DD295245A5 - PROCESS FOR THE PREPARATION OF CYCLOSPORIN A CONJUGATES - Google Patents
PROCESS FOR THE PREPARATION OF CYCLOSPORIN A CONJUGATES Download PDFInfo
- Publication number
- DD295245A5 DD295245A5 DD34152390A DD34152390A DD295245A5 DD 295245 A5 DD295245 A5 DD 295245A5 DD 34152390 A DD34152390 A DD 34152390A DD 34152390 A DD34152390 A DD 34152390A DD 295245 A5 DD295245 A5 DD 295245A5
- Authority
- DD
- German Democratic Republic
- Prior art keywords
- csa
- conjugates
- cyclosporin
- epoxide
- macromolecules
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims abstract description 10
- 229930105110 Cyclosporin A Natural products 0.000 title claims abstract description 9
- 108010036949 Cyclosporine Proteins 0.000 title claims abstract description 9
- 229960001265 ciclosporin Drugs 0.000 title claims abstract description 9
- PMATZTZNYRCHOR-CGLBZJNRSA-N Cyclosporin A Chemical compound CC[C@@H]1NC(=O)[C@H]([C@H](O)[C@H](C)C\C=C\C)N(C)C(=O)[C@H](C(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)N(C)C(=O)CN(C)C1=O PMATZTZNYRCHOR-CGLBZJNRSA-N 0.000 title claims abstract description 8
- 238000002360 preparation method Methods 0.000 title claims abstract description 6
- 229920002521 macromolecule Polymers 0.000 claims abstract description 8
- 108090000790 Enzymes Proteins 0.000 claims abstract description 5
- 102000004190 Enzymes Human genes 0.000 claims abstract description 5
- 150000001720 carbohydrates Chemical class 0.000 claims abstract description 4
- 235000014633 carbohydrates Nutrition 0.000 claims abstract description 4
- 102000004169 proteins and genes Human genes 0.000 claims description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 6
- 150000001875 compounds Chemical class 0.000 claims description 5
- 102000002260 Alkaline Phosphatase Human genes 0.000 claims 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 claims 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims 1
- 108020005199 Dehydrogenases Proteins 0.000 claims 1
- 108010015776 Glucose oxidase Proteins 0.000 claims 1
- 239000004366 Glucose oxidase Substances 0.000 claims 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 claims 1
- 102000003855 L-lactate dehydrogenase Human genes 0.000 claims 1
- 108700023483 L-lactate dehydrogenases Proteins 0.000 claims 1
- 108090000854 Oxidoreductases Proteins 0.000 claims 1
- 102000004316 Oxidoreductases Human genes 0.000 claims 1
- 108700020962 Peroxidase Proteins 0.000 claims 1
- 102000003992 Peroxidases Human genes 0.000 claims 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 claims 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 claims 1
- 229920002684 Sepharose Polymers 0.000 claims 1
- 229940098773 bovine serum albumin Drugs 0.000 claims 1
- 229920002678 cellulose Polymers 0.000 claims 1
- 239000001913 cellulose Substances 0.000 claims 1
- 229940088598 enzyme Drugs 0.000 claims 1
- 229940116332 glucose oxidase Drugs 0.000 claims 1
- 235000019420 glucose oxidase Nutrition 0.000 claims 1
- 229960000814 tetanus toxoid Drugs 0.000 claims 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 abstract description 5
- 239000000872 buffer Substances 0.000 abstract description 5
- 238000001514 detection method Methods 0.000 abstract description 5
- 230000001900 immune effect Effects 0.000 abstract description 5
- 150000002118 epoxides Chemical class 0.000 abstract description 4
- 230000002163 immunogen Effects 0.000 abstract description 4
- 238000005227 gel permeation chromatography Methods 0.000 abstract description 3
- 238000000746 purification Methods 0.000 abstract description 3
- 238000011084 recovery Methods 0.000 abstract description 3
- 238000003018 immunoassay Methods 0.000 abstract description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical class CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 abstract 1
- -1 Cyclosporin A epoxide Chemical class 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 6
- 239000000126 substance Substances 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- KFSLWBXXFJQRDL-UHFFFAOYSA-N Peracetic acid Chemical compound CC(=O)OO KFSLWBXXFJQRDL-UHFFFAOYSA-N 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 239000007790 solid phase Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 108010010803 Gelatin Proteins 0.000 description 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000012089 stop solution Substances 0.000 description 2
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 1
- NHQDETIJWKXCTC-UHFFFAOYSA-N 3-chloroperbenzoic acid Chemical compound OOC(=O)C1=CC=CC(Cl)=C1 NHQDETIJWKXCTC-UHFFFAOYSA-N 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000002764 solid phase assay Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
Die Erfindung betrifft ein Verfahren zur Herstellung von Cyclosporin-A-Konjugaten mit Makromolekuelen, die als Komponenten fuer immunologische Nachweisverfahren von Cyklosporin A bzw. als Immunogen fuer die Gewinnung von CsA-Antiseren oder als Hilfsmittel fuer deren Aufreinigung geeignet sind. Erfindungsgemaesz wird diese Aufgabenstellung dadurch geloest, dasz CsA-Epoxid 1 in waeszriger ethanolischer Loesung in Gegenwart von Carbonatpuffer bei p H 9,6 mit Makromolekuelen wie Eiweiszverbindungen, Enzymen oder Kohlehydraten zur Reaktion gebracht wird. Die so gebildeten Konjugate lassen sich durch Gelchromatographie reinigen. Nicht vorhersehbar war der Befund, dasz auf diesem Wege hergestellte Konjugate monoklonale Anti-CsA-Antikoerper binden koennen.{Cyclosporin-A-Konjugate; Nachweisverfahren, immunologische; Immunogen; Antiseren; Makromolekuele; Cyclosporin-A-Epoxid; Enzymimmunassay; Antikoerperfestphasentest}The invention relates to a process for the preparation of cyclosporin A conjugates with macromolecules, which are suitable as components for immunological detection of cyclosporin A or as immunogen for the recovery of CsA antisera or as an aid for their purification. According to the invention, this object is achieved by reacting CsA epoxide 1 in aqueous ethanolic solution in the presence of carbonate buffer at pH 9.6 with macromolecules, such as egg white compounds, enzymes or carbohydrates. The conjugates thus formed can be purified by gel chromatography. Unforeseeable was the finding that conjugates prepared in this way can bind monoclonal anti-CsA antibodies. {Cyclosporin A conjugates; Detection method, immunological; immunogen; antisera; macromolecule; Cyclosporin A epoxide; Enzyme immunoassay; Antikoerperfestphasentest}
Description
Hierzu 3 Seiten ZeichnungenFor this 3 pages drawings
Die Erfindung betrifft ein Verfahren zur Herstellung von Cyclosporin A-Konjugaten mit Makromolekülen, die als Komponenten für immunologische Nachweisverfahren von Cyclosporin A bzw. als Immunogen für die Gewinnung von CsA-Antiseren oder als Hilfsmittel für deren Aufreinigung geeignet sind.The invention relates to a process for the preparation of cyclosporin A conjugates with macromolecules, which are suitable as components for immunological detection of cyclosporin A or as an immunogen for the recovery of CsA antisera or as an aid for their purification.
Charakteristik des bekannten Standes der TechnikCharacteristic of the known state of the art
Die chemische Verknüpfung des cyclischen Undecapeptides CsA mit Makromolekülen ist aufgrund der Besonderheiten seiner Struktur außerordentlich erschwert. Sie erfordert in der Regel beachtliche Veränderungen der Struktur <>s CsA, was mit großem Arbeitsaufwand und häufig mit dem Verlust an immunologischer Identität verbunden ist (PCT, WO 86,'0zu80). Die chemische Bindung von CsA über das Epoxid 1 (Abb. 1) an Proteine oder andere makromolekulare Trägerstoffe ist bisher nicht beschrieben. Die Herstellung von 1 gelang P.L.Durette u. Mitarb. (Transplantation Proceedings Vol.XX, No 2, Suppl.2,51-57 [1988]) durch Reaktion von 3'-0-Acetyl-CsA mit 3-Chlorperbenzoesäure.The chemical linkage of the cyclic undecapeptide CsA with macromolecules is extremely difficult because of the peculiarities of its structure. It usually requires considerable changes in the structure of CsA, which involves a great deal of work and often the loss of immunological identity (PCT, WO 86, 080). The chemical bonding of CsA via the epoxide 1 (Figure 1) to proteins or other macromolecular carriers has not previously been described. The preparation of 1 was successful for P.L.Durette et al. Al. (Transplantation Proceedings Vol.XX, No 2, Suppl. 2, 51-57 [1988]) by reaction of 3'-O-acetyl-CsA with 3-chloroperbenzoic acid.
Ziel der ErfindungObject of the invention
Ziel der Erfindung ist es, durch die Entwicklung einfacher chemischer Verfahren zur Herstellung von CsA-Konjugaten die Voraussetzungen zu schaffen, die kosten- und arbeitsaufwendigen Radioimmunoassays zum CsA-Nachweis durch einfache Enzymimmunoassays zu ersetzen.The aim of the invention is to create by the development of simple chemical methods for the preparation of CsA conjugates the conditions to replace the costly and laborious radioimmunoassays for CsA detection by simple enzyme immunoassays.
Darlegung des Wesens der ErfindungExplanation of the essence of the invention
Der Erfindung liegt die Aufgabe zugrunde, die chemische Bindung von CsA an Makromoleküle unter Bildung von Konjugaten zu ermöglichen, die geeignet sind, als Komponenten immunologischer Nachweisverfahren für CsA bzw. als Immunogen für die Gewinnung von CsA-Antiseren oder als Hilfsmittel für deren Aufreinigung zu dienen. Erfindungsgemäß wird diese Aufgabenstellung dadurch gelöst, daß CsA-Epoxid (Abb. 1) in wäßrig ethanolischer Lösung in Gegenwart von Carbonatpuffer bei pH9,6 mit Makromolekülen wie Eiweißverbindungen, Enzymen oder Kohlehydraten zur Reaktion gebracht wird. Die so gebildeten Konjugate lassen sich durch Gelchromatographie reinigen. Nicht vorhersehbar war der Befund, daß auf diesem Wege hergestellte Konjugate monoclonale Anti-CsA-Antikörper binden können. Die Erfindung wird anhand von 3 Ausführungsbeispielen näher erläutertThe invention has for its object to enable the chemical bonding of CsA to macromolecules to form conjugates which are suitable to serve as components of immunological detection methods for CsA or as an immunogen for the recovery of CsA antisera or as an aid for their purification , According to the invention this object is achieved in that CsA-epoxide (Fig. 1) is reacted in aqueous ethanolic solution in the presence of carbonate buffer at pH9.6 with macromolecules such as protein compounds, enzymes or carbohydrates. The conjugates thus formed can be purified by gel chromatography. It was not foreseen that conjugates prepared in this way could bind to monoclonal anti-CsA antibodies. The invention will be explained in more detail with reference to 3 embodiments
Die dazugehörigen Abbildungen zeigen The accompanying pictures show
Abb.1: 3'-0-Acetyl-CsA-EpoxidFig.1: 3'-O-Acetyl CsA epoxide
Abb.2a: 3'-0-Acetyl-CsAFig.2a: 3'-O-acetyl-CsA
Abb.2b: restliches 3'-0-Acetyl-CsA nach Reaktion mit Peressigsäure Abb.2c: restliches 3'-0-Acetyl-Epoxid nach Reaktion mit RSA.Fig.2b: residual 3'-O-acetyl-CsA after reaction with peracetic acid Fig.2c: residual 3'-O-acetyl-epoxide after reaction with RSA.
3'-0-ACeIyI-CtA-EpOxId(D3'-0-ACeIyI CTA epoxide (D
Das aus 1,8mg (1,5 pmol) CsA (AWD) nach der Methode von R.Traber u. Mitarb. (HeIv. Chim. Acta 65,1655-1677,1982) in einer Ausbeute von 92% erhaltene 3'-0-Acetyl-CsA wird in 1 ml 70% wäßrigem Ethanol gelöst. Nach Zugabe von 0,3ml Peressigsäure (2%) wird das Gemisch 16Std. im verschlossenen Gefäß stehen gelassen. Nach Zufügen von 1 ml H2O und 5ml CCI4 (p. a., dest.). Rühren und Zentrifugieren, wird die wäßrige Phase abgesaugt und verworfen. Die organische Phase wird zweimal mit je 1 ml H2O gewaschen und anschließend bei 4O0C unter N2 zur Trockne eingeengt. Die Umsetzung des 3'-0-Acetyl-CsA zu 1 läßt sich durch physikalisch-chemische Methoden kontrollieren. Nach Quantifizierung durch HPLC erfolgt sie zu 70-80% (Abb. 2a und 2 b).That of 1.8mg (1.5 pmol) CsA (AWD) according to the method of R. Traber u. Al. (HeIv. Chim. Acta 65, 16565-1677, 1982) in a yield of 92% 3'-O-acetyl-CsA is dissolved in 1 ml of 70% aqueous ethanol. After addition of 0.3 ml of peracetic acid (2%), the mixture is 16h. left in the closed container. After adding 1 ml H 2 O and 5 ml CCI 4 (pa, dist.). Stirring and centrifuging, the aqueous phase is filtered off with suction and discarded. The organic phase is washed twice with 1 ml H 2 O and then concentrated at 4O 0 C under N2 to dryness. The conversion of 3'-O-acetyl-CsA to 1 can be controlled by physical-chemical methods. After quantification by HPLC, it is 70-80% (Fig. 2a and 2 b).
3'-0-AcOtVl-CsA-RSA (2)3'-O-AcOtVl-CsA-RSA (2)
Das nach Beispiel 1 erhaltene Rohprodukt 1 wird in 0,5ml 70% Ethanol gelöst. Dazu gibt man eine Lösung von 2mg RSA (PPH Polskie Odczynniki Chemiczne) in 0,3ml H2O und fügt 0,2 ml Carbonatpuffer (0,1 mol/l, pH 9,6) hinzu. Das Gemisch läßt man unter gelegentlichem Rühren 20Std. bei Raumtemperatur stehen. Nach Quantifizierung durch HPLC erfolgt die Umsetzung von 1 zum Konjugat 2 zu etwa 80% (Abb. 2c). Die Reinigung von 2 erfolgt durch Gelchromatographie über eine Sephadex G-25-Säule 35cm x 1,4cm I. D.The crude product 1 obtained according to Example 1 is dissolved in 0.5 ml of 70% ethanol. Add 2mg RSA (PPH Polskie Odczynniki Chemiczne) solution in 0.3 ml H 2 O and add 0.2 ml carbonate buffer (0.1 mol / l, pH 9.6). The mixture is left under occasional stirring for 20h. stand at room temperature. After quantification by HPLC, the conversion of 1 to conjugate 2 is about 80% (Figure 2c). Purify 2 by gel chromatography on a Sephadex G-25 column 35cm x 1.4cm ID
Material und ReagenzienMaterial and reagents
0,01 mg 2 (bez. auf RSA)AnI Carbonatpuffer (0,1 mol/l, pH 9,6).0.01 mg 2 (based on RSA) AnI carbonate buffer (0.1 mol / l, pH 9.6).
Anti-CsA-Antikorper, monoclonal nonspezifisch (Lyophilisat, Sandoz LTD, Basel, Schweiz).Anti-CsA antibody, monoclonal non-specific (lyophilisate, Sandoz LTD, Basel, Switzerland).
Anti Maus (Ziege) POD (Lyophilisat, SIFIN, Berlin, DDR).Anti mouse (goat) POD (lyophilisate, SIFIN, Berlin, DDR).
Waschlösung: PBS (Phosphat 0,01 mol/l, NaCI 0,3mol/l), Tween 20 0,1 Vol.-%.Washing solution: PBS (phosphate 0.01 mol / l, NaCl 0.3 mol / l), Tween 20 0.1 vol .-%.
Medium 1:Medium 1:
Carbonatpuffer (0,1 mol/l, pH 9,6)/Gelafusal® = 98/2 [v/v].Carbonate buffer (0.1 mol / l, pH 9.6) / Gelafusal® = 98/2 [v / v].
Medium 2:Medium 2:
PBS/Gelafusal«/Tween 20 = 95/5/0,1 [v/v].PBS / Gelafusal "/ Tween 20 = 95/5 / 0.1 [v / v].
1.2-Phenylendiamindihydrochlorid-Lösung (1 mg/ml CitratpufferO,1 mol/l, pH5,0)/H2O2-Lösung (30%) = 1/0,001 [v/v].1.2-phenylenediamine dihydrochloride solution (1 mg / ml citrate buffer O, 1 mol / l, pH 5.0) / H 2 O 2 solution (30%) = 1 / 0.001 [v / v].
Stop-Losung: Na2SO3 (0,5mol/l H2SO4,1 molar).Stop solution: Na 2 SO 3 (0.5 mol / l H 2 SO 4 , 1 molar).
Festphase:Solid phase:
Mikrotestplatten (Scopyrol, VEB MLW Polyplast, Halberstadt, DDR).Microplates (Scopyrol, VEB MLW Polyplast, Halberstadt, GDR).
Mikrotestplatten werden mit 0,05ml CsA-RSA-Lösung pro Well über Nacht bei 4°C in der feuchten Kammer beschichtet und anschließend dreimal mit Waschlösung gewaschen. Die Nachbenetzung der Platten erfolgt mit je 0,05ml Medium 1 innerhalb 2 Std. bei 370C. Danach wird dreimal mit Waschlösung gewaschen.Microtest plates are coated with 0.05ml CsA-RSA solution per well overnight at 4 ° C in the humidified chamber and then washed three times with wash solution. The subsequent wetting of the plates is carried out with 0.05 ml of medium 1 within 2 hrs. At 37 0 C then washed three times with washing solution.
Die beschichteten und nachbenetzten Platten werden mit 0,05ml Anti-CsA-Antikörper-Lösung (0,3mg Lyophilisat/ml Medium 2) pro Well 2 Std. bei Raumtemperatur inkubiert und dreimal mit Waschlösung gewaschen. Anschließend erfolgt Inkubation mit 0,05ml Anti-Maus-POD (0,017 mg/ml Medium 2) pro Well, 1,5Std. bei Raumtemperatur. Danach wird erneut dreimal mit Waschlösung gewaschen. Die Substratreaktion wird mit 0,05ml Substratmedium pro Well bei Raumtemperatur innerhalb von 30min durchgeführt und danach mit 0,05ml Stop-Lösung abgestoppt. Die Extinktionsmessung erfolgt bei 493nm/690nm.The coated and wetted plates are incubated with 0.05 ml of anti-CsA antibody solution (0.3 mg lyophilisate / ml medium 2) per well for 2 hours at room temperature and washed three times with washing solution. Subsequently, incubation with 0.05 ml of anti-mouse POD (0.017 mg / ml medium 2) per well, 1.5 h. at room temperature. Thereafter, it is washed again three times with washing solution. The substrate reaction is carried out with 0.05 ml substrate medium per well at room temperature within 30 min and then stopped with 0.05 ml stop solution. The absorbance measurement takes place at 493nm / 690nm.
Claims (4)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DD34152390A DD295245A5 (en) | 1990-06-11 | 1990-06-11 | PROCESS FOR THE PREPARATION OF CYCLOSPORIN A CONJUGATES |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DD34152390A DD295245A5 (en) | 1990-06-11 | 1990-06-11 | PROCESS FOR THE PREPARATION OF CYCLOSPORIN A CONJUGATES |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DD295245A5 true DD295245A5 (en) | 1991-10-24 |
Family
ID=5619083
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DD34152390A DD295245A5 (en) | 1990-06-11 | 1990-06-11 | PROCESS FOR THE PREPARATION OF CYCLOSPORIN A CONJUGATES |
Country Status (1)
| Country | Link |
|---|---|
| DD (1) | DD295245A5 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7361636B2 (en) | 2004-10-06 | 2008-04-22 | Amr Technology, Inc. | Cyclosporin alkynes and their utility as pharmaceutical agents |
| US7378391B2 (en) | 2004-09-29 | 2008-05-27 | Amr Technology, Inc. | Cyclosporin alkyne analogues and their pharmaceutical uses |
| US7511013B2 (en) | 2004-09-29 | 2009-03-31 | Amr Technology, Inc. | Cyclosporin analogues and their pharmaceutical uses |
| US7696166B2 (en) | 2006-03-28 | 2010-04-13 | Albany Molecular Research, Inc. | Use of cyclosporin alkyne/alkene analogues for preventing or treating viral-induced disorders |
-
1990
- 1990-06-11 DD DD34152390A patent/DD295245A5/en not_active IP Right Cessation
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7378391B2 (en) | 2004-09-29 | 2008-05-27 | Amr Technology, Inc. | Cyclosporin alkyne analogues and their pharmaceutical uses |
| US7511013B2 (en) | 2004-09-29 | 2009-03-31 | Amr Technology, Inc. | Cyclosporin analogues and their pharmaceutical uses |
| US7361636B2 (en) | 2004-10-06 | 2008-04-22 | Amr Technology, Inc. | Cyclosporin alkynes and their utility as pharmaceutical agents |
| US7632807B2 (en) | 2004-10-06 | 2009-12-15 | Albany Molecular Research, Inc. | Cyclosporin alkynes and their utility as pharmaceutical agents |
| US7696166B2 (en) | 2006-03-28 | 2010-04-13 | Albany Molecular Research, Inc. | Use of cyclosporin alkyne/alkene analogues for preventing or treating viral-induced disorders |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| DE69423601T2 (en) | Electrochemical determination method and new p-phenylenediamine compound | |
| DE69121844T2 (en) | Immunoassay for cyclosporin | |
| Bayer et al. | Biocytin hydrazide—a selective label for sialic acids, galactose, and other sugars in glycoconjugates using avidin-biotin technology | |
| DE2656155C2 (en) | ||
| DE69132055T2 (en) | LOCALLY CONJUGATE FORMATION BETWEEN IMMUNOGLOBULINES AND DETECTABLE MARKING SUBSTANCES | |
| DE2910998A1 (en) | PROCESSES FOR THE MANUFACTURE OF BOUND ENZYME CONTAINING PRODUCTS AND THEIR USE | |
| DE2206103B2 (en) | Method for detecting and determining a component of the reaction between specifically binding proteins and the substances which are specifically bound by these proteins | |
| JPH06504364A (en) | A method for diagnosing prehypertension, hypertension, white coat hypertension, congestive cardiomyopathy, renal failure, salt sensitivity and adenoma, and endocrine cell hyperplasia; a method for monitoring the treatment of hypertension, congestive cardiomyopathy, renal failure and adenoma, and endocrine cell hyperplasia; Method for treating hypertension and antibodies having binding specificity for ouabain | |
| EP0491362B1 (en) | Use of peptide pairs with extremely high mutual specific affinity in the domain of in-vitro diagnostics | |
| EP0794196A2 (en) | Antibody for the selective immuno assay of bile in biological matrices | |
| DE69737298T2 (en) | CONJUGATES OF POLYSACCHARIDES AND BIOMOLECULES | |
| Singer et al. | [142] Flavin peptides | |
| JPS63196596A (en) | Glucooligosaccharide with inositol residue attached to the terminal and method for producing the same | |
| DE3882727T2 (en) | Mycoplasma membrane antigens and their use. | |
| DD295245A5 (en) | PROCESS FOR THE PREPARATION OF CYCLOSPORIN A CONJUGATES | |
| Gregory et al. | Preparation and properties of purified antilactic dehydrogenase | |
| DE3150878A1 (en) | "METHOD AND REAGENT FOR DETERMINING CREATININ" | |
| DE3224217A1 (en) | ENZYME IMMUNOASSAY METHOD FOR QUALITATIVE AND QUANTITATIVE DETERMINATION OF SUBSTANCES | |
| DE69128127T2 (en) | NUCLEIC ACID SAMPLES with negatively charged haptens connected by hydrazine bonds | |
| DD144461A5 (en) | SPECIFIC BINDING ANALYSIS METHOD FOR DETERMINING LIGAND IN A LIQUID MEDIUM | |
| Jordan et al. | Detection and quantification of oxalic acid from the brown-rot decay fungus, Postia placenta | |
| Howe et al. | Immunochemical Studies on Blood Groups. XIII. The Action of Enzyme from Snail (Busycon) Liver on Blood Group A and O (H) Substances (Hog) 1a | |
| Shemin | 10 δ-Aminolevulinic Acid Dehydratase | |
| EP0326074A1 (en) | Haptene derivatives and haptene protein conjugates therefrom | |
| DE3533671A1 (en) | METHOD FOR ACCELERATING AN ANTIGO-ANTIBODY REACTION |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| ENJ | Ceased due to non-payment of renewal fee |