DE2426913A1 - Malonic or succinic acid di(imido ester) prodn. - by reacting dicarboxylic acid dinitriles with anhydrous lower alcohols - Google Patents
Malonic or succinic acid di(imido ester) prodn. - by reacting dicarboxylic acid dinitriles with anhydrous lower alcoholsInfo
- Publication number
- DE2426913A1 DE2426913A1 DE19742426913 DE2426913A DE2426913A1 DE 2426913 A1 DE2426913 A1 DE 2426913A1 DE 19742426913 DE19742426913 DE 19742426913 DE 2426913 A DE2426913 A DE 2426913A DE 2426913 A1 DE2426913 A1 DE 2426913A1
- Authority
- DE
- Germany
- Prior art keywords
- prodn
- dmd
- acid
- diimidates
- diimidate
- 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.)
- Pending
Links
- OFOBLEOULBTSOW-UHFFFAOYSA-N Malonic acid Chemical compound OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 title claims abstract description 7
- 150000001298 alcohols Chemical class 0.000 title claims abstract description 4
- 150000002463 imidates Chemical class 0.000 title description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 title 2
- 239000001384 succinic acid Substances 0.000 title 1
- 238000000034 method Methods 0.000 claims abstract description 9
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 6
- 239000000539 dimer Substances 0.000 claims abstract description 4
- KLZYRCVPDWTZLH-UHFFFAOYSA-N 2,3-dimethylsuccinic acid Chemical compound OC(=O)C(C)C(C)C(O)=O KLZYRCVPDWTZLH-UHFFFAOYSA-N 0.000 claims abstract description 3
- OREAFAJWWJHCOT-UHFFFAOYSA-N dimethylmalonic acid Chemical compound OC(=O)C(C)(C)C(O)=O OREAFAJWWJHCOT-UHFFFAOYSA-N 0.000 claims abstract description 3
- 230000004071 biological effect Effects 0.000 claims abstract 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 238000004519 manufacturing process Methods 0.000 claims description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 3
- CUONGYYJJVDODC-UHFFFAOYSA-N malononitrile Chemical compound N#CCC#N CUONGYYJJVDODC-UHFFFAOYSA-N 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 abstract description 9
- 102000004169 proteins and genes Human genes 0.000 abstract description 9
- 239000003431 cross linking reagent Substances 0.000 abstract description 3
- 108090000317 Chymotrypsin Proteins 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 229960002376 chymotrypsin Drugs 0.000 description 5
- 229940088598 enzyme Drugs 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 150000001408 amides Chemical class 0.000 description 1
- 125000000320 amidine group Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 230000006957 competitive inhibition Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- MUCZHBLJLSDCSD-UHFFFAOYSA-N diisopropyl fluorophosphate Chemical compound CC(C)OP(F)(=O)OC(C)C MUCZHBLJLSDCSD-UHFFFAOYSA-N 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000000058 esterolytic effect Effects 0.000 description 1
- 229960005051 fluostigmine Drugs 0.000 description 1
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 1
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 description 1
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 1
- HDJNHVNQRJMWSH-UHFFFAOYSA-N hydron;methyl benzenecarboximidate;chloride Chemical compound Cl.COC(=N)C1=CC=CC=C1 HDJNHVNQRJMWSH-UHFFFAOYSA-N 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 description 1
- SJFKGZZCMREBQH-UHFFFAOYSA-N methyl ethanimidate Chemical compound COC(C)=N SJFKGZZCMREBQH-UHFFFAOYSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- IAHFWCOBPZCAEA-UHFFFAOYSA-N succinonitrile Chemical compound N#CCCC#N IAHFWCOBPZCAEA-UHFFFAOYSA-N 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/006—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length of peptides containing derivatised side chain amino acids
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
Verfahren zu ihrer Herstellung und Anwendung in der Peptidchemie Die Erfindung betrifft die Dicarbonsäure-Diiinidate DMD und DSD, Verfahren zu ihrer Herstellung und Anwendung in der Peptidchemie.Process for their preparation and application in peptide chemistry The invention relates to the dicarboxylic acid diinidates DMD and DSD, processes for their Manufacture and application in peptide chemistry.
Es ist erwünscht bifunktionale Reagenzien, die unter milden Bedingungen zur Herstellung von Proteinderivaten verwendet werden können, zur Verfügung zu haben. Dabei ist es jedoch erforderlich, daß die Länge der Kohlenstoffketten, die eingefahrt werden, 3 -4 Kohlenstoffatome nicht übersteigt, da die Anwendung längerkettiger Verbindungen zur Herstellung von Peptidderivaten nicht zur Gewinnung stabilisierter Moleküle führen würde.It is desirable to use bifunctional reagents that operate under mild conditions for the production of protein derivatives can be used. However, it is necessary that the length of the carbon chains that are retracted does not exceed 3 -4 carbon atoms, since the application is longer-chain Compounds for the production of peptide derivatives not for the production of stabilized ones Molecules would lead.
Es ist bekannt zur Abwandlung von Protein-Aminogruppen Imidoester als Gruppen-spezifische Reagenzien zu verwenden, wobei Amidingruppen entstehen (vgl. Hunter, M.J. and Ludwig, M.L., J. Am. Chem. Soc. 84 (1962) 3491 - 3504). Nach dieser Arbeit wurden jedoch nur Monoimidate - wie salz saures Methyl-Benzimidat und Methyl-Acetimidat - hergestellt und verwendet.It is known that imidoesters are used to modify protein amino groups to be used as group-specific reagents, resulting in amidine groups (cf. Hunter, M.J. and Ludwig, M.L., J. Am. Chem. Soc. 84 (1962) 3491-3504). After this However, only monoimidates - such as hydrochloric acid methyl benzimidate and methyl acetimidate - were used - manufactured and used.
Der Erfindung liegt die Aufgabe zugrunde bifunktionale Reagenzien zu gewinnen, die zur Herstellung stabilisierter Proteinderivate unter milden Bedingungen geeignet sind.The invention is based on the object of bifunctional reagents to win the production of stabilized protein derivatives under mild conditions are suitable.
Diese Aufgabe wird erfindungsgemäß dadurch gelöst, daß man Dicarbonsäuredinitril mit einer Kohlenstoff-Kettenlänge von 3 - 4 mit wasserfreien kurzkettigen Alkoholen in Gegenwart von gasförmiger Salzsäure bei etwa Oo umsetzt.According to the invention, this object is achieved by using dicarboxylic acid dinitrile with a carbon chain length of 3 - 4 with anhydrous short-chain Reacts alcohols in the presence of gaseous hydrochloric acid at about Oo.
Nach diesem Verfahren wurden Malonsäuredinitril und Bernsteinsäuredinitril mit Methanol zu Dimethylmalonsäure-Diimidat und Dimethylbernsteinsäure-Diimidat - welche im folgenden mit DMD und DSD bezeichnet werden - umgesetzt.According to this procedure, malononitrile and succinic acid dinitrile were made with methanol to dimethylmalonic acid diimidate and dimethylsuccinic acid diimidate - which are referred to below as DMD and DSD - implemented.
Die nach der Erfindung erzielten Vorteile bestehen insbesondere darin, daß die Diimidate in großer Ausbeute zugänglich sind und sich das Verfahren zu ihrer Herstellung leicht automatisieren läßtO Die Anwendung der neuen Verbindungen DMD und DSD zur Herstellung neuer Proteinderivate hat den Vorteil, daß die Reaktion schnell und vollständig abläuft und eine hohe Selektivität besitzt. Dies wird auf die Kürze der eingeführten Kohlenstoffbrücke zurückgeführt, die besonders stabil und genau definiert ist. Sie ist auch ausreichend stabil für proteolytische Verfahren. Das Reagenz DMD und das gebildete Proteinderivat sind ferner stark wasserlöslich, und daher eignen sie sich für weitere Reaktionen und für optische Untersuchungen. Die Haltbarkeit der erfindungsgemäßen Diimidate entspricht etwa derjenigen des damit hergestellten Chymotrypsin-Dimers.The advantages achieved according to the invention are in particular: that the diimidates are accessible in large yield and the process to their Manufacturing can be easily automated. The application of the new DMD compounds and DSD for the production of new protein derivatives has the advantage that the reaction runs quickly and completely and has a high selectivity. This is going on the brevity of the carbon bridge introduced, which is particularly stable and is well defined. It is also sufficiently stable for proteolytic procedures. The reagent DMD and the protein derivative formed are also highly soluble in water, and therefore they are suitable for further reactions and for optical investigations. The shelf life of the diimidates according to the invention corresponds roughly to that of the diimidates according to the invention produced chymotrypsin dimer.
Es ist auch wahrscheinlich, daß die Reagenzien DMD und DSD benutzt werden können, um andere Mitglieder der Klasse von Enzymen zu aktivieren, zu der Chymotrypsin gehört.It is also likely that the reagents used DMD and DSD can be used to activate other members of the class of enzymes to which Chymotrypsin heard.
Beispiel Zu Malonsäuredinitril wird wasserfreies Methanol bei 0° hinzugefügt. Es wird langsam gasförmiger Chlorwasserstoff eingeleitet, sodaß die Temperatur wegen der stark exothermen Reaktion nicht ansteigt. Das Reaktionsprodukt DMD kristallisiert im Reaktionsgefäß aus und wird aus einer Mischung von Äthanol und Äther umkristallisiert. Ausbeute bis zu 80%; Fp = 980. Die Synthese wird unter wasserfreien Bedingungen ausgeführt, da das Produkt in Wasser langsam hydrolysiert und Nebenprodukte entstehen können, die schwer zu entfernen sind. Example Anhydrous methanol becomes malononitrile at 0 ° added. Gaseous hydrogen chloride is slowly passed in, so that the Temperature does not rise because of the strongly exothermic reaction. The reaction product DMD crystallizes out in the reaction vessel and is made from a mixture of ethanol and ether recrystallized. Yield up to 80%; M.p. = 980. The synthesis will under run anhydrous conditions as the product hydrolyzes slowly in water and by-products that are difficult to remove can arise.
Anwendung von DMD in der Peptidchemie DMD kann benutzt werden, um zwei Enzymmoleküle Chymotrypsin an ihren aktiven Stellen miteinander zu verbinden : es reagiert mit Methionyl- und Histidyl-Seitenketten, wobei covalente Brücken gebildet werden. Durch die Umsetzung von DMD mit Chymotrypsin wird 0 ) die akt-ive Stelle des Chymotrypsinmoleküls stabilisiert, da die Brücke kurz und fest ist, 2o) die spezifische esterolytische Aktivität des Enzyms um maximal das 500- bis 1000-fache erhöht, 3.) die Aktivität gegenüber kleinen spezifischen Amiden auf etwa*10°S des Ausgangswertes herabgesetzt und 4.) die frühere augenblickliche und irreversible Hemmung des Enzyms durch Diisopropylfluorphosphat zu einer schwachen konkurrierenden Hemmung verringert. Aus diesem Grunde können andere für dieses Gift empfindliche Enzyme in Gegenwart des modifizierten Chymotlypsin-dimers vollständig inaktiviert werden ohne Schädigung des letzteren. Die konkurrierende Hemmung kann durch Dialyse aufgehoben werden. Application of DMD in peptide chemistry DMD can be used to to connect two enzyme molecules chymotrypsin to each other at their active sites : it reacts with methionyl and histidyl side chains, with covalent bridges are formed. By implementing DMD with chymotrypsin, 0) becomes the active Location of the chymotrypsin molecule stabilized because the bridge is short and firm, 2o) the specific esterolytic activity of the enzyme by a maximum of 500 to 1000 times increased, 3.) the activity towards small specific amides to about * 10 ° S des Initial value reduced and 4.) the earlier instantaneous and irreversible Inhibition of the enzyme by diisopropyl fluorophosphate to a weak competitive Inhibition decreased. Because of this, others may be sensitive to this poison Enzymes completely inactivated in the presence of the modified chymotlypsin dimer will be without harming the latter. The competing inhibition can be through dialysis To get picked up.
Dieses Verfahren schärft die Spezifität durch begrenzte Proteolyse, was sowohl bei der Medizinischen wie auch der wissenschaftlichen Aewend.ng exstrebenswert ist.This procedure sharpens the specificity through limited proteolysis, something that is desirable in both medical and scientific research is.
PatentallsprüchePatent claims
Claims (1)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19742426913 DE2426913A1 (en) | 1974-06-04 | 1974-06-04 | Malonic or succinic acid di(imido ester) prodn. - by reacting dicarboxylic acid dinitriles with anhydrous lower alcohols |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19742426913 DE2426913A1 (en) | 1974-06-04 | 1974-06-04 | Malonic or succinic acid di(imido ester) prodn. - by reacting dicarboxylic acid dinitriles with anhydrous lower alcohols |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DE2426913A1 true DE2426913A1 (en) | 1975-12-11 |
Family
ID=5917265
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DE19742426913 Pending DE2426913A1 (en) | 1974-06-04 | 1974-06-04 | Malonic or succinic acid di(imido ester) prodn. - by reacting dicarboxylic acid dinitriles with anhydrous lower alcohols |
Country Status (1)
| Country | Link |
|---|---|
| DE (1) | DE2426913A1 (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4053498A (en) * | 1976-07-30 | 1977-10-11 | The United States Of America As Represented By The Secretary Of The Air Force | Perfluoroalkylene ether-imidate and -thioimidate esters |
| US4288383A (en) * | 1979-08-14 | 1981-09-08 | E. I. Du Pont De Nemours And Company | Propenimidates, their preparation and rearrangement to pyrimidines |
| US4301087A (en) * | 1980-08-18 | 1981-11-17 | Standard Oil Company | Manufacture of carbamates from cyanogen |
| US4824949A (en) * | 1986-12-16 | 1989-04-25 | Hoechst Aktiengesellschaft | Process for the preparation of pyrimidines |
| US4831138A (en) * | 1986-12-16 | 1989-05-16 | Hoechst Aktiengesellschaft | Process for the preparation of aminopyrimidines |
| US5070201A (en) * | 1989-10-24 | 1991-12-03 | Hoechst Ag | Process for the preparation of aminopyrimidines |
| US5208337A (en) * | 1990-09-15 | 1993-05-04 | Hoechst Aktiengesellschaft | Process for the preparation of aminopyrimidines |
| US7141670B2 (en) | 2001-10-15 | 2006-11-28 | Lonza Ltd. | Process for the preparation of 1-(pyrimidin-2-yl)propan-2-ones |
-
1974
- 1974-06-04 DE DE19742426913 patent/DE2426913A1/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4053498A (en) * | 1976-07-30 | 1977-10-11 | The United States Of America As Represented By The Secretary Of The Air Force | Perfluoroalkylene ether-imidate and -thioimidate esters |
| US4288383A (en) * | 1979-08-14 | 1981-09-08 | E. I. Du Pont De Nemours And Company | Propenimidates, their preparation and rearrangement to pyrimidines |
| US4301087A (en) * | 1980-08-18 | 1981-11-17 | Standard Oil Company | Manufacture of carbamates from cyanogen |
| US4824949A (en) * | 1986-12-16 | 1989-04-25 | Hoechst Aktiengesellschaft | Process for the preparation of pyrimidines |
| US4831138A (en) * | 1986-12-16 | 1989-05-16 | Hoechst Aktiengesellschaft | Process for the preparation of aminopyrimidines |
| US5070201A (en) * | 1989-10-24 | 1991-12-03 | Hoechst Ag | Process for the preparation of aminopyrimidines |
| US5208337A (en) * | 1990-09-15 | 1993-05-04 | Hoechst Aktiengesellschaft | Process for the preparation of aminopyrimidines |
| US7141670B2 (en) | 2001-10-15 | 2006-11-28 | Lonza Ltd. | Process for the preparation of 1-(pyrimidin-2-yl)propan-2-ones |
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