MXPA97002124A - Novedous sphingolipids and its procedimie - Google Patents
Novedous sphingolipids and its procedimieInfo
- Publication number
- MXPA97002124A MXPA97002124A MXPA/A/1997/002124A MX9702124A MXPA97002124A MX PA97002124 A MXPA97002124 A MX PA97002124A MX 9702124 A MX9702124 A MX 9702124A MX PA97002124 A MXPA97002124 A MX PA97002124A
- Authority
- MX
- Mexico
- Prior art keywords
- alkenyl
- straight
- alkyl
- formula
- branched chain
- Prior art date
Links
- 150000003408 sphingolipids Chemical class 0.000 title claims abstract description 70
- WWUZIQQURGPMPG-KRWOKUGFSA-N Sphingosine Chemical compound CCCCCCCCCCCCC\C=C\[C@@H](O)[C@@H](N)CO WWUZIQQURGPMPG-KRWOKUGFSA-N 0.000 claims abstract description 36
- 238000000034 method Methods 0.000 claims abstract description 26
- 241000233866 Fungi Species 0.000 claims abstract description 22
- 244000052616 bacterial pathogens Species 0.000 claims abstract description 3
- 125000003342 alkenyl group Chemical group 0.000 claims description 98
- 125000000217 alkyl group Chemical group 0.000 claims description 68
- 229910052757 nitrogen Inorganic materials 0.000 claims description 27
- -1 mercapto, amino, monosubstituted amino Chemical group 0.000 claims description 26
- VODZWWMEJITOND-OWWNRXNESA-N N-Stearoylsphingosine Chemical compound CCCCCCCCCCCCCCCCCC(=O)NC(CO)C(O)\C=C\CCCCCCCCCCCCC VODZWWMEJITOND-OWWNRXNESA-N 0.000 claims description 24
- 150000001875 compounds Chemical class 0.000 claims description 19
- 125000004435 hydrogen atoms Chemical group [H]* 0.000 claims description 19
- 125000003710 aryl alkyl group Chemical group 0.000 claims description 14
- 125000004415 heterocyclylalkyl group Chemical group 0.000 claims description 14
- 125000001424 substituent group Chemical group 0.000 claims description 14
- KUQZVISZELWDNZ-UHFFFAOYSA-N 3-aminopropyl dihydrogen phosphate Chemical compound NCCCOP(O)(O)=O KUQZVISZELWDNZ-UHFFFAOYSA-N 0.000 claims description 12
- 125000000623 heterocyclic group Chemical group 0.000 claims description 12
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 12
- 125000004122 cyclic group Chemical group 0.000 claims description 10
- 239000010452 phosphate Substances 0.000 claims description 10
- NJRWNWYFPOFDFN-UHFFFAOYSA-L phosphonate(2-) Chemical class [O-][P]([O-])=O NJRWNWYFPOFDFN-UHFFFAOYSA-L 0.000 claims description 10
- 150000003410 sphingosines Chemical class 0.000 claims description 10
- 125000005843 halogen group Chemical group 0.000 claims description 9
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 9
- 125000000304 alkynyl group Chemical group 0.000 claims description 8
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 8
- 229910052799 carbon Inorganic materials 0.000 claims description 7
- 125000003601 C2-C6 alkynyl group Chemical group 0.000 claims description 6
- 241000233654 Oomycetes Species 0.000 claims description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 6
- 125000000468 ketone group Chemical group 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 229940106189 Ceramides Drugs 0.000 claims description 5
- OVRNDRQMDRJTHS-DJLPIQJZSA-N N-[(2S,3S,4S,5R)-2,4,5-trihydroxy-6-(hydroxymethyl)oxan-3-yl]acetamide Chemical compound CC(=O)N[C@@H]1[C@@H](O)OC(CO)[C@H](O)[C@H]1O OVRNDRQMDRJTHS-DJLPIQJZSA-N 0.000 claims description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-K [O-]P([O-])([O-])=O Chemical class [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 5
- 150000001783 ceramides Chemical class 0.000 claims description 5
- 238000003306 harvesting Methods 0.000 claims description 5
- 150000003013 phosphoric acid derivatives Chemical class 0.000 claims description 5
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 claims description 4
- 238000006460 hydrolysis reaction Methods 0.000 claims description 4
- 150000002632 lipids Chemical class 0.000 claims description 4
- 125000000882 C2-C6 alkenyl group Chemical group 0.000 claims description 2
- CDAISMWEOUEBRE-GPIVLXJGSA-N Inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 claims description 2
- 229960000367 Inositol Drugs 0.000 claims description 2
- QCQYVCMYGCHVMR-AAZUGDAUSA-N N-[(2R,3R,4S,5R)-4,5,6-trihydroxy-1-oxo-3-[(2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyhexan-2-yl]acetamide Chemical compound CC(=O)N[C@@H](C=O)[C@H]([C@@H](O)[C@H](O)CO)O[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O QCQYVCMYGCHVMR-AAZUGDAUSA-N 0.000 claims description 2
- YHHSONZFOIEMCP-UHFFFAOYSA-O Phosphocholine Chemical compound C[N+](C)(C)CCOP(O)(O)=O YHHSONZFOIEMCP-UHFFFAOYSA-O 0.000 claims description 2
- 150000001413 amino acids Chemical class 0.000 claims description 2
- 125000000637 arginyl group Chemical group 0.000 claims description 2
- 125000001589 carboacyl group Chemical group 0.000 claims description 2
- 239000000969 carrier Substances 0.000 claims description 2
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 claims description 2
- 125000002519 galactosyl group Chemical group C1([C@H](O)[C@@H](O)[C@@H](O)[C@H](O1)CO)* 0.000 claims description 2
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 claims description 2
- 125000001288 lysyl group Chemical group 0.000 claims description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 2
- 239000002777 nucleoside Substances 0.000 claims description 2
- 150000003833 nucleoside derivatives Chemical class 0.000 claims description 2
- 229950004354 phosphorylcholine Drugs 0.000 claims description 2
- 229920000023 polynucleotide Polymers 0.000 claims description 2
- 239000002157 polynucleotide Substances 0.000 claims description 2
- 230000001681 protective Effects 0.000 claims description 2
- 239000000376 reactant Substances 0.000 claims description 2
- 125000004433 nitrogen atoms Chemical group N* 0.000 claims 17
- 125000000962 organic group Chemical group 0.000 claims 3
- 238000002955 isolation Methods 0.000 abstract description 7
- 239000000463 material Substances 0.000 abstract description 7
- 102000003923 Protein Kinase C Human genes 0.000 abstract description 3
- 108090000315 Protein Kinase C Proteins 0.000 abstract description 3
- 230000003110 anti-inflammatory Effects 0.000 abstract description 2
- 230000000259 anti-tumor Effects 0.000 abstract description 2
- 229930000044 secondary metabolites Natural products 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 24
- HEDRZPFGACZZDS-UHFFFAOYSA-N chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 22
- 238000004128 high performance liquid chromatography Methods 0.000 description 13
- 239000002609 media Substances 0.000 description 13
- 230000014759 maintenance of location Effects 0.000 description 7
- 150000002829 nitrogen Chemical group 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 7
- 241000196324 Embryophyta Species 0.000 description 6
- 240000004713 Pisum sativum Species 0.000 description 6
- 235000010582 Pisum sativum Nutrition 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 241000233622 Phytophthora infestans Species 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 239000007858 starting material Substances 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N HCl Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 241000233614 Phytophthora Species 0.000 description 4
- 241000233616 Phytophthora capsici Species 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 235000014113 dietary fatty acids Nutrition 0.000 description 4
- 239000000194 fatty acid Substances 0.000 description 4
- 150000004665 fatty acids Chemical class 0.000 description 4
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- WQZGKKKJIJFFOK-VFUOTHLCSA-N β-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 4
- 241000258957 Asteroidea Species 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N D-Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 240000002057 Secale cereale Species 0.000 description 3
- 235000007238 Secale cereale Nutrition 0.000 description 3
- 238000010192 crystallographic characterization Methods 0.000 description 3
- 230000001472 cytotoxic Effects 0.000 description 3
- 231100000433 cytotoxic Toxicity 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- GZCGUPFRVQAUEE-KCDKBNATSA-N D-(+)-Galactose Natural products OC[C@@H](O)[C@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-KCDKBNATSA-N 0.000 description 2
- 108060003339 GPLD1 Proteins 0.000 description 2
- 125000003047 N-acetyl group Chemical group 0.000 description 2
- 241001243925 Sia Species 0.000 description 2
- 238000005904 alkaline hydrolysis reaction Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 239000011152 fibreglass Substances 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 150000002339 glycosphingolipids Chemical class 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- 150000002402 hexoses Chemical class 0.000 description 2
- 238000000265 homogenisation Methods 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 238000006140 methanolysis reaction Methods 0.000 description 2
- 239000001965 potato dextrose agar Substances 0.000 description 2
- SQVRNKJHWKZAKO-OQPLDHBCSA-N sialic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)OC1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-OQPLDHBCSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 230000002194 synthesizing Effects 0.000 description 2
- 230000001225 therapeutic Effects 0.000 description 2
- 210000001519 tissues Anatomy 0.000 description 2
- ZMANZCXQSJIPKH-UHFFFAOYSA-N triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 2
- 239000007164 v-8 juice agar Substances 0.000 description 2
- QQVDJLLNRSOCEL-UHFFFAOYSA-L 2-phosphonatoethanamine Chemical compound NCCP([O-])([O-])=O QQVDJLLNRSOCEL-UHFFFAOYSA-L 0.000 description 1
- 244000075850 Avena orientalis Species 0.000 description 1
- 235000007319 Avena orientalis Nutrition 0.000 description 1
- 235000007558 Avena sp Nutrition 0.000 description 1
- 240000002791 Brassica napus Species 0.000 description 1
- 235000004977 Brassica sinapistrum Nutrition 0.000 description 1
- 210000003169 Central Nervous System Anatomy 0.000 description 1
- 235000008733 Citrus aurantifolia Nutrition 0.000 description 1
- 238000005952 Cope rearrangement reaction Methods 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 240000002860 Daucus carota Species 0.000 description 1
- 235000002243 Daucus carota subsp sativus Nutrition 0.000 description 1
- 206010012601 Diabetes mellitus Diseases 0.000 description 1
- 241000235426 Lagenidiales Species 0.000 description 1
- QQHVEZXEGWKJEP-UHFFFAOYSA-N OCC#P=O Chemical compound OCC#P=O QQHVEZXEGWKJEP-UHFFFAOYSA-N 0.000 description 1
- 240000000129 Piper nigrum Species 0.000 description 1
- 241000233656 Saprolegniales Species 0.000 description 1
- 240000001016 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 239000004809 Teflon Substances 0.000 description 1
- 235000015450 Tilia cordata Nutrition 0.000 description 1
- 235000011941 Tilia x europaea Nutrition 0.000 description 1
- 240000001417 Vigna umbellata Species 0.000 description 1
- 235000011453 Vigna umbellata Nutrition 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 125000004946 alkenylalkyl group Chemical group 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 150000001784 cerebrosides Chemical class 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 235000005824 corn Nutrition 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 229910001873 dinitrogen Inorganic materials 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- HZAXFHJVJLSVMW-UHFFFAOYSA-N ethanolamine Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 230000002538 fungal Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000003365 glass fiber Substances 0.000 description 1
- 239000011491 glass wool Substances 0.000 description 1
- 239000001963 growth media Substances 0.000 description 1
- 239000008079 hexane Substances 0.000 description 1
- 230000003301 hydrolyzing Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- KFZMGEQAYNKOFK-UHFFFAOYSA-N iso-propanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 1
- 230000004301 light adaptation Effects 0.000 description 1
- 239000004571 lime Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 1
- 229930014565 phytoalexin Natural products 0.000 description 1
- 239000000280 phytoalexin Substances 0.000 description 1
- 150000001857 phytoalexin derivatives Chemical class 0.000 description 1
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- 201000003068 rheumatic fever Diseases 0.000 description 1
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Abstract
The present invention relates to a process for novel sphingolipids and other secondary compounds, based on sphingosine, which comprises the cultivation of an appropriate fungus, the removal of mycelium from the fungus and the isolation of the sphingolipid from the mycelium. Sphingosine-based materials inhibit protein kinase C, provide an anti-inflammatory effect and anti-tumor effect, and these sphingosine-based materials provide protection to plants against pathogenic microorganisms
Description
NOVEDOUS SPHINGOLIPIDS AND THEIR PROCEDURE
The present invention relates to a process for the preparation of sphingolipids. In addition, some of the sphingolipids of this invention possess a polyunsaturated, branched, very unusual sphingosine half that has been previously reported (in the Kenneth L. Rinehart et al., "Ophidiacerebrosides: Cytotoxic Glycosphingolipids Containing a Novel Sphingosine from a Sea Star" (Ofidiacerebrosides: Cytotoxic glycosphingolipids containing a novel Sphingosine from a Starfish), J. Org Chem .1994 (59), pp. 144-147) which is present in the cytotoxic cerebrosides of starfish. Sphingosines and sphingolipids exert many effects on mammary cells [reviewed by S. Spiegel and S. Milstein, J. Membrane Biol. 146, 225 (1995)] and are useful as inhibitors of protein kinase C (US Patent 4,937,232) where said sphingosines and sphingolipids can be of therapeutic value in a number of disease states such as cancer, rheumatic arthritis, complication diabetic, diseases of the central nervous system, etc. In addition, inesitol sphingophospholipids and ceramide inoethylphosphonate represent two types of sphingolipids, which have been described in fungi of the Oomycetos class and which demonstrate biological activity. The sphingophospholipids of Phytophthora capsici have a protective effect on pepper plants against pathogenesis by P. Capsici (European Journal Qf Bíochsmístry) (European Journal of Biochemistry, 1990 (191), pp. 203-209). The ceramide aminoethyl phosphonate of Phytophthora infestans is reported to cause the accumulation of phytoalexins (Physiological and Molecular Plant Pathology) in the plant tissue, an abundance of the plant's tissue and plant pathology, 1986 (28), pp. 215-225). response frequently associated with the activation of disease resistance mechanisms. The present invention relates to a novel sphingolipid that has not been previously described. The process of this invention for the preparation of sphingolipids is novel and considerably more convenient than the isolation of said sphingolipids from starfish, or alternatively, by means of a laborious total synthetic procedure such as that described in the Richard. R. Schmidt et al., "Sphingosines - an Oxa-Cope Rearrangement Route for Their Synthesis", (Sphingosines - an Oxa-Layer Adaptation Route ", Synthesis, pp. 868-876 (July, 1995) This invention employs fungi , preferably Oomycete fungi, as a convenient source of desired sphingolipids After the isolation of the desired sphingolipid, the sphingolipid can be conveniently modified, if desired, in a sphingolipid child, from which the sphingolipid stem is subjected to hydrolytic conditions to remove the original head group and / or the fatty acid half, to produce a ceramide or a sphingosine phosphorylethanolamine. Amide or sphingosine phosphorylethanolamine can then be modified using techniques that are well known to a person skilled in the art, to produce a wide variety of compounds based on sphingosine daughters. The fungi Oo icetos can be subdivided into six orders according to "The Present Status of Classification of Biflagellate Fungi" (The Present State of Classification of the Biflagellate Fungi), p. 213-222 (1976) by F.K. Sparrow in Recent Advances in Aquatic Mycology (Recent Advances in Aquatic Mycology), E.B. Gareth Jones, editor, published by John Wiley (New York): Euricasmales, Saprolegniales, Lagenidiales, Peronsoporales, Traustoquitriales and Labirintulales. Said fungi Oomycetes can be cultured using various liquid or solid growth media, defined naturally or chemically, such as those described in "A Source Book of the Genus Phytophthora" (A Source Book of the Phytophthora Genus), by O.K. Ribeiro, published by A.R. Gantner Verlag, Vaduz, Germany. Examples of such natural media are the medium of the V-8 juice, the medium of the lime seed, the dextrose medium of the potato, the medium of the oat grain, the medium of the pea juice, the modified medium of the carrot, means of flour of a red bean, means of flour of corn, means of extract of a seed of rapeseed and means of extract of a seed of rye. Examples of chemically defined media include those described by Ribeiro in the same reference (pp. 79-99). One embodiment of this invention is a process for preparing a novel sphingolipid of the formula:
OH N s. H R4a (I)
wherein R4a is C (0) R8a; R8a is (Cn-C23) alkyl, (C13-C23) alkenyl or poly (Ci3-C23) alkenyl, all of which may be straight or branched chain; X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; and the enantiomorphs and structural isomers thereof; comprising the following steps: (i) cultivating an appropriate fungus (ii) collecting the mycelial harvest in said fungus, and In a preferred embodiment, the process for preparing a sphingolipid of the formula (I) wherein X is alkyl (C? 0-C20), alkenyl (C? 0-C20) or poly (C? 0-C2o) alkenyl, all these may be straight or branched chain; comprises cultivating an appropriate Oomycete fungus in step (i). In a more preferred embodiment, the process produces a sphingolipid of the formula (I), wherein X is alkenyl (C? 0-C20) or poly (C? 0-C2o) alkenyl, all these can be straight or branched chain . In an even more preferred embodiment, the process produces a sphingolipid of the formula (I), wherein X is a branched-chain alkanaryl (Ci4-C? 8) or a straight-chain (C11-C15) alkenyl and R8a is a (C13-C23) alkyl, (C? 3-C23) alkenyl or straight chain poly (C? 3-C23) alkenyl all. The following examples are intended to illustrate, but not limit, the scope of this embodiment of the invention.
Example 1: Isolation and Characterization of Sphingolipids from Pynthium ulumum Pynthiumumumumum (ATCC 26083) was obtained from the "American Type Culture Collection" (Collection of Cultures of Pynthiumumumum (ATCC 2 083) obtained from the "American Type Culture Collection" and maintained on a potato dextrose agar (Difco Laboratories) Petri dishes of 9 cm in diameter, containing 20 ml of a medium Sucrose-asparagine liquid [Erwin, DC and Katznelsoi, K., (1971), "Canadian Journal of My Crobiol ogy 7" (Canadian Journal of Microbiology 7), page 15] were inoculated with a 7 mm mycelium plug. in diameter, taken from the growing edge of a culture grown on a potato dextrose agar The discs were covered for 48 hours at 25 ° C, by shaking on a rotary shaker at 60 rpm The mycelium was then lifted by Filtration in df fiberglass filters, washed with water and dried by The largest sphingolipid of Pynthium ulumum was extracted and isolated as described in Example 4 and is shown in FIG.
Formula (IA). The sphingolipid structure of the Formula
(IA) was determined by mass spectrometry and nmr using methods well known to those skilled in the art. Heterogeneity was found in the fatty acid component of the sphongolipid.
Sphingolipid of Formula A:
Example 2: Isolation and characterization of sphingolipids from Phytoph thora infestans: Phytophthora infestans (Stretching) was maintained in the medium of pea juice or rye A agar [EC. Caten and J.L. Jinks, (1968), "Canadian Journal of Botany" (Canadian Journal of Botany) 46, page 329]. The medium of the pea juice was prepared by means of the autoclave of 283 gr. of frozen peas in one liter of distilled water, the peas were vigorously shaken in an autoclave and sieved through a thin cloth. 20 gr. of the agar to the filtrate, which was then autoclaved again, before emptying the culture plates. The petri disks of 9 cm. in diameter, which contained 20 ml. of the sucrose-aspargine liquid medium were inoculated with a 7 mm mycelium plug. in diameter, taken from the growing edge of a culture grown in the middle of pea juice or rye A agar. The discs were incubated for 20 days at 25 ° C by shaking on a rotary shaker at 60 rpm. The mycelium was then lifted by filtration on fiberglass filters, washed with water and dried by freezing. The sphingolipid major from the Phytophthora infestans was extracted and isolated as described in Example 4 and shown in Formula (IB). Heterogeneity was found in the fatty acid component of the heterogeneity in the fatty acid component of the sphingolipid. The structure of the sphingolipid of Formula (IB) was determined by mass spectrometry and nmr using methods well known to those skilled in the art.
Esfingolipido of the Formula IB:
Example 3: Isolation and characterization of sphingollids from Phytoph thora capsi ci Phytoph thora capsi ci (ATCC 15399) was maintained on a V-8 juice agar with a pH of 7.0, containing 200 ml. of juice V-8, 4 gr. of CaC03 and 20 gr. of agar per liter. The petri disks of 9 cm. in diameter, which contained 20 ml. of the sucrose-aspargine liquid medium were inoculated with a 7 mm mycelium plug. in diameter, taken from the growth edge of a culture grown on V-8 juice agar. The discs were incubated for 96 hours at 25 ° C with agitation on the rotary shaker at 60 rpm. The mycelium was then lifted by filtration in the glass fiber filters, washed with water and dried by freezing.
Two larger sphingolipids identical to the above Formulas (IA) and (IB) were isolated from Phytophthora capsici, as described in Example 4. Example 4: Lipid Extraction and Isolation Procedure by Chromatography. The sphingolipids were extracted from the lyophilized fungal cells (20 mg.) Prepared in Examples 1, 2 and 3, using chloroform (8 ml.), Methanol (16 ml.), Water (4.6 ml.) By homogenization for 30 minutes. seconds with a Polytron Homogenizer (Brinkman) in a tube with the threaded top, 20 x 200 mm., fitted with a Teflon cap. After homogenization, additional chloroform (8 ml.) And water (8 ml.) Were added. The extracts were mixed by inverting them 30 times and the extract was centrifuged at 70 x g for 10 minutes, twice, to separate the phases. The lower phase of chloroform was removed, evaporated under a stream of nitrogen gas, redissolved in 1 ml. of chloroform / methanol (85/15, v / v) and filtered through glass wool. All the lipid extract (6-20 mg in 1 ml.) Was separated by semi-preparative HPLC. The column (10 x 250 mm.) Contained LiChrosorb Silica, 10 microns, 60 Angstroms, and the mobile phase gradient was: A) hexane, B) isopropanol and C) 0.04% triethylamine in water at a flow rate of 5. ml./min. The time of the linear gradient was: 0 minutes, 100/0/0; 8 minutes, 100/0/0; 13 minutes, 95/5/0; 18 minutes, 85/15/0; 23 minutes, 40/60/0, 61 minutes, 40/51/9; 76 minutes, 40/51/9; 81 minutes, 40/60/0; 86 minutes, 100/0/0; 100 minutes, 100/0/0; for% A /% B /% C, respectively. The HPLC system of the quaternary gradient was a Model 1050 from Hewlett Packard. The injector was a model 7125 Rheodine, adjusted with a fixed magnifying glass of 1.0 ml. The column jet was broken with a Valco "T", 97% of the sample was collected in a test tube and 3% of the sample was detected by means of a Varex Mark II Evaporable Light Scattering Detector operated at 40 ° C, with nitrogen acting as an atomizing gas at 20 psi. Under these conditions, the phosphatidylethanolamine was eluted at a retention time of 48 minutes, the ceramide-PE (the sphingolipid of Formula B) from the Phytophthora infestans was collected in approximately 49 minutes and the ceramide-PE (the sphingolipid from Formula IA) from Pynthium ul timum was collected in approximately 50 minutes. The lipids from Phytophthora capsici contained two peaks of ceramide-PE, one (Formula IB) was collected in 49 minutes and the second (Formula IA) in 50 minutes. A second embodiment of the present invention concerns a process for the preparation of sphingosines, sphingosinephosphorylethanolamines and ceramides from the novel sphingolipids produced by the process described in the previous embodiment. These sphingosines, described in the previous modality. These sphingosines, sphingosinephosphorylethanolamines and ceramides are extremely useful as compounds used in the preparation of other subsequent sphingolipids, which are novel, as well as known and useful. Thus, this embodiment provides a method for preparing sphingosines, sphingosinephosphorylethanolamines and ceramides, comprising the following steps: (i) cultivating an appropriate fungus (ii) raising the mycelium harvest of said fungus, (iii) isolating the sphingolipid from said mycelium. the formula
where R < a is C (0) R8a; R8a is (C? 3-C23) alkenyl (C? 3-C23) or poly (C13-C23) alkenyl alkyl, all of these may be straight or branched chain; X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; and (iv) modifying said sphingolipid by selective hydrolysis to form (a) a sphingosine of the formula (II)
wherein X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; (b) a sphingosine phosphorylethanolamine of the formula
(neither:
(neither)
wherein X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; or (c) a ceramide of the formula (IV)
(IV) R4a is C (0) R8a; R8a is (C13-C23) alkyl, (C3-C23) alkenyl or (C13-C23) alkenyl poly, all these may be straight or branched chain; X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; and the enantiomorphs and structural isomers of these. In a preferred embodiment, the process for preparing a sphingosine of the formula (II), a sphingosinephosphorylethanolamine of the formula (III) or a ceramide of the formula (IV), wherein X is (C10-C20) alkyl, alkenyl (C) ? 0-C20) or poly (C? 0-C2o) alkenyl, all these may be straight or branched chain; includes cultivating an appropriate Oomycete fungus at the stage
(i) In a more preferred embodiment, the process produces a sphingosine of the formula (II), a sphingosinephosphorylethanolamine of the formula (III) or a ceramide of the formula (IV), wherein X is (C10-C20) alkenyl or poly (C) ? 0-C20) alkenyl, all these can be straight or branched chain. In an even more preferred embodiment, the process produces a sphingosine of the formula (II), a sphingosinephosphorylethanolamine of the formula (III) or a ceramide of the formula (IV), wherein sphingosinephosphorylethanolamine of the formula (III) * or a ceramide of the formula (IV), wherein X is a branched chain (C14-C? 8) alktrienyl or a straight chain (C11-C15) alkenyl and R8a is a (C13-C23) alkyl, alkenyl (C? 3 -C23) or straight chain poly (Ci3-C23) alkenyl all. The following compounds can be made and are intended to illustrate, but not limit, the scope of this embodiment of the invention:
Sphingosine of Formula IIA
Sphingosine of Formula IIB
Sphingosinephosphorylethanolamine of the Formula IIIA
Sphingosinephosphorylethanolamine of Formula IIIB
Ceramide of Formula IVB
Sphingosines IIA and HB, sphingosine phosphorylethanol inas IIIA and IIIB and ceramides IVA and IVB above, can be made from sphingolipids IA and IB using the procedures described in Examples 5-10, which are intended to illustrate, but not limit , the scope of the invention.
Example 5: Conversion of IA to IIA Sphingolipid IA is converted to sphingosine IIA by methanolysis in methanolic HCl (1N) according to the method of Gaver, R.C. and Sweeley, C.C. (1965) J. Am. Oil Chem. Soc, 42: 294-298. After the methanol, the methanol and HCl are removed by evaporation in a stream of nitrogen at room temperature. Espingosine IIA is purified from chloroform / methanol, 85/15, v / v, and injected into the semi-prepared HPLC system described for the initial purification of sphingolipid IA. In this HPLC system, espingosine IIA escapes in a retention time of approximately 34 minutes.
Example 6: Conversion of IB to IIB Sphingolipid IB is converted to sphingosine IIB by hydrolysis in methanolic HCl (IN) according to the method of Gaver, R.C. and Sweely, C.C. (1965) J. Am. Oil Chem. Soc. 42: 294-298. After methanolysis, the methanol and HCl are removed by evaporation under a stream of nitrogen at room temperature. Sphingosine IIB is purified from the starting material and any other contaminating material by re-dissolving the hydrolyzate in 1 ml. of chloroform / methanol, 85/15, v / v and injecting it into the semi-prepared HPLC system described for the initial purification of sphingolipid IA. In this HPLC system, sphingosine IIB escapes in a retention time of approximately 36 minutes. Example 7: Conversion of IA to IIA Sphingolipid IA becomes sphingosinephosphorylethanolamine IIA or by a) alkaline hydrolysis according to the Neuenhofer method, S., Schwarsmann, G., Egge, H., Snadhoff, K. (1985) "Biochemistry" (Biochemistry) 24: 525-529 or by b) enzymatic hydrolysis with N-diacylase of sphingolipid ceramide, according to with the method of Ito, M., Kurita, T. and Kita, K. (1995) J. Biol. Quim 270: 24370-3 '4. Sphingosinephosphorylethanolamine IIIA is purified from the starting material and any other contaminating material by re-dissolving the hydrolyzate in 1 ml. of chloroform / methanol, 85/15, v / v and injecting it into the semi-prepared HPLC system described for the initial purification of sphingolipid IA. In this HPLC system, sphingosine phosphorylethanolamine IIIA escapes in a retention time of approximately 54 minutes.
Example 8: Conversion of the IB into the IIIB
The sphingolipid IB is converted into sphingosine phosphorylethanolamine IIIB or by a) alkaline hydrolysis, according to the method of Neuenhofer, S., Schwarzmann, G., Egge, H., Sandhoff, K. (1985) "Bi ochemistry" (Biochemistry ) 24: 525-529 or by b) enzymatic hydrolysis with N-diacylase from sphingolipid ceramide, according to the method of Ito, M., Kurita, T. and Kita, K. (1995) J. Bi ol. Chem 270: 24370-374. Sphingosine phosphorylethanolamine IIIB is purified from the starting material and any other contaminating material by re-dissolving the hydrolyzate in 1 ml. of chloroform / methanol, 85/15, v / v and injecting it into the semi-prepared HPLC system described for the initial purification of sphingolipid IA. In this HPLC system, sphingosine phosphorylethanolamine IIIB escapes in a retention time of approximately 55 minutes.
Example 9: Conversion of the AI to the VAT The sphingolipid IA is converted into the ceramide IVA by means of the treatment with phospholipase C, according to the method of Morrison, W.R. (1969) Bi oquim. Biofis Minutes 176: 537-546. VAT ceramide is purified from the starting material and any other contaminating material by re-dissolving the hydrolyzate in 1 ml. of chloroform / methanol, 85/15, v / v and injecting it into the semi-prepared HPLC system described for the initial purification of sphingolipid IA. In this HPLC system the VAT ceramide escapes in a retention time of approximately 22 minutes.
Example 10: Conversion of IB to IVB Sphingolipid IB is converted to IVB ceramide enzymatically by treatment with phospholipase C, according to the method of Morrison, W.R. (1969) Biochim. Biophys. Minutes 176: 537-546. Ceramide IVB is purified from the starting material and any other contaminating material by re-dissolving the hydrolyzate in 1 ml. of chloroform / methanol, 85/15, v / v and injecting it into the system by re-dissolving the hydrolyzate in 1 ml. of chloroform / methanol, 85/15, v / v and injecting it into the semi-prepared HPLC system described for the initial purification of sphingolipid IA. In this HPLC system, the IVB ceramide escapes in a retention time of approximately 23 minutes. In a third embodiment of this invention, there is provided a method for preparing secondary sphingolipids or children of formula V
wherein R5 is a hydrogen atom, hydroxyl, raercapto, amino, monosubstituted amino, disubstituted or a disubstituted amino in which the two substituents, together with the nitrogen atom, to which they are attached, form a nitrogen atom containing a heterocyclic ring with 3 to 7 members, 0C (0) R6 or OR6, or together with the carbon, to which it is attached, forms a keto group; R6 is alkyl (Ci-Cs), alkenyl (C2-C6). (C2-C6) alkynyl, aralkyl or heterocyclylalkyl;
R3 and R4 are, independently, a hydrogen atom, lower alkyl, C (0) R8, C (0) OR8 or together with the nitrogen atom, to which they are attached, form a nitro group or a hydrogen atom. nitrogen containing a heterocyclic ring with 3 to 7 members; R8 is (C1-C23) alkyl, (C2-C23) alkenyl, poly (C4-C23) alkenyl, (C2-C23) alkynyl, aralkyl or heterocyclylalkyl, which is cyclic, straight or branched chain, and which can be substituted by one or more conventional pharmaceutically acceptable substituents, such as halo, nitro hydroxyl and the like, and the enantiomorphs and structural isomers thereof; comprising steps (i), (ii), (iii) and (iv) of the second embodiment and (v) reacting a sphingosine of the formula (II), a sphingosinephosphorylethanolamine of the formula (III) or a ceramide of the formula (IV) with one or more organic reactants suitable for forming the sphingolipid of the formula (V). In a preferred embodiment, the process produces a sphingolipid child of the formula (V), wherein R5 is a hydrogen atom, hydroxyl, mercapto, amino, monosubstituted amino, disubstituted amino, OC (0) R6 or OR6, or together with the carbon, to which it is attached, forms a keto group;
R6 is (C6C6) alkyl, (C2-C6) alkenyl, (C2-C6) alkynyl, aralkyl or heterocyclylalkyl, which is cyclic, straight or branched chain, and which can be substituted by one or more conventional substituents, pharmaceutically acceptable, such as halo, nitro, hydroxyl and the like; Q is a hydrogen atom, hydroxy, mercapto, NR1R2, formyl, alkanoyl, OC (0) R6, OR6, a substituted phosphate or phosphate, a substituted phosphonate or phosphonate, a nucieotide, a nucleoside, a polynucleotide, a polynucleoside, a amino acid, a peptide, a saccharidase or a polysaccharidase; R1 and R2 are, independently, a hydrogen atom, lower alkyl, C (0) R7, C (0) OR7 or together with the nitrogen atom, to which they are attached, form a nitro group or a nitrogen atom containing a heterocyclic ring with 3 to 7 members; R7 is alkyl, alkenyl, alkynyl, aralkyl or heterocyclylalkyl, which is cyclic, straight or branched chain, and which can be substituted by one or more conventional, pharmaceutically acceptable substituents, such as halo, nitro hydroxyl and the like; R3 and R4 are, independently, a hydrogen atom, lower alkyl, C (0) R8, C (0) OR8 or together with the nitrogen atom, to which they are attached, form a nitro group or a nitrogen atom containing a heterocyclic ring with 3 to 7 members;
R8 is alkyl (C? -C23), alkenyl (C2-C23), poly (C4-C23) alkenyl, (C2-C23) alkynyl, aralkyl or heterocyclylalkyl, which is cyclic, straight or branched chain, and which can substituted by one or more conventional, pharmaceutically acceptable substituents, such as halo, nitro, hydroxyl and the like; and the enantiomorphs and structural isomers of these. In a more preferred embodiment, the process produces a sphingolipid child of formula (V), wherein Q is selected from the group consisting of hydroxyl, glycyl, arginyl, lysyl, galactosyl, sulfogalactosyl, glucosyl, inositol, lactosyl, trihexosyl, phosphorylcholine. , phosphorylethanolamine, GalNAc-Gal-Glc, Gal-Gal-Glc, Sia-Gal-Glc, Gal-GalNAc GalNAc I and I Sia-Gal-Glc Sia-Gal-Glc.
In an even more preferred embodiment, the process produces a sphingolipid child of the formula (V), wherein Q is glucosyl, R8 is CH (OH) - (CH2) nCH3 and n is 11-21. By GalNAc is meant N-acetyl gallatosamine; Glc is understood to mean glucose; by Gal is meant galactose and by Sia sialic acid. By trihexosyl is meant polysaccharidase composed of three hexoses such. By GalNAc is meant galatose ina N-acetyl; Glc is understood to mean glucose; by Gal is meant galactose and by Sia sialic acid. By trihexosyl is meant polysaccharidase composed of three hexoses such as galactoa, glucose, mannose, etc. Both the D and L isomers are contemplated. Examples of compounds that can be made by the method of the latter mode include, but are not limited to the following:
In a fourth embodiment of this invention there is provided a method comprising contacting a mammary cell with a therapeutic amount of a compound of the formula (I), (II), (111), (IV) or (V) in the presence of a pharmaceutically acceptable carrier for Innibir protein kinase C, to provide anti-inflammatory or anti-tumor effect In a fifth embodiment of the present invention, a method is provided. op aa-l'j ^ j treatment of a plant with na i quantity) r »ncm? Effective amen of a compound of the formula (i, (II, tlll), (IV) or (V) in the presence of an agronomically acceptable carrier to provide a protothelial artifact of pathogenic microorganisms. in The plant.
Claims (17)
1. A process for preparing a sphingolipid of the formula: wherein R4a is C (0) R8a; Rβa is (C ?3-C23) alkyl, (C ?3-C23) alkenyl or poly (C ?3-C23) alkenyl, all of which may be straight or branched chain; X is a straight or branched chain alkyl, alkenyl or polyalkenyl; and the enantiomorphs and structural isomers thereof; comprising the following steps: (i) cultivating an appropriate fungus, (ii) raising the mycelium harvest of said fungus, and (iii) isolating the sphingolipid from said mycelium.
2. The process according to claim 1, for preparing a sphingolipid of the formula (I), wherein X is (C10-C20) alkyl, (C ?O-C20) alkenyl or (C10-C2o) alkenyl (C C) alkenyl, all these can be straight or branched chain; comprising cultivating an appropriate Oomycete fungus in step (i).
The process according to claim 2, for a sphingolipid of the formula (I), wherein: X is alkenyl (C? O-C2u) or poly (C? 0-C20) alkenyl, all these can be Straight or branched chain.
The process according to claim 3, for a sphingolipid of the formula (I), wherein: X is a branched-chain alkanthienyl (Ci-C 8 8) or a straight-chain achenyl (Cn-Cis), and R8a is a straight-chain alkyl (C? 3-C2j), alkenyl (C? 3-C23) or poly (C13-C23) alkenyl.
5. A compound of the formula (1) wherein R4a is C (0) R8a; R4a is C (0) R8a; R8a is (C13-C23) alkyl, (C3-C23) alkenyl or poly (CX3-C23) alkenyl, all of which may be straight or branched chain; X is a straight or branched chain alkyl, alkenyl or polyalkenyl; and the enantiomorphs and structural isomers thereof;
6. The compound according to claim 5, wherein X is (C10-C20) alkyl, (C? 0-C20) alkenyl or (C? 0-C2o) alkenyl poly, all these may be straight or branched chain .
7. The compound according to claim 6, wherein: X is (C10-C20) alkenyl or poly (? O_C2o) alkenyl, all of these may be straight or branched chain.
The compound according to claim 7, wherein: X is a branched-chain (C1-C18) alktrienyl or straight-chain (C11-C15) alkenyl, and R8a is an alkyl (C? 3-C23) , straight alkenyl (C? 3-C23) or poly (Ci3-C23) alkenyl straight chain.
9. A process for preparing sphingosines, sphingosinephosphorylethanolamines and ceramides, comprising the following steps: (ii) raising the mycelium harvest of said fungus, (iii) isolating the sphingolipid of the formula from said mycelium. wherein R 4a is C (0) R 3 - R 8a is (C 13 -C 23) alkyl, alkyl (C 3 -C63) or poly (C 3 3 C23) alkenyl, all these can be straight or branched chain; X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; and (iv) modifying said sphingolipid by selective hydrolysis to form (a) a sphingosine of the: ormula (II) wherein X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; (b) a sphingosine phosphorylethanolamine of the formula (III) wherein X is a straight or branched alkyl, alkenyl or polyalkenyl all; or (c) a ceramide of the formula (IV) wherein Ra is C (0) R8a; R8a is alkyl (C -C-, (C13-C23) alkenyl or poly (C3-C23) alkenyl, all of which may be straight or branched chain, X is a straight chain alkyl, alkyl or polyalkenyl or all the branched, and the enantiomorphs and structural isomers thereof 10. The process according to claim 9, for preparing a sphingosine of the formula (II), a
10. The process according to claim 9, for preparing a sphingosine of the formula (II), a sphingosinephosphorylethanolamine of the formula (II) or a ceramide of the formula (IV), wherein X is alkyl (C? 0-C20) , (C10-C2o) alkenyl or poly (C? 0-C2o) alkenyl, all these may be straight or branched chain; which comprises cultivating an appropriate Oomycete fungus in step (i).
11. The procedure according to the claim 10, for preparing a sphingosine of the formula (II), a sphingosine phosphorylethanolamine of the formula (III) or a ceramide of the formula (IV), wherein X is (C10-C20) alkenyl or (C10-C20) alkenyl poly (C10-C20) alkenyl, all these can be straight or branched chain.
12. The procedure according to the claim 11, to prepare a sphingosine of the formula (II), a sphingosinephosphorylethanolamine of the formula (III) or a ceramide of the formula (IV), wherein X is a branched chain alkanthienyl (Ci-C? 8) or an alkenyl (Cn-C? 5) straight chain, and R8a is a (C13-C23) alkyl, (C? 3-C23) alkenyl or straight-chain poly (C13-C23) alkenyl all.
13. A process for preparing daughter or daughter sphingolipids of the formula V (V) wherein R5 is a hydrogen atom, hydroxyl, mercapto, amino, monosubstituted amino, disubstituted amino or a disubstituted amino in which the two substituents, together with the nitrogen atom, to which they are attached, form a nitrogen atom containing a heterocyclic ring with 3 to 7 members, OC (0) R6 or OR6, or together with the carbon, to which it is attached, forms a group keto R6 is alkyl (d-C?), Alkenyl (C2-C6), (C2-C6) alkynyl, aralkyl or heterocyclylalkyl; Q is any organic group, a substituted phosphate or phosphate, or a substituted phosphonate or phosphonate, and is not necessarily more limited; R3 and R4 are, independently, a hydrogen atom, lower alkyl, C (0) Rs, C (0) 0Rfi or, together, with the nitrogen atom, to which they are attached, form a nitro group or a nitrogen atom containing a heterocyclic ring with 3 to 7 members; R8 is (C1-C23) alkyl, (C2-C23) alkenyl, poly (Ct-C23) alkenyl, (C2-C23) alkynyl, aralkyl or heterocyclylalkyl, which is cyclic, straight or branched chain, and which can be substituted by one or more conventional pharmaceutically acceptable substituents, such as halo, nitro hydroxyl and the like, and the enantiomorphs and structural isomers thereof; comprising the following steps: (i) cultivating an appropriate fungus, (ii) raising the mycelium harvest of said fungus, (iii) isolating said sphingolipid from said mycelium. wherein R4a is C (0) R8a; R8a is (C13-C23) alkyl, (C3-C23) alkenyl or poly (C3-C2) alkenyl, all of which may be straight or branched chain; X is a straight-chain or straight-chain alkyl, alkenyl or polyalkenyl all; (iv) modifying said lipid by selective hydrolysis to form (a) a sphingosine of the formula (II) wherein X is a straight chained or branched alkyl, achanyl or polyalkenyl all; (b) a sphingosine phosphorylethanolamine of the formula (III) wherein X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; or (c) a ceramide of the formula (IV) (IV) wherein R4a is C (0) R8a; wherein R4a is C (0) R8a; R8a is (C13-C23) alkyl, (C3-C23) alkenyl or (C13-C23) alkenyl-poly, all these may be straight or branched chain; X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; and the enantiomorphs and structural isomers thereof; and (v) reacting a sphingosine of the formula (II), a sphingosinephosphorylethanolamine of the formula (III) or a ceramide of the formula (IV) with one or more suitable organic reactants to form the desired sphingolipid child of the formula (V) ).
14. The process according to claim 13, for producing a sphingolipid child of the formula (V), wherein R5 is a hydrogen atom, hydroxyl, mercapto, amino, monosubstituted amino, disubstituted amino, OC (0) R6 or OR6 or together with the carbon, to which it is attached, forms a keto group; R6 is (C6-6) alkyl, (C2-C6) alkenyl, (C2-C6) alkynyl, aralkyl or heterocyclylalkyl, which is cyclic, straight-chain or branched, and which may be substituted by one or more conventional substituents, pharmaceutically acceptable, such as halo, nitro, hydroxyl and the like; Q is a hydrogen atom, hydroxy, mercapto, NRXR2, formyl, alkanoyl, OC (0) R6, OR6, a substituted phosphate or phosphate, a substituted phosphonate or phosphonate, a nucieotide, a nucleoside, a polynucleotide, a polynucleoside, a amino acid, a peptide, a saccharidase or a polysaccharidase; R1 and R2 are, independently, a hydrogen atom, lower alkyl, C (0) R7, C (0) 0R7 or together with the nitrogen atom, to which they are attached, form a nitro group or a nitrogen atom containing a heterocyclic ring with 3 to 7 members; R7 is alkyl, alkenyl, alkynyl, aralkyl or heterocyclylalkyl, which is cyclic, straight or branched chain, and which can be substituted by one or more conventional, pharmaceutically acceptable substituents, such as halo, nitro, hydroxyl and the like; R3 and R4 are, independently, a hydrogen atom, lower alkyl, C (0) R8, C (0) OR8 or together with the nitrogen atom, to which they are attached, form a nitro group or a nitrogen atom containing a ring with 3 to 7 members; R8 is (C? -C23) alkenyl (C2-C23) alkenyl, poly (C4-C23) alkenyl, (C2-C23) alkynyl, aralkyl or heterocyclylalkyl, which is cyclic, straight-chain or
15. The procedure according to the claim 14, to produce a sphingolipid child of formula (V), wherein: Q is selected from the group consisting of hydroxyl, glycyl, arginyl, lysyl, galactosyl, sulfogalactosyl, glucosyl, inositol, lactosyl, trihexosyl, phosphorylcholine, phosphorylethanolamine, GalNAc -Gal-Glc, Gal-Gal-Glc Sia-Gal-Glc, Gal-GalNAc GalNAc I and I Sia-Gal-Glc Sia-Gal-Glc.
16. The procedure according to the claim 15, to produce a sphingolipid child of the formula (V), wherein Q is glucosyl, R8 is CH (OH) - (CH2) nCH3 and n is 11-21.
17. A method comprising contacting a cell with a pharmaceutically effective amount of (i) a compound of the formula (I) (I) wherein R a is C (0) R 8 a; R8a is (C13-C23) alkyl, (C3-C23) alkenyl or poly (C3-C23) alkenyl, all of which may be straight or branched chain; X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; and the enantiomorphs and structural isomers thereof; (ii) a compound of the formula (II) wherein X is a straight or branched chain alkyl, alkemyl or polyalkenyl all; and the enantiomorphs and structural isomers thereof; (iii) a compound of the form (III) wherein X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; and the enantiomorphs and structural isomers thereof; (iv) a compound of the formula (IV) wherein R4a is C (0) Rfla; R8a is (C13-C23) alkyl, (C3-C23) alkenyl or poly (Ci3-C23) alkenyl, all of which may be straight or branched chain; X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; and the enantiomorphs and structural isomers thereof; or (v) a compound of the formula IV) wherein R5 is a hydrogen atom, hydroxyl, mercapto, amino, amino-unsubstituted amino, disubstituted amino or a disubstituted amino in which the two substituents together with the nitrogen atom, to which they are substituted, form a hydrogen atom. nitrogen containing a heterocyclic ring with from 3 to 7 R5 is a hydrogen atom, hydroxyl, mercapto, amino, amino monosubstituted amino, disubstituted amino or a disubstituted amino in which the two substituents together with the nitrogen atom, to which they are attached , they form a nitrogen atom containing a heterocyclic ring with 3 to 7 members, OC (0) R6 or OR6, or together with the carbon, to which it is attached, forms a keto group; R6 is alkyl (C6C6), alkenyl (C2-C6), (C2-C6) alkynyl, aralkyl or heterocyclylalkyl; Q is any organic group, a substituted phosphate or phosphate, or a substituted phosphonate or phosphonate, and is not necessarily more limited; R3 and R4 are, independently, a hydrogen atom, lower alkyl, C (0) R8, C (0) 0R8 or together with the nitrogen atom, to which they are attached, form a nitro group or a nitrogen atom containing a heterocyclic ring with 3 to 7 members; R8 is (C? -C23) alkyl, (C2-C23) alkenyl, poly (C-C23) alkenyl, (C2-C23) alkynyl, aralkyl or heterocyclylalkyl, which is cyclic, straight or branched chain, and which can be substituted with one or more conventional, pharmaceutically acceptable substituents, such as halo, nitro, hydroxyl and the like; and the enantiomorphs and structural isomers thereof; wherein R4a is C (0) RR "; R8a is (C ?3-C23) alkyl, (C13-C23) alkenyl or poly (Ca3-C23) alkenyl, all of which may be straight or branched chain; a straight or branched chain alkyl, alkenyl or polyalkenyl all, and the enantiomorphs and structural isomers thereof, (ii) a compound of the formula (II) wherein X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; and the enantiomorphs and structural isomers thereof; (iii) a compound of the formula (III) wherein X is a straight chain or branched alkyl, alkenyl c-polyalkenyl all; and the enantiomorphs and structural isomers thereof; (iv) a compound of the formula (IV) wherein Ra is C (0) R8a; R8a is (C ?3-C23) alkyl, (C13-C23) alkenyl or poly (Ca3-C23) alkenyl, all of which may be straight or branched chain: X is a straight or branched chain alkyl, alkenyl or polyalkenyl all; and the enantiomorphs and structural isomers thereof; or (v) a compound of the formula (V) (V) wherein R5 is a hydrogen atom, hydroxyl, mercapto, amino, monosubstituted amino, disubstituted amino or a disubstituted amino in which the two substituents together with the nitrogen atom, to which they are attached, form a nitrogen atom containing a heterocyclic ring with 3 to 7 members, OC (0) R6 or OR6, or together with the carbon, to which it is attached, forms a keto group; R6 is alkyl (C6C6), alkenyl (C2-C6), (C2-C6) alkynyl, aralkyl or heterocyclylalkyl; Q is any organic group, a substituted phosphate or phosphate, or a substituted phosphonate or phosphonate, and is not necessarily more limited; R3 and R4 are, independently, a hydrogen atom, lower alkyl, C (0) R8, C (0) 0R8 or together with the nitrogen atom, to which they are attached, form a nitro group or a nitrogen atom containing a heterocyclic ring with 3 to 7 members; R8 is (C?-C23) alkyl, (C2-C23) alkenyl, (C4-C23) alkenyl (C2-C23) alkynyl, aralkyl or heterocyclylalkyl, which is cyclic, straight or branched chain, and which could be substituted by one or more conventional pharmaceutically acceptable substituents, such as halo, nitro, hydroxyl and the like, and the enantiomorphs and structural isomers thereof; in the presence of an agronomically acceptable carrier to provide a protective effect against pathogenic microorganisms in the plant.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US1432696P | 1996-03-29 | 1996-03-29 | |
| US014326 | 1996-03-29 |
Publications (2)
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|---|---|
| MXPA97002124A true MXPA97002124A (en) | 1998-04-01 |
| MX9702124A MX9702124A (en) | 1998-04-30 |
Family
ID=21764809
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| MX9702124A MX9702124A (en) | 1996-03-29 | 1997-03-20 | Novel sphingolipids and a process thereto. |
Country Status (9)
| Country | Link |
|---|---|
| US (1) | US5958426A (en) |
| EP (1) | EP0821068A3 (en) |
| JP (1) | JPH1033193A (en) |
| KR (1) | KR970065722A (en) |
| AU (1) | AU734622C (en) |
| BR (1) | BR9701587A (en) |
| CA (1) | CA2200267A1 (en) |
| CO (1) | CO4761070A1 (en) |
| MX (1) | MX9702124A (en) |
Families Citing this family (25)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060094089A1 (en) * | 1988-09-07 | 2006-05-04 | Martek Biosciences Corporation | Process for the heterotrophic production of microbial products with high concentrations of omega-3 highly unsaturated fatty acids |
| US7033584B2 (en) * | 1988-09-07 | 2006-04-25 | Omegatech, Inc. | Feeding Thraustochytriales to poultry for increasing omega-3 highly unsaturated fatty acids in eggs |
| US6977167B2 (en) * | 1988-09-07 | 2005-12-20 | Martek Biosciences Corporation | Mixtures of omega-3 and omega-6 highly unsaturated fatty acids from euryhaline microorganisms |
| US6451567B1 (en) * | 1988-09-07 | 2002-09-17 | Omegatech, Inc. | Fermentation process for producing long chain omega-3 fatty acids with euryhaline microorganisms |
| US5340742A (en) * | 1988-09-07 | 1994-08-23 | Omegatech Inc. | Process for growing thraustochytrium and schizochytrium using non-chloride salts to produce a microfloral biomass having omega-3-highly unsaturated fatty acids |
| US6720184B1 (en) | 1991-08-05 | 2004-04-13 | Emory University | Method of altering sphingolipid metabolism and detecting fumonisin ingestion and contamination |
| US20080175953A1 (en) * | 1995-06-07 | 2008-07-24 | Martek Biosciences Corporation | Process for the Heterotrophic Production of Microbial Products with High Concentrations of Omega-3 Highly Unsaturated Fatty Acids |
| US6800661B1 (en) | 1997-04-15 | 2004-10-05 | Board Of Trustees Of The University Of Illinois | Spisulosine compounds |
| USRE38793E1 (en) | 1997-04-15 | 2005-09-06 | Rinehart Kenneth L | Spisulosine compounds |
| US6107520A (en) * | 1997-04-15 | 2000-08-22 | The Board Of Trustees Of The University Of Illinois | Spisulosine compounds |
| WO1999041266A1 (en) | 1998-02-12 | 1999-08-19 | Emory University | Sphingolipid derivatives and their methods of use |
| WO1999066936A1 (en) | 1998-06-24 | 1999-12-29 | Emory University | Use of 3'-azido-2',3'-dideoxyuridine in combination with further anti-hiv drugs for the manufacture of a medicament for the treatment of hiv |
| EP2341127B1 (en) * | 2000-01-28 | 2015-05-27 | DSM IP Assets B.V. | Enhanced production of lipids containing polyenoic fatty acids by high density cultures of eukaryotic microbes in fermentors |
| DE60140964D1 (en) * | 2001-09-26 | 2010-02-11 | Yissum Res Dev Co | Sphingolipiden |
| KR100917302B1 (en) * | 2001-12-10 | 2009-09-11 | 가오 가부시키가이샤 | Ceramide emulsions |
| US7645873B2 (en) | 2003-03-20 | 2010-01-12 | The Scripps Research Institute | 6″-amino-6″-deoxygalactosylceramides |
| AU2007234753B2 (en) * | 2006-04-07 | 2013-02-21 | Brigham Young University | Modified -galactosyl ceramides for staining and stimulating Natural Killer T cells |
| WO2007129729A1 (en) * | 2006-05-10 | 2007-11-15 | Kyushu University, National University Corporation | Process for production of fish-derived ceramide, and ceramide-containing composition |
| US8916164B2 (en) * | 2007-08-29 | 2014-12-23 | Abivax | Methods of enhancing adjuvaticity of vaccine compositions |
| EP2058011A1 (en) * | 2007-11-07 | 2009-05-13 | Wittycell | Nkt cell activating gycolipids covalently bound antigens and/or drug |
| EP2231145B1 (en) * | 2007-12-05 | 2014-09-17 | Wittycell | Use of glycosylceramides for enhancing the immune response to antigens |
| WO2010040710A1 (en) * | 2008-10-08 | 2010-04-15 | Wittycell | Vaccine composition for use against influenza |
| US10227290B2 (en) | 2012-02-07 | 2019-03-12 | The Regents Of The University Of California | Glycosphingolipids for use in modulating immune responses |
| US10329315B2 (en) | 2012-10-12 | 2019-06-25 | The Brigham And Women's Hospital, Inc. | Glycosphingolipids and methods of use thereof |
| JOP20190254A1 (en) | 2017-04-27 | 2019-10-27 | Pharma Mar Sa | Antitumoral compounds |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GR78246B (en) * | 1981-01-23 | 1984-09-26 | Ciba Geigy Ag | |
| US4816450A (en) * | 1986-09-15 | 1989-03-28 | Duke University | Inhibition of protein kinase C by long-chain bases |
| US4937232A (en) * | 1986-09-15 | 1990-06-26 | Duke University | Inhibition of protein kinase C by long-chain bases |
| FR2637598B1 (en) * | 1988-10-06 | 1991-10-11 | Agronomique Inst Nat Rech | NOVEL INHOSITOL SPHINGOPHOSPHOLIPIDS, THEIR OBTAINMENT, AND THEIR APPLICATION AS INDUCERS OF RESISTANCE TO VARIOUS CRYPTOGAMIC DISEASES IN PLANTS |
| IT1260155B (en) * | 1992-08-03 | 1996-03-28 | Fidia Spa | THERAPEUTIC USE OF PHOSPHORYL-L-SERINE-N-ACYL-SPHINGOSINE |
-
1997
- 1997-03-11 EP EP97301610A patent/EP0821068A3/en not_active Withdrawn
- 1997-03-17 AU AU16289/97A patent/AU734622C/en not_active Ceased
- 1997-03-18 CA CA002200267A patent/CA2200267A1/en not_active Abandoned
- 1997-03-20 MX MX9702124A patent/MX9702124A/en not_active IP Right Cessation
- 1997-03-24 US US08/824,646 patent/US5958426A/en not_active Expired - Fee Related
- 1997-03-28 JP JP9095311A patent/JPH1033193A/en not_active Withdrawn
- 1997-03-29 KR KR1019970011427A patent/KR970065722A/en not_active Application Discontinuation
- 1997-03-31 CO CO97015714A patent/CO4761070A1/en unknown
- 1997-03-31 BR BR9701587A patent/BR9701587A/en not_active Application Discontinuation
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