Isomeric aminophytosphingosines, a method of their production and use

Abstract

Stereoselective synthesis of 3,4-diamino-1,2-diols possessing a long hydrocarbon chain was carried out via olefin cross-metathesis between a commercial tetradec-1-ene and known diastereoisomeric trichloroacetamides prepared in our laboratory. Subsequent modification of the obtained coupling products used an approach based on functional group interconversions, which was accompanied by the rational choice of deprotection protocols. The resulting molecules represent novel isomeric amino analogues of natural D-ribo-phytosphingosine, which on the basis of their structure could have the status of skin sphingomimetics, i.e. substances that can affect the properties of the skin and dermis in terms of improving the quality of the lipid matrix (e.g. change in permeability, hydration), or stimulating the biosynthesis of procollagen in deeper structures. The cytotoxic effect of the prepared aminoisophytosphingosines was examined on two human non-malignant cell lines (BJ and HaCaT) and the screening results confirmed the safety in at least three representatives of the prepared series of compounds. In addition, the ability of the synthesized derivatives to inhibit cancer cell proliferation was also investigated. The results of preliminary anticancer screening showed weaker antiproliferative potential of these unusual sphingoid bases.

Description

[0001] Isomeric aminophytosphingosines, a method of their production and use

[0002] Technical field

[0003] This invention belongs to the fields of organic synthesis, pharmacology, and dermatology.

[0004] Background art

[0005] D-ribo-Phytosphingosine, or (2S,3S,4R)-2-aminooctadecane-l,3,4-triol (4-D- hydroxysphinganine, PHS, 1, Fig. 1), is a well-known long-chain amino alcohol that was first isolated from the mushroom (Amanita muscaria) in 1911 by the German chemist Zellner (1), and was therefore also for a time considered exclusively a product of the plant kingdom and fungi. It was later confirmed to be present across the entire spectrum of living organisms (2), either as a classical sphingoid base or as a structural component of various phytoceramides or more complex glycosphingolipids (3), such as compound 2 (4) (Fig. 1). PHS is implicated in several biological processes, including signalling processes (5) and cytotoxic effects against several types of human cancer cell lines (6). As mentioned above, N-acyl derivatives of 1, also called phytoceramides, were identified in the uppermost layer of the epidermis (Stratum corneum, SC) as essential constituents of the lipid architecture (2b, 7). The family of epidermal lipids is highly heterogeneous; apart from different ceramides derived from three archetypal sphingoid bases: D-ribo-PHS 1, D-erythro-sphingosine 3 (2, 3) and D-erythro- dihydrosphingosine 4 (2, 3) (Fig. 1), it also contains free fatty acids and cholesterol (7). All these components form a sophisticated, highly organised matrix that protects a body from excessive water loss (2b, 7). On the other hand, N-acetyl- D-ribo- phytosphingosine 5 and its fully protected derivative 6 are also part of this defence system and both demonstrate moderate anti-aging activity (8), however they also exhibit cytotoxicity at high concentrations (9). Surprisingly, their synthetic analogue 7 has been described as stimulating procollagen-I biosynthesis in human fibroblasts, and thus this molecule could be a novel agent for the repair pf photoaged skin (10). The aim of the present invention is to provide synthesized isomeric aminophytosphingosines that can be used as novel dermal sphingomimetics with applications in dermatology, and which could improve properties of the deeper layers of skin in terms of stimulating procollagen biosynthesis.

[0006] Summary of the invention

[0007] The subject matter of the present invention are the isomeric aminophytosphingosines of formula (I) where

[0008] R = H or (CH3)2C=

[0009] RI = NH2or NHCOCCl3

[0010] R2= NH2or H or NHCOCCl3

[0011] R3= H or NH2or NHCOCCl3or isomeric aminosphingosines of formula (II) where

[0012] R = H or (CH3)2C=

[0013] R1= NHAc or NH2.HCI

[0014] R2= NHAc or H or NH2.HCI

[0015] R3= H or NHAc or NH2. HCI.

[0016] Specific compounds of aminosphingosines of formula (I and II) are:

[0017] N,N '-[(1R,2R,E )-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadec-3-en-1,2- diyl]bis(2,2,2-trichloroacetamide) 16, N,N '-[(1R,2S,E)-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadec-3-en-1,2- diyl]bis(2,2,2-trichloroacetamide) 17,

[0018] N,N '-[(1R,2R)-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadecane-1,2- diyl]diacetamide 18,

[0019] N,N '-[(1R,2S )-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadecane-1,2- diyl]diacetamide 19,

[0020] N,N '-[(2S,3R,4R)-1 ,2-Dihydroxyoctadecane-3,4-diyl]diacetamide 8,

[0021] N,N '-[(2S,3R,4S)-1,2-Dihydroxyoctadecane-3,4-diyl]diacetamide 9, (2S,3R,4R)-3,4-Diamino-octadecane-1,2-diol-dihydrochloride 10, (2S,3R,4S)-3,4-Diamino-octadecane-1,2-diol-dihydrochloride 11,

[0022] N,N '-[(2S,3R,4R,E)-1,2-Dihydroxyoctadec-5-en-3,4-diyl]bis(trichloroacetamide) 20,

[0023] N,N '-[(2S,3R,4S,E)-1,2-Dihydroxyoctadec-5-en-3,4-diyl]bis(trichloroacetamide) 21,(2S,3R,4R,E)-3,4-Diaminooctadec-5-en-1,2-diol 12, or (2S,3R,4S,E)-3,4-Diaminooctadec-5-en-1,2-diol 13.

[0024] Another aspect of the present invention is a method of making the compound N,N '- [1R,2R,E)-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadec-3-en-1,2-diyl]bis(2,2,2- trichloroacetamide) 16, or N,N '-[(1R,2S,E)-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'- yl]hexadec-3-en-1,2-diyl]bis(2,2,2-trichloroacetamide) 17 where the compound of formula

[0025] / where R' =NHCOCCI3and R1'=H, or where R1'=NHCOCCI3and R'=H, is treated with tetradec-1-ene, in the presence of the the second-generation Grubbs catalyst in anhydrous CH2CI2to form a corresponding diastereoisomer.

[0026] Another aspect of the present invention is a method of making the compound N,N '- [(1R,2R)-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadecane-1,2-diyl]diacetamide 18, or N,N '-[(1R,2S)-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadecane-1,2- diyl]diacetamide 19, where compound 16 or 17 is treated with Et3N, 10% Pd / C, in anhydrous MeOH under H2atmosphere to form a corresponding diastereoisomer.

[0027] Another aspect of the present invention is a method of making the compound [(2S,3R,4R,E)-1,2-Dihydroxyoctadec-5-en-3,4-diyl]bis(trichloroacetamide) 20 or

[0028] N,N '-[(2S,3R,4S,E)-1,2-Dihydroxyoctadec-5-en-3,4-diyl]bis(trichloroacetamide) 21, where compound 16 or 17 is treated with p-Toluenesulfonic acid in MeOH, at room temperature to form a corresponding diastereoisomer.

[0029] Another aspect of the present invention is a method of making compound(2S,3R,4R,E)-3,4-Diaminooctadec-5-en-1,2-diol 12 or (2S,3R,4S,E)-3,4- Diaminooctadec-5-en-1,2-diol 13, where compound 20 or 21 is treated with a 2 M aqueous solution of NaOH, EtOH at 80 °C to form a corresponding diastereoisomer.

[0030] Another aspect of the present invention is a method of making the compound N,N '- [(2S,3R,4R)-,1E),2-Dihydroxyoctadecane-3,4-diyl]diacetamide 8 or N,N '-[(2S,3R,4S)- 1,2-Dihydroxyoctadecane-3,4-diyl]diacetamide 9, where compound 18 or 19 is treated with a 10% aqueous solution of HCI, in THF, at room temperature to form a corresponding diastereoisomer.

[0031] Another aspect of the present invention is a method of making the compound (2S,3R,4R) ,E-3),4-Diaminooctadecane-1,2-diol-dihydrochloride 10 or (2S,3R,4S),E-3),4- Diaminooctadecane-1,2-diol-dihydrochloride 11, where compound 18 or 19 is treated with 6 M HCI, at 65 °C to form a corresponding diastereoisomer.

[0032] Isomeric aminophytosphingosines are suitable for use in medicine as sphingomimetics.

[0033] In the context of the present invention, we have prepared a small library of 3,4- diamino-1,2-diols with a lipophilic long chain (Scheme 1), which are analogues of the natural D-ribo-phytosphingosine 1 (Fig. 1). We assume that the mentioned aminoisophytosphingosines possess the status of skin sphingomimetics, which means that they mimic the structure of classical sphingolipids present in the epidermis or in other skin layers. Given their similarity to 1, it is possible that they can have direct effects on membrane organisation and permeability or have other benefits that are not directly related to the SC lipids and membrane biophysics. As such, they may therefore find applications in dermatology. 12

[0034] Scheme 1. Synthetic plan for the preparation of unusual sphingoid bases 8-13

[0035] The synthesis of the target compounds presented in Scheme 1 utilised the olefin crossmetathesis reaction of amides 14 and 15, prepared by the inventors, with commercially available tetradec-1-ene in the presence of the second-generation Grubbs catalyst, providing corresponding alkenes 16 (90%) and 17 (72%) (Scheme 2). Saturation of the double bond coupled with reduction of trichloroacetyl to acetyl led to the formation of derivatives 18 (77%) and 19 (75%). Subsequent acid- catalyzed deprotection of acetonide in 18 and 19 delivered key ceramides 8 (79%) and 9 (86%). On the other hand, the conversion of aminoisophytosphingosines 18 and 19 to the corresponding HCI salts 10 (90%) and 11 (91%) was achieved by treatment with 6 M HCI at 65 °C. The preparation of unsaturated amines 12 (86%) and 13 (81%) was accomplished via the production of diols 20 and 21 and their subsequent saponification with 2 M aqueous solution of NaOH in ethanol at 80 °C.

[0036] The used olefin cross-metathesis and the commercial availability of alkenes with terminal double bond allows to obtain a variety of derivatives differing in a length of the basic carbon skeleton, which may be beneficial for the study of structure-activity relationships. The differential stereochemistry in the obtained diastereoisomeric products may ultimately have an impact on the observed activity.

[0037] According to the inventors of the present invention, based on a search of relevant chemical databases such as Reaxys and SciFinder, these are new chemical entities that have not been described in the literature so far. By comparing their structure with related substances, it can be stated that their synthesis is less demanding, does not require application of complicated reaction conditions and steps, we are able to obtain pairs of diastereoisomerically and enantiomerically pure compounds, and we are able to prepare the considered sphingoid bases on a gram scale, something which was also verified by the preparation of ceramide 9 (Scheme 2) in a larger quantity for further testing of its biological properties. These derivatives, because of their structure, can be further modified, for example by analogue synthesis. Another potential application is their implementation into the structure of hyaluronic acid with the intention of preparing conjugates (polymers) that could improve the properties of deeper layers of skin (coriurri), e.g. in terms of stimulation of procollagen biosynthesis. Scheme 2. Reagents and conditions: (a) tetradec-1-ene, Grubbs II, CH2CI2, reflux; (b) H2, Et3N, 10% Pd / C, MeOH; (c) 10% aq HCI, THF, rt; (d) 6 M HCI, 65 °C; (e) p-TsOH, MeOH, rt; (f) 2 M aq NaOH, EtOH, 80 °C.

[0038] Brief description of the drawings

[0039] Figure 1 shows D-ribo-phytosphingosine 1 and its analogues. Figure 2 shows1H NMR (400 MHz, CDCI3) of compound 16. Figure 3 shows13C NMR (101 MHz, CDCI3) of compound 16. Figure 4 shows1H NMR (400 MHz, CDCI3) of compound 17. Figure 5 shows13C NMR (101 MHz, CDCI3) of compound 17. Figure 6 shows1H NMR (400 MHz, CDCI3) of compound 18. Figure 7 shows13C NMR (101 MHz, CDCI3) of compound 18. Figure 8 shows1H NMR (400 MHz, CDCI3) of compound 19. Figure 9 shows13C NMR (101 MHz, CDCI3) of compound 19. Figure 10 shows1H NMR (400 MHz, CD3OD) of compound 8. Figure 11 shows13C NMR (101 MHz, CD3OD) of compound 8. Figure 12 shows1H NMR (400 MHz, CD3OD) of compound 9. Figure 13 shows13C NMR (101 MHz, CD3OD) of compound 9. Figure 14 shows1H NMR (400 MHz, CD3OD) of compound 10. Figure 15 shows13C NMR (101 MHz, CD3OD) of compound 10. Figure 16 shows1H NMR (400 MHz, CD3OD) of compound 11. Figure 17 shows13C NMR (101 MHz, CD3OD) of compound 11. Figure 18 shows1H NMR (400 MHz, CDCI3) of compound 20. Figure 19 shows13C NMR (101 MHz, CDCI3) of compound 20. Figure 20 shows1H NMR (400 MHz, CDCI3) of compound 21. Figure 21 shows13C NMR (101 MHz, CDCI3) of compound 21. Figure 22 shows1H NMR (400 MHz, CD3OD) of compound 12. Figure 23 shows13C NMR (101 MHz, CD3OD) of compound 12. Figure 24 shows1H NMR (400 MHz, CD3OD) of compound 13. Figure 25 shows13C NMR (101 MHz, CD3OD) of compound 13. Examples of embodiment

[0040] The HaCaT (human keratinocytes) cell line was provided by RNDr. Peter Gal, PhD., Centre of Clinical and Preclinical Research MEDIPARK, Faculty of Medicine, Pavol Jozef Safarik University in Kosice , and the Department of Burns Medicine, Third Faculty of Medicine, Charles University, Prague, Czech Republic.

[0041] Cell lines MCF-7 (human breast cancer cell line), HeLa (human cervical cancer cell line), Jurkat (human acute T-lymphoblastic leukemia cell line), HCT-116 (human colorectal cancer cell line), BLM-9 (human skin melanoma cell line) and BJ-5ta (human skin fibroblasts) were provided by the Institute of Pharmacology of the Faculty of Medicine of the Pavol Jozef Safarik University in Kosice, where the respective screening of the prepared compounds was also carried out. The safety of the prepared compounds was evaluated in normal (non-malignant) BJ and HaCaT cell lines. Their potential to affect cancer cell viability was tested on the malignant cell lines MCF-7, HeLa, Jurkat, HCT- 116, and BLM-9.

[0042] General methods

[0043] Preparative column chromatography was performed on Kieselgel 60 silica gel (0.040- 0.063 mm, 230-400 mesh, Merck) under pressure, with solvents that were distilled prior to use. Thin-layer chromatography was performed on Merck 60 F254 aluminium plates, and visualization was performed either using UV light (254 nm) or using detection reagents (phosphomolybdic acid solution), followed by heating. NMR spectra were recorded on a Varian Mercury Plus 400 FT NMR (400.13 MHz for1H and 100.61 MHz for13C) spectrometer. For1H, chemical shifts are given in units of ppm (parts per million) and the reference is given either relative to TMS (δ = 0.0 ppm) as an internal standard or to the signals of CDCI3solvent (δ = 7.26 ppm) and CD3OD solvent (5 = 3.31 ppm); and for13C relative to CDCI3(5 = 77.16 ppm) and CD3OD (5 = 49.00 ppm). The multiplicity of13C NMR signals was determined by the HSQC method. Chemical shifts (in ppm) and interaction constants (in Hz) were obtained by the first order analysis; using COSY and H / C correlation spectra. Infrared spectra were measured on a Nicolet 6700 FT-IR instrument, using the Smart Orbit method, and the relevant values are given in units of cm-1High-resolution mass spectra (HRMS) were recorded on a micrOTOF-Q II quadrupole-time of flight hybrid mass spectrometer (Bruker Daltonics). The specific rotation was measured on a Jasco P-2000 polarimeter. The melting point was determined on a Stuart SMP 10 device and are uncorrected. All commercial reagents were purchased from suppliers such as Sigma-Aldrich or Acros Organics. Solvents were purified according to standard procedures prior to their use. Table 1 gives1H and13C NMR spectra of compounds 8-21.

[0044] Table 1 and13C NMR spectra of compounds 8-21

[0045] Example 1

[0046] N,N '-[ (lR,2R,E)-1-[ (s) -2,'2'-Dimethyl1- ',3 '-ioxolane-4'yl]hexadec-3-en-1,2- diyl]bis(2,2,2-trichloroacetamide) 16

[0047] Tetradec- 1-ene (1 mL, 4.0 mmol) and the second-generation Grubbs catalyst (20.4 mg, 0.02 mmol) were successively added to a solution of compound 14 (0.38 g, 0.80 mmol) in anhydrous CH2CI2(13.6 mL). After 2 hours of stirring under reflux, the mixture was cooled to room temperature, the solvent was evaporated under reduced pressure, and the residue was purified through a short silica gel column (n- hexane / EtOAc, 7:1). This procedure provided 0.46 g (90%) of derivative 16 as a white solid; mp 93-96 °C; [a]D20-5.5 (c 0.98, CHCI3). IR (neat) v3284, 2915, 2848, 1685, 1530 cm1;1H NMR (400 MHz, CDCI3) δ0.88 (t, 3H, J = 6.8 Hz, CH3), 1.26-1.30 (m, 20H, 10 x CH2), 1.39 (s, 3H, CH3), 1.49 (s, 3H, CH3), 2.06-2.12 (m, 2H, CH2), 3.72 (dd, 1H, J= 6.2, 8.8 Hz, H-5'), 4.07-4.16 (m, 2H, H-l, H-5'), 4.54-4.61 (m, 1H, H-4'), 4.77-4.85 (m, 1H, H-2), 5.46 (dd, 1H, J= 5.6, 15.5 Hz, H-3), 5.75-5.84 (m, 1H, H-4), 7.14 (d, 1H, J= 8.8 Hz, NH), 7.83 (d, 1H, J= 8.9 Hz, NH);13C NMR (101 MHz, CDCI3) δ 14.3 (CH3), 22.8 (CH2), 24.7 (CH3), 26.5 (CH3), 29.0 (CH2), 29.3 (CH2), 29.5 (CH2), 29.6 (CH2), 29.7 (CH2), 29.8 (3 x CH2), 32.0 (CH2), 32.4 (CH2), 53.4 (C-l), 55.5 (C- 2), 66.6 (C-5'), 73.7 (C-4'), 92.3 (CCI3), 92.9 (CCI3), 110.6 (OCO), 123.8 (C-3), 135.7 (C-4), 161.7 (C=O), 162.4 (C=O). ESI-HRMS: m / z calculated for C25H40C16NaN2O4[M + Na]+665.1011, found 665.1023.

[0048] Example 2

[0049] N,N '-[ (1R,2R,E)-1-[ (S)-2,'2'-Dimethyl-1- ',3li'-oxolane-4'yl]hexadec-3-en-1,2- diyl]bis(2,2,2-trichloroacetamide) 17

[0050] Tetradec- 1-ene (0.9 mL, 3.65 mmol) and the second-generation Grubbs catalyst (19 mg, 0.02 mmol) were successively added to a solution of 15 (0.3 g, 0.73 mmol) in anhydrous CH2CI2(12.5 mL). After stirring for one hour under reflux, the mixture was cooled to room temperature, concentrated and the residue was purified by chromatography on silica gel ( / 7-hexane / EtOAc, 7:1) to give 0.39 g (82%) of compound 17 as a white solid; mp 55-56 °C; [a]D20+3.4 (c0.49, CHCI3). IR (neat) v3362, 2918, 2845, 2364, 1691, 1505 cm1;1H NMR (400 MHz, CDCI3) δ 0.88 (t, 3H, J = 6.7 Hz, CH3) , 1.25-1.30 (m, 20H, 10 x CH2), 1.35 (s, 3H, CH3), 1.49 (s, 3H, CH3), 2.07-2.12 (m, 2H, CH2), 3.64 (dd, 1H, J = 6.8, 8.6 Hz, H-5'), 3.97-4.02 (m, 1H, H-l), 4.08 (dd 1H, J= 6.9, 8.6 Hz, H-5'), 4.41-4.45 (m, 1H, H-4'), 4.63-4.69 (m, 1H, H-2), 5.38 (dd, 1H, J= 8.1, 15.3 Hz, H-3), 5.91-5.98 (m, 1H, H-4), 7.10 (d, 1H, J= 8.3 Hz, NH), 7.23 (d, 1H, J = 9.2 Hz, NH );13C NMR (101 MHz, CDCI3) δ 14.3 (CH3), 22.8 (CH2), 24.6 (CH3), 26.5 (CH3), 28.9 (CH2), 29.2 (CH2), 29.5 (CH2), 29.6 (CH2), 29.8 (4 x CH2), 32.1 (CH2), 32.5 (CH2), 54.9 (C-l), 56.6 (C-2), 66.4 (C-5'), 73.2 (C-4'), 92.2 (CCI3) 92.5 (CCI3), 110.2 (OCO), 123.9 (C-3), 138.8 (C-4), 161.9 (C=O), 163.9 (C=O). ESI-HRMS: m / z calculated for C25H40C16NaN2O4[M + Na]+665.1011, found 665.1012.

[0051] Example 3 N,N '-[ (1R,2R)-1-[rS)-2,'2'-Dimethyl-l, '3'-dioxolane-4'-yl]hexadecane-1,2- diyl]diacetamide 18

[0052] 10% Pd / C (0.29 g) and Et3N (0.30 mL, 2.20 mmol) were successively added to a solution of compound 16 (0.27 g, 0.42 mmol) in anhydrous MeOH (6 mL). The reaction mixture was stirred for 30 minutes at room temperature under an atmosphere of hydrogen, which was introduced under the pressure of 1 atm. Subsequently, the content of the flask was filtered through a small pad of Celite, and the filtrate was concentrated under reduced pressure. Chromatography of the residue on silica gel (n- hexane / EtOAc, 1:6) provided 0.14 g (77%) of compound 18 as a white solid; mp 158- 161 °C; [a]D20+20.6 (c0.40, CHCI3). IR (neat) v3295, 2911, 2848, 1640, 1565, 1410, 1022 cm1;1H NMR (400 MHz, CDCI3) δ0.88 (t, 3H, J = 6.6 Hz, CH3), 1.25-1.30 (m, 26H, 13 x CH2), 1.36 (s, 3H, CH3), 1.43 (s, 3H, CH3), 2.00 (s, 3H, CH3), 2.03 (s, 3H, CH3), 3.62-3.66 (m, 1H, H-5'), 4.01-4.14 (m, 3H, H-l, H-2, H-5'), 4.32-4.39 (m, 1H, H-4'), 5.93 (d, 1H, J= 8.8 Hz, NH), 6.18 (d, 1H, J = 9.3 Hz, NH);13C NMR (101 MHz, CDCI3) δ 14.2 (CH3), 22.8 (CH2), 23.5 (CH3), 23.7 (CH3), 25.1 (CH3), 26.3 (CH2), 26.5 (CH3), 29.5 (CH2), 29.6 (CH2), 29.7 (2 x CH2), 29.8 (5 x CHx), 32.0 (CH2), 32.4 (CH2), 51.8 (C-l), 52.6 (C-2), 66.9 (C-5'), 74.6 (C-4'), 109.6 (OCO), 170.3 (C=O), 170.7 (C=O). ESI-HRMS: m / z calculated for C25H48NaN2O4 [M + Na]+463.3506, found 463.3515.

[0053] Example 4

[0054] N,N'-[(lR,2S)-1-[(S)-2,'2'-Dimethyl-l,'3'-dioxolane-4'-yl]hexadecane-l,2- diyl]diacetamide 19

[0055] 10% Pd / C (0.3 g) and Et3N (0.32 mL, 2.31 mmol) were successively added to a solution of derivative 17 (0.28 g, 0.44 mmol) in anhydrous MeOH (6.2 mL). The resulting mixture was stirred for a period of 30 minutes at room temperature under an atmosphere of hydrogen, which was introduced under the pressure of 1 atm. After completion of the reaction, the mixture was filtered through a small pad of Celite, and the filtrate was concentrated under reduced pressure. Chromatography of the residue on silica gel (n-hexane / EtOAc, 1:6) provided 0.15 g (75%) of compound 19 as a white solid; mp 177-181 °C; [a]D20-4.4 (c l.l, CHCI3). IR (neat) v3287, 2912, 2845, 2365, 2335, 1663, 1557, 1365, 1071 cm1;1H NMR (400 MHz, CDCI3) δ0.88 (t, 3H, J = 6.8 Hz, CH3), 1.25-1.30 (m, 26H, 13 x CH2), 1.34 (s, 3H, CH3), 1.43 (s, 3H, CH3), 1.96 (s, 3H, CH3), 2.00 (s, 3H, CH3), 3.59 (dd, 1H, J= 6.8, 8.2 Hz, H-5'), 3.87 (td, 1H, J= 2.0, 9.4 Hz, H-l), 4.02 (dd, 1H, J = 6.9, 8.2 Hz, H-5'), 4.08-4.18 (m, 1H, H-2), 4.29-4.33 (m, 1H, H -4'), 5.71 (d, 1H, J = 9.5 Hz, NH), 6.04 (d, 1H, J = 9.5 Hz, NH);13C NMR (101 MHz, CDCI3) δ 14.3 (CH3), 22.8 (CH2), 23.4 (CH3), 23.5 (CH2), 24.8 (CH3), 25.7 (CH3), 26.5 (CH3), 29.5 (CH2), 29.6 (2 x CH2), 29.7 (CH2), 29.8 (5 x CH2), 31.9 (CH2), 32.1 (CH2), 51.6 (C-2), 53.8 (C-l), 66.7 (C-5'), 74.0 (C-4'), 109.5 (OCO), 170.9 (C=O),

[0056] 171.6 (C=O). ESI-HRMS: / 77 / zcalculated for C25H48NaN2O4[M + Na]+463.3506, found 463.3518.

[0057] Example 5

[0058] N,N '-[r2S,3R,4R)-1,2-Dihydroxyoctadecane-3,4-diyl]diacetamide 8

[0059] A 10% aqueous solution of HCI (1.5 mL) was added to a solution of compound 18 (0.13 g, 0.30 mmol) in THF (1.5 mL) and mixture was stirred for one hour at room temperature. After that time the reaction was quenched by neutralisation with solid NaHCO3. The insoluble material was removed by filtration and the filtrate was concentrated under reduced pressure. The residue was purified by chromatography on silica gel (CH2CI2 / MeOH, 12:1) to afford 94 mg (79%) of compound 8 as white crystals; mp 190-193 °C; [a]D21+5.2 (c 0.27, MeOH). IR (neat) v 3287, 2923, 2855, 2353, 2341, 1618, 1555, 1375 cm1;1H NMR (400 MHz, CD3OD) δ 0.90 (t, 3H, J = 6.2 Hz, CH3), 1.29-1.45 (m, 25H, 12 x CH2, H-CH2), 1.59-1.67 (m, 1H, H-CH2), 1.98-2.00 (m, 6H, 2 x CH3), 3.35-3.44 (m, 2H, 2 x H-l), 3.74-3.77 (m, 1H, H-2), 3.86 (dd, 1H, J = 1.5, 9.1 Hz, H-3), 3.94-3.99 (m, 1H, H-4);13C NMR (101 MHz, CD3OD) δ 14.3 (CH3), 22.5 (CH3), 22.6 (CH3), 23.6 (CH2), 27.0 (CH2), 30.4 (2 x CH2), 30.6 (2 x CH2), 30.7 ( 5 x CH2), 32.3 (CH2), 33.0 (CH2), 51.6 (C-4), 54.5 (C-3), 64.5 (C-l), 71.3 (C-2),

[0060] 173.6 (C=O), 174.1 (C=O). ESI-HRMS: m / z calculated for C22H44NaN2O4[M + Na]+423.3193, found 423.3200. Example 6

[0061] N,N '-[ (2S,3R,4S)-Dihydroxyoctadecane-3,4-diylJdiacetamide 9

[0062] Using the same procedure as described for the preparation of compound 8, derivative 19 (0.42 g, 0.95 mmol) was modified to substance 9 (white crystals, 0.33 g, 86%, CH2CI2 / MeOH, 12:1); mp 144-145 °C; [a]D21-29.7 (c 0.37, MeOH). IR (neat) v3282, 2917, 2850, 2359, 2342, 1617, 1549, 1375 cm1;1H NMR (400 MHz, CD3OD) δ 0.90 (t, 3H, J = 6.7 Hz, CH3), 1.29-1.47 (m, 25H, 12 x CH2, H-CH2), 1.59-1.69 (m, 1H, H- CH2), 1.94 (s, 3H, CH3), 1.98 (s, 3H, CH3), 3.40-3.49 (m, 2H, 2 x H-l), 3.74 (td, 1H, J= 3.0, 6.2 Hz, H-2), 3.94 (dd, 1H, J= 3.0, 7.5 Hz, H-3), 4.07-4.13 (m, 1H, H-4);13C NMR (101 MHz, CD3OD) δ 14.5 (CH3), 22.6 (CH3), 22.7 (CH3), 23.8 (CH2), 27.0 (CH2), 30.5 (CH2), 30.7 (2 x CH2), 30.8 (6 x CH2), 32.8 (CH2), 33.1 (CH2), 51.9 (C-4), 55.3 (C-3), 64.7 (C-l), 71.9 (C-2), 173.5 (C=O), 173.9 ( C=O). ESI-HRMS: m / z calculated for C22H44NaN2O4[M + Na]+423.3193, found 423.3200.

[0063] Example 7

[0064] (2S,3R, 4R)-3, 4-Diamino-octadecane-l,2-diol-dihydrochloride 10

[0065] Compound 18 (33 mg, 0.08 mmol) was treated with 6 M aqueous HCI solution (3.5 mL) and the resulting mixture was stirred and heated for 8 hours at 65 °C. After completion of the reaction, the mixture was concentrated and the isolated solid fraction was washed with anhydrous Et2O (3 x 10 mL). After decantation, the obtained material was dried for 6 hours under vacuum. The above procedure provided 26 mg (90%) of derivative 10 in the form of a pale yellow amorphous solid; [a]D21+ 15.8(c 0.52, MeOH). IR (neat) v3344, 2930, 1630, 1461 cm1;1H NMR (400 MHz, CD3OD) δ0.90 (t, 3H, J= 6.4 Hz, CH3), 1.28-1.56 (m, 24H, 12 x CH2), 1.75-1.87 (m, 2H, CH2), 3.63- 3.70 (m, 1H, H-4), 3.72-3.80 (m, 3H, 2 x H-l, H-3), 4.04-4.09 (m, 1H, H-2);13C NMR (101 MHz, CD3OD) δ 14.5 (CH3), 23.7 (CH2), 26.4 (CH2), 30.3 (CH2), 30.4 (CH2), 30.5 (CH2), 30.7 (2 x CH2), 30.8 (5 x CH2), 31.1 (CH2), 54.0 (C-4), 54.8 (C-3), 64.8 (C-l), 67.4 (C-2). ESI-HRMS: m / z calculated for C18H42CI2N2O2[M - HCI - Cl]+317.3163, found 317.3164. Example 8

[0066] (2S,3R,4S)-3,4-Diamino-octadecane-1,2-diol-dihydrochloride 11

[0067] Compound 19 (25 mg, 0.06 mmol) was treated with 6 M aqueous HCI solution (2.6 mL) and the resulting mixture was stirred and heated for 9.5 hours at 65 °C. After completion of the reaction, the mixture was concentrated under reduced pressure and the residue was washed with anhydrous Et20 (3 x 10 mL). The product was then dried under vacuum for 6 hours. The above procedure provided 19 mg (91%) of compound 11 as a pale yellow amorphous solid; [a]D20-7.3(c 0.38, MeOH). IR (neat) v 3357, 2921, 2851, 1631, 1446 cm1;1H NMR (400 MHz, CD3OD) δ 0.90 (t, 3H, J = 6.7 Hz, CH3), 1.29-1.55 (m, 24H, 12 x CH2), 1.73-1.98 (m, 2H, CH2), 3.60-3.66 (m, 1H, H- 4), 3.69-3.82 (m, 3H, 2 x H-l, H-3), 4.00-4.05 (m, 1H, H-2);13C NMR (101 MHz, CD3OD) δ14.4 (CH3), 23.7 (CH2), 26.7 (CH2), 30.1 (CH2), 30.4 (CH2), 30.5 (2 x CH2), 30.7 (2 x CH2), 30.8 (4 x CH2), 33.1 (CH2), 54.4 (C-4), 55.0 (C-3), 65.2 (C-l), 68.8 (C-2). ESI-HRMS: m / z calculated for C18H42CI2N2O2[M - HCI - Cl]+317.3163, found 317.3174.

[0068] Example 9

[0069] N,N'-[(2S,3R,4R,E)-1,2-Dihydroxyoctadec-5-en-3,4-diyl]bis(trichloroacetamide) 20 p-Toluenesulfonic acid (8.4 mg, 0.04 mmol) was added to a solution of compound 16 (70 mg, 0.11 mmol) in MeOH (1.2 mL), and the resulting mixture was stirred for 23 hours at room temperature. The reaction was quenched by neutralisation with Et3N, and then the mixture was concentrated under reduced pressure. Chromatography of the residue through a short silica gel column (n-hexane / EtOAc, 3:1) gave 58 mg (86%) of derivative 20 as a white solid; mp 111-112 °C; [a]D21-17.4 (c 0.52, CHCI3). IR (neat) v 3297, 2922, 2852, 1681, 1526, 1455, 1310, 1055 cm1;1H NMR (400 MHz, CDCI3) δO.88 (t, 3H, J = 6.7 Hz, CH3), 1.25-1.30 (m, 20H, 10 x CH2), 2.03-2.08 (m, 2H, CH2), 2.49 (br s, 1H, OH), 3.53 (br s, 1H, OH), 3.60-3.61 (m, 2H, 2 x H-l), 4.07- 4.13 (m, 2H, H-2, H-3), 4.63-4.71 (m, 1H, H-4), 5.48 (dd, 1H, J= 6.9, 15.3 Hz, H-5), 5.79-5.88 (m, 1H, H-6), 7.22 (d, 1H, J = 9.1 Hz, NH), 7.81 (d, 1H, J = 8.6 Hz, NH);13C NMR (101 MHz, CDCI3) δ 14.3 (CH3), 22.8 (CH2), 29.0 (CH2), 29.4 (CH2), 29.5 (CH2), 29.6 (CH2), 29.7 (CH2), 29.8 (3 x CH2) 32.1 (CH2), 32.5 (CH2), 54.0 (C-3), 55.4 (C-4), 63.4 (C-l), 69.9 (C-2), 92.3 (CCI3), 92.4 (CCI3), 123.7 (C-5), 137.4 (C-6), 162.6 (C=0), 162.9 (C=0). ESI-HRMS: m / z calculated for C22H36C16N2NaO4[M + Na] 625.0698, found 625.0676.

[0070] Example 10

[0071] N,N'-[(2S,3R,4S,E)-1,2-Dihydroxyoctadec-5-en-3,4<liyl]bis(trichloroacetamide) 21 p-TsOH (6.8 mg, 0.04 mmol) was added to a solution of compound 17 (60 mg, 0.09 mmol) in MeOH (1.1 mL), and the resulting mixture was stirred for a period of 21 hours at room temperature. After the indicated time, the reaction was quenched by the careful addition of Et3N and then the mixture was concentrated under reduced pressure. The residue was purified by chromatography on silica gel (n-hexane / EtOAc, 3:1) to provide 44 mg (81%) of derivative 21 as a white solid; mp 107-108 °C; [a]D21+4.8 (c O.61, CHCI3). IR (neat) v 3295, 2922, 2852, 2360, 1682, 1526, 1465, 1311, 1055 cm1;1H NMR (400 MHz, CDCI3) δ0.88 (t, 3H, J = 6.8 Hz) 1.25-1.38 (m, 20H, 10 x CH2), 2.02-2.12 (m, 2H, CH2), 2.41 (br s, 1H, OH), 3.29 (br s, 1H, OH), 3.56- 3.63 (m 2H, 2 x H-l), 4.03-4.13 (m, 2H, H-2, H-3), 4.75-4.85 (m, 1H, H-4), 5.43 (dd, 1H, J= 8.2, 15.3 Hz, H-5), 5.91-6.02 (m, 1H, H-6), 7.28 (d, 1H, J= 8.0 Hz, NH), 7.51 (d, 1H, J = 9.0 Hz, NH);13C NMR (101 MHz, CDCI3) δ 14.3 (CH3), 22.8 (CH2), 28.9 (CH2), 29.3 (CH2), 29.5 (CH2), 29.6 (CH2), 29.8 (4 x CH2), 32.1 (CH2), 32.6 (CH2), 55.2 (C-3), 55.8 (C-4), 64.0 (C-l), 69.6 (C-2), 92.2 (CCI3), 92.4 (CCI3), 124.1 (C-5), 139.0 (C-6), 162.3 (C=O), 164.0 (C=O) ESI-HRMS: m / z calculated for C22H36C16N2NaO4[M + Na]+625.0698, found 625.0676.

[0072] Example 11

[0073] N,N '-[ (2S,3R,4R,E)-3,4-Diaminooctadec-5-en-l,2-diol 12

[0074] 2 M aqueous solution of NaOH (1.9 mL) was added to a solution of compound 20 (58 mg, 96 pmol) in EtOH (1.9 mL). After 30 minutes of stirring at 80 °C, the mixture was cooled to room temperature and concentrated. The residue was extracted with CH2CI2(2 x 7 mL), the combined organic layers were dried over anhydrous Na2SO4, and the solvent was evaporated under vacuum. Chromatography of the crude product through a short column of silica gel in the (EtOAc / MeOH, 8:1) setup provided 26 mg (86%) of derivative 12 as a white waxy compound; [a]D21-23.6 (c 0.25, MeOH). IR (neat) v 3224, 2918, 2850, 1677, 1663, 1469, 1056 cm1;1H NMR (400 MHz, CD3OD) δ 0.90 (t, 3H, J= 6.9 Hz, CH3), 1.29-1.43 (m, 20H, 10 x CH2), 2.06-2.11 (m, 2H, CH2), 3.48 (dd, 1H, J = 5.6, 11.3 Hz, H-l), 3.56-3.64 (m, 2H, H-l, H-2), 3.82 (dd, 1H, J = 6.2,

[0075] 8.5 Hz, H-3), 4.21-4.25 (m, 1H, H-4), 5.64 (dd, 1H, J= 8.4, 15.4 Hz, H-5), 5.69-5.76 (m, 1H, H-6);13C NMR (101 MHz, CD3OD) δ 14.4 (CH3), 23.7 (CH2), 30.3 (2 x CH2),

[0076] 30.5 (CH2), 30.6 (CH2), 30.7 (CH2), 30.8 (3 x CH2), 33.1 (CH2), 33.3 (CH2), 58.9 (C- 4), 59.2 (C-3), 64.7 (C-l), 71.9 (C-2), 127.5 (C-5), 136.7 (C-6). ESI-HRMS: m / z calculated for C18H39N2O2[M + H]+337.2826, found 337.2828.

[0077] Example 12

[0078] (2S,3R,4S,E)-3,4-Diaminooctadec-5-en-1,2-diol 13

[0079] 2 M aqueous solution of NaOH (1.5 mL) was added to a solution of compound 21 (46 mg, 76 umol) in EtOH (1.5 mL). After 30 minutes of stirring at 80 °C, the mixture was cooled to room temperature and concentrated. The resulting residue was extracted with CH2CI2(2 X 7 mL), the combined organic layers were dried over anhydrous Na2SO4, and the solvent was evaporated under reduced pressure. Chromatography of the crude product on silica gel (EtOAc / MeOH, 8:1) provided 20 mg (81%) of derivative 13 as a white waxy compound; [a]D21+26.4 (c0.17, MeOH). IR (neat) v3277, 2918, 2849, 1690, 1660, 1440, 1376, 1255 cm1;1H NMR (400 MHz, CD3OD) δ0.90 (t, 3H, J = 6.8 Hz, CH3), 1.29-1.44 (m, 20H, 10 x CH2), 2.04-2.09 (m, 2H, CH2), 3.46-3.51 (m, 2H, H-2, H-3), 3.52-3.60 (m, 2H, 2 x H-l), 4.08-4.15 (m, 1H, H-4), 5.47 (dd, 1H, J = 7.7, 15.3 Hz, H-5), 5.67-5.76 (m, 1H, H-6);13C NMR (101 MHz, CD3OD) δ 14.5 (CH3), 23.8 (CH2), 30.3 (CH2), 30.5 (CH2), 30.6 (CH2), 30.8 (5 x CH2), 33.1 (CH2), 33.2 (CH2) 58.5 (C-4), 62.3 (C-3), 64.5 (C-l), 73.0 (C-2), 131.1 (C-5), 134.9 (C-6). ESI- HRMS: m / z calculated for C18H38N2NaO2[M + Na]+315.3006, found 315.3006.

[0080] Example 13

[0081] Cytotoxic activity

[0082] Cell culture

[0083] Cell lines of Jurkat (human acute T-lymphoblastic leukaemia), HeLa (human cervical cancer), HCT-116 (human colon cancer) were maintained in RPMI 1640 medium, MCF- 7 (human mammary gland adenocarcinoma), BLM (human melanoma cells) were maintained in DMEM medium, and BJ-5ta hTERT (immortalised foreskin fibroblast cells) were maintained in DMEM : M199 medium (4:1) (Biosera, Kansas City, MO, USA), were supplemented with 10% fetal bovine serum and lx HyClone™ antibiotic / antimycotic solution (PNC / STR / AMB) (GE Healthcare, Little Chalfont, UK) in humidified air atmosphere at 37 °C containing 5% CO2. Cell viability was determined by MTT assay.

[0084] Cytotoxicity assay

[0085] The cytotoxic and antiproliferative properties of the tested compounds (Table 1) were determined by colorimetric microculture assay with the MTT end-point. The amount of tetrazolium dye MTT 3-(4,5dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide, reduced to formazan was proportional to the number of living cells. The cells were seeded at density of 5 x 103cells / well in a 96-well polystyrene microplate (SARSTEDT, Numbrecht, Germany). Tested compounds were added at concentrations of 0.1-100 μM, 24 hours after cell seeding. After 72 hours of incubation, 10 μL MTT (5 mg / mL) was added to each well and incubated for an additional 4 hours at 37 °C in 5% CO2, during which the insoluble formazan was produced. To dissolve the formazan, 100 μL of 10% sodium dodecyl sulfate was added to each well and incubated for another 12 hours. The absorbance was measured at 540 nm using a Cytation™ 3 Cell Imaging Multi-Mode Reader (Biotek, Winooski, VT, USA).

[0086] The cytotoxic profile of final compounds 8-13 was evaluated in vitro against two human non-malignant cell lines: BJ and HaCaT. At the same time, the potential of the synthesized derivatives to inhibit the proliferation of cancer cell lines (MCF-7, HeLa, Jurkat, HCT-116, BLM) was also tested. The assay results are expressed as IC50values determined after 72 hours of cell incubation using the MTT assay (Table 1). Sphingoid bases 8, 9, 10 and 13 demonstrated low cytotoxicity against normal fibroblasts (BJ) and keratinocytes (HaCaT) with IC50values ranging from 82.71-111.6 μM (Table 1). These data indicate that the above-mentioned derivatives have a good in vitro safety profile, especially compounds 9 and 13 with IC50> 100 μM. On the other hand, substances 11 (IC50= 61.23 μM for HaCaT) and 12 ( IC50= 65.71 μM for BJ) showed higher cytotoxic effect on non-malignant cells. It seems that diastereoisomers 9 and 13 with (3R,4S)-stereochemistry, were shown to be less toxic in comparison with their (3R,4R) -congeners 8 and 12, with the exception of the pair of stereoisomers 10 and 11.

[0087] The results of preliminary anticancer screening showed weaker antiproliferative potential of these unusual sphingoid bases. Among all the aminoisophytosphingosines tested, analogue 11 was selected as the most active on leukaemia (IC50= 24.61 μM for Jurkat) as well as on solid tumour cells (IC50= 31.19 μM for HCT-116, IC50= 43.73 μM for HeLa).

[0088] Table 1. Cytotoxic activity of substances 8 - 13 on non-malignant cell lines (BJ, HaCaT) and 5 cancer cell lines (BLM, MCF-7, HCT- 116, Jurkat, and HeLa)

[0089] Compound Cell line, ICsoa± SD (pmol x L’1) no.

[0090] Industrial applicability

[0091] The present invention is industrially applicable in the field of consumer chemistry, cosmetics, and dermatology, e.g. in ingredients of preparations for skin and hair care such as creams, body lotions, and hair cosmetics.

[0092] REFERENCIES:

[0093] 1. Zellner, J. Monatschr. Chem. 1911, 36, 133-142.

[0094] 2. (a) Pruett, S.H.; Bushnev, A.; Hagedorn, K.; Adiga, M.; Haynes, C.A.; Sullards, M.C.; Liotta, D.C.; Merrill, Jr. A.H. J. Lipid Res. 2008, 49, 1621-1639 and papers cited therein; (b) Kolter, T.; Sandhoff, K. Angew. Chem. Int. Ed. 1999, 38, 1532-1568.

[0095] 3. Wennekes, T.; van den Berg, R.J.B.H.N.; Boot, R.G.; van der Marel, G.A.; Overkleeft, H.S.; Aerts, J.M.F.G. Angew. Chem. Int. Ed. 2009, 48, 8848-8868 and the papers cited therein.

[0096] 4. C. Romano, C.; Calusen, M.H. Eur. J. Org. Chem. 2022, e202200246 and papers cited therein.

[0097] 5. (a) Wells, G.B.; Dickson, R.C.; Lester, R.L. J. Bioi. Chem. 1998, 273, 7235- 7243; (b) Chung, N.; Jenkins, G.; Hannun, Y.A.; Heitman, J.; Obeid, L.M. J. Bioi. Chem. 2000, 275, 17229-17232.

[0098] 6. (a) Tamiya-Koizumi, K.; Muratte, T.; Suzuki, M.; Simbulan, C.M.G.; Nakagawa, M.; Takemura, M.; Furuta, K.; Izuta, S.; Yoshida, S. Biochem. Moi. Bioi. Int. 1997, 41, 1179-1189.; (b) Jeon, H.; Bae, H.; Baek, D.J. Kwak, Y.-S.; Kim, D.; Kim, S. Org. Biomol. Chem. 2011, 9, 7237-7242] (c) Kang, H.-M.; Son, H.-S. Cui, Y.-H.; Youn, B.; Son, B.; Kaushik, N.K.; Uddin, N.; Lee, J.-S; Song, J.-Y ; Kaushik, N.; Lee, S.-J. Oncotarget 2017, 8, 77794-77808.

[0099] 7. Skolova, B.; Kovacik, A.; Tesar, 0.; Opalka, L.; Vavrova, K. Biochim. Biophys. Acta 2017, 1859, 824-834 and papers cited therein.

[0100] 8. Cho, S.; Kim, H.H.; Lee, M.J.; Lee, S.; Park, C.-S.; Nam, S.-J. Han, J. -J.; Kim, J.-W.; Chung, J.H. J. Lipid Res. 2008, 49, 1235-1245.

[0101] 9. (a) Han, Y; Kim, Y.; Kang, H.; Hong, S.H.; Kim, Y.H.; Lim, D.-S.; Park, C.; Yun, Y.-S; Song, J.-Y. Apoptosis 2006, 11, 581-588; (b) Kim, B.-H.; Lee, J. M.; Jung, Y.-G.; Kim, S.; Kim, T.-Y. J. Invest. Dermatol. 2014, 134, 1023-1032.

[0102] 10. Farwick, M.; Watson, R.E.B.; Rawlings, A.V.; Wollenweber, U.; Lersch, P.; Bowden, J.J.; Bastrilles, J.Y.; Griffiths, C.E.M. Int. J. Cosmet. Sci. 2007, 29, 319-329.

Claims

C L A I M S1. Isomeric aminophytosphingosines of formula (I)whereR = H or (CH3)2C=R1= NH2or NHCOCCI3R2= NH2or H or NHCOCCI3R3= H or NH2or NHCOCCI3or isomeric aminophytosphingosines of formula (II)whereR = H or (CH3)2C=RI = NHAc or NH2.HCIR2= NHAc or H or NH2.HCIR3= H or NHAc or NH2.HCI.

2. Isomeric aminophytosphingosines according to claim 1, which are:N,N '-[(1R,2R,E)-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadec-3-en-1,2- diyl]bis(2,2,2-trichloroacetamide) 16,N,N '-[(1R,2S,E)-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadec-3-en-1,2- diyl]bis(2,2,2-trichloroacetamide) 17,N,N '-[(1R,2R)-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadecane-1,2- diyl]diacetamide 18,N,N '-[(1R,2S)-1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadecane-1,2- diyl]diacetamide 19,N,N '-[(2S,3R,4R -1,2-Dihydroxyoctadecane-3,4-diyl]diacetamide 8,N,N '-[(2S,3R,4S)-1,2-Dihydroxyoctadecane-3,4-diyl]diacetamide 9, (2S,3R,4R) -3),4-Diamino-octadecane-1,2-diol-dihydrochloride 10, (2S,3R,4S)-3,4-Diamino-octadecane-1,2-diol-dihydrochloride 11,N,N '-[(2S,3R,4S,E)-1,2-Dihydroxyoctadec-5-en-3,4-diyl]bis(trichloroacetamide) 20,N,N '-[(2S,3R,4S,E))-1,2-Dihydroxyoctadec-5-en-3,4-diyl]bis(trichloroacetamide) 21, (2S,3R,4S,E)-3,4-Diaminooctadec-5-en-1,2-diol 12, or (2S,3R,4RS,E)-3,4-Diaminooctadec-5-en-1,2-diol 13.

3. A method of production of the compound N,N '-[(1R,2R,E)-1-[(S)-2',2'-Dimethyl- 1',3'-dioxolane-4'-yl]hexadec-3-en-1,2-diyl]bis(2,2,2-trichloroacetamide) 16, or N,N '- [(1R,2S,E) -1-[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadec-3-en-1,2-diyl]bis(2,2,2- trichloroacetamide) 17 according to claim 2, where the compound of formula / where R' =NHCOCCI3and R1'=H, or where R1'=NHCOCCI3and R'=H, is treated with tetradec-1-ene, in the presence of the second-generation Grubbs catalyst in anhydrous CH2CI2to form a corresponding diastereoisomer.

4. The method of production according to claim 3 of the compound[(S)-2',2'-Dimethyl-1',3'-dioxolane-4'-yl]hexadecane-1,2-diyl]diacetamide 18, or[(1R,2R)-1-[(S)-2',2'-Dimethyl- 1',3'-dioxolane-4'-yl]hexadecane-1,2-diyl]diacetamide 19 according to claim 2, where compound 16 or 17 is treated with Et3N, 10% Pd / C, in anhydrous MeOH under H2atmosphere to form a corresponding diastereoisomer.

5. The method of production according to claim 3 of the compound N,N'- [(2S,3R,4R,E)-1,2-Dihydroxyoctadec-5-en-3,4-diyl]bis(trichloroacetamide) 20 orN,N '-[(2S,3R,4S,E )-1,2-Dihydroxyoctadec-5-en-3,4-diyl]bis(trichloroacetamide) 21 according to claim 2, where compound 16 or 17 is treated with p-Toluenesulfonic acid in MeOH at room temperature to form a corresponding diastereoisomer.

6. The method of production according to claim 5 of the compound (2S,3R,4R,E)_3,4-Diaminooctadec-5-en-1,2-diol 12 or (2S,3R,4S,E)_3,4_Diaminooctadec-5-en-1,2- diol 13 according to claim 2, where compound 20 or 21 is treated with a 2 M aqueous solution of NaOH, EtOH at 80 °C to form a corresponding diastereoisomer.

7. The method of production according to claim 4 of the compound N,N'- [(2S,3R,4R) -1,2-Dihydroxyoctadecane-3,4-diyl]diacetamide 8 or N,N['-(2S,3R,4RS)- 1,2-Dihydroxyoctadecane-3,4-diyl]diacetamide 9 according to claim 2, - where compound 18 or 19 is treated with 10% aqueous solution of HCI in THF at room temperature to form a corresponding diastereoisomer.

8. The method of production according to claim 5 of the compound (2S,3R,4R)-3, 4- Diaminooctadecane-1,2-diol-dihydrochloride 10 or (2S,3R,4R)_3),4_Diaminooctadecane-1,2-diol-dihydrochloride 11 according to claim 2, where compound 18 or 19 is treated with 6 M HCI at 65 °C to form a corresponding diastereoisomer.

9. The isomeric aminophytosphingosines according to claim 1 or 2 for use as sphingomimetics.

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