WO1996037465A1 - Chiral isothiocyanates and their use as chiral derivatising agents - Google Patents

Chiral isothiocyanates and their use as chiral derivatising agents Download PDF

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WO1996037465A1
WO1996037465A1 PCT/EP1996/002258 EP9602258W WO9637465A1 WO 1996037465 A1 WO1996037465 A1 WO 1996037465A1 EP 9602258 W EP9602258 W EP 9602258W WO 9637465 A1 WO9637465 A1 WO 9637465A1
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chiral
groups
compounds
group
optionally substituted
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PCT/EP1996/002258
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Wolfgang Lindner
Oliver Paul Kleidernigg
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Wolfgang Lindner
Oliver Paul Kleidernigg
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B57/00—Separation of optically-active compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D207/16—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D215/48—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
    • C07D215/50—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen attached in position 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D219/00—Heterocyclic compounds containing acridine or hydrogenated acridine ring systems
    • C07D219/04—Heterocyclic compounds containing acridine or hydrogenated acridine ring systems with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the ring system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
    • C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
    • C07D243/10—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
    • C07D243/14—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines
    • C07D243/16—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals
    • C07D243/18—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals substituted in position 2 by nitrogen, oxygen or sulfur atoms
    • C07D243/24—Oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/02—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings
    • C07D307/26—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
    • C07D307/30—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D307/32—Oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D307/78—Benzo [b] furans; Hydrogenated benzo [b] furans
    • C07D307/82—Benzo [b] furans; Hydrogenated benzo [b] furans with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the hetero ring
    • C07D307/83—Oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
    • C07F7/02—Silicon compounds
    • C07F7/08—Compounds having one or more C—Si linkages
    • C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
    • C07F7/1804—Compounds having Si-O-C linkages

Definitions

  • the present invention relates to new chiral isothiocyanates their preparation and their application as chiral derivatizing agents (CD A), which may also be termed carefulreactive chiral selector", for the indirect analytical and preparative liquid chromatographic resolution of chiral amino compounds such as amines, amino acids, amino alcohols, peptides and thiol compounds as diastereomeric derivatives. Similarly, such diastereomers may also be separated by capillary electrochromatographic and capillary electrophoretic methods.
  • the new chiral isothiocyanates may also serve as building blocks for chiral selectors and chiral stationary phases (CSPs), to be used for the liquid phase resolution of chiral compounds.
  • the new CD As comprise three parts: 1) The reactive part (isothiocyanate moiety) for the chemically selective derivatization of primary and secondary amino- compounds and thiols.
  • the detecto ⁇ hore for the selective and sensitive detection of above mentioned diastereomeric compounds at trace levels (pM-aM range).
  • This feature is realized by the introduction of e.g. a strong fluorophore (e.g. methoxy-quinolinic acid) or a potent chromophore like 3,5-dinitro benzoic acid.
  • the detecto ⁇ horic group may also be considered as rationally designed substituent of the chiral selector to introduce specific intra- and intermoleCular interactions with substituents stemming from the analyte. Accordingly, the detecto ⁇ horic group may also be considered as selecto ⁇ horic group in the case of chiral selectors or CSPs derived from the new CD As.
  • Other demands considered to be important for a useful CDA were the chemical and slereochemical stability, its enantiomeric purity (> 99.9 ee) and high diastereoselectivity.
  • CDAs in stereoselective analysis of chiral amino and thiol compounds
  • various CDAs with different functional groups are known [W. Lindner, C. Pettersson in Liquid Chromatography in Pharmaceutical Development, I. Wainer, Ed., Part 1, Aster, Springfield 1985]. Focusing only on isothiocyanate based CDAs the following examples should be mentioned:
  • optically active and fluorescence active isothiocyanates are the benzoxadiazole derivatives namely 4-(3- isothiocyanato-2-pyrrolidin - 1-yl) -7-nitro-2,l,3- benzoxadiazole (NBD-PyNCS) [ T. Toyooka et.al., Analyst, 120 (1995) 385.] and 4-(3- isothiocyanato- pyrrolidin-l-yl)-7-(N,N-dimethylaminosulfonyl)- 2,1,3- benzoxadiazole (DBD-PyNCS) [ T. Toyooka et.al., Analyst, 120 (1995) 385.].
  • NBD-PyNCS 4-(3- isothiocyanato-2-pyrrolidin - 1-yl) -7-nitro-2,l,3- benzoxadiazole
  • DBD-PyNCS 4-(3- isothiocyanato
  • CDAs Like the carbohydrate based isothiocyanates latter CDAs react selectively with primary and secondary amines forming the corresponding diastereomeric thiourea derivatives. These diastereomers then can be separated into their optically isomeric forms, preferably by liquid chromatographic [ T. Toyooka et.al., Analyst, 120 (1995) 385.] or electrochromatographic [ I. S. Lurie, J. Chromatogr. A, 605 (1992) 269.] methods. However, CDAs with more then one stereogenic center preferably integrated in a conformational rigid ring system seem to be superior in terms of stereoselectivity of the resulting derivatives. Compared to the isothiocyanates based on proline and developed by Toyooka the new class of chiral CDAs fullfill this element of improvement.
  • CDAs is the OPA reaction, which only can be applied to primary amines.
  • CDAs used to date may be their relatively low chemical and chiral stability in basic (pH>8.5) aqueous media. Furthermore the diastereoselectivity of the above cited CDAs is also not always satisfying.
  • the object underlying the present invention is to provide chiral isothiocyanate type compounds and chiral derivatizing agents as defined in claims 1-5.
  • Preferred compounds have the general formula
  • the present invention comprises the synthesis of subtypes of CDAs which can be ionized and their application to electrophoretic separation of the resulting diastereomeric reaction products with chiral amines and thiols.
  • salt e.g. hydrochloride
  • Metoprolol DDITC A 5.33 7.83 1.47 6.67 GITC A 3.81 4.62 1.21 2.58
  • CH3CN/NH4AC (20mM) 45/55, apparent pH of rthe mobile phase 7.30;: flow: lml/min; detection UV 254nm;
  • a portion of 100 ⁇ l (0.862mM) aqueous D- or L- Pro solution was transferred into a 2ml vial.
  • the solution was alkalized with 5 ⁇ l 5 % aqueous Na2CO3 solution, diluted with 100 ⁇ l H2O and 100 ⁇ l ACN.
  • a 100 ⁇ l aliquot of the derivatizing reagent stock solution in ACN (1.724 ⁇ M) was added to the reaction vial, which got tightly capped, vortexed and stored at 50°C over a period of 90 minutes.
  • the cooled reaction mixture was diluted and neutralized with 200 ⁇ l acidic mobile phase and 10 ⁇ l of this solution were directly injected onto the reversed phase column
  • Example 12 as chiral stationary phase (CSP) in HPLC

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  • Organic Chemistry (AREA)
  • Plural Heterocyclic Compounds (AREA)
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Abstract

The invention relates to a chiral compound of general formula (I), wherein the groups R are the same or different but preferentially the same and are aliphatic C1-10 groups or optionally substituted aromatic groups or that both groups R together with the C-atoms marked with * form a carbocyclic or heterocyclic or anellated ring system (with 3-10, preferably 5-8 ring members) of the formula (II): (CR'R')n-, wherein R' and R'' are the same or different and are hydrogen atoms, aliphatic C1-10 groups, optionally substituted aromatic groups or optionally substituted hetero aromatic groups or anellated groups, and n is an integer from 1-10, and R1 is a group of general formula (III, IV or V), wherein R4, R5 or R6 are aliphatic C1-18 groups or optionally substituted aromatic groups or optionally substituted hetero aromatic groups or methyl-aryl groups, the preparation of such compounds as well as their use.

Description

CHIRAL ISOTHIOCYANATES AND THEIR USE AS CHIRAL DERIVATISING AGENTS
The present invention relates to new chiral isothiocyanates their preparation and their application as chiral derivatizing agents (CD A), which may also be termed „reactive chiral selector", for the indirect analytical and preparative liquid chromatographic resolution of chiral amino compounds such as amines, amino acids, amino alcohols, peptides and thiol compounds as diastereomeric derivatives. Similarly, such diastereomers may also be separated by capillary electrochromatographic and capillary electrophoretic methods. The new chiral isothiocyanates may also serve as building blocks for chiral selectors and chiral stationary phases (CSPs), to be used for the liquid phase resolution of chiral compounds.
In general terms, the new CD As comprise three parts: 1) The reactive part (isothiocyanate moiety) for the chemically selective derivatization of primary and secondary amino- compounds and thiols.
2) The chiral template realized by trans- (R,R)- or (S,S)-l,2-diaminocyclohexane (DACH), trans- (R.R)- or (S,S)- 1,2-diphenylethanediamine (DPEDA) or similar 1,2- trans diamines. Similar chiral templates may also be introduced by taking a meso (R,S)- or (S,R)- 1,2 - cis diamine and substituting it asymmetrically.
3) The detectoφhore for the selective and sensitive detection of above mentioned diastereomeric compounds at trace levels (pM-aM range). This feature is realized by the introduction of e.g. a strong fluorophore (e.g. methoxy-quinolinic acid) or a potent chromophore like 3,5-dinitro benzoic acid. The detectoφhoric group may also be considered as rationally designed substituent of the chiral selector to introduce specific intra- and intermoleCular interactions with substituents stemming from the analyte. Accordingly, the detectoφhoric group may also be considered as selectoφhoric group in the case of chiral selectors or CSPs derived from the new CD As. Other demands considered to be important for a useful CDA were the chemical and slereochemical stability, its enantiomeric purity (> 99.9 ee) and high diastereoselectivity.
The concept of using CDAs in stereoselective analysis of chiral amino and thiol compounds is well established and various CDAs with different functional groups are known [W. Lindner, C. Pettersson in Liquid Chromatography in Pharmaceutical Development, I. Wainer, Ed., Part 1, Aster, Springfield 1985]. Focusing only on isothiocyanate based CDAs the following examples should be mentioned:
In the category of carbohydrate based chiral isothiocyanates the most important representatives are 2,3,4,6-tetra-O- acetyl-β- D- glucopyranosyl isothiocyanate (GITC) [ T. Kinoshita et.al., J. Chromatogr., 210 (1981) 77.], 2,3,4-tri- O-acetyl-α- D-arabinopyranosyl isothiocyanate (AITC) [ T. Kinoshita et.al., J. Chromatogr., 210 (1981) 77.] and 2,3,4,6-tetra-O- benzoyl-β- D- glucopyranosyl isothiocyanate (BGIT) [ M. Lobell et.al., J. Chromatogr., 633 (1993) 287.]. These CDAs were and are used quite often for the indirect resolution of chiral amino compounds [ J. Gal, J. Chromatogr., 307 (1984) 220.; j. Gal, J. Chromatogr., 331 (1985) 349.; J. Gal, J. Liquid Chromatogr., 9 (1986) 673.; M. Fujimaki et.al., J. Pharm. Sci., 79 (1990) 568.; I. S. Lurie, J. Chromatogr. A, 605 (1992) 269.] and thiols [ S. Ito et.al., J. Chromatogr., 626 (1992) 187.]. One disadvantage of these sugar based isothiocyanates and of the corresponding thioureas is their relative low chemical stability in basic aqueous media due to the easily hydrolizable ester functionalities, leading to a loss in diastereoselectivity and/or unclear mixture of reaction and decomposition compounds. This is, however, a major drawback in the field including bioanalysis, amino acid- and peptide- analysis etc.. Another disadvantage is the relatively low stereoselectivity, making optical trace analysis and applications, where high selectivity is a must, almost impossible. The enantiomeric forms of these CDAs are also not available due to the lack of the corresponding carbohydrate moieties. Last but not least the detectability - only depending on the thiocarbonyl absoφtion in UV - is very limited, because of absence of a potent chromophore (with the exception of BGIT). Therefore trace analysis in the pM-aM range may not be managed.
Another group of optically active and fluorescence active isothiocyanates are the benzoxadiazole derivatives namely 4-(3- isothiocyanato-2-pyrrolidin - 1-yl) -7-nitro-2,l,3- benzoxadiazole (NBD-PyNCS) [ T. Toyooka et.al., Analyst, 120 (1995) 385.] and 4-(3- isothiocyanato- pyrrolidin-l-yl)-7-(N,N-dimethylaminosulfonyl)- 2,1,3- benzoxadiazole (DBD-PyNCS) [ T. Toyooka et.al., Analyst, 120 (1995) 385.]. Like the carbohydrate based isothiocyanates latter CDAs react selectively with primary and secondary amines forming the corresponding diastereomeric thiourea derivatives. These diastereomers then can be separated into their optically isomeric forms, preferably by liquid chromatographic [ T. Toyooka et.al., Analyst, 120 (1995) 385.] or electrochromatographic [ I. S. Lurie, J. Chromatogr. A, 605 (1992) 269.] methods. However, CDAs with more then one stereogenic center preferably integrated in a conformational rigid ring system seem to be superior in terms of stereoselectivity of the resulting derivatives. Compared to the isothiocyanates based on proline and developed by Toyooka the new class of chiral CDAs fullfill this element of improvement.
Besides chiral isothiocyanates also other classes of CDAs and methods to be applied for the derivatization of amines have been reported in literature: N-(5-fluoro-2,4-dinitrophenyl)-L- alanine amide (Marfey's reagent) [ H. Bruckner et.al., J. Chromatogr., 555 (1991) 81.], (+) and (-)-l-(9-fluorenyl)-ethyl chloroformate ((+)- and (-)-FLEC) [ S. Einarsson et.al., Anal Chem., 59 (1987) 1191.] and the achiral ortho-phthaldialdehyde (OP A) [ A. L. L. Duchateau et.al., J.
Chromatogr., 623 (1992) 237.] together with chiral thiols which provide the chiral information.
All these CDAs react more or less selectively with chiral primary and / or secondary amino functions forming the corresponding diastereomers. The only exception of the above mentioned
CDAs is the OPA reaction, which only can be applied to primary amines.
One inherent disadvantage of the approach in general and most of the chiral derivatizing agents
(CDAs) used to date may be their relatively low chemical and chiral stability in basic (pH>8.5) aqueous media. Furthermore the diastereoselectivity of the above cited CDAs is also not always satisfying.
A new class of chiral isothiocyanates has now been discovered which can function as particularly efficient chiral derivatizing agents.
1.) The object underlying the present invention is to provide chiral isothiocyanate type compounds and chiral derivatizing agents as defined in claims 1-5. Preferred compounds have the general formula
Figure imgf000006_0001
Figure imgf000006_0002
Figure imgf000006_0003
and are recovered from symmetric or unsymmetric but preferentially symmetric (R,R)-, (S,S)-, (S,R)- and (R,S)-l,2-diamines wherein by cyclization of the above mentioned 1,2- diamines with CS2 in accordance with Davies et al. [ S. G. Davies et.al., Tetrahedron, 49 (1993) 4419, S.G. Davies, European Patent Application (03.05.1991) EP 0 457 469 Al] forming the corresponding imidazolidine-2-thiones of the general formula
Figure imgf000007_0002
and subsequent selective monoacylation of the above mentioned imidazolidine thiones with acid chlorides. These cyclic thioureas undergo monoacylation suφrisingly followed by a thermal ring opening and proton migration to the desired isothiocyanates under neutral and acidic conditions, preferably in protic and aprotic media in a temperature range from 35 - 180° C depending on the choice of the solvents. The above mentioned ring opening of the cyclic thioureas was carried out in accordance to Ulrich et al. [Ulrich, H. et.al., J. Org. Chem. 43 (8) 1978] describing a thermal ring opening of macrocyclic ureas to isocyanates, but not thioureas to isothiocyanates, and, definitely excluding, however, the ring opening of 5- and 6- membered ring systems. Along this line Davies et al. [ S. G. Davies et.al., Tetrahedron, 49 (1993) 4419, S.G. Davies, European Patent Application (03.05.1991) EP 0 457 469 Al] described the bisacylation of imidazolidine thiones and their application as chiral auxiliaries, however, also in these studies ring opening reaction has not been reported.
Furthermore, the present invention comprises the synthesis of subtypes of CDAs which can be ionized and their application to electrophoretic separation of the resulting diastereomeric reaction products with chiral amines and thiols.
Figure imgf000008_0001
Figure imgf000008_0002
2.) The tagging of primary and secondary chiral amines, amino alcohols, amino acids, thiol compounds and their combinations - even with the simultaneous presence of more than one of the above mentioned functionalities - with the new chiral isothiocyanates, mentioned under point 1.), forming the corresponding diastereomeric thioureas, Edman chemistry corresponding anilinothiazolinones (ATZ), thiohydantoines, dithiocarbamates as well as thiazolidines. However, this principal application of chiral isothiocyanates is in accordance with citations in literature related to amino alcohols [ M. Fujimaki et.al., J. Pharm. Sci., 79 (1990) 568.], amino acids [ M. Lobell et.al.,, J. Chromatogr., 633 (1993) 287.] and thiols [ S. Ito et.al., J. Chromatogr., 626 (1992) 187.]. 3.) Taking advantage of the introduced deiectophore within tne new CDAs mentioned under point 1.) with respect to the following detection principles a) electrochemical detection b) UV-VIS detection c) fluorescence detection d) chemiluminescence detection
also in combination with all kinds of liquid phase separation systems.
4.) Application of the new chiral isothiocyanate type CDAs mentioned under point 1.) for the indirect resolution of chiral analytes as diastereomeric derivatives by all liquid phase separation systems.
5.) Application of the new chiral isothiocyanate type CDAs mentioned under point 1.) for the synthesis of chiral selectors or chiral stationary phases (CSPs) to be used for the resolution of chiral compounds by any liquid phase separation technique.
The invention will now be described in more detail with reference to the following examples:
Examples:
Example 1
Synthesis of (1R,2R)- and (1S.2S) N-[(2-isothiocyanato)-cyclohexyl]-(3,5)-dinitrobenzoyl amide ((1R,2R)-DNB-DACH-ITC and (1S,2S)-D B-DACH-ITC) DDITC:
Figure imgf000009_0001
a) Cyclization of trans 1,2 (R,R)- or (S,S)-diaminocy ohexane (DACH) with carbon disulfide , forming trans 4,5- Tetramethyleneimidazolidine -2- thione , was accomplished according to a method described by Davies et. al. [ S. G. Davies et.al., Tetrahedron, 49 (1993) 4419, S.G. Davies, European Patent Application (03.05.1991) EP 0 457 469 Al] . Spectroscopic data, yields and melting points were almost identical as described by Davies. b) For the monoacylation of the cyclic thiourea 6g (38.4 mM) of 4,5- tetramethyleneimidazolidine -2- thione were dissolved in 120 ml dichloroethane in presence of a catalytic amount of 4-(dimethylamino)-pyridine and added to a suspension of 7.97g (34.58 mM) 3,5 - dinitrobenzoylchloride in 30 ml dichloroethane at a temperature of 50°C over a period of 15 minutes. During the reaction all reactants dissolved and the solution was refluxed (90°C) for 2n. A slightly yellowish crystalline solid (1st crop) was filtered uncooled, washed and the mother liquor was evaporated to dryness. The residue was redissolved in 60 ml dichloroethane and refluxed for another 16n to thermally dispropose the 5-membered ring product to the open isothiocyanate. f ,R.-enantiomer : C14H14N4O5S = 350.35, yellowish crystals; m.p.: >250°C; total yield of both fractions 6.9g (51.2%);1H- MR (ppm) in DMSO-d6 1.15-2.29 (8H, m, cyclohexyl), 3.90 (1H, m CH-NCS), 4.10 (1H, iό, CH-NH), 9.01 (1H, m, dinitrobenzoyl. 9.09 (1H, m dinitrobenzoyl) 9.30 (1H, d -NH); IR (KBr): 3254 (NH), 2953 (CH), 2055 (-
NCS, strong), 1650 (-CO-), 1540 (-NO2), 1343 cm"1 (-NO2). [α]546=-133°.c=l in. acetonitrile, Elemental analysis calculated: C (48%), H (4.03%), N (15.99%); found: C (47.76%), H (3.99%), N (15.62%). (S.S , enantiomer : C14H14N4O5S = 350.35; all physicochemical properties except the specific rotation were identical to the (R,R)-enantiomer. [α]546= +135° c=l in acetonitrile; Elemental analysis calculated: C (48%), H (4.03%), N (15.99%); found: C (47.68%), H (4.07%), N (15.92%).
Example 2
Preparative scale derivatization of (R)- and (S)-Propranolol with (R,R)-DDITC:
Figure imgf000011_0001
200 mg (0.77 mM) optically pure (R)- and (S)-propranolol and 240 mg (0.70 mM) of
(R,R)-DDITC were dissolved in 7 ml acetonitrile and kept at 60°C for one hour. The reaction solution was evaporated under reduced pressure and the residue redissolved in 15 ml dichloromethane. The organic phase was acidified with aqueous 0.2 m HC1. This two phase system was stirred intensively in order to extract the excess propranolol into the aqueous phase. After separation of the phases the organic layer was washed with 15 ml water, dried over Na2SO4 and evaporated to dryness. The crude product was purified via flash chromatography (eluent toluene/acetone = 3/1).
_R.R.R.-diastereomer: C30H36N5O7S = 610.31; yellow crystals; mp. 168°C; yield 380 mg (91%); 1H-NMR (ppm) in CDCI3 1.02 (3H, d, -CH3), 1.24 (3H, d, -CH3), 1.31-1.55 (4H, m, cyclohexyl cβ), 1.69-2.39 (4H, m, cyclohexyl Cα), 3.55(2H, m, -CHfc-N), 3.71 (IH, s, -OH), 3.88 (IH, m, -CH-NH-CO-), 4.11 (2H, m, -O-CH2-), 4.30 (IH, m, -CH- OH), 4.82 (IH, m, -CH-NH-CS-), 5.50 (IH, m, N-CH-), 6.73 (IH, d, Aromat), 7.22-7.52 (4H, m, Aromat), 7.77 (IH, m, Aromat), 8.11 (IH, m, Aromat), 8.62 (IH, d, -NH), 8.95 (IH, m, DNB-Aromat), 9.05 (2H, m, DNB-Aromat); IR (KBr) 3255 (-NH), 3074 (Aromat), 2933 (-CH-), 1652 (-CO-), 1580 (-NH), 1539 (-NO2), 1344 cm"1 (-NO2); [α]546=-202°, c=1.03 in CH2CI2.
(S.R.R.-diastereomer: C30H36N5O7S = 610.31; yellow crystals; mp. 172°C; yield 370 mg (89%); 1H-NMR (ppm) in CDCI3 1.17-1.23 (6H, m, -CH(M_θ2), 1.32-1.59 (4H, m, cyclohexyl cβ), 1.72-2.40 (4H, m, cyclohexyl Cα), 3.55(2H, m, -CH2-N), 3.65 (IH, s, - OH), 3.73 (IH, m, -CH-NH-CO-), 4.16 (2H, m, -O-CH2-), 4.29 (IH, m, -CH-OH), 4.75 (IH, m, -CH-NH-CS-), 5.55 (IH, m, N-CH-), 6.77 (IH, d, Aromat), 7.30-7.40 (4H, m, Aromat), 7.72 (IH, d, Aromat), 8 11 (IH, d, Aromal), 8.55 '(IH, ά, -NH)/S.94 (IH, m, DNB-Aromat), 8.97 (2H, m, DNB-Aromat), 9.25 (IH, d, -NH); IR (KBr) 3265 (-NH),
3055 (Aromat), 2927 (-CH-), 1650 (-CO-), 1579 (-NH), 1541 (-NO2), 1343 cm"1 (- NO2); [α]546=-l l*5°, c=1.07 in CH2CI2.
Example 3
Synthesis of (1S,2S) N-[(2-isothiocyanato)-cyclohexyl]-pivalinyl amide ((S,S)-PDITC):
Figure imgf000012_0001
a) Cyclization of trans 1,2 diaminocyclohexane (DACH) with carbon disulfide , forming trans 4,5- Tetramethyleneimidazolidine -2- thione , was accomplished according to a method described by Davies et. al. [ S. G. Davies et.al., Tetrahedron, 49 (1993) 4419, S.G. Davies, European Patent Application (03.05.1991) EP 0 457 469 Al] . b) For the monoacylation of the cyclic thiourea 2g (12.8 mM) of 4,5- tetramethyleneimidazolidine -2- thione were dissolved in 50 ml dichloroethane in presence of a catalytic amount of 4-(dimethylamino)-pyridine and added to a suspension of 2.09 g
(17.33 mM) pivalinic acidchloride in 10 ml dichloroethane at a temperature of 50°C over a period of 15 minutes. During the reaction all reactants dissolved and the solution was refluxed (90°C) for 2n. The reaction mixture was evaporated to dryness. The residue was redissolved in 60 ml dichloromethane and extracted twice with diluted aqueous HC1. The organic phase was washed with brine, dried over Na2SO4 and evaporated to dryness. The crude product was purified via flash chromatography (eluent toluene/acetone = 9.5/0.5).
C12H20N2OS = 240.36, white fluffy solid; m.p.: 152°C; yield: 1.21 g (39.5%);1H-NMR (ppm) in CDC13 1.21 (9H, s, t-butyl) 1.25-2.18 (8H, m, cyclohexyl), 3.61 (IH, m CH- NCS), 3.87 (IH, m, CH-NH), 5.65 (IH, d, -NH-); IR (KBr): 3316 (NH), 2943 (CH), 2085
(-NCS, strong), 1633 (-CO-) cm"1. [α]546=-123°.c=l in acetonitrile Example 4
Synthesis of (1R,2R)- and (lS,2S)-N-[(2-isothiocyanato)-cyclohexyl]-6-methoxy-4- quinolinylamide ((1R.2R)- and (1S,2S)-CDITC):
Figure imgf000013_0001
a) Cyclization of trans 1,2 diaminocyclohexane (DACH) with carbon disulfide , forming trans 4,5- Tetramethyleneimidazolidine -2- thione , was accomplished according to a method described by Davies et. al. [ S. G. Davies et.al., Tetrahedron, 49 (1993) 4419, S.G. Davies, European Patent Application (03.05.1991) EP 0 457 469 Al] . b) Synthesis of 6-methoxy-4-quinolinic acid was accomplished according to Skraup et al. [Skraup Monatshefte Chem. 2 (1881) 592 ]. c) For the monoacylation of the cyclic thiourea 2g (12.8 mM) of 4,5- tetramethyleneimidazolidine -2- thione were dissolved in 50 ml dichloroethane in presence of a catalytic amount of 4-(dimethylamino)-pyridine and added to a suspension of 3g (11.7 mM) 6-methoxy-4-quinolinic acidchloride hydrochloride in 30 ml dichloroethane at a temperature of 50°C over a period of 15 minutes. During the reaction all reactants dissolved and the solution was refluxed (90°C) for 2n. A slightly yellowish crystalline solid
(1st crop of isothiocyanate) was filtered, washed and the mother liquor was evaporated to dryness (2nd crop, mixture of isothiocyanate, ringclosed product and 4,5- tetramethyleneimidazolidine -2- thione). 1st and 2nd crop were separately dissolved in 50 ml DCM and extracted with 50 ml 6.5 % NaHCO3 solution. The organic phases were washed with brine, dried over Na2SO4 and evaporated to dryness. The residue of the 2nd crop was redissolved in 60 ml dichloroethane and refluxed for another 20" to thermally dispropose (fig. 1) the 5-membered ring product to the open isothiocyanate. The crude fractions were recrystallized in toluene. . R.-enantiomer: C18H18N3O2S = 341.42, yellowish crystals; m.p.: 190°C; total yield of both fractions 2.9g (65.2%);1H-NMR (ppm) in CDC13: 1.25-2.25 (8H, m, cyclohexyl), 3.62 (IH, m CH-NCS), 3.95 (3H, s, -OCHj), 4.22 (IH, m, CH-NH), 6.39 (IH, d, -NH), 7.36-7.41 (2H, m, quinolinic H3 and H7), 7.51 (IH, d, quinolinic H5), 7.94 (IH, d, quinolinic H8), 8.70 (IH, d, quinolinic H2); IR (KBr): 3260 (NH), 2927 (CH), 2100 (-
NCS, strong), 1640 (-CO-) cm"1; [α]546=-H2°.c=0.98 in DCM
(S.S . enantiomer: C18H18N3O2S = 341.42; all physicochemical properties except the specific rotation were identical to the (R,R)-enantiomer. [α]546= +115° c= 1.04 in DCM
Example 5
Synthesis of (lR,2R)-N-[(2-isothiocyanato)-cyclohexyl]-6-methoxy-N-methyl-4- chinolinylamide hydrochloride ((lR,2R)-N-methyl-CDITC):
Figure imgf000014_0001
500 mg (1.46 mM) of (1R,2R)-CDITC were dissolved in 50 ml acetonitrile. This solution was cooled to 5°C, the 5 ml CH3I were added dropwise. The reaction mixture was stirred 2 h at 5°C and then heated to 30°C for 48 h. The solution was evaporated to dryness and stirred in 20 ml Et2O (abs.) to cystallize the product. The residue was filtered in vaccuo, dissolved in 50 ml MeOH and purified via an ion exchanger (Cl'-form).
C19H22N3O2SCI = 391.91, resulting 380 mg (65%) yellowish crystals; m.p.: 224°C; ]H- NMR (ppm) in DMSO-d6:1.21-2.28 (8H, m, cyclohexyl), 3.92 (IH, m CH-NCS), 4.02 (3H, s, -OCHj), 4.15 (IH, m, CH-NH), 4.68 (3H, s, N-CHj), 7.73 (IH, d, quinolinic H3), 8.01 (IH, m, quinolinic H7), 8.13 (IH, d, quinolinic H8), 9.53 (IH, d, quinolinic H2), 9.81 (IH, d, -NH); IR (KBr): 3286 (NH), 2949 (CH), 210S (-NCS, strong), 1640 (-CO-) cm"1; [α]546=-108°,c=0.98 in ACN
Example 6
General derivatization protocol for primary and secondary amines, amino alcohols (e.g. β- blockers) and thiol-compounds
A 0.01 mM amount of the amino-compound, in free base form or as salt (e.g. hydrochloride), was dissolved in 0.5ml of acetonitrile in a test tube. If the compound was in salt form 0.25 ml of 5% Na2CO3 was added. Then 0.5 ml acetonitrile containing 0.05 mM DDITC was added, the test tube tightly capped, vortexed for 1 minute and stored in an oven at 60°C. After a period of 120 minutes (or less, depending on the CDA and analytes) the reaction was stopped and the excess of CDA was quenched by the addition of 0.1 mM (L)-Proline and the reaction mixture put into the oven for another 30 minutes. Then an lOOμl aliquot was taken out, diluted with mobile phase, neutralized with acetic acid and lOμl of this mixture were directly injected onto the HPLC-column.
Indirect resolution of chiral amino alcohols using (R,R)- and (S,S)-DDITC as CDA
Figure imgf000016_0001
Indirect resolution of chiral amino alcohols using (R,R)- and (S,S)-DDITC as CDA in comparison to GITC Influence of column surface and CDA on the resolution of amino alcohols and amines deriv. conditions k'ι' compound k'21 α*
Metoprolol DDITC A 5.33 7.83 1.47 6.67 GITC A 3.81 4.62 1.21 2.58
Normetoprolol DDITC B 4.47 4.69 1.05 0.67 GITC B 2.47 - 1.00 0.00
Carvedilol DDITC A 66..1111 77..3355 11..2200 3.23
GITC A 55..4411 55..9988 11..1111 1.05
General HPLC conditions:
A: Column = Lichrospher 60 RP select B (125 X 4 mm i.d., 5μm); mobile phase composition: CH3CN/NH4AC (20mM) = 55/45, apparent pH of rthe mobile phase
7.30;: flow: lml/min; detection UV 254nm;
B: Column = Hypersil ODS(125 X 4 mm i.d., 5μm : mobile phase composition:
CH3CN/NH4AC (20mM) = 45/55, apparent pH of rthe mobile phase 7.30;: flow: lml/min; detection UV 254nm;
*α=k's/k'R ; ** Rs=1.18 X t2-tl/w(0.5)ι+w(0.5)2
Indirect resolution of chiral thiol compounds using (R,R)- and (S,S)-DDITC as CDA
compound mobile phase ki ' k2' α s
(R, S)-phenylethylmercaptan A 14.60 20.86 1.43 5.18
(R, S)-phenylethylamine A 14.75 16.23 1.10 1.58
(R,S)-2-mercapto succinc B 1.90 2.51 1.32 1.37 acid
(R,S)- 2-mercapto proprionic B 3.19 4.53 1.42 3.28 acid
*** B 8.46 11.90 1.41 4.78
(R,S)-N-acetyl cystein B 2.04 2.28 1.12 0.57
General HPLC conditions: flow: lml min; detection UV 254nm; mobile phase composition A: CH3CN/NH4AC (20mM) = 45/55, pH= =7.30; B:
CH3OH/NH4AC (20mM) = 55/45, pH=3.38 adjusted with acetic acid;
Column: Hypersil ODS (125 X 4 mm i.d., 5μm); *α=k'2/k'l ; ** Rs= =1.18 X t2- t]/wι+w2 *** the corresponding thiazolidines of 2- mercaptoproprionic acid via intramolecular condensation at acidic conditions Example 7
General derivatization procedure for the derivatization of - and β- amino acids and peptides by using (R,R)- or (S,S)- CDITC as CDA
A portion of 100 μl (0.862mM) aqueous D- or L- Pro solution was transferred into a 2ml vial. The solution was alkalized with 5 μl 5 % aqueous Na2CO3 solution, diluted with 100 μl H2O and 100 μl ACN. Then a 100 μl aliquot of the derivatizing reagent stock solution in ACN (1.724 μM) was added to the reaction vial, which got tightly capped, vortexed and stored at 50°C over a period of 90 minutes. The cooled reaction mixture was diluted and neutralized with 200 μl acidic mobile phase and 10 μl of this solution were directly injected onto the reversed phase column
Indirect resolution of chiral α-amino acids using (R,R)- and (S,S)-CDITC as CDA
Figure imgf000019_0001
Indirect resolution of chiral β-amino acids using (R,R)- and (S,S)-CDITC as CDA
Chromatographic data of (R,R)-CDITC derivatized ] DL - β - amino acids racemic β-amino acids mobile k'ι k'2 Re e.o. phase β-amino isobutyric acid D 34.35 42.60 1.24 3.20 D<L β-amino butyric acid D 21.88 27.38 1.25 2.88 D<L
2-methyl-taurine D 4.87 6.04 1.24 1.48 D<L
General HPLC conditions: Column: Hvpersil ODS (125 x 4 mm i.d.. 5 μm). Flow: lml/min: Detection Fluorescence λκ> = 325 nm. λ m = 420 nm: Mobile phase compositions: D: MeOH/ ammonium acetate (20 mM) = 40/60 apparent pH = 4.22; Example 8
General derivatization procedure for the derivatization of primary and secondary amines, amino alcohols, α- and β- amino acids and peptides by using (R,R)- or (S,S)- PDITC as CDA; derivatization protocols see also examples 6 and 7.
Indirect resolution of chiral α-amino acids, amines, amino alcohols and thiols using (S,S)- PDITC as CDA in comparison to (S,S)-CDITC and the well established GITC
Figure imgf000021_0001
Example 9
Micellar Electrochromatographic resolution of (R,S)-Leucine derivative using (R,R)-CDITC as CDA
Figure 1/4
Example 10
Synthesis of (lR,2R)-N-[(2-isothiocyanato)-cyclohexyl]-undecenylamide ((lR,2R)- UDITC):
Figure imgf000022_0001
a) Cyclization of trans 1,2 diaminocyclohexane (DACH) with carbon disulfide , forming trans 4,5- Tetramethyleneimidazolidine -2- thione , was accomplished according to a method described by Davies et. al. [ S. G. Davies et.al., Tetrahedron, 49 (1993) 4419, S.G. Davies, European Patent Application (03.05.1991) EP 0 457 469 Al]. b) For the monoacylation of the cyclic thiourea 0.7 mg (4.5 mM) of 4,5- tetramethyleneimidazolidine -2- thione and 0.68 g (6.7 mM) triethylamine were dissolved in 40 ml dichloroethane in presence of a catalytic amount of 4-(dimethylamino)-pyridine, heated to 90° C and a suspension of 1.34 g (6.64mM) undecenoyl chloride in 4 ml dichloroethane was added at a temperature of 90°C over a period of 15 minutes. The solution was refluxed for 14n. The residue was extracted twice with diluted aqueous 40 ml 0.12 n HC1. The organic phase was washed with brine, dried over Na2SO4 and evaporated to dryness. The crude product was purified via flash chromatography (eluent toluene).
C18H29 2OS = 322.42, yellow oil; yield: 0.52 g (52%);1H-NMR (ppm) in CDC13 1.31 (14H, s, -(CH2)7-) 1.50-2.00 (8H, m, cyclohexyl), 2.10 (2H, d, -CO-CH2-), 3.15 (IH, m CH-NCS), 3.52 (IH, m, CH-NH), 4.95 (2H, in, CH2-CH-), 5.80 (IH, m, CH2-CH-); IR (KBr): 3293 (NH), 3076 (double bond), 2076 (-NCS, strong), 1708 (-CO-) cm'1.
Example 11
Synthesis of ( lR,2R)-N-[(2-isothiocyanato)-cyclohexyl]-6-methoxy-N-allyl-4- chinolinylamide hydrochloride ((lR,2R)-N-allyl-CDITC):
Figure imgf000023_0001
500 mg (1.46 mM) (1R,2R)-CDITC were dissolved in 50 ml acetonitrile. This solution was cooled to 5°C, the 5 ml allyl bromide were added dropwise. The reaction mixture was stirred 2 h at 5°C and then heated to 30°C for 48 h. The solution was evaporated to dryness and stirred in 20 ml Et2O (abs.) to crystallize the product. The residue was filtered in vaccuo, dissolved in 50 ml MeOH and purified via an ion exchanger (Cl'-form).
C21H24N3O2SCI = 417.91, resulting 415 mg (68%) yellowish crystals; d.p.: 196°C; 1H- NMR (ppm) in DMSO-d6: 1.21-2.30 (8H, m, cyclohexyl), 3.15 (IH, m CH-NCS), 3.52 (IH, m, CH-NH), 4.05 (3H, s, -OCH ), 4.58 (3H, s, N-Cϊfc), 4.95 (2H, m, CH-z-CH-), 5.80 (IH, m, CH2-CH-); 7.75 (IH, d, quinolinic H3), 8.02 (IH, m, quinolinic H7), 8.14 (IH, d, quinolinic H8), 9.55 (IH, d, quinolinic H2), 9.82 (IH, d, -NH); IR (KBr): 3286 (NH), 3075 (double bond), 2086 (-NCS, strong), 1708 (-CO-) cm-I.[α]546=-H7θ,c=0.98 in ACN Example 12
Synthesis of (lR,2R)-l-[3-trimethoxysilyl-propyl]-N-[(cyclohexyl)-2-(3,5)-dinitrobenzoyl amide]-thiourea and its immobilization onto silica
Figure imgf000024_0001
2 g (5.71 mM) of (R,R)-DDITC was dissolved in 100 ml absolute dioxane, heated up to 40°C, and 1.13 g (6.27 mM) 3-aminopropyl-trimethoxysilane were added. Then the solution was refluxed for 4h. The residue was evaporated to dryness and this crude product purified via flash chromatography (eluent EE/DCM=3/6). C20H3 lNsOsSSi = 529.81, yellow oil; yield 3.0 g (99%);1H-NMR (ppm) in CDC13 0.58 (2H, t, Si-CH2), 1.5-2.45 (10H, m, 4 x CH2 cyclohexyl, 1 x CH2 propyl), 3.30 (2H, m, CH2-NH-CS-, 3.52 (9H, s, 3 x -O- CH3), 3.88 (IH, m CH-NH-CS-), 4.65 (IH, m, CH-NH-CO-), 5.96 (IH, d, NH), 6.38 (IH, t, NH), 8.75 (IH, d, -NH-CO-), 9.14 (3H, m, dinitrobenzoyl; IR (KBr): 3311 (NH), 3097 (Ar), 2953 (CH), 1648 (-CO-), 1536 (-NO2), 1345 (-NO2), 1079 (-0 CH3) cm"*.
3g of the chirally modified siloxane and 5g silica (Kromasil, 100 A, 5μm) were refluxed 19 h in absolute toluene in the presence of a catalytic amount of p-toluene sulfonic acid. Elemental analysis of the modified silica: C (10.58%), H (1.62%), N (3.14%); leading to calculated surface coverage of 440 μM/g silica. 6g of the chirally modified silica were suspended in 150 toluene (abs.) and endcapped with 3 ml bistrimethylsilyl-acetamide by refluxing the supension for 3.5h. Elemental analysis: C (11.48%), H (1.90%), N (2.98%). This mentioned CSP was slurry packed in a HPLC stainless steel column (150 x 40 mm i.d.) which served as „chiral column". Example 13
Application of Example 12 as chiral stationary phase (CSP) in HPLC
Figure imgf000025_0001
Example 14
Capillary electrophoretic migration of (R,R)-CDITC at pH 3
Figure 2/4
Example 15
Synthesis of (lR,2R)-l-acridinoyl-trans-4,5-tetramethylenimidazolidine-2-thione and in situ reaction with (RS)-propranolol to the corresponding diastereomeric thioureas
Figure imgf000026_0001
(R,S)-PropranoIol
Cyclization of trans 1,2 diaminocyclohexane (DACH) with carbon disulfide , forming trans 4,5- Tetramethyleneimidazolidine -2- thione , was accomplished according to a method described by Davies et. al. [ S. G. Davies et.al., Tetrahedron, 49 (1993) 4419; S.G. Davies, European Patent Application (03.05.1991) EP 0 457 469 Al] .
For the monoacylation of the cyclic thiourea 0.76 g (4.9 mM) of 4,5- tetramethyleneimidazolidine -2- thione were dissolved in 20 ml dichloroethane in presence of a catalytic amount of 4-(dimethylamino)-pyridine and added to a suspension of 1.5 g (5.4 mM) acridinic acidchloride hydrochloride in 20 ml dichloroethane at a temperature of
50°C over a period of 15 minutes. During the reaction all reactants dissolved and the solution was refluxed (90°C) for 2n. A orange-yellowish crystalline solid was filtered containing the 5-membered ringproduct. The residue was dissolved in 50 ml DCM and extracted with 50 ml 6.5 % NaHCO3 solution. The organic phase was washed with brine, dried over Na2SO and evaporated to dryness. C21H19N3OS = 361.46, orange-yellowish crystals; m.p.: 195°C; yield: 1.07 g
(55.2%);1H-NMR (ppm) in CDC13: 1.1525-2.29 (8H, m, cyclohexyl), 3.88 (2H, m, 2 x CH- ), 6.99 (IH, d, -NH), 7.56-8.38 (8H, m, Ar); IR (KBr): 3175 (NH), 2931 (CH), 1634 (-
CO-) cm'1;
In an analytical scale reaction the monoacylated 5-membered ring product was heated with (R,S)-propranolol in o-dichlorobenzol to yield the diastereomeric thioureas. Chromatogram of the indirect resolution shown in figure 3/4.
Example 16
Synthesis of (1R,2R) DADEA mono Quininolinoy amide isothiocyanate and in situ reaction with (R,S)-propranolol to the corresponding diastereomeric thioureas
Syntheses of (R,R)-DADEA was accomplished according to [Brienne, M.-J. et.al., Bull. Soc. Chim. Fr., (1973) 190; Trost, B. M. et. al., J. Am. Chem. Soc. 114 (1992) 9327]. The monoacylation and in situ reaction was performed as described in example 15. The result of the derivatization yielding the diastereomeric thioureas is illustrated in figure 4/4.

Claims

PATENT CLAIMS
1. Compound of the general formula (I)
Figure imgf000028_0001
wherein the groups R are the same or different but preferentially the same and are aliphatic d-io groups or optionally substituted aromatic groups or that both groups R together with the C- atoms marked with *form a carbocyclic or heterocyclic or anellated ring system (with 3-10, preferably 5-8 ring members) of the formula (II)
-(CR'R")n-
(H)
wherein R' and R" are the same or different and are hydrogen atoms, aliphatic Cl-10 groups, optionally substituted aromatic groups or optionally substituted hetero aromatic groups or anellated groups, and n is an integer from 1-16, preferably from 1-10 and RΪ is a group of the general formula (III):
Figure imgf000029_0001
wherein t are aliphatic CMS groups or optionally substituted aromatic groups or optionally substituted hetero aromatic groups or aralkyl groups or Ri is a group of the general Formula IV
Figure imgf000029_0002
wherein R5 is C1-C10 aliphatic group or optionally substituted aralkyl groups selected from groups consisting of
Figure imgf000029_0003
or Ri is a group of the general formula V
Figure imgf000030_0001
wherein ; is an optionally substituted aromatic group an optionally substituted heteroaromatic group like e.g. the dansyl residue
Figure imgf000030_0002
2. Compound according to claim 1, wherein the groups R are the same or different and are selected from the group consisting of
R=
Figure imgf000030_0003
wherein m is an integer from 1-6,
Figure imgf000031_0001
wherein R' and R" in the group of formula (II) are the same or different, are hydrogen atoms, aliphatic Cl-10 groups, optionally substituted aromatic groups or optionally substituted hetero aromatic groups or anellated groups and n is an integer from 1-6, and wherein Rt is selected from the group consisting of
Figure imgf000032_0001
Figure imgf000032_0002
— C(CH3)3 -(CH2)8— CH=CH2
wherein R3 ins an aliphatic CMO group or an optionally substituted methylaryl, or an optionally substituted benzyl group.
3. Compound according to claim 2 wherein R3 is a group selected from the formulae:
CH3 } — H5
Figure imgf000032_0003
Compounds according to any of the claims 1-3, which are optically pure. 5. Compounds according to any one of claims 1-4, namely (1R,2R)- and (l'S,2S) N-[(2- isothiocyanato)-cyclohexyl]-(3,5)-dinitrobenzoyl amide.
6. Compounds according to any one of claims 1-4, namely (1R,2R)- and (1S,2S) N-[(2- isothiocyanato)-cyclohexyl]-pivalinyl amide.
7. Compounds according to any one of claims 1-4, namely (1R,2R)- and (lS,2S)-N-[(2- isothiocyanato)-cyclohexyl]-6-methoxy-4-quinolinylamide.
8. Compounds according to any one of claims 1-4, namely (1R,2R)- and (1S,2S) - N-[(2- isothiocyanato)-cyclohexyl]-6-methoxy-N-methyl-4-chinolinylamide.
9. Compounds according to any one of claims 1-4, namely (1R.2R)- and (1S,2S)-N- [(2-isothiocyanato)-cyclohexyl]-undecenylamide.
10. Compounds according to any one of claims 1-4, namely (1R,2R)- and (lS,2S)-N-[(2- isothiocyanato)-cyclohexyl]-6-methoxy-N-allyl-4-chinolinylamide.
11. Compounds according to any one of claims 1-4, namely (1R,2R)- and (lS,2S)-l-[3- trimethoxysilyl-propyl]-N-[(cyclohexyl)-2-(3,5)-dinitrober_zoyl amide]-thiourea.
12. Compounds according to any one of claims 1-4, namely (1R2R)- and (lS,2S)-N-[(2- isothiocyanato)- 1 ,2-diphenylethyl]-6-methoxy-4-chinolinylamide.
13. Compounds according to any one of claims 1-4, namely (1R,2R)- and (lS,2S)-N-[(2- isothiocyanato)-cyclohexyl]-4-nitro-benzyloxy amide.
14. Method for the preparation of a compound according to any one of claims 1-4, wherein a compound of the general formula (VI)
Figure imgf000034_0001
which may be optically pure and wherein R and Ri are as defined in claims 1-4, is subjected to a ring opening process.
15. Method according to claim 14, wherein the ring opening process is a thermal ring opening process.
16. Method according to claims 14 or 15 using an aprotic or protic solvent for the ring opening process.
17. Use of a compound according to any one of claims 1-13 for the specific tagging of compounds which contain functional groups which react with isothiocyanates.
18. Use of a compound according to any one of claims 1-13 for the tagging and separation of chiral compounds.
19. Use according to claims 17 or 18, wherein the chiral compounds are selected from the group consisting of chiral thiols, chiral amines and mixtures thereof each of which may have one or more of their respective functionalities.
20. Use according to claim 19 wherein the chiral amines are selected from the group consisting of primary and secondary amines, amino alcohols and mixtures thereof.
21. Use according to claim 17 or 18 wherein the chiral compounds are selected from the group consisting of amino acids, peptides and proteins.
22. Use according to claim 21 wherein the chiral amines are selected from amino acids whereby the resulting chiral thiourea derivatives may be converted to the corresponding chiral thiohytantoins and/or the corresponding openchained products. 23. Use according to claim 19 wherein the chiral thiols are selected from thiol group containing acids whereby the resulting chiral dithiocarbamate derivatives may be converted to the corresponding chiral thiazolidines.
24. Use of a compound according to any one of claims 1-13 for the tagging of amino groups in peptides or proteins and subsequent selective cleavage of the N-terminal peptide bond resulting in the corresponding chiral thiohydantoins and/or openchained products of the tagged N-terminal amino acid moieties.
25. Use of compound according to any one of claims 1-13 as building block for the synthesis of chiral selectors or chiral stationary phases (CSPs) to be used for the liquid phase separations of chiral compounds
26. Chiral selector type products obtainable by reacting of a compound according to any of claims 1-13 with a chiral or nonchiral anchoring group to be immobilized to silica or any other adsorbens.
27. Use of a compound according to any of claims 1-4, wherein at least one of the compounds of the general formula III, IV, V, or VI reacts in situ with compounds containing functional groups, which react with isothiocyanates.
28. Process for the production of diastereomeric derivatives from mixtures of chiral compounds which comprises reacting a mixture of chiral compounds with at least one compound according to any one of claims 1-13.
29. Process according to claim 28, wherein the mixture of chiral compounds is comprised of thiols, primary and secondary amines, amino alcohols, peptides and/or amino acids.
30. Process for the separation of the products obtainable according to any one of claims 28 or 29 which comprises subjecting the products to liquid phase separation systems.
31. Process according to claim 30, wherein the liquid phase separation systems are normal or reversed LC or any type of electrochromatography. 32. Process according to claim 30, wherein the separation system is an electrophoretic process.
33. Process for the specific detection of the products according to claims 1-13 and / or 17, optionally separated by a process according to any one of claims 30-32, which comprises subjecting the products to an electrochemical detection UV/VIS detection fluorescence detection chemiluminescence detection conductivity detection
34. Process according to to claim 33 wherein the detection method is specifically selected for the product to be detected.
PCT/EP1996/002258 1995-05-26 1996-05-24 Chiral isothiocyanates and their use as chiral derivatising agents WO1996037465A1 (en)

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CN107602666A (en) * 2017-09-30 2018-01-19 中国人民解放军第二军医大学 A kind of thioesters stapler peptide and preparation method and application
US10017463B2 (en) 2014-07-21 2018-07-10 Brandeis University Inhibitors of deubiquitinating proteases
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US10640494B2 (en) 2014-05-19 2020-05-05 Northeastern University N-acylethanolamine hydrolyzing acid amidase (NAAA) inhibitors and their use thereof

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CN108956823A (en) * 2018-09-18 2018-12-07 延边大学 Chiral sulfhydryl compound splits novel mass spectrum derivatization reagent

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Cited By (6)

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Publication number Priority date Publication date Assignee Title
US20160090351A1 (en) * 2013-04-18 2016-03-31 Brandeis University Inhibitors of deubiquitinating proteases
US10640494B2 (en) 2014-05-19 2020-05-05 Northeastern University N-acylethanolamine hydrolyzing acid amidase (NAAA) inhibitors and their use thereof
US10017463B2 (en) 2014-07-21 2018-07-10 Brandeis University Inhibitors of deubiquitinating proteases
CN107602666A (en) * 2017-09-30 2018-01-19 中国人民解放军第二军医大学 A kind of thioesters stapler peptide and preparation method and application
WO2019217977A1 (en) * 2018-05-08 2019-11-14 Northeastern University N-acylethanolamine hydrolyzing acid amidase (naaa) inhibitors and use thereof
US10689357B2 (en) 2018-05-08 2020-06-23 Northeastern University N-acylethanolamine hydrolyzing acid amidase (NAAA) inhibitors and use thereof

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