CN101289486A - Process for synthesizing RNA monomer - Google Patents
Process for synthesizing RNA monomer Download PDFInfo
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- CN101289486A CN101289486A CNA2007100395684A CN200710039568A CN101289486A CN 101289486 A CN101289486 A CN 101289486A CN A2007100395684 A CNA2007100395684 A CN A2007100395684A CN 200710039568 A CN200710039568 A CN 200710039568A CN 101289486 A CN101289486 A CN 101289486A
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- Prior art keywords
- rna
- monomer
- synthetic
- purification
- protecting group
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- 239000000178 monomer Substances 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title abstract description 7
- 230000002194 synthesizing effect Effects 0.000 title abstract 4
- 108091034117 Oligonucleotide Proteins 0.000 claims abstract description 3
- 238000011084 recovery Methods 0.000 claims abstract description 3
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 claims description 12
- DRTQHJPVMGBUCF-XVFCMESISA-N Uridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-XVFCMESISA-N 0.000 claims description 10
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 claims description 10
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 claims description 6
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 claims description 6
- 229940029575 guanosine Drugs 0.000 claims description 6
- UHDGCWIWMRVCDJ-UHFFFAOYSA-N 1-beta-D-Xylofuranosyl-NH-Cytosine Natural products O=C1N=C(N)C=CN1C1C(O)C(O)C(CO)O1 UHDGCWIWMRVCDJ-UHFFFAOYSA-N 0.000 claims description 5
- 239000002126 C01EB10 - Adenosine Substances 0.000 claims description 5
- UHDGCWIWMRVCDJ-PSQAKQOGSA-N Cytidine Natural products O=C1N=C(N)C=CN1[C@@H]1[C@@H](O)[C@@H](O)[C@H](CO)O1 UHDGCWIWMRVCDJ-PSQAKQOGSA-N 0.000 claims description 5
- 229960005305 adenosine Drugs 0.000 claims description 5
- DRTQHJPVMGBUCF-PSQAKQOGSA-N beta-L-uridine Natural products O[C@H]1[C@@H](O)[C@H](CO)O[C@@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-PSQAKQOGSA-N 0.000 claims description 5
- UHDGCWIWMRVCDJ-ZAKLUEHWSA-N cytidine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O1 UHDGCWIWMRVCDJ-ZAKLUEHWSA-N 0.000 claims description 5
- DRTQHJPVMGBUCF-UHFFFAOYSA-N uracil arabinoside Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-UHFFFAOYSA-N 0.000 claims description 5
- 229940045145 uridine Drugs 0.000 claims description 5
- 238000010189 synthetic method Methods 0.000 claims description 4
- 125000006239 protecting group Chemical group 0.000 abstract description 4
- 239000002253 acid Substances 0.000 abstract description 2
- 230000000295 complement effect Effects 0.000 abstract description 2
- 239000007791 liquid phase Substances 0.000 abstract description 2
- 238000010532 solid phase synthesis reaction Methods 0.000 abstract description 2
- 238000000746 purification Methods 0.000 abstract 5
- 230000015572 biosynthetic process Effects 0.000 abstract 1
- 238000003786 synthesis reaction Methods 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 5
- 0 CC*OC(C1OC(CC2)(CCN2c2cc(Cl)ccc2)OC)[C@@](CO*)O[C@@]1N1C=CC(NC(c2ccccc2)=O)=NC1O Chemical compound CC*OC(C1OC(CC2)(CCN2c2cc(Cl)ccc2)OC)[C@@](CO*)O[C@@]1N1C=CC(NC(c2ccccc2)=O)=NC1O 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 238000009833 condensation Methods 0.000 description 2
- 230000005494 condensation Effects 0.000 description 2
- 238000010511 deprotection reaction Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- PNPCRKVUWYDDST-UHFFFAOYSA-N 3-chloroaniline Chemical compound NC1=CC=CC(Cl)=C1 PNPCRKVUWYDDST-UHFFFAOYSA-N 0.000 description 1
- PZAWPYOUTRTPAA-VVSUAJNRSA-N CC(C)N(C(C)C)P(OCCC#N)OC([C@@H](COC(c1ccccc1)(c(cc1)ccc1OC)c(cc1)ccc1OC)O[C@H]1C(CC2)C=CC(NC(c3ccccc3)=O)=NC2=O)C1OC(CC1)(CCN1c1cc(Cl)ccc1)OC Chemical compound CC(C)N(C(C)C)P(OCCC#N)OC([C@@H](COC(c1ccccc1)(c(cc1)ccc1OC)c(cc1)ccc1OC)O[C@H]1C(CC2)C=CC(NC(c3ccccc3)=O)=NC2=O)C1OC(CC1)(CCN1c1cc(Cl)ccc1)OC PZAWPYOUTRTPAA-VVSUAJNRSA-N 0.000 description 1
- RNPXJLCFYHCLII-XLHDYKNESA-O CC[SH+](CC)(OC1C2O)O[Si](C)(C(C)C)OC[C@H]1O[C@H]2N(C=CC(NC(c1ccccc1)=O)=N1)C1=O Chemical compound CC[SH+](CC)(OC1C2O)O[Si](C)(C(C)C)OC[C@H]1O[C@H]2N(C=CC(NC(c1ccccc1)=O)=N1)C1=O RNPXJLCFYHCLII-XLHDYKNESA-O 0.000 description 1
- ZXDWVUFLACRIQN-MKMLSURGSA-N COC(CC1)(CCN1c1cc(Cl)ccc1)OC1[C@H](N(C=CC(NC(c2ccccc2)=O)=N2)C2=O)O[C@H](CO)C1O Chemical compound COC(CC1)(CCN1c1cc(Cl)ccc1)OC1[C@H](N(C=CC(NC(c2ccccc2)=O)=N2)C2=O)O[C@H](CO)C1O ZXDWVUFLACRIQN-MKMLSURGSA-N 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000002841 Lewis acid Substances 0.000 description 1
- UHDGCWIWMRVCDJ-AYZDMWBASA-N NC(C=CN1[C@@H](C2O)O[C@H](CO)C2O)=NC1=O Chemical compound NC(C=CN1[C@@H](C2O)O[C@H](CO)C2O)=NC1=O UHDGCWIWMRVCDJ-AYZDMWBASA-N 0.000 description 1
- CMLKTGCCWMTTEQ-MMSDAREKSA-N OC[C@H](C(C1O)O)O[C@H]1C(CC1)C=CC(NC(c2ccccc2)=O)=NC1=O Chemical compound OC[C@H](C(C1O)O)O[C@H]1C(CC1)C=CC(NC(c2ccccc2)=O)=NC1=O CMLKTGCCWMTTEQ-MMSDAREKSA-N 0.000 description 1
- BNXBRFDWSPXODM-DPBOGPHSSA-N OC[C@H](C(C1O)O)O[C@H]1N(C=CC(NC(c1ccccc1)=O)=N1)C1=O Chemical compound OC[C@H](C(C1O)O)O[C@H]1N(C=CC(NC(c1ccccc1)=O)=N1)C1=O BNXBRFDWSPXODM-DPBOGPHSSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 150000007517 lewis acids Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- AAAQKTZKLRYKHR-UHFFFAOYSA-N triphenylmethane Chemical compound C1=CC=CC=C1C(C=1C=CC=CC=1)C1=CC=CC=C1 AAAQKTZKLRYKHR-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
The invention discloses a method for synthesizing monomers of RNA. The unique 2'-protecting groups of the synthesized monomers of the RNA adopted in the method are regarded as starting points and the structure of the synthesizing monomers of the RNA is different from the structure of previous synthesizing monomers of the RNA. The method is characterized in that the protecting groups are stable in acid de-protection conditions and alkaline de-protection conditions, and after solid phase synthesis is finished, on condition that the protection group of 5'-DMT is removed and the 2'-protecting group is still on the chain, high-pressured liquid phase purification can be carried out. Regarding the products after first grade purification, the 2'-protecting groups can be completely removed in proper conditions, which can be annealed to become a double strand with another complementary strand after further purification, and the purity of the RNA oligonucleotide produced after the synthesis and purification can be above 97 percent; furthermore the purification recovery rate can be above 55 percent.
Description
The invention discloses a kind of synthetic RNA monomer methods.The RNA synthon that is adopted in this method is a starting point with 2 ' of its uniqueness-protecting group, the RNA synthon that RNA synthon structure is different from the past.Its characteristics are that this protecting group is stable under acid deprotection condition, and are also very stable under alkali deprotection condition.After solid phase synthesis is finished; 2 '-protecting group still can be carried out the high-pressure liquid phase purifying under the situation on the chain in that 5 '-DMT protecting group is taken off; product behind the one-level purifying can take off 2 '-protecting group under suitable condition fully, can be annealed into two strands with another complementary strand after being further purified.RNA oligonucleotide purity synthetic thus and that purifying obtains can reach more than 97%, and the purifying rate of recovery can reach more than 55%.
Technical field
The present invention relates to the monomeric preparation method of a kind of RNA.
Background technology
The RNA monomer is at present by Glen Research, Transgenomic, and a few major company's monopolization such as ChemGene is produced, domestic no any RNA monomer manufacturer; The RNA monomer mass index that we rely on own technology to produce is higher than existing manufacturer production product;
Summary of the invention
The objective of the invention is to adopt 2 ' unique-protecting group is starting point, synthetic RNA monomer.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
A. prepare 2 '-protecting group 1-(3-chlorobenzene)-4-piperidines alkene (CPP);
B. the uridine monomer is synthetic;
C. the cytidine monomer is synthetic;
D. the adenosine monomer is synthetic;
E. the guanosine monomer is synthetic;
Described RNA monomer synthetic method is characterized in that:
Adopting unique 2 '-protecting group 1-(3-chlorobenzene)-4-piperidines alkene (CPP) is starting point, synthetic RNA monomer.
Description of drawings
Fig. 1 is a CPP preparation method structure iron:
Fig. 2 is a uridine monomer synthetic schemes:
Fig. 3 is a cytidine monomer synthetic schemes:
Fig. 4 is an adenosine monomer synthetic schemes:
Fig. 5 is a guanosine monomer synthetic schemes:
Embodiment
The preparation method of 2 '-protecting group CPP:
3-chlorpromazine chloride and ethene produce 1 under the effect of lewis acid, 5-dichloro pentanone, form N-(3-chlorobenzene)-piperidone with the condensation of 3-chloroaniline again, the latter generates dimethyl-N-(3-chlorobenzene)-piperidines ketal with the methyl alcohol condensation again, takes off a part methyl alcohol again under the decompression heating condition at last and generates desired 1-(3-chlorobenzene)-4-piperidines alkene (CPP);
The preparation of RNA synthon:
Uridine monomer: 3 '; 5 '-hydroxy is protected with the Marcwicz protecting group; under acidic conditions, introduce 2 '-CPP protecting group then; slough the Marcwicz protecting group subsequently; 5 '-hydroxy is with 1; 4-dimethoxy tritane chlorine (DMTrCl) is protected, and carry out phosphoramiditeization in 3 '-position with phosphorylation agent at last, makes the uridine monomer.
Cytidine monomer, adenosine monomer, the preparation of guanosine monomer: the base of cytidine/adenosine/guanosine is protected with benzoyl earlier, and subsequent step is identical with guanosine.
Claims (2)
1, the monomeric synthetic method of a kind of RNA is characterized in that 2 ' unique-protecting group is a starting point, the RNA synthon that RNA synthon structure is different from the past.Synthetic method comprises the steps:
A. prepare 2 '-protecting group 1-(3-chlorobenzene)-4-piperidines alkene (CPP);
B. the uridine monomer is synthetic;
C. the cytidine monomer is synthetic;
D. the adenosine monomer is synthetic;
E. the guanosine monomer is synthetic.
2, the RNA monomer of RNA monomer synthetic method acquisition according to claim 1 is characterized in that, RNA oligonucleotide purity synthetic and that purifying obtains can reach more than 97%, and the purifying rate of recovery can reach more than 55%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100395684A CN101289486B (en) | 2007-04-18 | 2007-04-18 | Process for synthesizing RNA monomer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100395684A CN101289486B (en) | 2007-04-18 | 2007-04-18 | Process for synthesizing RNA monomer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101289486A true CN101289486A (en) | 2008-10-22 |
| CN101289486B CN101289486B (en) | 2012-05-30 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007100395684A Expired - Fee Related CN101289486B (en) | 2007-04-18 | 2007-04-18 | Process for synthesizing RNA monomer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101289486B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103492405A (en) * | 2011-03-28 | 2014-01-01 | 波恩莱茵弗里德里希·威廉大学 | Purification of triphosphorylated oligonucleotides using capture tags |
| CN104703996A (en) * | 2012-09-27 | 2015-06-10 | 波恩莱茵弗里德里希·威廉大学 | Novel RIG-I ligands and methods for producing them |
-
2007
- 2007-04-18 CN CN2007100395684A patent/CN101289486B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103492405A (en) * | 2011-03-28 | 2014-01-01 | 波恩莱茵弗里德里希·威廉大学 | Purification of triphosphorylated oligonucleotides using capture tags |
| CN103492405B (en) * | 2011-03-28 | 2017-02-15 | 波恩莱茵弗里德里希·威廉大学 | Purification of triphosphorylated oligonucleotides using capture tags |
| CN104703996A (en) * | 2012-09-27 | 2015-06-10 | 波恩莱茵弗里德里希·威廉大学 | Novel RIG-I ligands and methods for producing them |
| CN106279300A (en) * | 2012-09-27 | 2017-01-04 | 波恩莱茵弗里德里希·威廉大学 | RIG I part and production method thereof |
| CN104703996B (en) * | 2012-09-27 | 2018-01-12 | 波恩莱茵弗里德里希·威廉大学 | New RIG I parts and its production method |
| CN106279300B (en) * | 2012-09-27 | 2020-07-10 | 波恩莱茵弗里德里希·威廉大学 | RIG-I ligands and methods for producing same |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101289486B (en) | 2012-05-30 |
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| ASS | Succession or assignment of patent right |
Owner name: SHANGHAI ZHENGTI BIOTECHNOLOGY CO., LTD. Effective date: 20110824 |
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Free format text: CORRECT: ADDRESS; FROM: 201203 PUDONG NEW AREA, SHANGHAI TO: 200086 HONGKOU, SHANGHAI |
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Effective date of registration: 20110824 Address after: 200086 Shanghai city Hongkou District Feihong Road No. 528 building 403 room four Applicant after: Shanghai whole biological technology Co.,Ltd. Address before: 619-21 room 680, 201203 Guiping Road, Shanghai Applicant before: SHANGHAI GENECHEM Co.,Ltd. |
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Owner name: SHANGHAI JIKAI GENE CHEMICAL TECHNOLOGY INC. Free format text: FORMER OWNER: SHANGHAI ZHENGTI BIOTECHNOLOGY CO., LTD. Effective date: 20120221 |
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Effective date of registration: 20120221 Address after: 200233 room 619-21, Guiping Road, Shanghai, Xuhui District, 680 Applicant after: SHANGHAI GENECHEM Co.,Ltd. Address before: 200086 Shanghai city Hongkou District Feihong Road No. 528 building 403 room four Applicant before: Shanghai whole biological technology Co.,Ltd. |
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Address after: 200233, room 680, 619-21 Guiping Road, Shanghai, Xuhui District Patentee after: Shanghai Jikai gene Medical Technology Co.,Ltd. Address before: 200233, room 680, 619-21 Guiping Road, Shanghai, Xuhui District Patentee before: SHANGHAI GENECHEM Co.,Ltd. |
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