PREPARATION COMPRISING OMEGA-3 FATTY ACID SALTS AND EXTRACTS OF GUM RESINS FROM BOSWELLIA SPECIES

Abstract

Preparations may include at least one extract of gum resins from Boswellia species and at least one omega-3 fatty acid salt selected from eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Surprisingly, such combinations of extracts of gum resins from Boswellia species and polyunsaturated fatty acid salt can cause a marked and unexpected increase in the biosynthesis of specialized pro-resolving mediators (SPM) in a synergistic manner, proposing benefit for resolution of inflammatory conditions.

Claims

1. A preparation, comprising an extract of one or more gum resins from Boswellia species as a Boswellia extract; and a polyunsaturated fatty acid salt comprising (i) an omega-3 fatty acid comprising eicosapentaenoic acid (EPA) and/or docosahexaenoic acid (DHA) and (ii) a basic amino acid.

2. The preparation of claim 1, wherein the Boswellia species comprises Boswellia serrata, Boswellia carterii, Boswellia papyrifera, Boswellia ameero, Boswellia bullata, Boswellia dalzielii, Boswellia dioscorides, Boswellia elongata, Boswellia frereana, Boswellia nana, Boswellia neglecta, Boswellia ogadensis, Boswellia pirottae, Boswellia popoviana, Boswellia rivae, Boswellia sacra, and/or Boswellia socotrana.

3. The preparation of claim 1, wherein the extract is prepared by hydro-distillation, steam distillation, extraction by percolation, extraction under ultrasonic waves, solvent extraction, Soxhlet's extraction, supercritical fluid extraction, or membrane nanofiltration.

4. The preparation of claim 1, comprising a boswellic acid.

5. The preparation of claim 1, further comprising: acid resin, gum; tetratriterpene acid; Pentacyclic triterpene; incensole acetate; phellandrene; (+)-cis-olibanic acid; and/or (+)-trans-olibanic acid.

6. The preparation of claim 1, wherein the omega-3 fatty acid salts comprises an organic counter ion comprising lysine, arginine, ornithine, and/or choline.

7. The preparation of claim 1, comprising at least 10 weight-% of the Boswellia extract.

8. The preparation of claim 1, comprising at least 10 weight-% of the polyunsaturated fatty acid salt.

9. The preparation of claim 1, comprising: a phospholipid.

10. The preparation of claim 9, wherein the phospholipid is present in a mass ratio to fatty acid salt in greater than 0.01.

11. The preparation of claim 1, wherein the Boswellia extract is present in a mass ratio relative to the polyunsaturated fatty acid salt in a range of from 0.5:1 to 1:0.5.

12. The preparation of claim 1, further comprising: a targeted-release formulation.

13. The preparation of claim 1, further comprising: an anthocyanin; vitamin; minerals; fiber; fatty acid; amino acid; and/or a proteins.

14. A tablet, pellet, microparticle, or microparticulate composition, or capsule, comprising: the preparation the preparation of claim 1.

15. A feed, food supplement, pharmaceutical product, or topical application, comprising: the preparation of claim 1.

16. The preparation of claim 1, formulated for use in treating or preventing one or more chronic inflammatory diseases.

17. The preparation of claim 1, comprising beta-boswellic acid, acetyl-beta-boswellic acid, 11-keto-beta-boswellic acid, 3-O-acetyl-11-keto-beta-boswellic acid (AKBA), alpha-boswellic acid, 3-O-acetyl-alpha-boswellic acid, and/or 3-O-acetyl-beta-boswellic acid.

18. The preparation of claim 1, comprising at least 20 weight-% of the Boswellia extract.

Description

WORKING EXAMPLES

[0059] Isolation and Incubations of Macrophages, and LM Metabololipidomics

[0060] Leukocyte concentrates from freshly withdrawn peripheral blood of healthy adult human donors were provided by the Institute of Transfusion Medicine, University Hospital Jena, Germany. The experimental protocol was approved by the ethical committee of the University Hospital Jena. All methods were performed in accordance with the relevant guidelines and regulations. Peripheral blood mononuclear cells (PBMC) were isolated using dextran sedimentation and Ficoll-Histopaque 1077-1 (Sigma-Aldrich, Taufkirchen, Germany) centrifugation. For differentiation and polarization towards M1 and M2, published criteria were used [32]. Thus, M1 were generated by incubating monocytes with 20 ng/ml GM-CSF (Peprotech, Hamburg, Germany) for 6 days in RPMI 1640 supplemented with 10% fetal calf serum, 2 mmol/L 1-glutamine (Biochrom/Merck, Berlin, Germany), and penicillin-streptomycin (Biochrom/Merck), followed by 100 ng/ml LPS (Sigma-Aldrich) and 20 ng/ml INF-γ (Peprotech) treatment for another 48 h. M2 were incubated with 20 ng/ml M-CSF (Peprotech) for 6 days of differentiation plus 20 ng/ml IL-4 (Peprotech) for additional 48 h of polarization.

[0061] Macrophages (2×10.sup.6/ml) were incubated in PBS containing 1 mM CaCl.sub.2). Extracts of Boswellia serrata or vehicle control (0.1% DMSO) were applied 15 min prior to stimulation with E. coli (serotype O6:K2:H1) at a ratio of 1:50 (M1/M2:E. coli) for 180 min at 37° C. Supernatants were transferred to 2 ml of ice-cold methanol containing 10 μl of deuterium-labeled internal standards (200 nM da-55-HETE, d.sub.4-LTB.sub.4, d.sub.5-LXA.sub.4, d.sub.5-RvD2, d.sub.4-PGE.sub.2 and 10 μM d.sub.8-AA) to facilitate quantification. Deuterated and non-deuterated LM standards were purchased from Cayman Chemical/Biomol GmbH (Hamburg, Germany). Sample preparation was conducted by adapting published criteria [33]. In brief, samples were kept at −20° C. for 60 min to allow protein precipitation. After centrifugation (1200 g, 4° C., 10 min) 8 ml acidified H.sub.2O (pH 3.5) was added and subjected to solid phase extraction. Solid phase cartridges (Sep-Pak® Vac 6 cc 500 mg/6 ml C18; Waters, Milford, Mass.) were equilibrated with 6 ml methanol and 2 ml H.sub.2O before samples were loaded onto columns. After washing with 6 ml H.sub.2O and additional 6 ml n-hexane, LM were eluted with 6 ml methyl formiate. Finally, the samples were brought to dryness using an evaporation system (TurboVap LV, Biotage, Uppsala, Sweden) and resuspended in 100 μl methanol-water (50/50, v/v) for UPLC-MS-MS automated injections. LM profiling was analyzed with an Acquity™ UPLC system (Waters, Milford, Mass.) and a QTRAP 5500 Mass Spectrometer (ABSciex, Darmstadt, Germany) equipped with a Turbo VTM Source and electrospray ionization (ESI). LM were eluted using an ACQUITY UPLC® BEH C18 column (1.7 μm, 2.1×100 mm; Waters, Eschborn, Germany) at 50° C. with a flow rate of 0.3 ml/min and a mobile phase consisting of methanol-water-acetic acid of 42:58:0.01 (v/v/v) that was ramped to 86:14:0.01 (v/v/v) over 12.5 min and then to 98:2:0.01 (v/v/v) for 3 min (Table 51). The QTrap 5500 was operated in negative ionization mode using scheduled multiple reaction monitoring (MRM) coupled with information-dependent acquisition. The scheduled MRM window was 60 sec, optimized LM parameters (CE, EP, DP, CXP) were adopted [33], and the curtain gas pressure was set to 35 psi. The retention time and at least six diagnostic ions for each LM were confirmed by means of an external standard (Cayman Chemicals). Quantification was achieved by calibration curves for each LM. Linear calibration curves were obtained for each LM and gave r.sup.2 values of 0.998 or higher (for fatty acids 0.95 or higher).

[0062] Polyunsaturated Fatty Acid Compositions

[0063] In the examples for the present invention, different polyunsaturated fatty acid compositions were used. Different omega-3 fatty acid salts having an organic counter ion selected from the basic amino acids lysine, arginine and ornithine were prepared. The omega-3 fatty acids Eicosapentaenoic acid (C20:5w3c) (EPA) and Docosahexaenoic acid (C22:6w3c) (DHA) are present in a ratio of around 2:1 (ratio EPA: DHA).

[0064] The omega-3 lysine salt (AvailOm®) contains around 32 weight-% of L-lysine and around 65 weight-% of polyunsaturated fatty acids. The major polyunsaturated fatty acids in the composition are the omega-3 fatty acids Eicosapentaenoic acid (C20:5w3c) (EPA) and Docosahexaenoic acid (C22:6w3c) (DHA), summing up to around 58 weight-% of the composition. The composition also contains minor amounts of Docosaenoic acid isomer (incl. erucic acid) (C22:1), Docosapentaenoic acid (C22:5w3c) and of the omega-6 fatty acids Arachidonic acid (C20:4w6) and Docosatetraenoic acid (C22:4w6c).

[0065] The omega-3 arginine salt (omega-3-arg) contains around 35 weight-% of L-arginine and around 64 weight-% of polyunsaturated fatty acids. The major polyunsaturated fatty acids in the composition are the omega-3 fatty acids Eicosapentaenoic acid (C20:5w3c) (EPA) and Docosahexaenoic acid (C22:6w3c) (DHA), summing up to around 49 weight-% of the composition. The composition also contains minor amounts of Docosaenoic acid isomer (incl. erucic acid) (C22:1), Docosapentaenoic acid (C22:5w3c) and of the omega-6 fatty acids Arachidonic acid (C20:4w6) and Docosatetraenoic acid (C22:4w6c).

[0066] The omega-3 ornithine salt (omega-3-orn) contains around 29 weight-% of L-ornithine and around 70 weight-% of polyunsaturated fatty acids. The major polyunsaturated fatty acids in the composition are the omega-3 fatty acids Eicosapentaenoic acid (C20:5w3c) (EPA) and Docosahexaenoic acid (C22:6w3c) (DHA), summing up to around 54 weight-% of the composition. The composition also contains minor amounts of Docosaenoic acid isomer (incl. erucic acid) (C22:1), Docosapentaenoic acid (C22:5w3c) and of the omega-6 fatty acids Arachidonic acid (C20:4w6) and Docosatetraenoic acid (C22:4w6c).

Example 1: Extracts of Gum Resins from Boswellia Species Stimulate LM Biosynthesis Formation in E. coli-Stimulated Human Monocyte-Derived M1 and M2 Macrophages

[0067] Human monocyte-derived macrophages were polarized for 48 hrs to M1 (FIG. 1) or to M2 (FIG. 2) subtypes and subsequently incubated with extracts of gum resins from Boswellia species (Boswellia extracts) BS (Boswellin Super®, Sabinsa Corporation, USA, a standardized extract from the gum resin of B. serrata containing min. 10% AKBA, total identified boswellic acids min. 20%), CAS (Casperome®, Indena, a purified extract, obtained from the gum resin of Boswellia serrata, containing ≥25% boswellic acids), and AUR (Aureliasan extract, a Boswellia carterii extract) (50 μg/mL, each) or AKBA (10 μM) supplemented with (grey bars) or without AvailOm® (3 μg/mL, black bars). After 180 min incubation at 37° C., lipid mediators were isolated by solid phase extraction and analyzed by UPLC-MS-MS. Data are means±S.E.M., n=3. One way ANOVA with log transformed data was performed (Tukey post-hoc test; *, # p<0.05; **, ## p<0.01; ***, ### p<0,005).

[0068] Formation of the LM RvD2, RvD4, RvD5, PDX, PD1, MaR1, 17-HDHA, 14-HDHA, 18-HEPE and LTB.sub.4 and PGE.sub.2 in M1 macrophages is shown in FIG. 1 and in M2 macrophages in shown in FIG. 2.

[0069] In pro-inflammatory M1 macrophages that usually produce low amounts of SPM, the addition of AvailOm® caused only weak increases in LM formation, and also exposure to Boswellia extracts (AUR, BS or CAS) or AKBA did not significantly elevate LM biosynthesis. However, the combination of AvailOm® with Boswellia extracts or AKBA induced significant elevation of EPA- and DHA-derived LM, particularly of RvD5, PD1, PDX, MaR1 and 18-HEPE. In contrast, AA-derived pro-inflammatory LM (PGE2 and LTB4) were not increased under any condition. These data suggest that AvailOm® causes a switch of LM formation in M1 from pro-inflammatory to pro-resolving character.

[0070] More striking effects by the combination of AvailOm® and Boswellia extracts (AUR, BS or CAS) or

[0071] AKBA on EPA- and DHA-derived LM production was evident in M2 (FIG. 2). Surprisingly, while either the Boswellia extracts or AvailOm® had moderate effects, the combination clearly yielded synergistic elevation of lipid mediators, for example for RvD5, PD1, PDX and the precursor 17-HDHA; the same applies to MaR1 and 14-HDHA. The data suggest a synergistic mechanism of SPM formation where Boswellia extracts activate the key enzyme 15-LOX-1 and where AvailOm® serves as available substrate. Supplementation of EPA and/or DHA as substrate (AvailOm®) alone is however not sufficient to induce substantial SPM formation as compared to the combination with Boswellia extracts.

[0072] It can be concluded that supplementation of human M1 and M2 macrophages with AvailOm® in vitro promotes the formation of SPM and their precursors, particularly when cells were activated with AKBA, a pharmacological anti-inflammatory agent, or with Boswellia extracts. These data strongly suggest to combine AKBA or parental Boswellia extracts with AvailOm® to promote formation of pro-resolving LM (i.e. SPM) and consequently to resolve inflammatory disorders.

Example 2: Effects of EPA/DHA Lys-Salt and Free Fatty Acids on Lipid Mediator Biosynthesis Formation in Human Monocyte-Derived M1 and M2 Macrophages

[0073] Human monocyte-derived macrophages were polarized for 48 hrs to M1 (FIG. 3) or to M2 (FIG. 4) subtypes and subsequently incubated with different sources for omega-3 fatty acids: Omegatex (Omegatex5723, 57% EPA, 23% DHA), Omega3 (Omega-3-fatty acid, 57% EPA, 23% DHA), AvailOm®, and liposomal AvailOm® (corresponding to 3 μg/ml EPA plus DHA) supplemented with (grey bars) or without Boswellia extract AUR (50 μg/mL, black bars). After 180 min incubation at 37° C., lipid mediators were isolated by solid phase extraction and analyzed by UPLC-MS-MS. Data are means±S.E.M., n=3. One way ANOVA with log transformed data was performed (Tukey post-hoc test; *, # p<0.05; **, ## p<0.01; ***, ### p<0,005).

[0074] Formation of the LM RvD2, RvD4, RvD5, PDX, PD1, MaR1, 17-HDHA, 14-HDHA, 18-HEPE and LTB.sub.4 and PGE.sub.2 in M1 macrophages is shown in FIG. 3 and in M2 macrophages in shown in FIG. 4.

[0075] Comparison of various sources of DHA and EPA as substrates for SPM/precursors upon supplementation of macrophages showed that in M1, AvailOm® in combination with Boswellia extract AUR caused most prominent elevation of RvD5, PD1, PDX, MaR1, and 14-HDHA, followed by liposomal AvailOm® that gave highest 17-HDHA formation with AUR (FIG. 3). Again, AvailOm® and AUR in combination caused synergistic effects for RvD5, PD1, PDX, MaR1, 17-HDHA and 14-HDHA but no stimulatory effects were evident for AA-derived PGE2 and LTB4.

[0076] In M2 that in general possess higher capacities for SPM production due to the high expression levels of SPM-biosynthetic key enzyme 15-LOX-1, Boswellia extract AUR strongly elevated all investigated SPM and precursors (FIG. 4) in combination with Omega3, AvailOm®, or liposomal AvailOm®. Again, only moderate effects of either AUR or Omega3, AvailOm®, and liposomal AvailOm® or combinations thereof were evident for LTB4 and PGE2 formation.

Example 3: Effects of EPA/DHA Lys-Salt and Free Fatty Acids on Lipid Mediator Biosynthesis Formation in Human Monocyte-Derived M1 and M2 Macrophages

[0077] Human monocyte-derived macrophages were polarized for 48 hrs to M2 subtypes and subsequently incubated with different omega-3 fatty acid salts: omega-3 lysine salt (AvailOm®), omega-3 arginine salt and omega-3 ornithine salt and supplemented with (grey bars) or without Boswellia extract AUR (50 μg/mL, black bars). After 180 min incubation at 37° C., lipid mediators were isolated by solid phase extraction and analyzed by UPLC-MS-MS. Data shown in FIGS. 5 and 6 are means±S.E.M., n=3. One way ANOVA with log transformed data was performed (Tukey post-hoc test; *, # p<0.05; **, ## p<0.01; ***, ### p<0,005).

[0078] Table 1 summarizes the values for the stimulation of LM biosynthesis formation in human M2 macrophages by Boswellia extract AUR and lysine salts of EPA and DHA (AvailOm®) in μg/million cells, table 2 for the arginine salts of EPA and DHA and table 3 for the ornithine salts of EPA and DHA. The values “-fold” refer to the relative fold increase in comparison to the Boswellia extract.

[0079] Those values clearly show that for all three omega-3 salts, there is a synergistic effect with the Boswellia extract on the production of SPM, as the measured values for the combination product is much higher than the sum of the values for the single substances. For example, for all the amino acid salts this effect is drastic referring to the SPMs 17-HDHA, 14-HDHA, 7-HDHA, 4-HDHA, 18-HEPE, 15-HEPE, 12-HEPE, 11-HEPE, 5-HEPE.

TABLE-US-00001 TABLE 1 Stimulation of LM biosynthesis formation in human M2 macrophages by Boswellia extract AUR and lysine salts of EPA and DHA (AvailOm ®), values correspond to pg/million cells Bosw. extr. + veh. lysine salt Bosw. extr. lysine salt -fold RvD1 1.0 ± 0.3   7 ± 2.6 2.0 ± 0.2   8 ± 2.7 3.8 RvD2 0.4 ± 0.1  4.4 ± 2.2 0.8 ± 0.1   5 ± 2.4 6.9 RvD5 1.0 ± 0.3  73 ± 57 3.7 ± 3.5  151 ± 111 40.8 RvD6 .sup. 7 ± 0.6 .sup. 13 ± 3.9 .sup. 6 ± 0.2 23 ± 5 3.8 MaR1 0.3 ± 0.1  28 ± 16 0.3 ± 0.1  44 ± 17 136.6 PD1 1.4 ± 0.3 144 ± 36 2.2 ± 1.5 253 ± 27 116.8 AT-PD1 2.5 ± 0.3 134 ± 13 3.3 ± 0.5 204 ± 7  62.2 PDX 0.7 ± 0.0  27 ± 10 1.0 ± 0.3 40 ± 6 40.9 RvE3 1.8 ± 0.8 85 ± 9 2.6 ± 0.9 .sup. 94 ± 3.0 36.0 LXA4 0.3 ± 0.1  4.5 ± 1.3 2.9 ± 0.8   8 ± 1.8 2.7 5.15-diHETE 4.4 ± 0.5 29 ± 9  10 ± 4.3  49 ± 24 5.1 LTB4 3.4 ± 0.4 .sup. 11 ± 3.9 4.5 ± 0.6 .sup. 17 ± 3.8 3.8 t-LTB4 3.5 ± 0.8 22 ± 5 4.6 ± 0.5 .sup. 34 ± 1.9 7.4 20-OH-LTB4 0.7 ± 0.1 .sup. 14 ± 0.3 3.5 ± 0.2 .sup. 19 ± 0.4 5.3 PGD2 4.0 ± 1.3 11 ± 5  13 ± 2.1 27 ± 7 2.0 PGE2  11 ± 2.0  36 ± 10 38 ± 6   97 ± 15 2.6 PGF2a  21 ± 3.1  43 ± 10  50 ± 3.2 68 ± 7 1.4 TXB2 268 ± 129  421 ± 250 569 ± 243  486 ± 274 0.9 17-HDHA 54 ± 38  3424 ± 1985 188 ± 144  6047 ± 2171 32.1 14-HDHA .sup. 8 ± 2.8 1054 ± 477 31 ± 14 2141 ± 410 69.1 7-HDHA .sup. 7 ± 1.2 1053 ± 149  10 ± 2.2 1402 ± 23  134.5 4-HDHA .sup. 6 ± 2.0  663 ± 163  13 ± 3.7 853 ± 7  64.5 18-HEPE .sup. 8 ± 2.6 14340 ± 1429  14 ± 2.1 18472 ± 371  1325.6 15-HEPE 15 ± 9   4557 ± 3167 37 ± 28  9114 ± 4078 244.1 12-HEPE 1.9 ± 0.9  2927 ± 1199 .sup. 8 ± 2.1 6038 ± 907 782.9 11-HEPE 2.5 ± 1.3 3644 ± 800 .sup. 7 ± 1.5 6556 ± 118 934.4 5-HEPE 3.8 ± 1.3 5272 ± 867 .sup. 7 ± 0.9 7802 ± 379 1090.2 15-HETE 21 ± 12  925 ± 695 206 ± 155  2064 ± 1159 10.0 12-HETE .sup. 7 ± 1.7 174 ± 90  64 ± 3.4  334 ± 132 5.3 11-HETE 3.4 ± 1.4 223 ± 47 26 ± 6  426 ± 13 16.4 AA 2552 ± 733   65339 ± 11504 18222 ± 3471  115317 ± 9345  6.3 EPA 2834 ± 1328 57083 ± 5768 5039 ± 1237  79805 ± 14232 15.8 DHA 8242 ± 3481 71168 ± 8521 15765 ± 4058  107948 ± 13000 6.8

TABLE-US-00002 TABLE 2 Stimulation of LM biosynthesis formation in human M2 macrophages by Boswellia extract AUR and arginine salts of EPA and DHA, values correspond to pg/million cells Bosw. extr. + veh. arginine salt Bosw. extr. arginine salt -fold RvD1 1.0 ± 0.3 2.3 ± 0.4 2.0 ± 0.2 4.6 ± 1.2 2.3 RvD2 0.4 ± 0.1 0.8 ± 0.1 0.8 ± 0.1 2.9 ± 1.5 3.7 RvD5 1.0 ± 0.3 1.4 ± 0.4 3.7 ± 3.5 67 ± 64 18.1 RvD6 .sup. 7 ± 0.6 7.6 ± 0.3 .sup. 6 ± 0.2 .sup. 8 ± 2.1 1.3 MaR1 0.3 ± 0.1 0.4 ± 0.2 0.3 ± 0.1 12 ± 11 38.6 PD1 1.4 ± 0.3 .sup. 7 ± 2.1 2.2 ± 1.5 26 ± 17 12.2 AT-PD1 2.5 ± 0.3 .sup. 7 ± 0.8 3.3 ± 0.5 18 ± 5  5.6 PDX 0.7 ± 0.0 1.3 ± 0.3 1.0 ± 0.3 .sup. 6 ± 4.1 6.3 RvE3 1.8 ± 0.8  29 ± 2.5 2.6 ± 0.9  23 ± 2.5 9.0 LXA4 0.3 ± 0.1 1.2 ± 0.1 2.9 ± 0.8 4.9 ± 1.3 1.7 5.15-diHETE 4.4 ± 0.5 .sup. 6 ± 1.1  10 ± 4.3 32 ± 24 3.3 LTB4 3.4 ± 0.4 4.3 ± 0.4 4.5 ± 0.6 10 ± 6  2.3 t-LTB4 3.5 ± 0.8 4.9 ± 0.4 4.6 ± 0.5  11 ± 3.3 2.4 20-OH-LTB4 0.7 ± 0.1 3.0 ± 0.3 3.5 ± 0.2 .sup. 6 ± 0.2 1.7 PGD2 4.0 ± 1.3 3.3 ± 0.4  13 ± 2.1 20 ± 8  1.5 PGE2  11 ± 2.0  14 ± 2.1 38 ± 6  54 ± 15 1.4 PGF2a  21 ± 3.1 20 ± 5   50 ± 3.2 62 ± 10 1.2 TXB2 268 ± 129 173 ± 43  569 ± 243 548 ± 282 1.0 17-HDHA 54 ± 38 188 ± 33  188 ± 144 1185 ± 935  6.3 14-HDHA .sup. 8 ± 2.8 64 ± 9  31 ± 14 310 ± 220 10.0 7-HDHA .sup. 7 ± 1.2 84 ± 15  10 ± 2.2 198 ± 49  19.0 4-HDHA .sup. 6 ± 2.0 139 ± 20   13 ± 3.7 223 ± 27  16.8 18-HEPE .sup. 8 ± 2.6 941 ± 140  14 ± 2.1 1499 ± 149  107.6 15-HEPE 15 ± 9  74 ± 16 37 ± 28 1401 ± 1209 37.5 12-HEPE 1.9 ± 0.9 115 ± 20  .sup. 8 ± 2.1 563 ± 292 73.0 11-HEPE 2.5 ± 1.3 135 ± 27  .sup. 7 ± 1.5 520 ± 78  74.1 5-HEPE 3.8 ± 1.3 513 ± 124 .sup. 7 ± 0.9 1103 ± 85  154.2 15-HETE 21 ± 12 39 ± 19 206 ± 155 897 ± 793 4.4 12-HETE .sup. 7 ± 1.7  23 ± 1.8  64 ± 3.4 139 ± 81  2.2 11-HETE 3.4 ± 1.4  19 ± 3.3 26 ± 6  87 ± 29 3.3 AA 2552 ± 733  17169 ± 2186  18222 ± 3471  69716 ± 12350 3.8 EPA 2834 ± 1328 41966 ± 1104  5039 ± 1237 64639 ± 9735  12.8 DHA 8242 ± 3481 44473 ± 1803  15765 ± 4058  71498 ± 11435 4.5

TABLE-US-00003 TABLE 3 Stimulation of LM biosynthesis formation in human M2 macrophages by Boswellia extract AUR and ornithine salts of EPA and DHA, values correspond to pg/million cells Bosw. extr. + veh. ornithine salt Bosw. extr. ornithine salt -fold RvD1 1.0 ± 0.3 2.1 ± 0.1 2.0 ± 0.2 2.2 ± 0.5 1.1 RvD2 0.4 ± 0.1 1.0 ± 0.0 0.8 ± 0.1 1.5 ± 0.6 1.9 RvD5 1.0 ± 0.3 1.2 ± 0.4 3.7 ± 3.5 26 ± 23 6.9 RvD6 .sup. 7 ± 0.6 .sup. 7 ± 0.4 .sup. 6 ± 0.2 .sup. 6 ± 0.4 1.0 MaR1 0.3 ± 0.1 0.1 ± 0.0 0.3 ± 0.1 3.2 ± 2.9 9.9 PD1 1.4 ± 0.3 2.3 ± 0.8 2.2 ± 1.5 9 ± 5 4.2 AT-PD1 2.5 ± 0.3 3.3 ± 0.1 3.3 ± 0.5 .sup. 8 ± 2.2 2.5 PDX 0.7 ± 0.0 0.9 ± 0.2 1.0 ± 0.3 2.7 ± 1.4 2.7 RvE3 1.8 ± 0.8 .sup. 8 ± 0.8 2.6 ± 0.9 .sup. 6 ± 1.4 2.2 LXA4 0.3 ± 0.1 0.4 ± 0.1 2.9 ± 0.8 3.6 ± 0.5 1.3 5.15-diHETE 4.4 ± 0.5 4.2 ± 0.8  10 ± 4.3 17 ± 9  1.8 LTB4 3.4 ± 0.4 3.9 ± 0.2 4.5 ± 0.6 .sup. 6 ± 1.9 1.4 t-LTB4 3.5 ± 0.8 4.8 ± 0.5 4.6 ± 0.5 .sup. 6 ± 1.2 1.4 20-OH-LTB4 0.7 ± 0.1 1.2 ± 0.1 3.5 ± 0.2 3.8 ± 0.4 1.1 PGD2 4.0 ± 1.3 3.4 ± 0.8  13 ± 2.1  21 ± 4.9 1.5 PGE2  11 ± 2.0  12 ± 2.2 38 ± 6  40 ± 7  1.1 PGF2a  21 ± 3.1 24 ± 8   50 ± 3.2 59 ± 13 1.2 TXB2 268 ± 129 220 ± 56  569 ± 243 627 ± 372 1.1 17-HDHA 54 ± 38 117 ± 38  188 ± 144 497 ± 354 2.6 14-HDHA .sup. 8 ± 2.8  31 ± 3.0 31 ± 14 120 ± 75  3.9 7-HDHA .sup. 7 ± 1.2  38 ± 3.8  10 ± 2.2 77 ± 12 7.4 4-HDHA .sup. 6 ± 2.0 35 ± 7   13 ± 3.7 62 ± 9  4.7 18-HEPE .sup. 8 ± 2.6 406 ± 56   14 ± 2.1 591 ± 44  42.4 15-HEPE 15 ± 9  57 ± 19 37 ± 28 551 ± 440 14.8 12-HEPE 1.9 ± 0.9 51 ± 8  .sup. 8 ± 2.1 204 ± 94  26.4 11-HEPE 2.5 ± 1.3 56 ± 12 .sup. 7 ± 1.5 176 ± 22  25.1 5-HEPE 3.8 ± 1.3 157 ± 34  .sup. 7 ± 0.9 296 ± 24  41.4 15-HETE 21 ± 12 22 ± 8  206 ± 155 358 ± 284 1.7 12-HETE .sup. 7 ± 1.7 21 ± 6   64 ± 3.4 55 ± 32 0.9 11-HETE 3.4 ± 1.4 .sup. 8 ± 1.9 26 ± 6  40 ± 11 1.5 AA 2552 ± 733  15574 ± 2895  18222 ± 3471  53632 ± 5295  2.9 EPA 2834 ± 1328 45097 ± 1406  5039 ± 1237 65206 ± 10780 12.9 DHA 8242 ± 3481 47473 ± 2492  15765 ± 4058  69161 ± 9918  4.4

[0080] The formation of the LM RvD2, RvD4, RvD5, PDX, PD1, MaR1, 17-HDHA, 14-HDHA, 18-HEPE and LTB.sub.4 and PGE.sub.2 in M2 macrophages for omega-3 lysine (AvailOm®), arginine and ornithine salts are shown in FIG. 5.

[0081] FIG. 6 shows the stimulation of LM biosynthesis formation in human M2 macrophages for omega-3 arginine and ornithine salts.

Example 4: Capsule Comprisinq EPA/DHA Lys-Salt and Boswellia Extract

[0082] The following components were filled in HPMC capsules:

TABLE-US-00004 TABLE 4 Preparations for filling into HPMC capsules. Compound Capsule I Capsule II Capsule III Omega-3 lysine salt 400 mg 200 mg 50 mg (AvailOm ®) Boswellia extract 400 mg 200 mg 50 mg

[0083] The capsules may further contain amino acids selected from L-ornithine, L-aspartate, L-lysine and L-arginine. The capsules may further contain further carbohydrate ingredients, selected from arabinoxylans, barley grain fibre, oat grain fibre, rye fibre, wheat bran fibre, inulins, fructooligosaccharides (FOS), galactooligosaccharides (GOS), resistant starch, beta-glucans, glucomannans, galactoglucomannans, guar gum and xylooligosaccharides.

[0084] The capsules may further contain one or more plant extracts, selected from ginger, cinnamon, grapefruit, parsley, turmeric, curcuma, olive fruit, panax ginseng, horseradish, garlic, broccoli, spirulina, pomegranate, cauliflower, kale, cilantro, green tea, onions, and milk thistle. The capsules may further contain charcoal, chitosan, glutathione, monacolin K, plant sterols, plant stanols, sulforaphane, collagen, hyalurone. The capsules may comprise further vitamins selected from biotin, vitamin A, vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B9 (folic acid or folate), vitamin C (ascorbic acid), vitamin D (calciferols), vitamin E (tocopherols and tocotrienols) and vitamin K (quinones) or minerals selected from sulfur, iron, chlorine, calcium, chromium, cobalt, copper, magnesium, manganese, molybdenum, iodine, selenium, and zinc.

Example 5: Enteric Delivery Capsule Comprising EPA/DHA Lys-Salt and Boswellia Extract

[0085] The capsules as prepared in example 3 were coated with an enteric coating composition:

TABLE-US-00005 TABLE 5 Coating composition Dry Content Content substance based on Weight based on Compound [g] coating [%] gain [%] capsule [%] EUDRAGUARD ® 40.8 36.9 8.2 6.7 biotic HPMC 43.1 39.0 8.6 7.1 Talc 20.4 18.4 4.0 3.3 Polyethylene 4.3 3.9 0.9 0.7 glycol Triethyl citrate 2.0 1.8 0.4 0.3

Example 6: Tablet Formulation Comprising EPA/DHA Lys-Salt and Boswellia Extract

[0086] A formulation (table 6) was prepared and used for tableting. The tablet core components (except magnesium stearate) at their corresponding mass in the respective formulation were blended by the use of a turbola blender. Magnesium stearate was added in a second blending step just before the compression process.

TABLE-US-00006 TABLE 6 Preparation for preparation of tablets Substance Content [weight-%] AvailOm ® 25 Phospholipids (Lipoid H 20) 12.9 Boswellia serrata extract (AureliaSan) 12.5 Dicalcium phosphate anh. (Cafos N201) 19 Micro crystalline cellulose (Avicel 200) 20 Croscarmellose-Na (Ac-Di-Sol SD 711) 2.6 Magnesium stearate 3 Crospovidone (Polyplasdone XL) 5 Sum total 100

[0087] Tablet compression was conducted on a Korsch XP 1 eccentric press. A 21 mm×9 mm oval biconvex tooling was used to gain tablets of a target weight of 1000 mg. A compression force of approx. 15 kN resulted in tablets of a hardness (resistance to crushing) of approx. 75N. The friability was below 1% and the uniformity of mass showed variation less than 5%.

[0088] As a final production step, the resulting tablets are coated with a EUDRUARD Natural based coating. The coating provides a taste- and odor masking as well as a barrier against moisture uptake and can improve photostability. The coating was conducted on a O'Hara Labcoat drum coater with a perforated 15″ drum. 4.0 mg/cm.sup.2 of the coating suspension consisting of EUDRAGUARD Natural, Talc, Glycerol and Chlorophyll E 141 ii were applied at an average spraying rate of 5.5 g/min/kg.

[0089] The resulting film coated tablets were intact and non-aggregated. The characteristic smell of AvailOm® and Boswelia serrata extract was significantly masked. The film coated tablets disintegrated in 0.1 N HCl pH 1.20 within 30 min, provided a uniformity of mass with a variation less than 5%, and the water content was between 4-7%.

[0090] The compressed tablets were stable for at least two months at 25° C. and 60% relative humidity and at 40° C. and 75% relative humidity.

[0091] The tablet may comprise further vitamins selected from biotin, vitamin A, vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B9 (folic acid or folate), vitamin C (ascorbic acid), vitamin D (calciferols), vitamin E (tocopherols and tocotrienols) and vitamin K (quinones) or minerals selected from sulfur, iron, chlorine, calcium, chromium, cobalt, copper, magnesium, manganese, molybdenum, iodine, selenium, and zinc.

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