EP3880662A1 - Neuartiger weg zur synthese von diazirinen, die mit wasserstoff-15 angereichert sein können oder nicht - Google Patents

Neuartiger weg zur synthese von diazirinen, die mit wasserstoff-15 angereichert sein können oder nicht

Info

Publication number
EP3880662A1
EP3880662A1 EP19804716.9A EP19804716A EP3880662A1 EP 3880662 A1 EP3880662 A1 EP 3880662A1 EP 19804716 A EP19804716 A EP 19804716A EP 3880662 A1 EP3880662 A1 EP 3880662A1
Authority
EP
European Patent Office
Prior art keywords
formula
nmr
aliphatic chain
mhz
diazirine
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP19804716.9A
Other languages
English (en)
French (fr)
Inventor
Vincent Reboul
Xavier Franck
Thomas GLACHET
Hamid MARZAG
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Centre National de la Recherche Scientifique CNRS
Universite de Caen Normandie
Institut National des Sciences Appliquees de Rouen
Universite de Rouen Normandie
Ecole Nationale Superieure dIngenieurs de Caen
Original Assignee
Centre National de la Recherche Scientifique CNRS
Universite de Caen Normandie
Institut National des Sciences Appliquees de Rouen
Universite de Rouen Normandie
Ecole Nationale Superieure dIngenieurs de Caen
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Centre National de la Recherche Scientifique CNRS, Universite de Caen Normandie, Institut National des Sciences Appliquees de Rouen, Universite de Rouen Normandie, Ecole Nationale Superieure dIngenieurs de Caen filed Critical Centre National de la Recherche Scientifique CNRS
Publication of EP3880662A1 publication Critical patent/EP3880662A1/de
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D229/00—Heterocyclic compounds containing rings of less than five members having two nitrogen atoms as the only ring hetero atoms
    • C07D229/02—Heterocyclic compounds containing rings of less than five members having two nitrogen atoms as the only ring hetero atoms containing three-membered rings

Definitions

  • the present invention relates to a new method for the synthesis of diazirines, enriched or not with nitrogen-15, from amino acids or imines via a synthesis method in a single pot.
  • the present invention also relates to the preparation of a solution of ammonia NH3 and 15 to novel compounds diazirines, enriched or not enriched in nitrogen-15.
  • These diazirines can be advantageously used in photoaffinity labeling ("photoaffinity labeling").
  • the 15-N2 diazirines hyperpolarization can be used.
  • Diazirines are three-membered heterocycles with a nitrogen-nitrogen double bond and an sp 3 carbon. They are generally obtained in several stages from carbonyl compounds.
  • diaziridines in 2 or 3 stages by passing through imines or oximes provided with leaving groups.
  • the corresponding diaziridines are then oxidized to diazirines via an oxidant (Hill, JR; Robertson, AABJ Med. Chem. 2018, 61, 6945-6963).
  • Diazirines have been known since the 1980s as being very reactive carbene precursors. This property has enabled the development of the use of diazirines as photoaffinity labeling reagents (“photoaffinity labeling” and “photo-crosslinking”), the carbene generated quickly forming a covalent bond with a nearby atom. These methods allow in particular to identify active sites and to study protein-protein interactions.
  • Diazirines as photoactive probes have been widely used, for example for the design of organic electronic components (WO2016-049123), in cosmetic methods of treating human body odors (WO2016-096897) and of treatment for attenuation of wrinkles (W02010-076490), of photolabile substances capturing perfumes (WO2017-045891) or also as bio-adhesives (W02014-081391).
  • Diazirines labeled with nitrogen-15, noted 15 N2-diazirines, can be used in hyperpolarization, a promising NMR technique for MRI imaging.
  • NMR Magnetic resonance
  • the hyperpolarization of 15 N 2 -diazirines uses the SABER-SHEATH technique (Signal Amplification by Reversible Exchange - Shield Enables Alignment Transfer to Heteronuclei) which is carried out using an organometallic catalyst based on iridium and parahydrogen (pF; available from hydrogen H 2 ).
  • This polarization is transferred to the 15 N 2 -diazirines thanks to the scalar couplings existing between the pF and the nitrogen atoms of the diazirine, following exchanges of reversible ligands. This then results in a spectacular increase in the NMR signal intensity of nitrogen-15 (up to approximately 15,000 times).
  • this hyperpolarization can also be transferred to the proton, which makes it possible to envisage its use in MRI, in particular in vivo [Shen, K .; Logan, AWJ; Colell, JFP; Bae, J .; Ortiz Jr., GX; Theis, T .; Warren, WS; Malcolmson, SJ; Wang, Q. Angew. Chem. Int. Ed.
  • diazirines The synthesis of diazirines is generally carried out in two or three stages, with low overall yields and has major drawbacks such as the use of dangerous reagents and restrictive reaction conditions such as the use of liquid ammonia (condensed ammonia gas at a temperature of -78 ° C).
  • Ls N-HOSA hydroxylamine-O-sulfonic acid
  • CISO 3 H chlorosulfonic acid
  • Diazirines are obtained with good yields of up to 99%. These methodologies could also be applied to the formation of diazirines enriched in nitrogen-15, or 15 N2-diazirines, also with high yields (ranging from 47% to 91%) using an ammonia solution enriched in nitrogen-15 ( 15 N HB) previously formed.
  • Zakarian, A. et al. J. Am. Chem. Soc. 2018, 140, 6027-6032 have developed a synthesis of 15 pure liquid NH3 from ammonium chloride, itself enriched in nitrogen-15 ( 15 NH 4 CI) .
  • This method nevertheless has the drawback of passing the ammonia to the gaseous state which means that it has to be recondensed at very low temperature (-78 ° C).
  • the inventors have also developed a synthesis of 15 NH3 in solution in an alcohol, typically methanol, from 15 NH 4 Cl, under very simple reaction conditions.
  • the subject of the present invention is a process for the synthesis in a single pot of a diazirine, in which the nitrogen atoms each correspond independently of each other to the 14 N isotope OR to the 1S N isotope, by reacting an amino acid or an imine, with ammonia of formula 14 NH3 or 15 NH3 and an oxidant containing a hypervalent iodine atom.
  • a second subject of the invention relates to a process for the synthesis of ammonia enriched with nitrogen-15, of formula 15 NH3, comprising the following steps:
  • the present invention also relates to diazirines derived from amino acids capable of being obtained or directly obtained by the process according to the invention.
  • Another subject of the invention relates to diazirines of formula (G) enriched with nitrogen-15 and their use in hyperpolarization.
  • the present invention also relates to the use of diazirines derived from amino acids capable of being obtained or directly obtained by the process according to the invention and of diazirines of formula (G) in photoaffinity labeling.
  • the present invention relates firstly to a process for the synthesis in a single pot of diazirine enriched or not with nitrogen-15, from an amino acid or an imine, comprising the reaction of the amino acid or of the imine with ammonia, enriched or not with nitrogen-15, and an oxidant containing a hypervalent iodine atom.
  • synthesis in a single pot is meant, within the meaning of the present invention, that the synthesis of the diazirines of the invention from amino acids or imines is carried out without isolation of the reaction intermediates, such as for example the corresponding diaziridines.
  • the synthesis of the diazirines of the invention is advantageously carried out in a single reaction solvent.
  • the term "diazirine" within the meaning of the present invention designates any three-membered heterocycle having a nitrogen-nitrogen double bond and an sp 3 carbon, substituted or not on this sp 3 carbon, enriched or not in nitrogen-15.
  • the term “enriched in nitrogen-15” means that the compound concerned comprises a proportion of isotope 1S N clearly greater than the natural isotopic abundance (0.36%).
  • the nitrogen-enriched compounds typically have an incorporation rate of between 60.0 and 99.9%. This incorporation rate depends on the percentage of nitrogen-15 enrichment of the nitrogen-15 source reagents. In the sense of the present invention, these are mainly commercial amino acids enriched in nitrogen-15 and commercial ammonium chloride enriched in nitrogen-15. These reagents are typically 99% enriched in nitrogen-15.
  • the terms “enriched with nitrogen-15”, “containing a nitrogen atom-15", "including a nitrogen atom is a 15 N atom "have the same meaning. Likewise, “nitrogen-15” and “ 15 N” are equivalent. If it is not specified that the compound is enriched in nitrogen-15, then it contains only nitrogen 14.
  • the diazirine obtained is enriched in nitrogen-15
  • Other nitrogen possibly present in diazirine outside the three-membered heterocycle can also be nitrogen-15.
  • amino acid is meant in the sense of the present invention any natural or unnatural amino acid, enriched or not with nitrogen-15, whatever their stereochemistry.
  • amino acid designates any chemical compound which has both at least one carboxylic acid function and at least one amine function.
  • the amino acid side chain can be protected or unprotected.
  • At least one carboxylic acid function and at least one amine function within the amino acid are unprotected.
  • it is advantageously an acid-a-amino, that is to say an amino acid in which the carboxylic acid function and the amine function are carried by the same carbon atom .
  • the natural amino acids correspond to the following compounds: /.-alanine, /.-asparagine, acid-Z.-aspartic, /.-glycine, /.-glutamine, L-leucine, L-isoleucine, L- phenylalanine, /. -tryptophan, /.-valine, /.-histidine, /.-tyrosine, /.-glutamic acid, L- arginine, /.-lysine, /.-sérine, /.-thréonine, /.-proline, /. -methionine, /.- cysteine.
  • the amino acid is enriched in nitrogen-15.
  • the amino acid is enriched in nitrogen-15
  • at least the nitrogen of the amine function linked to the carbon carrying the carboxylic acid function corresponds to the 15 N isotope.
  • Other nitrogen likely to be present on the side chain amino acid can also correspond to the 15 N isotope.
  • hypovalent iodine we mean an iodine atom within a molecule having more than eight electrons in its valence layer and therefore not respecting the byte rule.
  • the iodine is thus either in an oxidation state (+111), it is then of the 3- iodane type, or in an oxidation state (+ V), it is then of the 5- iodane type.
  • the oxidant used comprises an iodine atom advantageously of the 3- iodane type.
  • RECTIFIED SHEET (RULE 91) ISA / EP Said oxidant can be prepared and isolated upstream of the process or prepared in situ during the process of the invention from an iodine derivative in the presence of an oxidant or from an iodoso type derivative.
  • the oxidant containing a hypervalent iodine atom used during the process of the invention is obtained upstream of the process.
  • hypervalent iodine oxidant is meant, within the meaning of the present invention, a chemical compound having an oxidizing power and containing a hypervalent iodine atom.
  • the oxidant with hypervalent iodine has the role of oxidizing the reaction intermediates, such as diaziridines, to lead to the corresponding diazirines. It can also act as a catalyst for intermediate reactions taking place during the synthesis of the diazirines of the invention from amino acids or imines.
  • the oxidant containing a hypervalent iodine atom can be chosen from compounds of the 3- iodane type.
  • the hypervalent iodine oxidant can be chosen from the following compounds:
  • Ra and Rb each independently of one another represent a C -C alkyl, in particular a methyl or a tert-butyl, or a C -C haloalkyl, in particular a trifluoromethyl
  • Rc represents one or more substituents chosen from C 1 -C 6 alkyls, in particular a methyl group or an ethyl group
  • the halogens in particular Cl, Br or I
  • Rd represents a hydrogen, a C 1 -C 6 alkyl, such as methyl, or an acetate group.
  • the oxidant with hypervalent iodine can be chosen from the following compounds:
  • the oxidant used is PIDA (Phenyllodide DiAcetate; Phl (OAc) 2 ] of the following formula:
  • the ammonia source can be pure gaseous or liquid ammonia, ammonia in solution in a solvent such as methanol or water, or an ammonium of general formula NH 4 + X, where X advantageously represents a against ion chosen from the group consisting of:
  • the source of ammonia is ammonia in solution in a solvent, preferably ammonia in solution in methanol.
  • the ammonia used may or may not be enriched in nitrogen-15. According to a preferred embodiment, the ammonia used is enriched in nitrogen-15.
  • the diazirine obtained using the process of the invention does not contain nitrogen-15.
  • the diazirine obtained using the process of the invention is enriched in nitrogen-15 and contains one or two nitrogen-15.
  • the present invention relates to a process for the synthesis in a single pot of diazirine enriched or not with nitrogen-15, from an acid-a-amino or an imine, in particular an acid-a-amino , comprising the reaction of the amino-acid or the imine with ammonia, enriched or not with nitrogen-15, and an oxidant containing a hypervalent iodine atom of type 3- iodane.
  • the method of the invention makes it possible to synthesize a diazirine of formula (I) below:
  • RECTIFIED SHEET (RULE 91) ISA / EP or a physiologically acceptable salt or solvate thereof, a stereoisomer or a mixture of stereoisomers in any proportion, wherein the nitrogen atoms each independently correspond to the 14 N isotope or to the 'isotope 1S N, and in which R 1 represents H, V, W or VW where:
  • W represents a cycloalkyl, an aryl, a heterocycle or a heteroaryl; W is optionally substituted by 1 to 4 groups selected from halogens, CN, NO2, OH, NR'R "and an aliphatic chain, where up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O) , S, S (O), S (0) 2 , NR 'or SiR'R ",
  • R 2 represents H or an aliphatic chain, where up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O), S, S (O), S (0) 2 , NR 'or SiR'R ",
  • said method comprising the following steps:
  • A represents NH2
  • B represents COOH
  • R 1 and R 2 are as defined above
  • R 3 represents V, W or VW as defined above
  • Y and Z independently of one another represent OH, halogen, NH, CN, (Ci-Cio) alkyl, ((Ci-Cio) alkyl) -aryl or ((Ci-Cio) alkyl) - heteroaryl, where up to 4 methylene units of said alkyl are optionally replaced by O, C (O), S (0) 2 or NH, and in the presence of ammonia 14 NH or 15 NH, in a reaction solvent at a temperature advantageously between 0 ° C and room temperature, typically for a time between 1 h and 4 h.
  • Physiologically acceptable salts of the compounds of the present invention include conventional non-toxic salts of the compounds of the invention, such as those formed from organic or inorganic acids or organic or inorganic bases.
  • Physiologically acceptable salts of the compounds of the present invention include conventional non-toxic salts of the compounds of the invention, such as those formed from organic or inorganic acids or organic or inorganic bases.
  • the counter-ions chloride, bromide, fluoride, nitrate or bicarbonate forming ammonium salts and the counter-ions derived from alkali metals such as Na + , Li + , K + forming carboxylate salts.
  • alkali metals such as Na + , Li + , K + forming carboxylate salts.
  • Acceptable solvates for the compounds according to the present invention include conventional solvates such as those formed during the last step of the process for preparing the compounds according to the invention, with the reaction solvent (s).
  • aliphatic chain is meant, within the meaning of the present invention, a linear or branched hydrocarbon chain, completely saturated or containing one or more unsaturations, but not aromatic.
  • An aliphatic chain according to the present invention advantageously comprises from 1 to 18 carbon atoms, preferably from 1 to 12 carbon atoms, more preferably from 1 to 10 carbon atoms.
  • the term “aliphatic chain” according to the present invention includes alkyl, alkenyl or alkynyl groups, linear or branched, substituted or not.
  • alkyl group is meant, within the meaning of the present invention, a saturated, linear or branched hydrocarbon chain.
  • alkenyl group is meant, within the meaning of the present invention, a hydrocarbon chain, linear or branched, comprising one or more double bonds.
  • alkynyl group is meant, within the meaning of the present invention, a hydrocarbon chain, linear or branched, comprising at least one triple bond.
  • aryl is meant, within the meaning of the present invention, an aromatic hydrocarbon group, preferably comprising from 6 to 10 carbon atoms and comprising one or more contiguous rings. It will advantageously be a phenyl or naphthyl group.
  • cycloalkyl is meant, within the meaning of the present invention, a non-aromatic hydrocarbon ring, completely saturated or containing one or more unsaturations, advantageously comprising 3 to 10 members.
  • the term includes fused, spiro, or bridged polycycles.
  • Examples of cycloalkyl are cyclopropenyl, cyclohexyl or cyclobutyl.
  • heterocycle is meant, within the meaning of the present invention, a non-aromatic ring, completely saturated or containing one or more unsaturations, advantageously comprising 3 to 10 members in which one or more carbon atoms, advantageously 1 to 4 and again more advantageously 1 or 2, are each replaced by a heteroatom chosen from sulfur, nitrogen and oxygen atoms.
  • the term includes fused, spiro, or bridged polycycles.
  • it will be a unicycle or a bicycle in which 1 or 2 carbon atoms are each replaced by a heteroatom chosen from sulfur, nitrogen and oxygen atoms, preferably chosen from atoms. nitrogen and oxygen, such as nitrogen.
  • heterocycles are piperidinyl, piperizinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, azepanyl, thiazolidinyl, isothiazolidinyl, oxazocanyl, thiazepanyl, benzimidazolonyl.
  • heteroaryl is meant, within the meaning of the present invention, an aromatic group comprising one or more, in particular 1 or 2 attached hydrocarbon rings, in which one or more carbon atoms, advantageously 1 to 4 and even more advantageously 1 or 2, are each replaced by a heteroatom chosen from sulfur, nitrogen and oxygen atoms and in which each cycle advantageously comprises 5 to 7 links, preferably 5 or 6 links.
  • aromatic group comprising 1 or 2 attached hydrocarbon rings, each ring being 5 or 6 members, in which 1 or 2 carbon atoms are each replaced by a heteroatom chosen from sulfur atoms, d nitrogen and oxygen, preferably chosen from nitrogen and oxygen atoms, such as nitrogen.
  • heteroaryl groups are furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrididinyl quinol, pyrimidinyl quinol . It will especially be a pyridyl, indolyl or imidazolyl group.
  • halogen atom is meant, within the meaning of the present invention, fluorine, chlorine, bromine and iodine atoms.
  • unsaturated is meant, within the meaning of the present invention, that the hydrocarbon chain can comprise one or more unsaturation (s), advantageously one.
  • stereoisomer is meant, within the meaning of the present invention, a geometric isomer or an optical isomer.
  • the geometric isomers result from the different position of the substituents on a double bond which can then have a Z or E configuration.
  • optical isomers result in particular from the different position in the space of the substituents on a carbon atom comprising 4 different substituents. This carbon atom then constitutes a chiral or asymmetric center.
  • Optical isomers include diastereoisomers and enantiomers.
  • the optical isomers which are images of one another in a mirror but not superimposable are designated by "enantiomers”.
  • the optical isomers which are neither superimposable nor images of one another in a mirror are designated by “diastereoisomers”.
  • racemic mixture A mixture containing equal amounts of two individual enantiomeric forms of opposite chirality is referred to as a "racemic mixture".
  • chiral group is meant, within the meaning of the present invention, a group which is not superimposable on its image in a mirror.
  • Such a chiral group may in particular comprise an asymmetric carbon atom, that is to say a carbon atom substituted by four different substituents (including hydrogen).
  • the stereoisomerism of the compounds can be induced by the side chains R 1 and / or R 2 .
  • an aryl is said to be substituted by an aliphatic chain
  • the aliphatic chain can be located between said aryl and the rest of the molecule.
  • R 3 represents an aryl substituted by an aliphatic chain
  • this may mean that the aliphatic chain is directly linked to the nitrogen atom carrying the radical R 3 and said aliphatic chain carries an aryl.
  • the groups are benzyl and tosyle, where the methylene group of the aliphatic chain is replaced by a sulfone group.
  • An aryl substituted by an aliphatic chain also designates the case where the aryl is directly linked to the rest of the molecule and carries an aliphatic chain.
  • ambient temperature used subsequently denotes a temperature ranging from 18 ° C to 30 ° C, preferably ranging from 20 ° C to 25 ° C, more preferably being 22 ° vs.
  • R 1 represents H, V, W or VW where:
  • W represents a cycloalkyl, an aryl, a heterocycle or a heteroaryl; W is optionally substituted by 1 to 4 groups selected from halogens, CN, NO2, OH, NR'R "and an aliphatic chain, where up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O) , S, S (O), S (0) 2 , NR 'or SiR'R ", R' and R" being as defined above.
  • R 2 represents H or an alkyl group, preferably methyl.
  • R 2 is a hydrogen atom.
  • R 3 represents an aliphatic chain, a cycloalkyl or an aryl; in which up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O), S, S (O), S (O) 2 , NR 'or SiR'R ", and in which said aryl is optionally substituted with 1 to 4 groups selected from halogens, CN, NO2, OH, NR'R "and an aliphatic chain as defined above.
  • R 3 represents a benzyl, tosyl, cyclohexyl group, a hydroxyl or an alkyl such as a tert-butyl group.
  • R 3 is a tert-butyl or tosyl group.
  • X is an aryl, for example a phenyl, and / or Y and Z are preferably identical and even more preferably, Y and Z represent a halogen, for example a chlorine or a fluorine, an OH group, NH2, CN, 0- (Ci-C 6 ) alkyl, 0- (Ci-C 6 ) -perfluoroalkyle, O-aryl, 0-S (0) 2 - (Ci-Ce) alkyl, 0-S (0) 2 -heteroaryl, NH-S0 2 - (C I - Ce) alkyl, NH-S (0) 2 -heteroaryl, N (S0 2 - (Ci-Ce) alkyl) 2 , N (S (0) 2 -heteroaryl ) 2 or an ester group 0 (CO) (Ci-
  • the solvent used is a solvent advantageously chosen from alcohols, acetonitrile, THF, toluene, dichloromethane, ethyl acetate, DMSO, acetone, pyridine or DMF.
  • the reaction advantageously takes place in an alcohol such as methanol, ethanol, propan-l-ol, n-butanol, ferf-butanol, pentan-l-ol, hexan-l-ol, preferably in methanol.
  • reaction solvent corresponds to the solvent of the ammonia source.
  • the ammonia is used in excess relative to the compound of formula (II), preferably in an amount of 15 to 20 equivalents, preferably from 17 to 18 equivalents. In particular, ammonia is used in an amount of 17.5 equivalents relative to the compound of formula (II).
  • the compound of formula (II) is added to a solution of ammonia in a solvent, preferably to a solution of ammonia in methanol.
  • the oxidant comprising a hypervalent iodine atom is advantageously used in excess relative to the compound of formula (II), in a proportion of 2 to 5 equivalents, preferably 3 equivalents.
  • the oxidant comprising the hypervalent iodine atom is added to the reaction medium consisting of ammonia, the compound of formula (II) and the solvent, preferably when the reaction medium is at a temperature of 0 ° C.
  • a single addition of oxidant is advantageously carried out during the process for obtaining diazirines from the compounds of formula (II).
  • the temperature of the reaction medium is firstly maintained at 0 ° C. for a period of between 15 minutes and 45 minutes, preferably 30 minutes, after the addition of the oxidant comprising a hypervalent iodine atom, then it is brought up to ambient temperature, and typically maintained at this temperature typically for a time between 1 hour and 3 hours, preferably for 1 hour 30 minutes.
  • the diazirine obtained in step (a) is advantageously purified, preferably by chromatography or distillation.
  • chromatography is meant, within the meaning of the present invention, a technique for purifying the compounds based on the difference in affinity of the compounds with the mobile phase and the stationary phase.
  • chromatography a technique for purifying the compounds based on the difference in affinity of the compounds with the mobile phase and the stationary phase.
  • the compounds will be separated by adsorption chromatography.
  • the mobile phase can be a liquid, a gas or a supercritical fluid.
  • the mobile phase will be a liquid, preferably a mixture of solvents of different polarity, for example two solvents, chosen from pentane, ethyl acetate, methanol, dichloromethane, ethanol or diethyl ether.
  • the stationary phase can be paper, silica, modified or not, polymers, alumina, etc.
  • the stationary phase will be silica.
  • the chromatography will be chromatography on silica gel.
  • the compound of formula (II) is an amino acid of the following formula (IIa):
  • W represents a cycloalkyl, an aryl, a heterocycle or a heteroaryl; W is optionally substituted by 1 to 4 groups selected from halogens, CN, NO2, OH, NR'R "and an (Ci-Cio) aliphatic chain, where up to 4 methylene units of the (Ci-Cio) aliphatic chain are optionally replaced by O, C (O), S, S (O), S (0) 2 , NR 'or SiR'R ",
  • the compound (IIa) corresponds to the following amino acids: /.-aspartic acid, /.-asparagine, /.-glutamine, /.-glycine, /.-alanine, L-valine, L- isoleucine, /.-leucine, L-phenylalanine, /.-tryptophan, 4-methyl - / .- phenylalanine, 4-iodo - / .- phenylalanine /.-Histidine, /.-tyrosine, acid /.-glutamic acid.
  • the compound (11a) corresponds to natural amino acids.
  • the compound (IIa) corresponds to the amino acids making it possible to obtain non-volatile diazirines, such as the following natural amino acids: L-Histidine, /.-tyrosine or acid /.-glutamic acid.
  • the method comprises a preliminary step of protecting the compound (11a).
  • protection step means the establishment of a protective group on a chemical function present in the group R 1 of the compound (IIa).
  • the term “protection step” can also denote a reversible chemical reaction making it possible to inhibit the reactivity of a chemical function under the reaction conditions of the process of the invention, such as for example an oxidation, reduction or amidation.
  • protecting group is meant, for the purposes of the present invention, a group which makes it possible to protect a reactive chemical function against undesirable reactions such as the groups described by TW Greene, “Protective Groups In Organic synthesis”, (John Wiley & Sons, New York (1981)) and Harrison et al. "Compendium of Synthetic Organic Methods", Vols. 1 to 8 (J. Wiley & sons, 1971 to 1996).
  • the term “chemical function” designates, according to the present invention, any reactive chemical entity capable of reacting during the process of the invention. It can be an atom, a set of atoms, for example an unsaturated or aromatic saturated cycle or heterocycle.
  • the preliminary protection step is carried out on the compound (lia) in which R 1 comprises a chemical function incompatible with the reagents used in the process of the invention, such as OH, NH2, C (0) NH 2 , C (0) 0H, SH, a guanidine function, an indole ring or a thioether function.
  • the preliminary protection step is carried out on the compound (lia) in which R 1 corresponds to the side chain of the following amino acids: lysine, arginine, serine, threonine, methionine, tryptophan and cysteine.
  • the protective groups according to the present invention include in particular a tert-butoxycarbonyl group, commonly abbreviated Boc, a benzyl group (Bn), a trityl group (Tr), a carboxybenzyl group (CBz), acyl groups such as the acetate group ( Ac) or silylated protecting groups, such as ferf-butylediphenylsilyl (TBDPS).
  • the step of protecting a compound (IIa) whose group R 1 comprises a thioether function corresponds to an oxidation of this atom to sulfoxide.
  • a thiol function is protected in the form of thioethers, for example with a trityl.
  • an NH 2 function present in the group R 1 of the compound (IIa) is protected in the form of an amide or a carbamate, for example using an acetate group, a Boc group or a CBz group.
  • an OH function present in the group R 1 of the compound (IIa) is protected in the form of an ether, in particular using a benzyl, and more particularly of silylated ethers, for example using TBDPS .
  • an indole function in the group R 1 of the compound (IIa) is protected by the introduction of a group Boc.
  • the compound (11a) corresponds to the following protected amino acids: / V e -Acetyl-Z.-lysine, / V e -benzyloxycarbamate - / .- lysine, /.- methionine sulfoxide,
  • the compound (11a) undergoes a functional modification aimed at reducing the volatility of the corresponding diazirines.
  • Said functional modification typically corresponds to the introduction of groups which will increase the molecular weight and / or the polarity of the compound to make it less volatile.
  • the groups making it possible to reduce the volatility of the compounds include, for example, halogens, in particular iodine, or also isopropyl or tert-butyl groups.
  • the introduction of an iodine atom is particularly interesting because it makes it possible to carry out subsequent functionalizations by organometallic coupling in an efficient manner.
  • the compound (11a) corresponds to 4-iodophenyl-alanine.
  • the diazirine according to the invention advantageously of formula (I), enriched in nitrogen-15 is obtained using the process according to the invention from the amino acid ( 11a) enriched in nitrogen-15 and ammonia enriched in nitrogen-15.
  • the diazirine according to the invention, advantageously of formula (I), enriched in nitrogen-15 obtained from the amino acid (IIa) enriched in nitrogen-15 and ammonia enriched in nitrogen-15 has a degree of incorporation nitrogen-15 greater than 99%.
  • the diazirine according to the invention advantageously of formula (I), enriched in nitrogen-15 is obtained using the method according to the invention from the amino acid (lia) not enriched in nitrogen-15 and ammonia enriched in nitrogen-15.
  • the diazirine according to the invention, advantageously of formula (I), enriched in nitrogen-15 obtained from the amino acid (IIa) not enriched in nitrogen-15 and ammonia enriched in nitrogen-15 has a rate of incorporation of nitrogen-15 between 60% and 90%.
  • the compound of formula (II) is an imine of formula (Mb) below:
  • R 1 represents H, V, W or VW where:
  • W represents a cycloalkyl, an aryl, a heterocycle or a heteroaryl; W is optionally substituted by 1 to 4 groups selected from halogens, CN, NO2, OH, NR'R "and an aliphatic chain, where up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O) , S, S (O), S (0) 2 , NR 'or SiR'R ", R' and R” each independently of one another H or an aliphatic chain, where up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O), S, S (O), S (0) 2 , NR 'or SiR'R ",
  • R 2 represents H or an aliphatic chain, where up to 4 methylene units of the chain
  • aliphatics are optionally replaced by O, C (O), S, S (O), S (0) 2 , NR 'or SiR'R ", and R 3 represents V, W or VW as defined above.
  • R 1 represents an aliphatic (C 1 -C 4 ) chain as defined above.
  • R 1 comprises an acid function COOH carboxylic.
  • R 1 represents an alkyl-COOH group, for example CH2CH2COOH.
  • R 1 when R 1 is an aryl, it is advantageously unsubstituted or substituted by 1 to 4 groups selected from CN, NO2, OH, NR'R "and an aliphatic chain , where up to 4 methylene units of the aliphatic chain are optionally replaced by C (0) 0, S, S (O), S (0) 2 , NR 'or SiR'R ".
  • R 1 is an aryl, preferably a phenyl substituted by a group CN, NO2, C (O) - (Ci-C 4 alkyl) or C (0) 0- (Ci-C 4 alkyl). More preferably, R 1 is a phenyl substituted by a nitrile group (CN) or an NO2 group.
  • R 2 represents H or an alkyl, preferably a methyl.
  • R 3 represents an aliphatic chain, a cycloalkyl or an aryl; in which up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O), S, S (O), S (O) 2 , NR 'or SiR'R ", and in which said aryl is optionally substituted with 1 to 4 groups selected from halogens, CN, NO2, OH, NR'R "and an aliphatic chain as defined above.
  • R 3 represents a benzyl, tosyl, cyclohexyl group, a hydroxyl or an alkyl such as a tert-butyl group.
  • R 3 is a tert-butyl or tosyl group.
  • the imine of formula (Mb) is obtained beforehand from a carbonyl compound of formula (IV) below:
  • R 1 and R 2 are as defined above.
  • the reaction for obtaining the imine of formula (Mb) advantageously comprises the reaction of the carbonyl compound of formula (IV) with an amine of formula (V) below:
  • R 3 is as defined above.
  • R 3 represents a benzyl, tosyl, cyclohexyl group, a hydroxyl or an alkyl such as a tert-butyl group.
  • R 3 is a tert-butyl or tosyl group.
  • the diazirine according to the invention advantageously of formula (I), enriched in nitrogen-15 is obtained using the method according to the invention from imine (Mb) not enriched in nitrogen -15 and ammonia enriched in nitrogen-15.
  • the present invention also relates to a process for synthesizing ammonia enriched nitrogen-15, Formula 15 NH 3, comprising the steps of: (a ') reaction of the enriched ammonium chloride with nitrogen-15 formula 15 NH 4 CI with an alcoholate of formula RO-L in the corresponding anhydrous alcohol ROH, where
  • R x corresponds to an alkyl group, preferably a C1-C6 alkyl group, more preferably R x is methyl, and
  • L is a counterion chosen from Na + , K + and Li + , preferably Na + ,
  • the ammonium chloride used is typically 99% enriched in nitrogen-15.
  • the ammonium chloride enriched with nitrogen-15 and the alcoholate of formula RO-L are used in equivalent quantity.
  • the process temperature is preferably between 0 ° C and room temperature and the reaction time is between 2:30 and 3 hours.
  • the alcoholate of formula RO-L is added to a solution of ammonium chloride enriched in nitrogen-15 in the corresponding anhydrous alcohol ROH at a temperature of 0 ° C.
  • the alcoholate of formula RO-L is added to a solution of ammonium chloride enriched in nitrogen-15 in the corresponding anhydrous alcohol ROH in several portions, for example five portions, over a certain period of time typically included between 15 minutes and 1 hour, preferably over a period of 30 minutes.
  • reaction medium composed of the alcoholate of formula RO-L and of a solution of ammonium chloride enriched with nitrogen-15 in the corresponding anhydrous alcohol ROH is typically stirred at temperature advantageously ambient for a period of time between 1.5 hours and 3 hours, preferably for 2 hours.
  • the anhydrous alcohol used is chosen from methanol, ethanol, trifluoroethanol, propan-1-ol, propan-2-ol, hexafluoropropan-2-ol, n- butanol, tert-butanol, pentan-l-ol, pentan-2-ol, pentan-3-ol, hexan-l-ol, hexan- 2-ol and hexan-3- ol, preferably, the anhydrous alcohol according to the invention is methanol.
  • the alcoholate of formula RO-L used is advantageously chosen from lithium, sodium or potassium alcoholates of the anhydrous alcohols mentioned above, preferably the alcoholate according to the invention corresponds to sodium methanolate.
  • diazirines according to the invention advantageously of formula (I), not enriched in nitrogen-15, derived from amino acids, likely to be obtained or directly obtained via the process for obtaining diazirines according to the invention.
  • the diazirines of formula (I) not enriched in nitrogen-15 according to the invention come from natural or non-natural amino acids, protected or not.
  • diazirines according to the invention capable of being obtained or directly obtained via the process for obtaining the diazirines according to the invention are of formula (I) below:
  • W represents a cycloalkyl, an aryl, a heterocycle or a heteroaryl; W is optionally substituted by 1 to 4 groups selected from halogens, CN, NO2, OH, NR'R "and an aliphatic chain, where up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O) , S, S (O), S (0) 2 , NR 'or SiR'R ", R' and R" each independently of one another H or an aliphatic chain, where up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O), S, S (O), S (0) 2 , NR 'or SiR'R ", and
  • R 2 represents H or an aliphatic chain, where up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O), S, S (O), S (0) 2 , NR 'or SiR'R ".
  • R 2 represents a hydrogen or a (Ci-Ce) alkyl group, such as a methyl, preferably a hydrogen.
  • R 1 is preferably a phenyl substituted by a group CN, NO2, C (O) - (Ci-C 4 alkyl) or C (0) 0- (Ci-C 4 alkyl). More preferably, R 1 is a phenyl substituted by a nitrile (CN) or NO2 group.
  • diazirines of formula (I) not enriched in nitrogen-15 capable of being obtained or directly obtained by the process of the invention advantageously correspond to the following compounds:
  • the diazirines of formula (I) not enriched in nitrogen according to the invention are capable of being obtained or directly obtained by the process of the invention in which the compound (II) is of formula (Mb).
  • the diazirines of formula (I) according to the invention advantageously correspond to the following compounds:
  • the diazirines of formulas (I), not enriched in nitrogen-15, capable of being obtained or directly obtained via the process for obtaining the diazirines according to the invention are derived from amino acids, preferably natural amino acids.
  • the diazirines of formula (I) according to the invention derived from amino acids are capable of being obtained or directly obtained by the process of the invention in which the compound (II) is of formula (IIa).
  • the diazirines correspond to the following formula (1A):
  • R 1 is as defined above.
  • Another object of the present invention relates to diazirines enriched with nitrogen-15 of formula (G) below, capable of being obtained or directly obtained by the process for obtaining diazirines according to the present invention:
  • R 1 represents H, V, W or VW where:
  • R 2 represents H or an aliphatic chain, where up to 4 methylene units of the aliphatic chain are optionally replaced by O, C (O), S, S (O), S (0) 2 , NR 'or SiR'R ".
  • R 1 corresponds to the side chain of amino acids, natural or unnatural, protected or not.
  • R 2 represents a hydrogen or a (Ci-C 6 ) alkyl group, such as a methyl, preferably a hydrogen.
  • R 1 is preferably a phenyl substituted by a group CN, NO2, C (O) - (Ci-C 4 alkyl) or C (0) 0- (Ci-C 4 alkyl). More preferably, R 1 is a phenyl substituted by a nitrile (CN) or NO2 group.
  • diazirines of formula (I) capable of being obtained or directly obtained by the process of the invention advantageously correspond to the following compounds:
  • the diazirines of formula (G) enriched in nitrogen-15 according to the invention are capable of being obtained or directly obtained by the process of the invention in which the compound (II) is of formula ( Mb), preferably enriched in nitrogen-15.
  • the diazirines of formula (G) according to the invention advantageously correspond to the following compounds:
  • the diazirines of formula (G) are capable of being obtained or directly obtained according to the method of the invention from amino acids.
  • These amino acids can be natural or unnatural, preferably natural. Said amino acids may have been previously protected as described in the present application.
  • the amino acids may or may not be enriched in nitrogen-15, preferably, they are enriched in nitrogen-15.
  • the diazirines of formula (G) are derived from the following amino acids: L-aspartic acid, L-asparagine, Z.
  • the diazirines of formula (G) according to the invention derived from amino acids are capable of being obtained or directly obtained by the process of the invention in which the compound (II) is preferably of formula (IIa) enriched in nitrogen- 15.
  • the diazirines have the following formula (GA):
  • R 1 is as defined above.
  • diazirines of formula (G) derived from amino acids correspond to the following formulas:
  • the present application also relates to the use of diazirines 1a to 1s and optionally diazirines enriched in nitrogen-15 as defined above for their application in photoaffinity labeling.
  • An object of the present application also relates to the use of diazirines enriched in nitrogen-15 of formula (G) for their application in hyperpolarization, in particular in the field of medical imaging.
  • the level of 15 N 2 -diazirine is greater than 99%.
  • the medium is then filtered over Na 3 S0 4 and the residue is washed with ethyl acetate, then the solvent is evaporated under reduced pressure and the reaction crude is purified by chromatography on silica gel to provide the 3n imine or 3o.
  • R x 'Bu, Ts or S (Q) -pTol
  • N2-diazirine / ( 15 N 14 N) -diazirine ratio:> 99% from N-tBu imine 3a; 71% from N-Ts imine 3h (According to NMR measurements “N ⁇ H ⁇ ).
  • NMR “N ⁇ H ⁇ (60.8 MHz, CDCI 3 ): ⁇ 5 464.2 (broad s, 2 N, 15 N 2 -diazirine).
  • Diazirine la derived from tyrosine
  • UV irradiation 360 nm
  • the carbene generated in situ is then directly trapped by the reaction solvent, methanol, to lead to ether 6a (see figure below) with total conversion.
  • Molecular probes are generally complex molecules, having particular groups such as fluorescent markers or groups having a particular affinity with their biological target.
  • the inventors have shown that the diazirines according to the invention can be easily functionalized. Diazirine 1a therefore reacts with tert-butyl bromoacetate to lead to ester 7, then quickly hydrolyzed to obtain a carboxylic acid 8.
  • diazirine la O 0.5 mmol
  • acetonitrile 1.5 mL
  • potassium carbonate 0.75 mmol
  • potassium iodide 15 mol%
  • tert-butyl bromoacetate 0.625 mmol
  • the reaction medium is stirred for 24 h at room temperature, diluted with distilled water and then extracted with ethyl acetate.
  • the organic phase is dried over magnesium sulfate and then filtered. After evaporation under vacuum, diazirine 7 is obtained (131 mg; 99%) in the form of an orange oil.
  • diazirine 7 (0.4 mmol) in dichloromethane (1 ml) is added dropwise the trifluoroacetic acid (1 ml). After magnetic stirring of the reaction medium for 1 h at room temperature, it is evaporated and the residue is dissolved in diethyl ether. The organic phase is dried over magnesium sulfate and then filtered. After evaporation, diazirine 8 (82 mg; 99%) is obtained in the form of a beige solid.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
EP19804716.9A 2018-11-14 2019-11-14 Neuartiger weg zur synthese von diazirinen, die mit wasserstoff-15 angereichert sein können oder nicht Withdrawn EP3880662A1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR1860507A FR3088322A1 (fr) 2018-11-14 2018-11-14 Nouvelle voie de synthese de diazirines, enrichies ou non en azote-15
PCT/EP2019/081393 WO2020099596A1 (fr) 2018-11-14 2019-11-14 Nouvelle voie de synthese de diazirines, enrichies ou non en azote-15

Publications (1)

Publication Number Publication Date
EP3880662A1 true EP3880662A1 (de) 2021-09-22

Family

ID=65685751

Family Applications (1)

Application Number Title Priority Date Filing Date
EP19804716.9A Withdrawn EP3880662A1 (de) 2018-11-14 2019-11-14 Neuartiger weg zur synthese von diazirinen, die mit wasserstoff-15 angereichert sein können oder nicht

Country Status (5)

Country Link
US (1) US20220098155A1 (de)
EP (1) EP3880662A1 (de)
CA (1) CA3119871A1 (de)
FR (1) FR3088322A1 (de)
WO (1) WO2020099596A1 (de)

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2939657B1 (fr) 2008-12-15 2011-02-11 Oreal Composition cosmetique comprenant une polyamine portant des groupes diazirines et utilisation pour le photo-greffage d'un polymere non saccharidique different des polymeres polyamines
US10005936B2 (en) 2012-11-23 2018-06-26 Nanyang Technological University Photoactive bioadhesive compositions
TWI648271B (zh) 2014-09-23 2019-01-21 日商住友電木股份有限公司 二氮環丙烯化合物及由其衍生之組成物
FR3029788B1 (fr) 2014-12-16 2018-03-02 L'oreal Procede cosmetique pour attenuer les odeurs
DE102015217978A1 (de) 2015-09-18 2017-03-23 Henkel Ag & Co. Kgaa Diazirine als Photocages, die Doppelbindungen enthaltende Moleküle freisetzen

Also Published As

Publication number Publication date
US20220098155A1 (en) 2022-03-31
WO2020099596A1 (fr) 2020-05-22
CA3119871A1 (fr) 2020-05-22
FR3088322A1 (fr) 2020-05-15

Similar Documents

Publication Publication Date Title
EP0167459B1 (de) 2-Amino-oxazoline und Verfahren zu ihrer Herstellung
EP0528729A1 (de) Verfahren zur Herstellung von Beta-Phenylisoserinderivate und ihre Verwendung
EP0309324A1 (de) Verfahren zur Herstellung von N-alkylierten Aminosäuren und deren Estern, Verwendung zur Synthese von Carboxyalkyldipeptiden
FR2490491A1 (fr) Composition pharmaceutique analgesique contenant comme ingredient actif l'acide 3-amino-2-hydroxy-4 phenylbutanoique et ses derives et application
EP0007834B1 (de) Verfahren zur Herstellung von optisch aktiven Alpha-Aminosäuren und deren Derivate
WO2020099596A1 (fr) Nouvelle voie de synthese de diazirines, enrichies ou non en azote-15
EP0915858B1 (de) Zwischenprodukte um 2-imidazolin-5onederivate herzustellen.
EP0500443A1 (de) Phenylethanolamino- und Phenylethanolaminomethyltetraline, Verfahren zu deren Herstellung Zwischenprodukte davon und diese enthaltende pharmazeutische Zusammensetzungen
EP2084125B1 (de) Aminobenzocycloheptenderivate, verfahren zu deren herstellung und deren verwendung in der therapie
EP1140794B1 (de) Verfahren zur herstellung von chiralen aminosäuren
WO2005000823A1 (fr) Procede de preparation du cis-8b-methyldecahydro-2a,4a,6a,8a-tetraazacyclopenta[fg]acenaphthylene, du cis-decahydro-2a,4a,6a,8a-tetraazacyclopenta[fg]acenaphthylene, du cyclene, et de cyclenes fonctionnalises
JPH11512412A (ja) 光学活性α−アミノ酸およびα−アミノ酸誘導体の製造方法
EP4073080B1 (de) Verfahren zur industriellen herstellung des dinatriumsalzes von (2s)-3-([1,1'-biphenyl]-4-yl)-2-((hydroxy((1r)-1-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)ethyl)phosphoryl)methyl)propanoyl)-l-alanin
RU2824994C1 (ru) Способ получения дексмедетомидина и его фармацевтически приемлемых солей
FR2760452A1 (fr) Procede d'obtention d'enantiomeres d'acides alpha-amines et sels diastereoisomeriques intermediaires
EP0837843B1 (de) Verfahren zur herstellung von enantiomeren der aminoalkylaminophenylpropansäure
WO2003068140A2 (fr) Derives de la pyridazine, leur utilisation comme medicaments, et leur procede de preparation
WO1994006760A1 (fr) Derives d'(amino-3 phenyl)-1 ethanesulfonate d'alkylammonium optiquement actifs, leur preparation et leur utilisation
EP2300405B1 (de) Verfahren zur asymmetrischen synthese von 6-fluor-l-dopa und analoga davon
WO2001066530A1 (fr) URETHANE N-CARBOXYANHYDRIDES ISSUS DE β-AMINOACIDES, LEUR PROCEDE DE PREPARATION ET LEURS UTILISATIONS.
EP0925280A1 (de) N-geschützte alpha-aminosulfinsäuresalze, verfahren zu ihrer herstellung sowie die daraus abgeleiteten sulfonopeptiden
JPS6042359A (ja) α−置換−α−シアノメチルアルコ−ルの不整合成方法
CH482702A (fr) Procédé de préparation de dérivés d'acides aminés
EP0228356A1 (de) Hydroxy-4-indol-Derivate, Verfahren zur Herstellung und ihre Anwendung
FR2863609A1 (fr) Procede d'enrichissement enantiomerique d'un melange d'enantiomeres d'alpha-aminoacides

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: UNKNOWN

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20210614

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

RAV Requested validation state of the european patent: fee paid

Extension state: MD

Effective date: 20210614

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20230601