of chemical reagent.
Trifluoroacetylated propargylamine: 25g (0.45 mol,1 eq) propargylamine was weighed, dissolved in 600mL of methylene chloride, then 110g of trifluoroacetic anhydride (0.5 mol,1.1 eq) was slowly added at zero degree, after the addition was completed, the mixture was stirred at room temperature for half an hour, TLC was monitored for reaction, color development was performed with potassium permanganate, and the consumption of raw materials was completed. Saturated sodium bicarbonate was added to adjust ph=7, the aqueous phase was extracted twice, the organic phases were combined, dried over anhydrous sodium sulfate, and then concentrated, and the obtained crude product was distilled 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 process for the preparation of compound 2 comprising the steps of:
preparation of compound 2: compound 1 was dissolved with pyridine, followed by the addition of 1.2 equivalents of t-butyldimethylchlorosilane (TBSCl) and reaction overnight. TLC monitored the completion of the reaction (EA plate), post-treatment rotary evaporation removed most of the pyridine, which was then poured into a large amount of water, precipitating a white solid with a volume ratio of n-hexane (PE)/Ethyl Acetate (EA) of 10: pulping the compound 1, and filtering to obtain a compound 2;
The nuclear magnetic data of compound 2 are as follows:
1H NMR(d6 DMSO)δ0.21(s,6H,CH3),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
Example 2 provides a method for preparing a cytosine azide comprising the steps of:
S1, dissolving a compound 2 by using dimethyl sulfoxide (DMSO), adding acetic acid, and adding acetic anhydride (Ac 2 O) under ice bath, wherein the volume ratio of acetic acid/dimethyl sulfoxide/acetic anhydride is 1:2:3, a step of; the amino group on the pyrimidine ring is protected by acetyl at the same time, the reaction is carried out overnight, TLC monitors the completion of the reaction (PE/EA=1:1 climbing plate, rf=0.4), a large amount of water and ethyl acetate are added in the post-treatment, most of acetic acid is removed, then saturated sodium bicarbonate is added to adjust the pH to 7, the organic phases are combined, and the compound 3 is obtained by concentration;
The nuclear magnetic data of compound 3 are as follows:
1H NMR(d6 DMSO)δ0.21(s,6H,CH3),0.98(s,9H,t-Bu),2.10-2.33(m,2H),2.15(s,3H,SCH3),2.35(s,3H,COCH3),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 solids from the ammonia water, and filtering to obtain a white solid compound 4 with high purity;
the nuclear magnetic data of compound 4 are as follows:
1H NMR(d6 DMSO)δ0.21(s,6H,CH3),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, dissolving the compound 4 by using DMF, adding trifluoroacetyl propargylamine, replacing three times by using nitrogen, then adding CuI and Pd (PPh 3)4, replacing by using nitrogen after adding, finally adding triethylamine, stirring for 2-3 hours at 50 ℃, monitoring the reaction by using a TLC plate (EA climbing plate, rf=0.3), showing that the raw materials are basically consumed completely, pouring the reaction solution into a mixed liquid of water and EA after the reaction is cooled, fully stirring, washing once by using water, adding saturated ammonium chloride to complex copper ions, finally washing with saturated salt, drying by using anhydrous sodium sulfate, pulping the concentrated solid product by using diethyl ether, filtering the product to obtain white solid, obtaining 68.5% yield, directly pulping the crude product by using diethyl ether, and removing triphenylphosphine oxide to obtain a white solid compound 5 with high purity;
the nuclear magnetic data of compound 5 are as follows:
1H NMR(d6 DMSO)δ0.21(s,6H,CH3),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, dissolving the compound 5 by using Dichloromethane (DCM), protecting amino on pyrimidine ring by using 20 equivalents of acetic anhydride, and stirring for 3 hours at normal temperature. TLC plate monitored the reaction (DCM/meoh=20:1 plate, rf=0.3), indicating essentially complete consumption of starting material with main product above. And (3) post-reaction treatment: the reaction solution was poured into DCM, neutralized with saturated sodium bicarbonate to neutrality, the organic phase was extracted, washed twice with saturated brine, dried and concentrated. 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 of compound 6 are as follows:
1H NMR(d6 DMSO)δ0.12(s,6H,CH3),0.78(s,9H,t-Bu),2.03(s,3H,SCH3),1.96-2.20(m,2H),2.35(s,3H,COCH3),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, dissolving the compound 6 by using DMF, adding 1.5 equivalents of N-chlorosuccinimide (NBS-Cl), and stirring for 1h at normal temperature. Subsequently TLC (EA, rf=0.1) plates showed complete consumption of starting material, sodium azide was added under ice bath and stirred at room temperature for 1-2h after 10 min. TLC plate monitored the reaction (PE/ea=1:1 climbing plate, rf=0.3), indicating complete consumption of intermediate, product above. The reaction solution was poured into a mixed solution of water and EA, and stirred well. After washing twice, finally, saturated brine is washed, and dried over anhydrous sodium sulfate. The concentrate was purified on normal phase silica gel and the product was flushed out with PE/ea=2:1 and 1:1. The product was concentrated to give a pale yellow oil. Dissolved in DCM, followed by addition of n-hexane, a pale yellow solid was precipitated, the supernatant was decanted, and the solid was drained off in 60% yield.
The nuclear magnetic data for compound 7 are as follows:
1H NMR(d6 DMSO)δ0.12(s,6H,CH3),0.88(s,9H,t-Bu),2.15-2.25(m,1H),2.35(s,3H,COCH3),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, dissolving the compound 7 by using Tetrahydrofuran (THF), adding 10 equivalents of triethylamine hydrochloride (Et 3 N-3 HF) to remove TBS protective groups, reacting overnight, monitoring the completion of the reaction by TLC (EA climbing plate), performing post-treatment, steaming to remove the THF, adding ethyl acetate and saturated sodium bicarbonate to adjust the pH to 7, and merging organic phases to concentrate to obtain a light yellow solid compound 8 with high purity.
The nuclear magnetic data for compound 8 are as follows:
1H NMR(d6 DMSO)δ2.01(s,3H,CH3CO),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:
The present invention has been described in detail with reference to the above embodiments, but the present invention is not limited to the above embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the spirit of the present invention.
Claims (11)
1. A method for preparing a cytosine azide, comprising the steps of:
s1, adding a compound 2, an acidic reagent and an anhydride compound into dimethyl sulfoxide for reaction to obtain a compound 3;
s2, carrying out hydrolysis reaction on the compound 3 and ammonia water to obtain a compound 4;
s3, in a first solvent and an inert atmosphere, reacting the compound 4, trifluoroacetyl propargylamine, cuI, a palladium catalyst and triethylamine to obtain a crude product, and pulping and purifying the crude product by diethyl ether to obtain a compound 5;
S4, in a second solvent, reacting the compound 5 with acetic anhydride to obtain a compound 6;
s5, in a third solvent, reacting the compound 6 with a chloro reagent; adding azide compounds at 0-5 ℃ for continuous reaction to obtain a compound 7;
s6, in a fourth solvent, reacting the compound 7 with an organic fluoride to obtain a compound 8;
in the step S1, the volume ratio of the acidic reagent, dimethyl sulfoxide and anhydride compound is 1: (1.5-3): (2.5-4);
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 compound 2 is prepared by the steps of:
Reacting the compound 1, pyridine and tert-butyl dimethyl chlorosilane to obtain a compound 2;
The structural formula of the compound 1 is as follows:
3. The method for preparing a cytosine azide according to claim 1, wherein in step S3, the reaction includes at least the following conditions:
the temperature is 30-70 ℃; the time is 1 h-5 h.
4. The method for preparing cytosine azide according to claim 1, wherein in step S3, the molar ratio of the compound 4 to trifluoroacetylated propargylamine to CuI to palladium catalyst to triethylamine is 1: (2-3): (0.1-0.2): (0.05-0.1): (2-3).
5. The method for producing a cytosine azide according to claim 1, wherein in step S4, the reaction temperature is 20 to 30 ℃.
6. The method for preparing cytosine azide according to claim 1, wherein in step S5, the molar ratio of the compound 6, the chloro reagent and the azide is 1: (1.2-1.5): (4-10).
7. The method for preparing cytosine azide according to claim 1, wherein in step S6, the molar ratio of the compound 7 to the organofluoride is 1: (8-15).
8. The method for producing a cytosine azide compound according to any one of claims 1 to 7, wherein the first solvent is selected from at least one of N, N-dimethylformamide or dimethylsulfoxide.
9. The method for producing a cytosine azide compound according to claim 1, wherein the second solvent is selected from at least one of dichloromethane and pyridine.
10. The method for preparing cytosine azide compounds according to claim 1, wherein the third solvent is selected from the group consisting of N, N-dimethylformamide.
11. The method for producing a cytosine azide compound according to claim 1, wherein the fourth solvent is at least one selected from tetrahydrofuran and ethyl acetate.
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