Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C215/00—Compounds containing amino and hydroxy groups bound to the same carbon skeleton
  • C07C215/02—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton
  • C07C215/22—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being unsaturated
  • C07C215/24—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being unsaturated and acyclic
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C213/00—Preparation of compounds containing amino and hydroxy, amino and etherified hydroxy or amino and esterified hydroxy groups bound to the same carbon skeleton
  • C07C213/08—Preparation of compounds containing amino and hydroxy, amino and etherified hydroxy or amino and esterified hydroxy groups bound to the same carbon skeleton by reactions not involving the formation of amino groups, hydroxy groups or etherified or esterified hydroxy groups
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
  • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
  • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
  • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more 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, directly attached to ring carbon atoms
  • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
  • C07D239/60—Three or more oxygen or sulfur atoms
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
  • Y02P20/00—Technologies relating to chemical industry
  • Y02P20/50—Improvements relating to the production of bulk chemicals
  • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

• the present invention relates to sphingolipids, and more particularly to a method for the production of sphingoid bases, especially sphingosine, and novel derivatives thereof.
• Sphingolipids are important structural components of plasma and organelle membranes of essentially all eukaryotic cells and play critical roles in many biological processes. They are distributed widely in plants, mammals and microbes. Sphingolipids constitute a diverse group of complex membrane lipids, all containing a long-chain aliphatic amino alcohol, usually having 18 carbons, connected by an amide linkage to a fatty acid to form ceramides. However, sphingoid bases having longer or shorter alkyl chains are also well- known constituents of sphingolipids of eukaryotic life.
• Sphingoid bases themselves, their O-glycosylated and O-phosphorylated forms, their N-acylated forms, furthermore their substituted/unsubstituted ceramide forms are the most abandoned sphingoid base molecules in nature.
• Phytoceramides, ceramides, dihydroceramides and 6-hydroxy- ceramides are the most significant ceramide structures in humans carrying phytosphingosine, sphingosine, dihydrosphingosine and 6-hydroxy-sphingosine, respectively. These different ceramide molecules are often referred to as ceramides.
• Ceramides are usually further modified in life chemistries by enzymatic cascades yielding phosphosphingolipids, glycosylinositol phosphoceramides, glycosphingolipids and their intermediates/metabolites.
• Sphingolipids - especially glycosphingolipids - have structural and biological functions in maintaining cell membrane integrity, controlling cell growth, apoptosis, signal transduction, etc. by acting as the main components of lipid rafts.
• a large number of sphingolipid molecules have been gaining interest for nutritional, cosmetic and therapeutic applications due to their high biological activities, excellent pharmacokinetics and their presence in human milk.
• access to human identical sphingolipids and their precursors/metabolites has been prevented by the lack of enabling synthetic production technologies.
• sphingolipid products commercialized mainly for pharmaceutical applications were obtained by extraction of animal brains, especially mammalian brains.
• the obtained sphingolipids were heterogenous mixtures of structurally different sphingolipids, which were potentially unsafe due to the possible presence of hazardous prion, viral and other microbiological entities.
• extraction and subsequent purification of sphingolipids from animal brain tissues is a laborious and expensive method, characterized by non-human chemical structures, low yields and undesired structural diversities ln order to meet the growing demand for sphingolipids, numerous attempts have been made to develop synthetic pathways for producing sphingolipids and their sphingoid base building blocks.
• One of the important factors for the successful synthesis of natural and non-natural sphingolipids is the synthetic access of the appropriate sphingoid bases, which are the principal backbones of sphingolipids.
• ⁇ -erythro-sphingosine (CAS: 123-78-4; C18H37NO2) (1), D-r/bo-phytosphingosine (CAS: 388566-94-7; C18H39NO3) (2), DL- erythro- Dihydrosphingosine (CAS: 3102-56-5; C18H39NO2) (3) and 6 -Hydroxy- D-ery thro - sphingosine (CAS: 566203-07-4; C18H37NO3) (4) are the most important and common species.
• D-r/bo-Phytosphingosine is readily obtainable on industrial scale from a yeast fermentation process.
• D-erythro-sphingosine is the naturally active configuration of sphingosine and is rather expensive and available only from laborious animal tissue extraction or chemical synthesis.
• a second popular approach involves the use of chiral starting materials, such as carbohydrates and amino acids.
• O-erythro-sphingosine has been synthesized starting from D-glucosamine in T. Murakami et. al. Tetrahedron Lett. 1994, 35(5), 745-748 and Y. Masaki et. al. Chem. Pharm. Bull. 1996, 44(5), 927-932, respectively.
• the relatively inexpensive D-r/ho-phytosphingosine 2 has also been employed as a starting material for the synthesis of D-e/ythro-sphingosine 1.
• a number of protecting group strategies have been described involving different steps such as azide formation, silylation, oxidation, sulfonylation, cyclic sulfate formation for the regio- and stereoselective transformation of the C-4 hydroxyl group of D-r/ho-phytosphingosine 2 into the characteristic 4,5 -trans double bond of D-erythro-sphingosine 1.
• EP 1767644 B1 describing microbial strains capable to convert D-r/ho-phytosphingosine 2 into D-erythro-sphingosine 1 with a poor conversion rate. This approach has never been industrialized and the related specific strains and enzymes have never been described in the scientific literature.
• Sphingoid bases especially D-erythro-sphingosine 1
• Novel large volume, low cost production methods are required for sphingoid base, especially D-erythro-sp ingosine 1, manufacturing to initiate commercial activities while also achieving high purities and other quality requirements of pharmaceutical, nutrition and cosmetic industries.
• the present invention provides an economically feasible process for the production Method for producing a sphingoid base of General Formula 1, or a salt thereof,
• R ' is H, aryl or an alkyl chain having 1-43 carbon atoms, preferably 5-25 carbon atoms, more preferably 10-20 carbon atoms, which may be a straight chain or branched, and/or which may be saturated or contain one or more double bonds, and/or which may contain one or more functional groups, the functional group being preferably selected from the group consisting of a hydroxyl group, an alkoxy group, a primary, secondary or tertiary amine, a thiol group, a thioether or a phosphorus containing functional group, starting from a compound of General Formula 11
• R’ is as defined in (1)
• Ri is selected from H, alkyl, substituted alkyl, aryl, substituted aryl,
• R2 and R3 are independently selected from alkyl, aryl, substituted alkyl, substituted aryl, O-alkyl, substituted O-alkyl, O-aryl, substituted O-aryl, NH2, NHR", NR"R”’, wherein R" and R’" are independently selected from the group consisting of alkyl and substituted alkyl, or might form a cyclic structure characterized by 5 - 8 ring sizes,
• the dashed line . represents a hydrogen bond.
• R’, Ri, R2 and R3 are as defined in (1) or (2),
• R 4 and R5 are independently selected from optionally substituted benzyl, optionally substituted alkoxymethyl, trialkylsilyl, trialylsilyl or wherein R4 and Rs form a cyclic structure,
• the dashed line . represents a hydrogen bond.
• R’, Ri, R2, R3, R4 and Rs are as defined in (1), (2) or (3),
• R6 is a halide or a sulfonic ester derivative
• the dashed line . represents a hydrogen bond.
• R’, Ri, R2, R3, R4 and Rs are as defined in (1), (2) or (3),
• the dashed line . represents a hydrogen bond.
• R’, Ri, R2 and R3 are as defined in (1) or (2), and
• the dashed line . represents a hydrogen bond.
• the vinylogous amide N-protection strategy according to the method of the present invention enables easy introduction and easy removal of the N-protecting group and prevention of deprotonation of N-H functionality in the basic conditions of the coming elimination step that enables the formation of the C4-C5 double bond.
• the intermediate compounds according to General Formulae Ill-Vll can be crystallized and have outstanding stabilities even in drastic conditions and have robust stabilities in the presence of secondary and tertiary organic bases.
• a major aim of the present invention is to identify the most efficient syntheses of sphingoid bases, especially D- erythro-s phingosine 1, and derivatives thereof, suitable for glycosylation reactions.
• substituted means that the group in question is substituted with a group which typically modifies the general chemical characteristics of the group in question.
• Preferred substituents include but are not limited to halogen, nitro, amino, azido, oxo, hydroxyl, thiol, carboxy, carboxy ester, carboxamide, alkylamino, alkyldithio, alkylthio, alkoxy, acylamido, acyloxy, or acylthio, each of 1 to 6 carbon atoms, preferably of 1 to 3 carbon atoms.
• the substituents can be used to modify characteristics of the molecule as a whole such as molecule stability, molecule solubility and an ability of the molecule to form crystals.
• Ci-e-alkyl-oxy C2-6-alkenyloxy, carboxy, oxo, Ci-e-alkoxycarbonyl, Ci-e-alkylcarbony!, formyl, aryl, aryloxycarbonyl, aryloxy, arylamino, arylcarbonyl, heteroaryl, heteroarylamino, heteroaryloxycarbonyl, heteroaryloxy, heteroarylcarbonyl, amino, mono- and di(Ci-6-alkyl)amino, carbamoyl, mono- and di(Ci-6-alkyl)aminocarbonyl, amino-Ci-6-alkyl-aminocarbonyl, mono- and di(Ci-6-alkyl)amino-Ci-6-alkyl- aminocarbonyl, Ci-6-alkylcarbonylamino, cyano, guanidino, carbamido, Ci-6-alkyl- sulphonyl-amino, aryl-
• leaving group means a group capable of being displaced by a nucleophile in a substitution chemical reaction or can promote elimination reaction.
• Common leaving groups include halides, triflates (OTf), diazonium salts (N2 + ), mesylates (OMs), tosylates (OTs), nosylates (ONs), imidazole-l-sulfonate (0S02lm) and other sulfonic esters.
• derivative refers to a modified form of a compound, having one or more substituents.
• sphingoid base derivative include, but not limited to forms of a sphingoid base, of sphingosine, and of phytosphingosine, respectively, that have been modified to contain an N- and/or an O-protecting group on an aminoalcohol molecular scaffold, wherein the N-protecting group is a vinylogous amide-type protecting group.
• General Formulae 1I1-V11, Illa-Vlla, lllb-Vllb and lllc-Vllc represent derivatives of a sphingoid base.
• General Formulae of llla-lVa and Illc-IVc, wherein R’ Ci3H27, represent derivatives of phytosphingosine;
• human sphingosine refers to the fact that D-erythro-sphingosine is a prevalent sphingoid base present in humans. ln the present invention, the terms “phytosphingosine” and “D-r/bo-phytosphingosine” are used interchangeably.
• cyclic structures characterized by 5-8 ring sizes refers to a 5-8- membered ring, which contain single bonds and/or double bonds, which may be aromatic or not aromatic, homoaromatic or heteroaromatic, which may contain carbon atoms in the ring structure or wherein one or more of the carbon atoms may optionally be substituted or replaced by an oxygen atom, a nitrogen atom or a sulfur atom.
• human identical sphingolipids refers to sphingolipids that are naturally present in humans. Sphingolipids that are naturally present in humans contain sphingoid bases selected from D-erythro-sphingosine, D-r/ho-phytosphingosine, DL- erythro- Dihydrosphingosine and e-Hydroxy-D-e/ythro-sphingosine. While the carbon chains of the fatty acyl moieties in human sphingolipids usually have an even number, the carbon chains of the fatty acyl moieties in other mammals may also have odd numbers. Accordingly, "human identical sphingolipids" especially refers to sphingolipids having sphingoid bases typically present in humans and having an even number of carbon chains of their fatty acyl moieties.
• the term "a” grammatically is a singular, but it may as well mean the plural of e.g. the intended compound.
• the production of a sphingoid base the production of not only one single sphingoid base, but of many sphingoid bases of the same type are meant.
• the present invention relates to a method for producing a sphingoid base according to General Formula 1, or a salt thereof,
• R ' is H, aryl or an alkyl chain having 1-43 carbon atoms, preferably 5-25 carbon atoms, more preferably 10-20 carbon atoms, which may be a straight chain or branched, and/or which may be saturated or contain one or more double bonds, and/or which may contain one or more functional groups, the functional group being preferably selected from the group consisting of a hydroxyl group, an alkoxy group, a primary, secondary or tertiary amine, a thiol group, a thioether or a phosphorus containing functional group, starting from a compound according to General Formula 11
• R’ is as defined above
• Ri is selected from H, alkyl, substituted alkyl, aryl, substituted aryl, R2 and R3 are independently selected from alkyl, aryl, substituted alkyl, substituted aryl, O-alkyl, substituted O-alkyl, O-aryl, substituted O-aryl, NH2, NHR", NR"R”’, wherein R" and R’" are independently selected from the group consisting of alkyl and substituted alkyl, or might form a cyclic structure characterized by 5 - 8 ring sizes,
• the dashed line . represents a hydrogen bond
• b protecting the hydroxyl (OH) groups at C-l and C-3 of a compound of General Formula 111 with an O-protecting group by addition of an O-protecting group reagent to form a compound according to General Formula IV:
• R’, Ri, R2 and R3 are as defined above,
• R 4 and R5 are independently selected from optionally substituted benzyl, optionally substituted alkoxymethyl, trialkylsilyl, trialylsilyl or wherein R4 and Rs form a cyclic structure,
• the dashed line . represents a hydrogen bond
• c introducing a leaving group R6 at C-4 position of a compound of General Formula IV, by substitution or replacement of the C-4 hydroxyl group to form a compound according to General Formula V:
• R’, Ri, R2, R3, R4 and Rs are as defined above,
• R6 is a halide, such as 1, Br, Cl or F, or a sulfonic ester derivative, such as mesylate (OMs), tosylate (OTs), triflate (OTf), nosylate (ONs), imidazole-l-sulfonate (OSCtelm) or chlorophosphite ester (OPCI2),
• OMs mesylate
• OTs tosylate
• OTf triflate
• nosylate ONs
• OSCtelm imidazole-l-sulfonate
• OPCI2 chlorophosphite ester
• the dashed line . represents a hydrogen bond
• R’, Ri, R2, R3, R4 and Rs are as defined above,
• the dashed line . represents a hydrogen bond, e. removing the O-protecting group of a compound of General Formula VI to form a compound according to General Formula Vll
• R’, Ri, R2 and R3 are as defined above,
• the dashed line . represents a hydrogen bond, f. removing the N-protecting group of a compound of General Formula Vll, to form a sphingoid base according to General Formula 1, or a salt thereof.
• the present invention provides a method for the production of a sphingoid base according to General Formula 1 or a salt thereof, starting from a compound according to General Formula 11 or a salt thereof, and having compounds according to General Formulae I1I-V11 as intermediate compounds.
• the method of the present invention is preferably a synthetic method.
• the salts of the compounds of General Formula 1 and General Formula 11 are preferably pharmaceutically acceptable salts or other generally acceptable salts, unless they would be excluded for chemical reasons, which the skilled person will readily understand.
• the stereochemistry of a compound according to General Formula 1 and 11, or of salts thereof, and accordingly of an intermediate compound according to General Formula 111-V11 corresponds to the stereochemistry of D -erythro- sphingosine and of D-r/ho-phytosphingosine, respectively
• the stereochemical configuration of the compounds of General Formulae 1 to VII preferably equals the stereochemical configuration of ⁇ -erythro-sphingosine (2S,3R,4E) and of D- r/ho-phytosphingosine (2S,3S,4R), respectively.
• stereochemical configuration of the compounds of General Formulae 11 to V is (2S,3S,4R) and the stereochemical configuration of the compounds of General Formulae 1, VI and VII is (2S,3R,4E).
• Those preferred compounds are represented by General Formulae llla to Vlla.
• R’, Ri, R2, R3, R4, R5 and R6 are as defined above,
• R’ of General Formulae 1 to Vll or of General Formulae la to Vlla is an alkyl chain having 13 carbon atoms. Even more preferably, R’ of General Formulae I to VII is C13H27 or CH(OH)Ci2H25, especially -C13H27 or -CH(OH)Ci2H25.
• the starting compound of General Formula 11 defines the possible General Formulae of the following steps of the method of the present invention lf e.g. a compound of General Formula la is used as a starting compound for the method, a compound of General Formula llla (and not a compound of General Formula Illb) may be formed in step (a) of the method of the present invention.
• step (a) of the method of the present invention the NH2 (amino) group of a compound represented by General Formula 11, preferably of phytosphingosine 2, is protected with an N-protecting group by addition of an N-protecting group reagent being a vinylogous reagent to form a compound according to General Formula 111, preferably General Formula llla.
• the vinylogous reagent may be a vinylogous acid, a vinylogous ester, a vinylogous amide or a vinylogous acid halide.
• the vinylogous reagent is an N,N-disubstituted vinylogous amide reagent, more preferably an N,N-dialkyl-barbituric acid-derived reagent.
• the vinylogous reagent is l,3-dimethyl-5- [(dimethylamino)methylene]-2,4,6(l//,3//,5//)-trioxopyrimidine (DTPM-reagent) (CAS: 35824-98-7; C9H13N3O3).
• R’ and Ri are as defined above,
• the dashed line . represents a hydrogen bond.
• the vinylogous reagent preferably has a cyclic structure providing a robust stability and crystalline properties.
• the preparation of vinylogous reagents are described in publications Tetrahedron Letters, 2001, 42, 3129-3132; WO 98/38197.
• the compound according to General Formula 11 and the vinylogous reagent may be mixed in water, organic solvents or in their aqueous mixtures.
• the reactions may optionally be catalyzed with organic or inorganic bases at temperatures ranges from 0 - 150 °C, preferably at temperatures ranging from 20 - 120 °C. More preferably, the reaction goes to completion at ambient temperature.
• the reactions are preferably carried out in organic solvents at room temperature (r.t.) or between 40 - 100 °C.
• a person skilled in art has the knowledge to conduct, isolate and purify the novel compounds by using standard methods of synthetic organic chemistry.
• the introduction of DTPM protecting group may be performed using the DTPM-reagent dissolved in H2O or in organic solvents, such as methanol and CH2CI2.
• the reaction does not require extreme conditions and affords high conversion.
• the DTPM- protected compounds of General Formula lllb and lllc may be precipitated directly from the reaction mixtures.
• Example 1 provides a representative experimental example of the described DTPM-protection.
• step (b) of the method of the present invention the hydroxyl groups at C-l and C-3 of a compound of General Formula 111, preferably of General Formula llla or General Formula lllb, more preferably of General Formula lllc, are protected with an 0- protecting group by addition of an O-protecting group reagent.
• the O-protecting groups at C-l and C-3 may be the same or different and are selected from optionally substituted benzyl, optionally substituted alkoxymethyl, trialkylsilyl, trialylsilyl or preferably one O-protecting group may form one cyclic structure with the hydroxyl groups of both C-l and C-3. More preferably, the O-protecting group forming the cyclic structure with the hydroxyl groups of both C-l and C-3 is an optionally substituted cyclic carbonate, an optionally substituted cyclic acetal or an optionally substituted cyclic ketal, even more preferably an optionally substituted cyclic acetal or an optionally substituted cyclic ketal. Especially, the optionally substituted cyclic acetal is an optionally substituted benzylidene and the optionally substituted cyclic ketal is an optionally substituted isopropylidene.
• O-protecting group used is a well-known process for a person skilled in art by reacting 1,3-diols of compounds characterized by General Formula 111, llla, lllb or lllc with aldehyde, ketone, acyclic dialkylacetals or acyclic dialkylketal, preferably dimethyl acetal and dimethyl ketal, reagents in the presence of protic acid or Lewis acid catalysts in organic solvents.
• the acetal or ketal formation is catalyzed by acid catalysts such as strong organic or inorganic acids, including protic acids such as tosic acid, champhorsulfonic acid, acidic ion-exchange resin, HC1, H2SO4, etc.
• acid catalysts such as strong organic or inorganic acids, including protic acids such as tosic acid, champhorsulfonic acid, acidic ion-exchange resin, HC1, H2SO4, etc.
• Example 2 provides one representative experimental example for step (b) of the method of the present invention.
• Ri, R2, Ri, R4 and Rs are as defined above,
• the dashed line . represents a hydrogen bond.
• R’, Ri, R2 and R3 are as defined above,
• R 4 and R5 form an optionally substituted cyclic acetal or an optionally substituted cyclic ketal
• a leaving group R6 is introduced at C-4 position of General Formula IV, preferably of General Formula lVa or of General Formula lVb, more preferably of General Formula lVc, by substitution or replacement of the C-4 hydroxyl group, to form a compound according to General Formula V, preferably General Formula Va or General Formula Vb, more preferably General Formula Vc, respectively.
• R6 may be a halide, such as 1, Br, Cl or F, or a sulfonic ester derivative such as mesylate (OMs), tosylate (OTs), triflate (OTf), nosylate (ONs), imidazole- 1-sulfonate (0S02lm) and chlorophosphite ester (OPCI2).
• OMs mesylate
• OTs tosylate
• OTf triflate
• nosylate ONs
• imidazole- 1-sulfonate (0S02lm) and chlorophosphite ester
• R6 When R6 is a halide, it may preferably be introduced by using triphenylphosphine (PPI13), imidazole and iodine or bromine, preferably iodine.
• PPI13 triphenylphosphine
• imidazole imidazole
• iodine or bromine preferably iodine.
• R6 is a sulfonic ester
• the respective sulfonic acid chlorides or anhydrides may be employed in the presence of a base, preferably in the presence of an organic base, more preferably in the presence of pyridine, triethylamine (TEA) or diisopropylethylamine (D1PEA) for the introduction of the sulfonic ester moiety.
• a base preferably in the presence of an organic base, more preferably in the presence of pyridine, triethylamine (TEA) or diisopropylethylamine (D1PEA) for the introduction of the sulfonic ester moiety.
• a base preferably in the presence of an organic base, more preferably in the presence of pyridine, triethylamine (TEA) or diisopropylethylamine (D1PEA) for the introduction of the sulfonic ester moiety.
• TAA triethylamine
• D1PEA
• Step (c) requires neutral or basic conditions to keep the acetal or ketal protection group at C-l and C-3 position of the compound stable.
• mesylation, tosylation, imidazoylsulfonylation can use simple organic bases like pyridine, triethylamine along with the required reagents of mesylchloride, tosylchloride, etc. on organic solvents like dichloromethane, toluene, etc.
• Deoxy iodination usually takes place in dichloromethane in the presence of triphenylphosphine/ /imidazole reagent combinations known by the person skilled in Art.
• Chlorophosphate esterification preferably requires pyridine/POCh reagent pair acting in dichloromethane, toluene or pyridine.
• Example 3 (see below under “Examples”) provides one representative representative example for step (c) of the method of the present invention.
• R’, Ri, R2, RI, R 4 , R5 and R6 are as defined above,
• step (d) of the method of the present invention an elimination reaction is induced to form a double bond between the C-4 and C-5 carbon atoms of a compound according to General Formula V, preferably General Formula Va or General Formula Vb, more preferably General Formula Vc, resulting in a compound according to General Formula VI, preferably General Formula Vla or General Formula Vlb, more preferably General Formula Vlc, respectively.
• Elimination reactions are often accompanied by nucleophilic substitution reactions as competing reactions. However, conditions could be found in the present invention to make the elimination reaction the main, dominant reaction; for example, strong bases and weak nucleophiles favor the elimination products and are therefore preferred. ln a preferred embodiment of the present invention, bulky, sterically hindered non- nucleophilic bases such as KOtBu, DBU or DBN are used to promote the elimination reaction. The elimination reaction appears to take place with virtually complete selectivity towards the desired trans-s phingoid base, which appeared to be virtually free of unwanted side products.
• KOtBu sterically hindered non- nucleophilic bases
• organic solvents such as acetonitrile, 2 -methyltetrahydrofuran, propionitrile, 1,4-dioxane, toluene, xylene or N,N-dimethylformamide may be used to perform the elimination reaction.
• the elimination reaction is preferably performed at a temperature range between 60 to 150 °C.
• Example 4 provides one representative experimental example for step (d) of the method of the present invention.
• R’, Ri, R2, Ri, R 4 , and Rs are as defined above,
• the dashed line . represents a hydrogen bond.
• step (e) of the method of the present invention the O-protecting group(s) is/are removed from a compound of General Formula VI, preferably from a compound according to General Formula Vla or General Formula Vlb, more preferably from a compound according to General Formula Vlc, to form a compound according to General Formula Vila or General Formula Vllb, more preferably from a compound according to General Formula Vllc, respectively.
• the O-protecting group (s) may be removed via deprotecting procedures carried out in acidic conditions.
• a typical example demonstrates the removal of 1,3-acetal or ketal groups in acidic conditions in the presence of water or alcohols catalyzed by inorganic or organic acid catalysts such as HC1, TsOH, champhorsulfonic acid, acidic ion exchange resin, etc. at temperatures ranging from 0 - 130 °C. Vinylogous amides are stable in such conditions.
• Example 5 provides one representative experimental example for step (e) of the method of the present invention.
• General Formula Vllb General Formula Vllc ln step (f) of the method of the present invention, the N-protecting group is removed from a compound of General Formula Vll, preferably from a compound according to General Formula Vlla or from a compound according to General Formula Vllb, more preferably from a compound according to General Formula Vile, to form a compound according to General Formula 1, preferably ⁇ -erythro-sphingosine 1, or a salt thereof.
• the reagent used for this step may be selected from an aqueous inorganic base, NH3, primary amines, hydrazine, hydrazine derivatives, hydroxylamine and hydroxylamine derivatives.
• the reaction may be performed in an organic solvent such as dichlormethane, acetonitrile, methanol, tetrahydrofuran or toluene.
• the preferred temperature range for the reaction is from 20 to 120 °C.
• Example 7 (see below under “Examples") provides a representative experimental example for step (f) of the method of the present invention.
• Steps (a) to (f) of the method of the present invention are performed in the order mentioned, i.e. beginning with step (a), continuing with step (b), (c), (d), (e) and ending with step (f).
• steps (e) and (f) of the method of the present invention may be swapped. That means that the removal of the N-protecting group may be performed before the removal of the O-protecting group.
• Example 7 provides a representative experimental example for this alternative embodiment, wherein step (f) is performed on a compound still carrying the O-protecting group, i.e. wherein step (e) has not been performed yet.
• the method of the present invention may as well comprise further steps in addition to steps (a) to (f), provided they do not negatively affect the reactions of steps (a) to (f), which the skilled person will easily determine.
• the present invention provides novel intermediate compounds of the method according to the present invention, represented by General Formulae 111 to Vll.
• the General Formulae of the (preferred) intermediate compounds are not illustrated again. A skilled person will be aware, however, that the General Formulae and the defined rests (R) as defined above in detail will also apply to the intermediate compounds.
• the intermediate compounds are represented by General Formulae llla to Vlla or by General Formulae lllb to Vllb. More preferably, the intermediate compounds are represented by General Formulae lllc to Vllc.
• R’ of General Formulae Ill to Vll (or the preferred versions of General Formulae llla to Vlla, lllb to Vllb or lllc to Vllc) is an alkyl chain having 13 carbon atoms. Even more preferably, R’ of General Formulae 111 to Vll (or the preferred versions of General Formulae llla to Vlla, lllb to Vllb or lllc to Vllc) is C13H27 or CH (OH)CI 2 H25, especially -C13H27 or -CH (OH)CI 2 H25.
• a compound represented by General Formula III, llla, lllb or lllc is especially obtained by step (a) of the method of the present invention.
• a compound represented by General Formula IV, lVa, lVb or lVc is especially obtained by steps (a) to (b) of the method of the present invention.
• a compound represented by General Formula V, Va, Vb or Vc is especially obtained by steps (a) to (c) of the method of the present invention.
• a compound represented by General Formula VI, Vla, Vlb or Vlc is especially obtained by steps (a) to (d) of the method of the present invention.
• a compound represented by General Formula Vll, Vlla, Vllb or Vllc is especially obtained by steps (a) to (e) of the method of the present invention.
• a compound of General Formulae 111 to Vll preferably of General Formulae llla to Vlla or of General Formulae lllb to Vllb, more preferably of General Formulae lllc to Vllc Vll, Vlla, Vllb or Vllc is suitable for glycosylation, phosphorylation and other nucleophilic 0- substitution reactions - due to its enhanced O-nucleophilicity.
• a compound of General Formulae III to VII preferably of General Formulae llla to Vlla or of General Formulae lllb to Vllb, more preferably of General Formulae lllc to Vllc Vll, may be used for the preparation of sphingolipids such as glycosphingolipids, glycosylsphingosines, sphingosine-l-O-phosphates, sphingomyelins, phosphosphingolipids, or glycosylinositol phosphoceramides; especially, a compound of General Formulae III to Vll, preferably of General Formulae llla to Vlla or of General Formulae lllb to Vllb, more preferably of General Formulae lllc to Vllc, may be used for the preparation of glycosphingolipids.
• sphingolipids such as glycosphingolipids, glycosylsphingosines, sphingosine-l-O-phosphates, sphingomy
• sphingolipids are optionally substituted compounds containing a sphingoid base characterized by the presence of a molecular motif of General Formula 1 or II.
• Preferred substituents are carbohydrate- and/or phosphate-containing moieties of naturally occurring sphingolipids.
• a compound of General Formula 1 obtainable by the method according to the present invention may be used for cosmetic, nutritional and/or pharmaceutical applications.
• a compound of General Formula 1 obtainable by the method according to the present invention may be used as a pharmaceutical agent and/or for the preparation of a pharmaceutical composition.
• a compound of General Formula 1 obtainable by the method according to the present invention may be used for the preparation of a nutritional formulation, e.g. a food supplement.
• a compound of General Formula 1 obtainable by the method according to the present invention may be used for the preparation of a cosmetic product.
• a compound of General Formula I, preferably of General Formula la, obtainable by the method according to the present invention, may also be used for the production of ceramides, phosphosphingolipids or glycosphingolipids.
• the production of ceramides, phosphosphingolipids or glycosphingolipids may be performed by N-acylation.
• N-acylation may be performed using an acyl moiety of a C12- C30 acyl group which can be saturated, unsaturated or optionally substituted.
• the acylation may be performed by both lipase-assisted biocatalysis or chemistry via the use of the corresponding carboxylic acid, acid chloride, ester or anhydride in the presence of a base, preferably in the presence of an organic base, more preferably in the presence of pyridine, triethylamine (TEA) or diisopropylethylamine (DIPEA).
• TAA triethylamine
• DIPEA diisopropylethylamine
• D-ribo-Phytosphingosine (4.8 g, 15.1 mmol) is added to methanol (150 mL) at room temperature (r.t). and heated to approx. 30 °C until complete dissolution of the solid.
• the solution is cooled to r.t, then DTPM-reagent (3.5 g, 16.6 mmol) is added in one portion, and the stirring is continued at r.t. for lh. (After approx. 5 min. crystallization of the product starts.)
• the slurry is cooled to approx. 5 °C, then kept at 5 °C for 2 h.
• lodine (1.64 g, 6.5 mmol) is added, and the reaction mixture is heated to 80-82 °C, and stirred for 12 h.
• the mixture is cooled to r.t, washed with aqueous solution of sodium thiosulfate (1 x 40 mL, 10 %), hydrochloric acid (2 x 40 mL, 0.1M), then sodium chloride (2 x 40 mL, 10 %, pH 1 st wash ⁇ 3, pH 2 nd wash ⁇ 6).
• the organic phase is then dried (MgS0 4 ), the solid is filtered off, and the filtrate is concentrated in vacuo to a syrup (approx. 4.3 g).
• (2S,3R,4E)-2-NHDTPM-l,3-(benzylidene)-octadec-4-ene (800 mg, 1.4 mmol) is dissolved in THF (10 mL), then methanol (20 mL) and CSA (200 mg) are added. The mixture is stirred at 45 °C for 2.5 h. The mixture is cooled to r.t. On cooling, crystallization started. The slurry is stirred at r.t. for 3 h. The solid is filtered off, and washed with methanol (5 mL, r.t), and dried in vacuum drying oven at 50°C to yield 250 mg white solid.

Landscapes

• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
• Cosmetics (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Priority Applications (6)

Applications Claiming Priority (2)

Publications (1)

Family

ID=67145768

Family Applications (1)

Country Status (7)

Cited By (6)

Families Citing this family (1)

Citations (4)

Family Cites Families (1)

• 2019
  • 2019-06-14 JP JP2020567066A patent/JP2021526528A/ja not_active Withdrawn
  • 2019-06-14 CA CA3101413A patent/CA3101413A1/en not_active Abandoned
  • 2019-06-14 EP EP19735495.4A patent/EP3807244B1/en active Active
  • 2019-06-14 CN CN201980040317.9A patent/CN112368261B/zh active Active
  • 2019-06-14 WO PCT/EP2019/065785 patent/WO2019238970A1/en not_active Ceased
  • 2019-06-14 US US17/251,978 patent/US12054462B2/en active Active
  • 2019-06-14 KR KR1020217001110A patent/KR20210021371A/ko not_active Ceased

Patent Citations (4)

Non-Patent Citations (19)

Also Published As

Similar Documents

Legal Events