DE10147238A1 - Preparation of carrier bound vinyl nucleobases and their polymers, useful, e.g. as antiviral and anticancer agents, or for diagnostics or purification, comprises reaction of vinyl nucleobases with a carrier - Google Patents
Preparation of carrier bound vinyl nucleobases and their polymers, useful, e.g. as antiviral and anticancer agents, or for diagnostics or purification, comprises reaction of vinyl nucleobases with a carrierInfo
- Publication number
- DE10147238A1 DE10147238A1 DE2001147238 DE10147238A DE10147238A1 DE 10147238 A1 DE10147238 A1 DE 10147238A1 DE 2001147238 DE2001147238 DE 2001147238 DE 10147238 A DE10147238 A DE 10147238A DE 10147238 A1 DE10147238 A1 DE 10147238A1
- Authority
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- Germany
- Prior art keywords
- vinyl
- nucleobases
- carrier
- support
- see formula
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 title claims abstract description 24
- 229920002554 vinyl polymer Polymers 0.000 title claims abstract description 20
- 238000002360 preparation method Methods 0.000 title claims description 3
- 239000003443 antiviral agent Substances 0.000 title claims 2
- 238000000746 purification Methods 0.000 title claims 2
- 238000006243 chemical reaction Methods 0.000 title description 2
- 230000000840 anti-viral effect Effects 0.000 title 1
- 239000002246 antineoplastic agent Substances 0.000 title 1
- 229920000642 polymer Polymers 0.000 title 1
- 238000000034 method Methods 0.000 claims abstract description 11
- 125000000623 heterocyclic group Chemical group 0.000 claims abstract description 7
- 125000006850 spacer group Chemical group 0.000 claims abstract description 6
- 229920001577 copolymer Polymers 0.000 claims description 12
- 150000001875 compounds Chemical class 0.000 claims description 7
- 230000000295 complement effect Effects 0.000 claims description 5
- 150000007523 nucleic acids Chemical group 0.000 claims description 3
- 229960001438 immunostimulant agent Drugs 0.000 claims description 2
- 239000003022 immunostimulating agent Substances 0.000 claims description 2
- 230000003308 immunostimulating effect Effects 0.000 claims description 2
- 229920002477 rna polymer Polymers 0.000 claims 3
- 150000001335 aliphatic alkanes Chemical class 0.000 claims 1
- 229940121357 antivirals Drugs 0.000 claims 1
- 238000001514 detection method Methods 0.000 claims 1
- 150000002170 ethers Chemical class 0.000 claims 1
- 239000012442 inert solvent Substances 0.000 claims 1
- 239000007858 starting material Substances 0.000 claims 1
- 125000000217 alkyl group Chemical group 0.000 abstract 6
- 125000001475 halogen functional group Chemical group 0.000 abstract 6
- 125000003118 aryl group Chemical group 0.000 abstract 5
- 125000004414 alkyl thio group Chemical group 0.000 abstract 3
- 125000005110 aryl thio group Chemical group 0.000 abstract 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 abstract 2
- JCXJVPUVTGWSNB-UHFFFAOYSA-N Nitrogen dioxide Chemical compound O=[N]=O JCXJVPUVTGWSNB-UHFFFAOYSA-N 0.000 abstract 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 abstract 1
- 125000003282 alkyl amino group Chemical group 0.000 abstract 1
- 125000001769 aryl amino group Chemical group 0.000 abstract 1
- 229910052717 sulfur Inorganic materials 0.000 abstract 1
- 239000011593 sulfur Substances 0.000 abstract 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 239000004480 active ingredient Substances 0.000 description 6
- 229920001519 homopolymer Polymers 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- -1 1-vinyl-2-hydroxy-4-mercaptopyrimidine (1-vinyl mercaptouracil) Chemical compound 0.000 description 4
- 102000004127 Cytokines Human genes 0.000 description 4
- 108090000695 Cytokines Proteins 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000009396 hybridization Methods 0.000 description 4
- 102000014150 Interferons Human genes 0.000 description 3
- 108010050904 Interferons Proteins 0.000 description 3
- 101710163270 Nuclease Proteins 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 239000013543 active substance Substances 0.000 description 3
- 239000012876 carrier material Substances 0.000 description 3
- 238000007334 copolymerization reaction Methods 0.000 description 3
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 3
- 239000010931 gold Substances 0.000 description 3
- 229910052737 gold Inorganic materials 0.000 description 3
- 229940079322 interferon Drugs 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000002519 immonomodulatory effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- AUHDWARTFSKSAC-HEIFUQTGSA-N (2S,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)-2-(6-oxo-1H-purin-9-yl)oxolane-2-carboxylic acid Chemical compound [C@]1([C@H](O)[C@H](O)[C@@H](CO)O1)(N1C=NC=2C(O)=NC=NC12)C(=O)O AUHDWARTFSKSAC-HEIFUQTGSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- GRSZFWQUAKGDAV-UHFFFAOYSA-N Inosinic acid Natural products OC1C(O)C(COP(O)(O)=O)OC1N1C(NC=NC2=O)=C2N=C1 GRSZFWQUAKGDAV-UHFFFAOYSA-N 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N adenyl group Chemical group N1=CN=C2N=CNC2=C1N GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000010668 complexation reaction Methods 0.000 description 1
- 125000002740 cytidyl group Chemical group 0.000 description 1
- 239000000032 diagnostic agent Substances 0.000 description 1
- 229940039227 diagnostic agent Drugs 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 231100000171 higher toxicity Toxicity 0.000 description 1
- 239000002955 immunomodulating agent Substances 0.000 description 1
- 229940121354 immunomodulator Drugs 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 229940028843 inosinic acid Drugs 0.000 description 1
- 235000013902 inosinic acid Nutrition 0.000 description 1
- 239000004245 inosinic acid Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 125000002652 ribonucleotide group Chemical group 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 125000002223 uridyl group Chemical group 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F8/00—Chemical modification by after-treatment
- C08F8/34—Introducing sulfur atoms or sulfur-containing groups
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F8/00—Chemical modification by after-treatment
- C08F8/30—Introducing nitrogen atoms or nitrogen-containing groups
Landscapes
- Chemical & Material Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Addition Polymer Or Copolymer, Post-Treatments, Or Chemical Modifications (AREA)
Abstract
Description
Die Erfindung betrifft Homo- und Copolymere der Vinylnukleobasen der allgemeinen Formel I
(s. Formelblatt I)
sowie Copolymere mit Vinylverbindungen der allgemeinen Formel II,
(s. Formelblatt 1I)
ihre Herstellung, Trägerfixierung und ihre Anwendung. Aus den Vinylmonomeren I und II
(s. Formelblätter I und II)
hergestellte und Träger-fixierte Homo- und Copolymere der allgemeinen Formel III,
(s. Formelblatt III)
sind bisher noch nicht bekannt.
The invention relates to homopolymers and copolymers of vinyl nucleobases of the general formula I.
(see formula sheet I)
as well as copolymers with vinyl compounds of the general formula II,
(see formula sheet 1I)
their manufacture, support fixation and their application. From vinyl monomers I and II
(see formula sheets I and II)
prepared and carrier-fixed homopolymers and copolymers of the general formula III,
(see formula sheet III)
are not yet known.
Als Immunostimulanzien wurden eine Reihe von nativen homopolymeren RNA-Komplexen synthetisiert und in zellulären Systemen auf die Cytokinantwort hin getestet (P. F. Torrence, E. De Clercq, Pharmac. Ther. 2 (1977) 1-88, S. Hoffmann, Z. Chem. 2 (1982) 357-367 sowie in B. Glück, W.-V. Meister, W. Witkowski, A. Stelzner und S. Hoffmann, J. Basic Micrbiol. 28 (1988) 8, 501-507. Einer medizinischen Anwendung von Hybriden der natürlichen Homopolymeren: Guanyl-, Inosin-, Cytidyl-, Adenyl- und Uridylsäure stehen neben der hohen Nukleaseempfindlichkeit und den zelltoxischen Nebeneffekten auch die hohen Herstellungskosten entgegen. So wurden zwei Wege zur Optimierung der biologischen Aktivität dieser polymeren Wirkstoffe beschritten. Der erste Schritt besteht in einer Aktivitätserhöhung durch perfekte Basenpaarmischungen, optimale Moleküllängen und einer geeigneten Komplexierung der Verbindungen gegen den enzymatischen Abbau. Die Toxizitätsminderung kann durch Divergenzen von therapeutischen und kinetischen Effekten erzielt werden (W.-V. Meister, B. Glück, W. Witkowski, A. Stelzner, S. Hoffmann, Cytokin(Interferon)induktoren, DD WP C 07 h/309 246/2, 20.11.1987 sowie W.-V. Meister, W. Römer, E. Tonew, S. Hoffmann, Cytokin(Interferon)induktoren, DD WP C 07 h/309 245/4, 20.11.1987). A number of native homopolymeric RNA complexes have been used as immunostimulants synthesized and tested for the cytokine response in cellular systems (P.F. Torrence, E. De Clercq, Pharmac. Ther. 2 (1977) 1-88, S. Hoffmann, Z. Chem. 2 (1982) 357-367 and in B. Glück, W.-V. Meister, W. Witkowski, A. Stelzner and S. Hoffmann, J. Basic Micrbiol. 28 (1988) 8, 501-507. A medical application of hybrids of natural homopolymers: guanyl, Inosinic acid, cytidyl acid, adenyl acid and uridyl acid stand alongside the high sensitivity to nucleases and the cell-toxic side effects also counter the high manufacturing costs. So two became Ways pioneered to optimize the biological activity of these polymeric active ingredients. The The first step is an increase in activity through perfect base pair mixtures, optimal Molecular lengths and a suitable complexation of the compounds against the enzymatic Degradation. The toxicity reduction can be caused by divergences from therapeutic and kinetic Effects are achieved (W.-V. Meister, B. Glück, W. Witkowski, A. Stelzner, S. Hoffmann, Cytokine (interferon) inductors, DD WP C 07 h / 309 246/2, 20.11.1987 and W.-V. Meister, W. Römer, E. Tonew, S. Hoffmann, cytokine (interferon) inductors, DD WP C 07 h / 309 245/4, 20.11.1987).
Überwunden wurde die Nukleaseempfindlichkeit von diesen Poly(desoxy)ribonukleotid-komplexen durch den Einsatz von artifiziellen Polyvinylnukleobasen (W.-V. Meister, H.-D. Gottstein, W. Witkowski, K. Waschke, S. Hoffmann, Verfahren zur Herstellung basenmodifizierter semiplastischer Cytokin(Interferon)induktoren und Aktivatoren natürlicher Killerzellen, DD WP C 08 f/319 957/7, 20.09.1988). Ein entscheidender Durchbruch wurde aber auch dadurch nicht erzielt. Neben einer erhöhten biologischen Wirksamkeit (Superinduktion) besitzen diese "semiplastischen" Immunomodulatoren (Viro- und Cancerostatica) jedoch den Nachteil einer erhöhten Toxizität im Organismus. The nuclease sensitivity of these poly (deoxy) ribonucleotide complexes was overcome through the use of artificial polyvinyl nucleobases (W.-V. Meister, H.-D. Gottstein, W. Witkowski, K. Waschke, S. Hoffmann, Process for the preparation of base-modified semi-elastic cytokine (interferon) inducers and activators of natural killer cells, DD WP C 08 f / 319 957/7, 20.09.1988). This did not make a decisive breakthrough achieved. In addition to an increased biological effectiveness (superinduction), these have However, "semi-plastic" immunomodulators (viro- and cancerostatica) have the disadvantage of one increased toxicity in the organism.
Die Erfindung hat das Ziel, therapeutische, insbesondere immunomodulatorische und träger-fixierte Wirkstoffe herzustellen, welche gegenüber den bekannten lösbaren Substanzen bei ihrer Anwendung eine vergleichbare Wirksamkeit und eine geringere Toxizität aufweisen. Diese neuen Wirkstoffe sollen als potentielle Wirkstoffe mit geringeren Nebenwirkungen für den Patienten geeignet sein. The aim of the invention is therapeutic, in particular immunomodulatory and carrier-fixed To produce active ingredients, which compared to the known soluble substances in their Application have comparable efficacy and lower toxicity. These new ones Active ingredients are said to be potential active ingredients with fewer side effects for the patient be suitable.
Die Zielstellung der Erfindung wird mit den Verbindungen der allgemeinen Formeln I und II (s. Anlage) und der Trägerfixierung der plastischen Homo- und Copolymere der allgemeinen Formel III (s. Anlage) erreicht. Durch die Fixierung dieser Homo- und Copolymerstränge kann deren unkontrollierbare und unerwünschte Wirkungsdauer und somit Toxizität im Organismus auf ein Minimum reduziert werden. Durch diese Erfindung können durch die Variationen der Nukleobasenfolge I eine große Zahl von Verbindungen III zur Verfügung gestellt werden, womit man eine Erhöhung der Selektivität dieser Wirkstoffe erreicht. Copolymerisate mit Vinylverbindungen der allgemeinen Formel II führen zu einer erhöhten Wasserlöslichkeit dieser Verbindungen. Eine weitere Erhöhung der biologischen Wirksamkeit dieser Verbindungen ist durch die Einführung von die Helix stabilisierenden Spacern an den Heterocyclen gegeben. Eine Substitution durch basische Gruppen an den Heterocyclen führt zu einer sinnvollen Erhöhung der thermischen Stabilität der semiplastischen Wirkstoffstränge. Durch den Einsatz kugelförmiger Trägermaterialien geeigneter Porengrößen kann die Wirkstoffbeladungsdichte wesentlich variiert werden. The object of the invention is achieved with the compounds of the general formulas I and II (see. Appendix) and the fixation of the plastic homo- and copolymers of the general formula III (see attachment) reached. By fixing these homo- and copolymer strands, their uncontrollable and undesirable duration of action and thus toxicity in the organism on one Minimum be reduced. Through this invention, the variations of Nucleobase sequence I a large number of compounds III are made available, with which an increase in the selectivity of these active ingredients is achieved. Copolymers with Vinyl compounds of the general formula II lead to an increased water solubility of these Links. A further increase in the biological effectiveness of these compounds is due to given the introduction of the helix-stabilizing spacers on the heterocycles. A Substitution by basic groups on the heterocycles leads to a meaningful increase in thermal stability of the semi-elastic active substance strands. By using spherical Carrier materials of suitable pore sizes can vary the active substance loading density considerably become.
Die Homo- und Copolymerisationsprodukte der Verbindungen I und II sind immunomo-dulatorisch wirksam. The homo- and copolymerization products of compounds I and II are immunomodulatory effective.
Eine sehr wichtige weitere Anwendungsmöglichkeit der erfindungsgemäßen trägerfixierten Homo- und Copolymerisationsprodukte aus I und II besteht in deren Einsatz als Diagnosemittel in der Medizin auf Grund ihrer Fähigkeit zur Bindung spezifischer Nukleinsäuresequenzen. A very important further application of the carrier-fixed homo- according to the invention and copolymerization products from I and II consists in their use as diagnostic agents in the Medicine due to its ability to bind specific nucleic acid sequences.
Die Herstellung solcher trägerfixierten Homo- und Copolymere erfolgt durch Umsetzung der
Vinylnukleobasen der allgemeinen Formel I mit Vinylverbindungen der allgemeinen Formel II
(siehe Formelblätter I und II).
Such carrier-fixed homopolymers and copolymers are prepared by reacting the vinyl nucleobases of the general formula I with vinyl compounds of the general formula II
(see formula sheets I and II).
Die Fixierung der ersten trägerterminalen Vinylnukleobase erfolgt hierbei durch Umsetzung des
jeweiligen N-vinylierten und C-mercaptolierten Heterocyclus mit dem vergoldeten Träger bei
Raumtemperatur.
(s. Formelblatt III)
The first carrier-terminal vinyl nucleobase is fixed here by reacting the respective N-vinylated and C-mercaptolated heterocycle with the gold-plated carrier at room temperature.
(see formula sheet III)
Die Erfindung soll nachstehend durch ein Ausführungsbeispiel näher erläutert werden. The invention will be explained in more detail below using an exemplary embodiment.
Für Proben mit gewünschten geringen Beladungsdichten an Homo- und Copolymeren wird eine Siliziumoberfläche im Hochvacuum mit Gold bedampft und sofort in eine 1 mM Lösung aus frisch umkristallisiertem 1-Vinylmercaptouracil in Ethanol : Wasser 1 : 1, v/v überführt. Nach 12 ständiger Reaktion bei Raumtemperatur wird die Goldoberfläche vorsichtig von den ungebundenen Nukleobasen mittels wässrigem Ethanol befreit, bei Raumtemperatur getrocknet und bis zur Polymerisation im Kühlschrank aufbewahrt. Die Beladungsdichten liegen hierbei bei ca. 2.5 nM 1- Vinylmercaptouracil pro cm2 glattem, porenfreiem Trägermaterial. Kontrollierbar ist der Beladungsprozeß durch eine Quarzwaage. Durch den Einsatz kugelförmiger Trägermaterialien geeigneter Porengrößen wird die Wirkstoffbeladungsdichte wesentlich variiert. For samples with the desired low loading densities of homo- and copolymers, a silicon surface is evaporated with gold in a high vacuum and immediately transferred to a 1 mM solution of freshly recrystallized 1-vinyl mercaptouracil in ethanol: water 1: 1, v / v. After 12 hours of reaction at room temperature, the gold surface is carefully freed from the unbound nucleobases using aqueous ethanol, dried at room temperature and kept in the refrigerator until polymerization. The loading densities are approximately 2.5 nM 1-vinyl mercaptouracil per cm 2 of smooth, non-porous carrier material. The loading process can be controlled by a quartz balance. The active substance loading density is varied considerably by using spherical carrier materials of suitable pore sizes.
Zur Kettenelongation wird der mit 1-Vinylmercaptouracil beladene Goldträger in einer entsprechenden 1.0 mM Mischung aus Vinylnukleobasen der allgemeinen Formel 1 und ggf. zusätzlich mit anderen Vinylverbindungen der allgemeinen Formel II in DMF bei Raumtemperatur und im Vacuum mittels Co60 (1.35 Mrd) bestrahlt. Im Anschluß daran wird die Trägeroberfläche mittels DMF sowie wässrigem Ethanol gewaschen und an der Luft getrocknet. For chain elongation, the gold carrier loaded with 1-vinyl mercaptouracil is irradiated in a corresponding 1.0 mM mixture of vinyl nucleobases of the general formula 1 and, if appropriate, additionally with other vinyl compounds of the general formula II in DMF at room temperature and in vacuo using Co 60 (1.35 billion). The carrier surface is then washed with DMF and aqueous ethanol and air-dried.
Vor dem Hybridisierungsverfahren wird der mit dem Homo- bzw. Copolymerisat beschichtete Träger mittels üblicher Verfahren von Nukleasen befreit. Danach wird der beladene Träger in einer geringen Menge an sterilem und destilliertem Wasser bei ca. 90°C mit einer 0.1 mM wässrigen Lösung an denaturierten komplementären natürlichen Nukleinsäuresträngen versetzt. Anschließend erfolgt eine langsame Abkühlung des Hybridisierungsansatzes auf Raumtemperatur und seine dreitägige Lagerung unter sterilen Bedingungen bei +4°C. Zur Beladungskontrolle wird die mittels sterilem destilliertem Wasser bei 4°C gewaschene feste Probe in einer mit 1xPBS gefüllten Küvette kontrolliert erwärmt. Nach vollständigem Denaturierungsprozeß der hybridisierten natürlichen komplementären Polyribonucleotide wird aus der jeweiligen UV-Absorption bei λmax die Beladungsdichte des Trägers bestimmt. Before the hybridization process, the carrier coated with the homopolymer or copolymer is freed from nucleases by means of customary processes. The loaded carrier is then mixed with a 0.1 mM aqueous solution of denatured complementary natural nucleic acid strands in a small amount of sterile and distilled water at approx. 90 ° C. The hybridization mixture is then slowly cooled to room temperature and stored for three days under sterile conditions at + 4 ° C. To check the loading, the solid sample, washed with sterile distilled water at 4 ° C, is heated in a controlled manner in a cuvette filled with 1 x PBS. After the complete denaturing process of the hybridized natural complementary polyribonucleotides, the loading density of the carrier is determined from the respective UV absorption at λ max .
Zur Garantie höchster Effektivität sollte erst kurz vor dem Einsatz die Hybridisierung der semiplastischen Wirkstoffe durchgeführt werden.. To guarantee maximum effectiveness, the hybridization of the semi-elastic active ingredients are performed ..
Eine weitere Variante der erfindungsgemäßen Umsetzung, bevorzugt beim Einsatz von
Vinylnukleobasen mit zusätzlichen Spacern am Heterocyclus, besteht darin, daß man die terminale
Vinylnukleobase über den Spacer mit dem Träger über eine Ester-, Ether- oder Amidbindung fixiert.
Von wesentlicher Bedeutung für den erfindungsgemäßen Erfolg ist hierbei die absolute
Wasserfreiheit.
Another variant of the implementation according to the invention, preferably when using vinyl nucleobases with additional spacers on the heterocycle, consists in fixing the terminal vinyl nucleobase via the spacer with the support via an ester, ether or amide bond. Absolute freedom from water is essential for the success according to the invention.
Claims (7)
(s. Formelblatt)
bevorzugt mit langen Spacern am Heterocyclus mit einem Träger umsetzt. 1. A process for the preparation of vinyl nucleobases fixed to the support, characterized in that a vinyl nucleobase of the general formula I
(see formula sheet)
preferably with long spacers on the heterocycle with a support.
(s. Formelblatt)
bei Raumtemperatur bis +100°C in einem inerten Lösungsmittel, vorzugsweise in Alkanen, Ethern oder Aromaten mit einem Überschuß an den jeweiligen Vinylnukleobasen der allgemeinen Formel (I) (s. Formelblatt) und Vinylverbindungen der allgemeinen Formel (II) (s. Formelblatt) in Gegenwart eines radikalischen Starters bzw. durch Bestrahlung mittels γ - Co(60) zur Reaktion gebracht wird. 2. The method according to claim 1, characterized in that the corresponding terminal vinyl nucleobases of the general formula I fixed to the support
(see formula sheet)
at room temperature to + 100 ° C in an inert solvent, preferably in alkanes, ethers or aromatics with an excess of the respective vinyl nucleobases of the general formula (I) (see formula sheet) and vinyl compounds of the general formula (II) (see formula sheet) is reacted in the presence of a radical starter or by irradiation with γ-Co (60) .
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE2001147238 DE10147238A1 (en) | 2001-09-25 | 2001-09-25 | Preparation of carrier bound vinyl nucleobases and their polymers, useful, e.g. as antiviral and anticancer agents, or for diagnostics or purification, comprises reaction of vinyl nucleobases with a carrier |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE2001147238 DE10147238A1 (en) | 2001-09-25 | 2001-09-25 | Preparation of carrier bound vinyl nucleobases and their polymers, useful, e.g. as antiviral and anticancer agents, or for diagnostics or purification, comprises reaction of vinyl nucleobases with a carrier |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DE10147238A1 true DE10147238A1 (en) | 2003-04-24 |
Family
ID=7700219
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DE2001147238 Withdrawn DE10147238A1 (en) | 2001-09-25 | 2001-09-25 | Preparation of carrier bound vinyl nucleobases and their polymers, useful, e.g. as antiviral and anticancer agents, or for diagnostics or purification, comprises reaction of vinyl nucleobases with a carrier |
Country Status (1)
| Country | Link |
|---|---|
| DE (1) | DE10147238A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022012799A1 (en) | 2020-07-15 | 2022-01-20 | Merck Patent Gmbh | Optically active devices |
| US11702396B2 (en) | 2017-02-15 | 2023-07-18 | Johnson & Johnson Surgical Vision, Inc. | Hydrophobic compounds for optically active devices |
| US11958819B2 (en) | 2015-08-21 | 2024-04-16 | Johnson & Johnson Surgical Vision, Inc. | Compounds for optically active devices |
-
2001
- 2001-09-25 DE DE2001147238 patent/DE10147238A1/en not_active Withdrawn
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11958819B2 (en) | 2015-08-21 | 2024-04-16 | Johnson & Johnson Surgical Vision, Inc. | Compounds for optically active devices |
| US11702396B2 (en) | 2017-02-15 | 2023-07-18 | Johnson & Johnson Surgical Vision, Inc. | Hydrophobic compounds for optically active devices |
| WO2022012799A1 (en) | 2020-07-15 | 2022-01-20 | Merck Patent Gmbh | Optically active devices |
| WO2022012798A1 (en) | 2020-07-15 | 2022-01-20 | Merck Patent Gmbh | Optically active devices |
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