ME03811B - Postupci za pripremu polinukleotida upotrebom kompozicija multivalentne katjonske soli - Google Patents
Postupci za pripremu polinukleotida upotrebom kompozicija multivalentne katjonske soliInfo
- Publication number
- ME03811B ME03811B MEP-2020-161A MEP2020161A ME03811B ME 03811 B ME03811 B ME 03811B ME P2020161 A MEP2020161 A ME P2020161A ME 03811 B ME03811 B ME 03811B
- Authority
- ME
- Montenegro
- Prior art keywords
- polynucleotide
- composition
- salt
- subunits
- independently
- Prior art date
Links
- 108091033319 polynucleotide Proteins 0.000 title claims 46
- 102000040430 polynucleotide Human genes 0.000 title claims 46
- 239000002157 polynucleotide Substances 0.000 title claims 46
- 239000000203 mixture Substances 0.000 title claims 25
- 238000000034 method Methods 0.000 title claims 20
- -1 CATION SALT Chemical class 0.000 title claims 3
- 238000002360 preparation method Methods 0.000 title claims 3
- 125000002091 cationic group Chemical group 0.000 claims 13
- 150000003839 salts Chemical class 0.000 claims 12
- 239000002777 nucleoside Substances 0.000 claims 11
- 150000003833 nucleoside derivatives Chemical class 0.000 claims 11
- KDCGOANMDULRCW-UHFFFAOYSA-N Purine Natural products N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 claims 6
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims 6
- 150000002500 ions Chemical class 0.000 claims 6
- 108010017842 Telomerase Proteins 0.000 claims 5
- 230000000295 complement effect Effects 0.000 claims 5
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 claims 4
- 238000010828 elution Methods 0.000 claims 4
- 150000002632 lipids Chemical class 0.000 claims 4
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims 3
- 229910052739 hydrogen Inorganic materials 0.000 claims 3
- 239000001257 hydrogen Substances 0.000 claims 3
- 239000011777 magnesium Substances 0.000 claims 3
- 229910052749 magnesium Inorganic materials 0.000 claims 3
- 239000002244 precipitate Substances 0.000 claims 3
- 238000004366 reverse phase liquid chromatography Methods 0.000 claims 3
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims 2
- 108091034117 Oligonucleotide Proteins 0.000 claims 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical group [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims 2
- 125000003545 alkoxy group Chemical group 0.000 claims 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims 2
- 229910052782 aluminium Inorganic materials 0.000 claims 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical group [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims 2
- 230000015572 biosynthetic process Effects 0.000 claims 2
- 239000011575 calcium Substances 0.000 claims 2
- 229910052791 calcium Inorganic materials 0.000 claims 2
- 125000001153 fluoro group Chemical group F* 0.000 claims 2
- 150000002431 hydrogen Chemical class 0.000 claims 2
- 238000002372 labelling Methods 0.000 claims 2
- 239000003550 marker Substances 0.000 claims 2
- 229910052760 oxygen Inorganic materials 0.000 claims 2
- 239000001301 oxygen Substances 0.000 claims 2
- 229920000642 polymer Polymers 0.000 claims 2
- 229920001184 polypeptide Polymers 0.000 claims 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims 2
- 102000004196 processed proteins & peptides Human genes 0.000 claims 2
- 108090000765 processed proteins & peptides Proteins 0.000 claims 2
- 125000000561 purinyl group Chemical group N1=C(N=C2N=CNC2=C1)* 0.000 claims 2
- 125000005415 substituted alkoxy group Chemical group 0.000 claims 2
- 229910052717 sulfur Chemical group 0.000 claims 2
- 239000011593 sulfur Chemical group 0.000 claims 2
- 238000003786 synthesis reaction Methods 0.000 claims 2
- 239000011701 zinc Substances 0.000 claims 2
- 229910052725 zinc Inorganic materials 0.000 claims 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims 1
- 238000005341 cation exchange Methods 0.000 claims 1
- 238000005119 centrifugation Methods 0.000 claims 1
- 239000003153 chemical reaction reagent Substances 0.000 claims 1
- JLYVRXJEQTZZBE-UHFFFAOYSA-N ctk1c6083 Chemical compound NP(N)(N)=S JLYVRXJEQTZZBE-UHFFFAOYSA-N 0.000 claims 1
- 238000001914 filtration Methods 0.000 claims 1
- 239000000047 product Substances 0.000 claims 1
- 238000000926 separation method Methods 0.000 claims 1
- 159000000000 sodium salts Chemical class 0.000 claims 1
- 239000007790 solid phase Substances 0.000 claims 1
- 239000002904 solvent Substances 0.000 claims 1
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- C12N15/09—Recombinant DNA-technology
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- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
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- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/101—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
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- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
- C12P19/28—N-glycosides
- C12P19/30—Nucleotides
- C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides
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- C12Y207/07—Nucleotidyltransferases (2.7.7)
- C12Y207/07049—RNA-directed DNA polymerase (2.7.7.49), i.e. telomerase or reverse-transcriptase
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Claims (28)
1. Postupak za pripremu polinukleotida, pri čemu postupak sadrži:a) dovođenje u kontakt prve polinukleotidne kompozicije sa multivalentnom katjonskom solju kako bi se istaložila prva polinukleotidna so;b) razdvajanje prve polinukleotidne soli od prve polinukleotidne kompozicije dovedene u kontakt, kako bi se dobila druga polinukleotidna kompozicija koja sadrži prvu polinukleotidnu so;gde prva polinukleotidna kompozicija sadrži:(i) polinukleotid koji ima sekvencu od 7 ili više nukleozidnih subjedinica i najmanje dve od nukleozidnih subjedinica su povezane N3’→P5’ tiofosforamidatnom vezom između subjedinica;i(ii) solubilne proizvode i reagense sinteze koji nisu cilj sintezec) dovođenje u kontakt druge polinukleotidne kompozicije iz koraka (b) sa nosačem za reverzno-faznu hromatografiju;id) eluciju sa nosača za reverzno-faznu hromatografiju treće polinukleotidne kompozicije koja sadrži drugu solubilnu polinukleotidnu so.
2. Postupak prema patentnom zahtevu 1, naznačen time što su sve nukleozidne subjedinice povezane vezama između subjedinica, nezavisno izabranim od N3’→-P5’ tiofosforamidatne veze između subjedinica i N3’→-P5’ fosforamidatne veze između subjedinica.
3. Postupak prema patentnom zahtevu 1 ili 2, naznačen time što je polinukleotid opisan Formulom (I): pri čemu: svako B je nezavisno purin, protektovani purin, pirimidin ili protektovani pirimidin, ili njihov analog; svako X je nezavisno kiseonik ili sumpor; svako R3 je nezavisno vodonik, fluoro, hidroksil, alkoksi, supstituisani alkoksi ili protektovani hidroksil; R6 je amino, hidroksil, protektovani amino, protektovani hidroksi, -O-T-Z ili -NH-T-Z; svako T je nezavisno opcioni linker;svako Z je nezavisno H, lipid, nosač, oligonukleotid, polimer, polipeptid, marker koji može da se detektuje ili grupa za obeležavanje; i n je ceo broj od 7 do 100.
4. Postupak prema bilo kom od patentnih zahteva 1-3, naznačen time što: polinukleotid sadrži sekvencu koja sadrži 13 ili više nukleozidnih subjedinica koje su komplementarne sa RNK komponentom humane telomeraze, ilipolinukleotid sadrži između 10 i 50 nukleozidnih subjedinica u neprekinutom nizu koje su komplementarne sa RNK komponentom humane telomeraze.
5. Postupak prema bilo kom od patentnih zahteva 1-4, naznačen time što su sve nukleozidne subjedinice povezane N3’→P5’ tiofosforamidatnom vezom između subjedinica.
6. Postupak prema bilo kom od patentnih zahteva 1-5, naznačen time što polinukleotid sadrži sekvencu odabranu iz grupe koja se sastoji od: GTTAGGGTTAG (SEQ ID NO:4), TAGGGTTAGACAA (SEQ ID NO:3) i CAGTTAGGGTTAG (SEQ ID NO:5).
7. Postupak prema bilo kom od patentnih zahteva 1-6, naznačen time što je polinukleotid konjugovan sa lipidnom komponentom preko opcionog linkera.
8. Postupak prema bilo kom od patentnih zahteva 1-7, naznačen time što druga polinukleotidna so ima strukturu: pri čemu je svako Mx+ nezavisno vodonik ili katjonski kontra jon, svako x je nezavisno 1, 2 ili 3 i n je ceo broj od 5 do 13.
9. Postupak prema bilo kom od patentnih zahteva 1-8, naznačen time što nakon koraka elucije d) druga polinukleotidna so predstavlja farmaceutski prihvatljivu so polinukleotida.
10. Postupak prema bilo kom od patentnih zahteva 1-9, naznačen time što nakon koraka elucije d), druga polinukleotidna so predstavlja monovalentnu katjonsku so polinukleotida, poželjno natrijumovu so polinukleotida.
11. Postupak prema bilo kom od patentnih zahteva 1-10, koji dodatno sadrži, pre koraka dovođenja u kontakt a), odvajanje polinukleotida sa nosača čvrste faze, kako bi se dobila prva polinukleotidna kompozicija u vidu sirove sintetičke preparacije polinukleotida.
12. Postupak prema bilo kom od patentnih zahteva 1-11, naznačen time što pre koraka dovođenja u kontakt a) prva polinukleotidna kompozicija sadrži monovalentnu katjonsku so polinukleotida.
13. Postupak prema bilo kom od patentnih zahteva 1-12, naznačen time što korak dovođenja u kontakt a) sadrži nanošenje i eluciju prve polinukleotidne kompozicije sa katjonskog izmenjivačkog nosača.
14. Postupak prema bilo kom od patentnih zahteva 1-13, naznačen time što korak razdvajanja b) sadrži centrifugiranje prve polinukleotidne kompozicije dovedene u kontakt, kako bi se oborio precipitat prve polinukleotidne soli, ili filtraciju prve polinukleotidne soli iz prve polinukleotidne kompozicije dovedene u kontakt.
15. Postupak prema bilo kom od patentnih zahteva 1-13, naznačen time što se druga polinukleotidna kompozicija iz koraka b) nanosi direktno na nosač za reverzno-faznu hromatografiju.
16. Postupak prema bilo kom od patentnih zahteva 1-15, koji dodatno sadrži, pre koraka dovođenja u kontakt c), rastvaranje druge polinukleotidne kompozicije u rastvaraču.
17. Postupak prema bilo kom od patentnih zahteva 1-16, naznačen time što:najmanje jedan multivalentni katjonski kontra jon je dvovalentan, poželjno odabran iz grupe koja se sastoji od magnezijuma, cinka i kalcijuma; ili najmanje jedan multivalentni katjonski kontra jon je trovalentan, poželjno aluminijum.
18. Postupak prema bilo kom od patentnih zahteva 1-17, naznačen time što prva polinukleotidna so dodatno sadrži monovalentni katjonski kontra jon.
19. Kompozicija koja sadrži:precipitat soli polinukleotida, koji sadrži najmanje jedan multivalentni katjonski kontra jon;pri čemu polinukleotid ima sekvencu od 7 ili više nukleozidnih subjedinica komplementarnih sa RNK komponentom humane telomeraze i najmanje dve od nukleozidnih subjedinica su povezane N3’→P5’ tiofosforamidnom vezom između subjedinica.
20. Kompozicija prema patentnom zahtevu 19, naznačena time što:najmanje jedan multivalentni katjonski kontra jon je dvovalentan, poželjno odabran iz grupe koja se sastoji od magnezijuma, cinka i kalcijuma, poželjnije magnezijum; ili najmanje jedan multivalentni katjonski kontra jon je trovalentan, poželjno aluminijum.
21. Kompozicija prema bilo kom od patentnih zahteva 19-20, naznačena time što polinukleotidna so sadrži 3 molska % ili više multivalentnog katjonskog kontra jona ili sadrži 1.0 %, po težini, ili više, multivalentnog katjonskog kontra jona.
22. Kompozicija prema bilo kom od patentnih zahteva 19-21, naznačena time što kompozicija predstavlja precipitat.
23. Kompozicija prema bilo kom od patentnih zahteva 19-22, naznačena time što je polinukleotid opisan Formulom (I): pri čemu: svako B je nezavisno purin, protektovani purin, pirimidin ili protektovani pirimidin, ili njihov analog; svako X je nezavisno kiseonik ili sumpor; svako R3 je nezavisno vodonik, fluoro, hidroksil, alkoksi, supstituisani alkoksi ili protektovani hidroksil; R6 je amino, hidroksil, protektovani amino, protektovani hidroksi, -O-T-Z ili -NH-T-Z; svako T je nezavisno opcioni linker;svako Z je nezavisno H, lipid, nosač, oligonukleotid, polimer, polipeptid, marker koji može da se detektuje ili grupa za obeležavanje; i n je ceo broj od 7 do 100.
24. Kompozicija prema bilo kom od patentnih zahteva 19-23, naznačena time što: polinukleotid sadrži sekvencu koja sadrži 13 ili više nukleozidnih subjedinica koje su komplementarne sa RNK komponentom humane telomeraze, ilipolinukleotid sadrži između 10 i 50 nukleozidnih subjedinica u neprekinutom nizu koje su komplementarne sa RNK komponentom humane telomeraze.
25. Kompozicija prema bilo kom od patentnih zahteva 19-24, naznačena time što su sve nukleozidne subjedinice povezane N3’→P5’ tiofosforamidnom vezom između subjedinica.
26. Kompozicija prema bilo kom od patentnih zahteva 19-25, naznačena time što polinukleotid sadrži sekvencu odabranu iz grupe koja se sastoji od: GTTAGGGTTAG (SEQ ID NO:4), TAGGGTTAGACAA (SEQ ID NO:3) i CAGTTAGGGTTAG (SEQ ID NO:5).
27. Kompozicija prema bilo kom od patentnih zahteva 19-26, naznačena time što je 5’ ili 3’ terminus polinukleotida konjugovan sa lipidnom komponentom preko opcionog linkera.
28. Kompozicija prema bilo kom od patentnih zahteva 19-27, naznačena time što polinukleotid ima strukturu: pri čemu je svako Mx+ nezavisno katjonski kontra jon, svako x je 1, 2 ili 3 i n je 5 do 12.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201562151891P | 2015-04-23 | 2015-04-23 | |
| PCT/US2016/028657 WO2016172346A1 (en) | 2015-04-23 | 2016-04-21 | Methods of polynucleotide preparation using multivalent cation salt compositions |
| EP16720636.6A EP3286203B1 (en) | 2015-04-23 | 2016-04-21 | Methods of polynucleotide preparation using multivalent cation salt compositions |
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| HRP20201218T1 (hr) | 2015-04-23 | 2020-12-11 | Geron Corporation | Postupci za pripremu polinukleotida uporabom pripravaka multivalentne kationske soli |
| US11384352B2 (en) | 2016-12-13 | 2022-07-12 | Modernatx, Inc. | RNA affinity purification |
| EP4060345A1 (en) | 2016-12-19 | 2022-09-21 | Ventana Medical Systems, Inc. | Peptide nucleic acid conjugates |
| EP3668977A4 (en) * | 2017-08-18 | 2021-04-21 | Modernatx, Inc. | HPLC ANALYTICAL PROCESSES |
| AU2018337854B2 (en) * | 2017-09-22 | 2025-01-16 | Katarzyna JASTRZEBSKA | Thiomorpholino oligonucleotides for the treatment of muscular dystrophy |
| WO2019120635A1 (en) * | 2017-12-18 | 2019-06-27 | Ventana Medical Systems, Inc. | Peptide nucleic acid conjugates |
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| DE69130333T2 (de) | 1990-08-31 | 1999-03-04 | Regents Of The University Of Minnesota, Minneapolis, Minn. | Harze für die Feststoff-Peptidsynthese |
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