Nutraceutical composition based on dry extract of boswellia serrata gum resin

Abstract

The object of the invention is a nutraceutical composition comprising dry extract of Boswellia Serrata gum resin in a quantity comprised between 50% and 90% by weight, stearic acid in a quantity comprised between 45% and 4% by weight and a source of Phosphadityl-Choline in a quantity comprised between 5% and 1 % by weight. The invention also relates to a process for the preparation of the nutraceutical composition. The invention is inter alia intended to increase the bioavailability of the product.

Description

[0001] TITLE: NUTRACEUTICAL COMPOSITION BASED ON DRY EXTRACT OF BOSWELLIA SERRATA GUM RESIN

[0002] Description Text

[0003] FIELD OF THE INVENTION

[0004] The present invention has as its object a nutraceutical composition based on dry extract of gum resin of Boswellia Serrata.

[0005] PRIOR ART

[0006] As is known, Boswellia Serrata is a plant of the Burseraceae family widely used for various therapeutic purposes ("Boswellia serrata, a potential antiinflammatory agent: an overview" Indian J Pharm Sci. 2011 May-Jun; 73(3): 255-261 ). The dry extract of the gum resin secreted by the plant is widely used in the formulation and packaging of nutraceuticals (also called food supplements). In fact, the use of the plant in food supplements is authorised by the E.F.S.A. (European Food Safety Authority), without daily dosage limits and without warnings (List of allowed plants and plant extracts, Annex I - Botanicals E.F.S.A.). To the dry extract of the gum resin, the E.F.S.A. attributes certain health claims, concerning the joint functionality, the digestive system, and the contrast of localised states of tension (List of allowed plants and plant extracts, Annex I - Botanicals E.F.S.A.). The dry extract of Boswellia Serrata gum resin has a defined percentage content in the pharmacologically active substances, Boswellic Acids, usually between 25 and 90%. The dry extract of the gum resin of Boswellia Serrata is typically processed alone or in a mixture with other active substances (those allowed for use in food supplements in general, and in particular those with similar health purposes recognized by the E.F.S.A., such as Bromelain, Turmeric, Glucosamine, Chondroitin, Horse Chestnut and others) and excipients of various kinds such as maltodextrins, microcrystalline cellulose, talc, magnesium salts of fatty acids, gum arabic, silicon dioxide and others.

[0007] The processing of the dry extract of gum resin does not present particular difficulties, either alone or in mixture with other substances, whether they are tablets or capsules, or whether they are granules or powders to be used in single-dose sachets. The effectiveness of dry gum resin extract has been proven by numerous published and indexed studies in the various health areas accredited by the E.F.S.A. (Inflammatory Bowel Disease, Alyssa M. Parian MD, Amy C. Brown PhD, in Integrative Medicine (Fourth Edition), 2018. “Effectiveness of Boswellia and Boswellia extract for osteoarthritis patients: a systematic review and meta-analysis” Yu et al. BMC Complementary Medicine and Therapies (2020) 20:225).

[0008] The disadvantages of the dry extract of Boswellia Serrata gum resin are not related to its preparation, nor to its processing alone or in a mixture, which, as stated in point 1 , can be made quickly and economically, whatever the percentage content in Boswellic Acids of the dry extract (from 25 to 90%), whatever the chosen pharmaceutical form (tablets, capsules, granules in sachets) for the manufacture of the finished nutraceutical product.

[0009] The disadvantages lie essentially in the poor bioavailability of the substance under conditions of intake on an empty stomach. It has been determined that the intake of the dry extract under conditions of an empty stomach (away from meals) results in an extremely low plasma concentration of the pharmacologically active compounds (Boswellic Acids, and in particular the most active ones, alpha-Boswellic Acid, beta- Boswellic Acid, AKBA and KBA, respectively Acetyl Keto Boswellic Acid and Keto Boswellic Acid). The plasma concentration of the active ingredients of the dry extract can be so low, under empty stomach conditions, that it is even below the limit of detectability of the analytical instruments used to measure it, in particular HPLC (High Performance Liquid Chromatography) with photodiode detector ("Effect of Food Intake on the Bioavailability of Boswellic Acids from a Herbal Preparation in Healthy Volunteers" Verk et al. Planta med 2004; 70: 1155-1160). Taking Boswellia dry extract on an empty stomach also leads, in addition to the insignificant plasma concentration, to a significant decrease in the plasma half-life, i.e. the time necessary to halve the concentration of the compounds analysed ("Effect of Food Intake on the Bioavailability of Boswellic Acids from a Herbal Preparation in Healthy Volunteers" Verk et al. Planta med 2004; 70: 1155-1160).

[0010] On the other hand, it has been determined that, instead, the intake of the dry extract of Boswellia Serrata gum resin on a full stomach entails a considerable increase not only in the plasma concentration of the active molecules (Boswellic Acids), but also a considerable increase in the plasma half-life. These effects, potentially very positive in terms of health benefits, were particularly decisive in the case of taking the dry extract of gum resin in conjunction with meals with a markedly fatty character, also defined in the literature as "fatty meals" ("Effect of Food Intake on the Bioavailability of Boswellic Acids from a Herbal Preparation in Healthy Volunteers" Verk et al. Planta med 2004; 70: 1155-1160).

[0011] The disadvantage of the poor bioavailability of the dry extract of gum resin on an empty stomach could, in principle, be circumvented by recommending the intake of the substance at meals (and it is in these conditions that the studies aimed at proving the effectiveness of the substance have been carried out).

[0012] However, the disadvantage persists for two different and equally critical reasons:

[0013] 1 . It is unreasonable to prescribe to the consumer of dry extract to place himself in the ideal conditions for the maximum bioavailability of the active substances, or to feed mainly with meals with a strong fat composition (fatty meal).

[0014] 2. Taking the dry extract of Boswellia Serrata gum resin on a full stomach would exclude, for obvious pharmacokinetic contrast, the association of this plant with any other substance authorised by the E.F.S.A. that must instead be taken on an empty stomach, away from meals. This evidence would exclude, for example, the possibility of associating the dry extract of gum resin with all that class of substances known as "proteolytic enzymes", which are instead strictly recommended to be taken on an empty stomach (characteristic example: Bromelain from Ananas Comosus).

[0015] On the other hand, the intake of the dry extract of gum resin on a necessarily full stomach (and better if in "fatty meal" conditions), would force the user to consume on a full stomach, at the meal, also any associations of Boswellia with substances whose intake is instead completely independent of meals (characteristic example: Glucosamine from shellfish exoskeleton or from bio-fermentation of plant substances), with the risk of a possible decrease in overall compliance.

[0016] The document WO 2021 / 019037 A1 describes a composition comprising a dry extract of the gum resin of Boswellia Serrata, Omega-3 polyunsaturated fatty acids with 20 and 22 Carbon atoms, and specifically EPA (Eicosapentaenoic Acid) and DHA (Docosahexaenoic Acid), salified with an amino acid base.

[0017] It is also known the commercial product called Nature's Way Standardized Boswellia which consists of a dry extract of the gum resin of Boswellia Serrata at a dosage of 307 mg per tablet, with a titer of 40% in boswellic acids, for a dosage of 122.8 mg per tablet. Other declared ingredients are cellulose, croscarmellose sodium, silica, stearic acid, organic maltodextrins, organic sunflower lecithin, organic palm oil, organic guar gum.

[0018] In the publication "Enhanced absorption of boswellic acids by a lecithin delivery form (Phytosome®) of Boswellia extract". Jan Husch et al. Fitoterapia 84 (2013) 89-98) it is described a phytosomal composition that has the purpose of increasing the bioavailability of pharmacologically active boswellic acids (BAs), contained in a dry gum-resin extract of the plant. This preparation is phytosomal, that is, the dry extract of the Boswellia Serrata gum resin is emulsified in Soy Lecithin and then finely divided to obtain spherical phytosomal particles, which mimic the double phospholipid layer of cell membranes.

[0019] An object of the present invention is therefore to overcome the aforementioned problems of the prior art, by providing a nutraceutical composition having high bioavailability of the substance under conditions of intake on an empty stomach.

[0020] A further object of the present invention is to obtain the aforesaid results in an economical and practical manner.

[0021] Other objects and advantages of the invention will become apparent from the following description.

[0022] BRIEF SUMMARY OF THE INVENTION

[0023] The described objects are obtained from a nutraceutical composition comprising dry extract of Boswellia Serrata gum resin in a quantity comprised between 50% and 90% by weight, stearic acid in a quantity comprised between 45% and 4% by weight and a source of Phosphadityl Choline in a quantity comprised between 5% and 1 % by weight. The invention also comprises a process for the preparation of the described nutraceutical composition, wherein the following steps are provided: definition of the relative proportions of dry extract of Boswellia Serrata gum resin, stearic acid and the source of Phosphadityl Choline; precise determination of the weights of the dry extract of Boswellia Serrata gum resin, of the stearic acid and of the Phosphadityl Choline source taken individually; sieving of the dry extract of Boswellia Serrata gum resin, of the stearic acid and of the Phosphadityl Choline source taken individually; mixing of the stearic acid with the Phosphadityl Choline source; addition in progressive amounts of the dry extract of Boswellia Serrata gum resin to the mixture of stearic acid and of the source of Phosphaditil-Choline; and - final sieving of the dry extract composition of Boswellia Serrata gum resin, stearic acid and the source of Phosphadityl Choline.

[0024] Further characteristics of the invention can be deduced from the dependent claims.

[0025] DESCRIPTION OF THE DRAWINGS figure 1 illustrates the plasma concentrations of [3-Boswellic Acid in the different treatments A and B described below; figure 2 illustrates the plasma concentrations of a-Boswellic Acid in the different treatments A and B; figure 3 illustrates the plasma concentrations of 11 -keto-[3-Boswellic Acid in the different treatments A and B; and figure 4 illustrates the plasma concentrations of acetyl-11 -keto-[3- Boswellic Acid in the different treatments A and B.

[0026] DETAILED DESCRIPTION OF THE INVENTION

[0027] According to a first aspect of the invention, it provides for the combination of the three following substances:

[0028] 1 ) Dry extract of Boswellia Serrata gum resin.

[0029] 2) Stearic Acid. 3) Sunflower Lecithin.

[0030] The three substances will be combined by mixing in the solid state, in a mutual relative percentage ratio within a range.

[0031] In particular, the composition comprises dry extract of Boswellia Serrata gum resin in a quantity comprised between 50% and 90% by weight, stearic acid in a quantity comprised between 45% and 4% by weight and a source of Phosphadityl Choline in a quantity comprised between 5% and 1 % by weight.

[0032] In particular, the source of Phosphadityl Choline can be chosen from sunflower lecithin, soya lecithin and pure Phosphadityl Choline or combinations thereof.

[0033] Two examples are now provided within the range defined above of possible composition of the combination preparation.

[0034] 1 ) Dry extract of Boswellia Serrata gum resin in a quantity of 92.5% by weight, stearic acid in a quantity of 6.5% and sunflower lecithin in a quantity of 1 % by weight.

[0035] 2) Dry extract of Boswellia Serrata gum resin in a quantity equal to 52% by weight, stearic acid in a quantity equal to 45% and sunflower lecithin in a quantity equal to 3% by weight.

[0036] The novelties introduced by the invention are therefore the combination, by solid state mixing, of the dry extract of Boswellia Serrata gum resin with Stearic Acid and Sunflower Lecithin, in the proportions described above, in order to improve the absorption, and therefore the plasma concentration and half-life, of Boswellia Serrata and its active ingredients even under conditions of an empty stomach. In fact, it has been shown that taking dry extract of Boswellia Serrata gum resin on a full stomach, and in particular at a meal with a high lipid content, greatly increases the absorption of the active ingredient ("Effect of Food Intake on the Bioavailability of Boswellic Acids from a Herbal Preparation in Healthy Volunteers" Verk et al. Planta med 2004; 70: 1155-1160).

[0037] The combination of the three substances indicated above is intended to mimic the intake of the pharmacologically active ingredient under conditions of a full stomach with high lipid foods, with the aim of achieving ideal absorption even on an empty stomach, and more generally regardless of meals. This will allow Boswellia Serrata to be suitably associated, in nutraceutical preparations, with any substance allowed by the E.F.S.A in food supplements, to be taken necessarily on an empty stomach (example: Bromelain from Ananas Comosus) or regardless of meals (example: Glucosamine).

[0038] According to a relevant aspect of the invention, the dry extract of Boswellia Serrata gum resin is mixed in the solid state with Stearic Acid, in the proportions indicated above, within the range defined above; Stearic Acid is a saturated carboxylic acid with 18 carbon atoms, with the I.U.P.A.C. (International Union of Pure and Applied Chemistry) name Octadecanoic Acid. Stearic Acid has a strong structural analogy with Oleic Acid (I.U.P.A.C. name cis-9-Octadecenoic Acid), a mono-unsaturated fatty acid which, in the form of triglyceride, makes up 75% of the composition of olive oil, giving this food its characteristic lipophilicity. Olive oil is in fact among the most lipid-rich foods, with a composition consisting of 98% fat ("Lipid Profiling and Stable Isotopic Data Analysis for differentiation of Extra Virgin Olive Oils Based on Their Origin" Mattivi et al. Molecules 2020, 25, 4). Among the fatty acids, Stearic Acid has been identified as the most suitable to play the role of camouflage of a fatty meal, for a series of chemical, physical, technical and economic requirements:

[0039] 1. It is a solid substance, white in color, with melting point at 68.8°C and boiling temperature at 350°C, these requirements are important for the mixing process with the dry extract of Boswellia Serrata gum resin (NCBI, National Center for Biotechnology Information, PUBCHEM databank).

[0040] 2. It has proved to be stable under the temperature and pressure conditions of the processing process necessary for the preparation of the mixture with dry extract of Boswellia gum resin and with Sunflower Lecithin, in the range of relative percentage proportions defined above.

[0041] 3. It is a substance allowed by the E.F.S.A. for use in food supplements, as an additive, without dosage restrictions and without specific warnings.

[0042] 4. It is a substance already widely used in the food sector, with widespread and consolidated practice, considered not only harmless, but even beneficial for cardiovascular health ("Short-term diets enriched in stearic or palmitic acids do not alter plasma lipids, platelet aggregation or platelet activation status" Kelly FD, Sinclair AJ, Mann NJ, Turner AH, Raffin FL, Blandford MV, Pike MJ, Eur J Clin Nutr. 2002 Jun;56(6):490-9. “A stearic acid-rich diet improves thrombogenic and atherogenic risk factor profiles in healthy males” FD Kelly 1 , AJ Sinclair, NJ Mann, AH Turner, L Abedin, D Li, Clinical Trial Eur J Clin Nutr 2001 Feb;55(2):88-96).

[0043] 5. It is an inexpensive and easily available substance on a continuous basis.

[0044] Another relevant aspect of the invention is the presence, in solid state mixture with dry extract of Boswellia Serrata gum resin and Stearic Acid, of Sunflower Lecithin, in the relative percentage proportions defined within the range specified above; the lecithins, in general, are mixtures of glycerophospholipids, including Phosphatidyl-Choline (PC), Phosphatidyl-ethanolamine (PE), Phosphatidyl-inositol (PI), Phosphatidyl-serine (PS) and Phosphatidic acid.

[0045] Phospholipids are an essential component of all cell membranes, both animal and plant. Among the plants with the highest Phospholipid content we include Soya, Sunflower, Walnuts, Wheat (Szuhaj, B. F. Lecithins. In Bailey's Industrial Oil and Fat Products, 6th ed.; Shahidi, F., Ed.; Wiley: New York, 2005; Vol. 2, pp. 361 -456). Phospholipids, along with proteins, are the main structural components of the membranes of all cell types. Phospholipids play an important role in different biological functions, such as maintaining the integrity of the cell membrane, preventing certain neurological diseases and regulating basic biological processes such as the transmission of signals between cells, which is essential for life (Guo, Z.; Vikbjerg, A.; Xu, X. Enzymatic Modification of Phospholipids for Functional Applications and Human Nutrition. Biotechnol Adv. 2005, 23, 203-259).

[0046] The key biological role of Phospholipids is the ability of the phosphate group to combine with water and hydrophilic molecules such as proteins, sugars and oxygen, while fatty acids interact with hydrophobic substances such as hormones and non- esterified fatty acids (Quezada, A. N. Synthesis of Structured Phospholipids with Conjugated Linolenic Acid, and Evaluation of Their Physical Properties. Master's Thesis, Texas A&M University, 2007). The beneficial effects of dietary Phospholipids (PLs) have been mentioned since the early 1900s in relation to several diseases and symptoms, such as coronary heart disease, inflammation and cancer (Kullenberg, D.; Taylor, L. A.; Schneider, M.; Massing, II. Health Effects of Dietary Phospholipids. Lipids Health Dis. 2012, 11 (3), 1-16).

[0047] For these reasons, Phospholipids have also been used as dietary supplements for health and therapeutic purposes. Lecithins are widely used in foods as emulsifying agents, and are not only harmless and well tolerated, and distinctly non-toxic (the F.D.A. classifies them as G.R.A.S., Generally Recognized As Safe), but thanks to their high content of Phospholipids they boast the health properties described above. The Lecithin chosen in this invention is that of Sunflower, easily found in the solid state as a powder. Compared to Soya Lecithin, it boasts a slightly higher content of Phosphatidyl-Choline (PC), estimated at 16% vs 15% (Sunflower Lecithin, E.N. Guiotto et al. Polar Lipids - Biology, Chemistry and Technology 2015, 57-75).

[0048] The association with dry extract of Boswellia Serrata gum resin arises from the hypothesis, confirmed by subsequent analytical determinations, that the Phospholipids of which it is the source, with their amphiphilic characteristics, can optimize the non- covalent interactions of the invented mixture with the double phospholipid layer of cell membranes, by chemical-physical affinity, thus determining an increase in the absorption of Boswellic Acids, compared to a dry extract of gum resin not associated with Sunflower Lecithin and Stearic Acid.

[0049] Sunflower Lecithin is stable under the pressure and temperature conditions of the processing process required to obtain the mixture with dry extract of Boswellia Serrata gum resin and Stearic Acid, in all possible proportions within the range defined above.

[0050] A possible variant of the combination described above would consist in the mixing, in the solid state, of dry extract of Boswellia Serrata gum resin (with a Boswellic Acid titre between 25 and 90%), Stearic Acid and Phosphatidyl-Choline, in the same range of relative proportions proposed for the invented mixture; pure Phosphatidyl-Choline would therefore replace Sunflower Lecithin, chosen precisely as a source of Phosphatidyl-Choline. This alternative solution did not determine, in the analytical measurements carried out, an increase in the bioavailability of Boswellic Acids such as to justify the replacement of an inexpensive and easily available natural substance, such as Sunflower Lecithin, with a pure substance that is much more complex to obtain and much more expensive.

[0051] Similarly, the hypothesis of adding Phosphatidyl-Choline to the invented combination was considered, but not realised, with the aim of further implementing the bioavailability of Boswellic Acids. This variant was not added as the results obtained with the invented mixture were already significantly improved compared to the dry extract of Boswellia alone, and therefore the possible variant by adding Phosphatidyl-Choline to the three basic components of the mixture was considered not strictly necessary.

[0052] For the preparation of the combination in the composition described above, the three substances are weighed individually, following the recipe given (i.e. once the relative proportions between the three substances are defined, within the range indicated above), on a technical balance under a laminar flow hood. After weighing, each of the three substances is then sieved, through manual sieves of 16 mesh / cm2 for the substances with the lowest percentage (Sunflower Lecithin and Stearic Acid), and through the automatic vibrating screen, for the substance present in the highest percentage (dry extract of the Boswellia Serrata gum resin).

[0053] Once sieved, the substances with lower relative weight in the composition of the combination, i.e. Stearic Acid and Sunflower Lecithin, are mixed together, until a homogeneous core is obtained (solid state mixture of Stearic Acid and Sunflower Lecithin). This homogeneous core is subsequently "diluted" with the addition of progressive, always equal, amounts of dry extract of Boswellia Serrata gum resin in granules, according to the "progressive dilution" technique (in the solid state).

[0054] Once the mixing of all three substances constituting the invention has been completed, a homogeneous solid is obtained, ready to be possibly combined with other substances (example, as described above: Bromelain, Glucosamine et al.), through the well-known "vacuum loading" system.

[0055] Before its possible use in mixture with other substances, or in any case before being further processed (in tablets, capsules, granules in sachets), the solid combination Boswellia Serrata dry extract / Stearic Acid / Sunflower Lecithin is subjected to a further sieving process on a 16 mesh / cm2 sieve, obtaining a homogeneous and finely divided compound.

[0056] As already specified above, a possible variant could be the use of pure Phosphaditil- Choline instead of its natural source, Sunflower Lecithin, to be preferred in any case for its costs, its availability, and for the increase in bioavailability that it already guarantees; another variant, as already mentioned above, could be the addition, to the mixture constituting the invention, of an aliquot of Phosphaditil-Choline, with the well- founded prediction of a further increase in the bioavailability of Boswellic Acids. In this case, a solid state mixture of the minority substances, i.e. Stearic Acid / Sunflower Lecithin / Phosphadityl Choline, would first be obtained as a homogeneous core, and progressive, and always equal, amounts of dry extract granules of the Boswellia Serrata gum resin would be added to it, according to the method of progressive dilutions.

[0057] From a technical-preparative point of view, there are no particular differences or complexity with respect to the combination of Dry extract of gum resin of Boswellia Serrata / Stearic Acid / Sunflower Lecithin, in the proportions defined above; the only difference would consist in the composition of the homogeneous core to which the dry extract of the gum resin of Boswellia Serrata would be added by the method of progressive dilutions (homogeneous core with three "minority" substances instead of two). The substantial difference would lie in a substantial increase in costs, as well as in the greater complexity of the preparatory process (four weighings instead of three, a more complex primordial core).

[0058] Another variant could be the choice, as a source of Phosphaditil-Choline, of Soya Lecithin. Soya Lecithin is a slightly less rich, but still comparable, source of Phosphaditil-Choline, compared to Sunflower Lecithin (Sunflower Lecithin, E.N. Guiotto et al. Polar Lipids - Biology, Chemistry and Technology 2015, 57-75).

[0059] SUPPORTING ANALYTICAL DATA

[0060] Criteria used for the collection of bioavailability data of the Boswellia Serrata / Stearic Acid / Sunflower Lecithin combination (within the relative ratios described in the patent application) versus dry extract of gum resin of Boswellia Serrata:

[0061] To assess whether and by how much the composition presented in the application increases the bioavailability of pharmacologically active Boswellic Acids compared to a dry gum resin extract as such, a method inspired by previous scientific literature was applied, and in particular to the publication "Effect of Food Intake on the Bioavailability of Boswellic Acids from a Herbal Preparation in Healthy Volunteers", Sterk et Al., Planta Med. 2004; 70: 1155-1160, with particular reference to the design and layout of study1, as summarized below.

[0062] Twelve healthy male volunteers were enrolled, without ongoing pharmacological therapies, with normal blood chemistry parameters, negative for HIV, Hepatitis B and Hepatitis C, with regular ECG traces and normal blood pressure values. In the aforementioned study, the 12 volunteers received the same administration of dry extract of gum resin of Boswellia Serrata, at the same dose, in two different conditions (empty stomach or full stomach for a fatty meal), after a wash out of a week, according to a monocentric "two ways cross-over" design.

[0063] In our case, the 12 healthy volunteers instead receive two different preparations, but in the same conditions, that is, always on an empty stomach, also here after a one- week wash out. The first administration (TREATMENT A) consisted of three capsules of dry extract of Boswellia Serrata 65% in boswellic acids (a-boswellic acid, [3-boswellic acid, 11 -keto-[3-boswellic acid, acetyl-11-keto-[3-boswellic acid), of 300 mg each, for a total of 900 mg, on an empty stomach, as a single oral dose. The second administration (TREATMENT B), after the one-week wash out, consisted of three capsules each containing 300 mg of the Boswellia Serrata / Stearic Acid / Sunflower Lecithin combination (within the relative ratios described in the patent application), for a total of 900 mg of preparation, as a single oral dose, for the purpose, as represented in the application, of mimicking the conditions of a fatty meal.

[0064] Treatment B (Boswellia Serrata / Stearic Acid / Sunflower Lecithin combination) always used the same relative ratio between the three substances, set within the range described in the patent application. In both treatment A and treatment B, volunteers were fasted for 10 hours before administration, and for the following 4 hours, limiting fluid intake to mineral water.

[0065] One hour before the intake of the preparations, both in treatment A and in treatment B, plasma samples were collected for all volunteers, immediately centrifuged and stored at -20°C until analysis, in order to build a baseline. Plasma samples were then collected at times 0.5 h, 1 h, 1 .5 h, 2 h, 3 h, 4 h, 8 h, 12 h, 18 h, 24 h, 36 h, 48 h, 60 h from the administration of the preparation, both in the case of treatment A and in the following case (after a week of washing out) of treatment B. The collected samples were similarly (as for the baseline) centrifuged at 546 x g for 20 minutes at 4 °C, and stored at -20 °C until quantitative analysis of Boswellic Acid concentrations, performed in reverse phase HPLC, as described below.

[0066] The procedure used for the analytical determination of the plasma concentrations of Boswellic Acids (i.e. the pharmacologically active species, as reported in the literature2) in the 12 healthy volunteers reproduced, both in the case of treatment A and in the case of treatment B, what was already standardized and published in the previous scientific literature3’4’5.

[0067] The extraction of the samples on Diatomaceous Earth and graphitized Carbon Black was followed by HPLC (High Performance Liquid Chromatography) analysis on reverse phase, with a photodiode array detector. The Boswellic Acids whose concentration was to be determined in the two cases A and B, i.e. [3-boswellic Acid, 11 -keto-[3-boswellic Acid, a-boswellic Acid, Acetyl-11 -keto-[3- boswellic Acid (AKBA), were identified by the peak retention time (which had to be within a range of 0.1 min from the peak of the corresponding standard), by the spectrum of the photodiode array detector, which had to overlap that of the corresponding standard, by the widening of the peak area at the retention time of the standard, by adding the standard itself to the real sample (method of standard additions). Selectivity, sensitivity, precision, accuracy and stability of the analytical method used were achieved by operating as already described and standardised in the literature1’4 5.

[0068] All 12 volunteers (males aged between 23 and 50 years) concluded the protocol without any deviation from the previously coded parameters and without any adverse effect, and the plasma concentration data of the 4 different Boswellic Acids collected in the two different treatments were subjected to Wilcoxon statistical analysis, and therefore represented as follows:

[0069] The plasma concentration determined at each plasma collection time (0.5 h, 1 h, 1.5 h, 2 h, 3 h, 4 h, 8 h, 12 h, 18 h, 24 h, 36 h, 48 h, 60 h from administration) was reported in nanograms per milliliter of plasma (ng / mL), as an average of the 12 subjects enrolled, with relative standard deviation, for each of the 4 Boswellic Acids measured, both in the case of treatment A (Boswellia Serrata dry extract taken on an empty stomach, 300 mg x 3 in a single dose) and in the case of treatment B (Boswellia Serrata / Stearic Acid / Sunflower Lecithin combination taken on an empty stomach, 300 mg x 3 in a single dose, with relative ratio between the substances within the range defined in the application).

[0070] Below we refer to the graphs with the plasma concentrations measured in the 2 different treatments A and B, at each sampling time, for each Boswellic Acid measured, in order of decreasing average concentration of the acid measured.

[0071] Figure 1 shows the plasma concentrations of [3-Boswellic Acid in the different treatments A and B.

[0072] These are in particular average concentrations (calculated therefore on the average of the 12 samples collected, with standard deviation from the average) at each time Tnof collection, for n ranging from 1 to 13. T1 will therefore be the sampling time 0.5 hours after taking treatment A (dry extract as such) or treatment B (combination submitted in the application), and so on according to the scheme, where h = 60 minutes: Concentrations are all expressed in nanograms of analyte per mL of plasma:

[0073] At time T7, i.e. 8 hours after taking the preparation, a peak concentration is observed for the Boswellia Serrata / Stearic Acid / Sunflower Lecithin combination (treatment B), higher than the corresponding peak of treatment A by a factor of approximately 5.3 (1 .5 x 103ng / mL vs 0.28 x 103ng / mL).

[0074] Figure 2 illustrates the plasma concentrations of a-Boswellic Acid in the different treatments A and B.

[0075] Also here, at time T7, i.e. 8 hours after taking the preparation, a peak concentration is observed for the Boswellia Serrata / Stearic Acid / Sunflower Lecithin combination (treatment B), higher than the corresponding peak of treatment A by a factor of about 9.1 (0.57 x 103ng / mL vs 0.062 x 103ng / mL). Therefore, already from the analytical results relating to the two most abundant Boswellic Acids it is possible to infer that the association Boswellia Serrata / Stearic Acid / Sunflower Lecithin (within the range of relative ratios between the 3 substances described in the application) effectively mimics, even on an empty stomach (common condition to both A and B), the bioavailability conditions of Boswellia Serrata dry extract as it is, on a full stomach for fatty meals, as described in literature1. Similarly, it is found for the two less abundant, but pharmacologically active species2, as reported below.

[0076] Figure 3 illustrates the plasma concentrations of 11 -keto-[3-Boswellic Acid in the different treatments A and B.

[0077] Figure 4 illustrates the plasma concentrations of acetyl-11-keto-[3-Boswellic Acid in the different treatments A and B.

[0078] In the case of 11 -keto-[3-Boswellic Acid, the analyte reaches a peak plasma concentration at T6, i.e. 4 h after taking treatment B (Boswellia Serrata / Stearic Acid / Sunflower Lecithin in relative ratios set within the range described in the application), where the peak for A is reached at T7= 8h. At the peak of B, the ratio of B to A concentrations is a factor of about 6.1 (0.32 x 103ng / mL vs 0.052 x 103ng / mL).

[0079] In the case of acetyl-11 -keto-[3-Boswellic Acid (AKBA), the analyte reaches a peak plasma concentration at T5, i.e. 3 h after taking treatment B (Boswellia Serrata / Stearic Acid / Sunflower Lecithin in relative ratios set within the range described in the application), as well as in the case of treatment A. At the peak of B, the ratio between the concentrations of B and A is equal to a factor of about 10 (0.123 x 103ng / mL vs 0.012 x 103ng / mL).

[0080] Therefore, even for the less abundant Boswellic Acids, the association of dry extract of gum resin of Boswellia Serrata with Stearic Acid and Sunflower Lecithin (in the relative ratios described in the application, and set at an ideal value in the analysis) seems to implement the absorption of Boswellic Acids on an empty stomach, compared to a dry extract as such, bringing the absorption on an empty stomach to levels comparable to those described in the literature for intake on a full stomach1. So much was to be expected, and the quantitative instrumental analysis confirms it (note the similarity of the concentration profiles graphed above with those reported in the literature for the 4 Boswellic Acids taken on a full stomach), due to the presence of a fatty substance (Stearic Acid) strongly related to the most lipophilic of meals, namely olive oil, composed mainly of oleic acid (as already specified in the application).

[0081] The combination in the solid state of the three substances therefore not only leads to an increase in the absorption of Boswellic Acids on an empty stomach, as reported above, but also extends the associability of Boswellia Serrata with other substances that must necessarily be taken on an empty stomach (example: Bromelain, as already mentioned in the application). Bibliography:

[0082] 1 ) Effect of Food Intake on the Bioavailability of Boswellic Acids from a Herbal Preparation in Healthy Volunteers, Sterk V et al., Planta Med. 2004; 70: 1155- 1160.

[0083] 2) Work-up dependent formation of5-lipoxygenase inhibitory boswellic acid analogues, Schweizer S et al. , J Nat Prod 2000; 63: 1058-1061 .

[0084] 3) Analysis of boswellic acids in human plasma after oral administration of a boswellic acid containing drug, Buchele B et al., Naunyn-Schmiedberg's Arch Pharmacol 2000; 361 (Suppl): 487.

[0085] 4) Analysis of pentacyclic triterpenic acids from Frankincense gum resins and related phytopharmaceuticals by high-performance liquid chromatography. Identification of lupeolic acid, a novel pentacyclic triterpene, Buchele B et al., J Chromatography B 2003; 791 : 21 -30.

[0086] 5) Analysis of 12 different pentacyclic triterpenic acids from Frankincense in human plasma by high-performance liquid chromatography and photodiode array detection, Buchele B et al., J Chromatography B 2003; 795: 355-362.

[0087] Naturally, amendments or improvements may be made to the invention as described, dictated by contingent or particular reasons, without thereby departing from the scope of the invention.

Claims

CLAIMS1. Nutraceutical composition comprising dry extract of Boswellia Serrata gum resin in a quantity comprised between 50% and 90% by weight, stearic acid in a quantity comprised between 45% and 4% by weight and a source of Phosphadityl Choline in a quantity comprised between 5% and 1 % by weight and wherein the percentage content in Boswellic Acids of the dry extract of Boswellia Serrata is between 25% and 90%.

2. Composition as in claim 1 , wherein the source of Phosphadityl Choline can be chosen from sunflower lecithin, soya lecithin and pure Phosphadityl Choline or combinations thereof.

3. Process for the preparation of the nutraceutical composition as claimed in any one of the preceding claims, wherein the following steps are provided:- definition of the relative proportions of dry extract of Boswellia Serrata gum resin, of the stearic acid and of the source of Phosphadityl Choline;- accurate determination of the weights of the dry extract of Boswellia Serrata gum resin, of the stearic acid and of the Phosphadityl Choline source taken individually;- sieving the dry extract of Boswellia Serrata gum resin, of the stearic acid and of the Phosphadityl Choline source taken individually;- mixing of the stearic acid with the source of Phosphadityl Choline;- addition in progressive amounts of the dry extract of Boswellia Serrata gum resin to the mixture of stearic acid and of the Phosphadityl Choline source; and- final sieving of the composition of the dry extract of Boswellia Serrata gum resin, stearic acid and the source of Phosphadityl Choline.

4. Process as in claim 3, wherein the sieving steps take place using a 16 mesh / cm2sieve.

5. Composition as in any one of claims 1 to 2 for uses concerning the treatment of joint functionality, digestive system functionality, and for countering localised states of tension.

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