Composition based on ubidecarenone

Abstract

A composition based on ubidecarenone, or coenzyme Q10 or CoQ10, comprises CoQ10, one or more specific hydrophylic carriers selected from the maltodextrins class and one or more adjuvant agents selected from the sucrose-esters class.

Claims

1. A ternary composition based on ubidecarenone, consisting of ubidecarenone, one or more maltodextrin and one or more sucrose-ester, wherein the weight percentage of the ubidecarenone is between 10% and 33.3%; the weight percentage of the sucrose-ester is between 10% and 33.3%; and wherein the ubidecarenone, the maltodextrin and the sucrose-ester are combined by co-grinding in a dry state.

2. The ternary composition of claim 1, wherein maltodextrin is present in the ternary composition in a weight adjusted to bring the percent total weight of all components to 100%.

3. The ternary composition of claim 1, wherein the percentages of ubidecarenone and sucrose-esters in the composition are the same.

4. The ternary composition of claim 1, wherein the percentages of ubidecarenone and sucrose-esters in the composition are different.

5. The ternary composition of claim 1, wherein the sucrose-ester is selected from the group consisting of saccharose monopalmitate, saccharose monostearate, saccharose dipalmatate, saccharose distearate, saccharose alkylate, and mixtures thereof.

6. The ternary composition of claim 1, wherein the weight percentage of the ubidecarenone is 33.3%; and the weight percentage of the sucrose-ester is 33.3%.

7. The ternary composition of claim 1, wherein the weight percentage of the maltodextrin is between 33.3% to 80%.

8. The ternary composition of claim 1, wherein the weight percentage of the ubidecarenone is between 5% and 30%; and the weight percentage of the sucrose-ester is between 5% and 30%.

9. The ternary composition of claim 1, wherein the weight percentage of the ubidecarenone is between 10% and 15%; and the weight percentage of the sucrose-ester is between 10% and 15%.

10. A formulation comprising the ternary composition of claim 1, mixed with vitamin A.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a graph illustrating a comparison of dissolution rates between ternary compositions configured in accordance with various embodiments of the present invention.

(2) FIG. 2 is a graph illustrating a comparison of capacity for intracellular penetration of ternary compositions configured in accordance with various embodiments of the present invention.

DETAILED DESCRIPTION OF SOME FORMS OF EMBODIMENT

(3) A specific form of embodiment of the present invention provides CoQ10 at 10% in weight, sucrose-ester at 10% in weight and maltodextrin at 80% in weight. This specific form of embodiment is also referred to as ternary composition of CoQ10/maltodextrin/sucrose-ester 1/8/1, or simply 1/8/1, referring to the ratios of weight to weight (w/w).

(4) Another specific form of embodiment of the present invention provides CoQ10 at 15% in weight, sucrose-ester at 5% in weight and maltodextrin at 80% in weight. This specific form of embodiment is also referred to as ternary composition of CoQ10/maltodextrin/sucrose-ester 1.5/8/0.5, referring to the ratios of weight to weight (w/w).

(5) A specific form of embodiment of the present invention provides CoQ10 at 0.5% in weight, sucrose-ester at 15% in weight and maltodextrin at 80% in weight. This specific form of embodiment is also referred to as ternary composition of CoQ10/maltodextrin/sucrose-ester 0.5/8/1.5, referring to the ratios of weight to weight (w/w).

(6) Another specific form of embodiment of the present invention provides CoQ10 at 5% in weight, sucrose-ester at 5% in weight and maltodextrin at 90% in weight. This specific form of embodiment is also referred to as ternary composition of CoQ10/maltodextrin/sucrose-ester 0.5/9/0.5, referring to the ratios of weight to weight (w/w).

(7) Another specific form of embodiment of the present invention provides CoQ10 at 15% in weight, sucrose-ester at 15% in weight and maltodextrin at 70% in weight. This specific form of embodiment is also referred to as ternary composition of CoQ10/maltodextrin/sucrose-ester 1.5/7/0.5, referring to the ratios of weight to weight (w/w).

(8) Another specific form of embodiment of the present invention provides CoQ10 at 20% in weight, sucrose-ester at 20% in weight and maltodextrin at 60% in weight. This specific form of embodiment is also referred to as ternary composition of CoQ10/maltodextrin/sucrose-ester 2/6/2, referring to the ratios of weight to weight (w/w).

(9) Another specific form of embodiment of the present invention provides CoQ10 at 25% in weight, sucrose-ester at 25% in weight and maltodextrin at 50% in weight. This specific form of embodiment is also referred to as ternary composition of CoQ10/maltodextrin/sucrose-ester 2.5/5/2.5, referring to the ratios of weight to weight (w/w).

(10) Another specific form of embodiment of the present invention provides CoQ10 at 30% in weight, sucrose-ester at 20% in weight and maltodextrin at 50% in weight. This specific form of embodiment is also referred to as ternary composition of CoQ10/maltodextrin/sucrose-ester 3/5/2, referring to the ratios of weight to weight (w/w).

(11) Another specific form of embodiment of the present invention provides CoQ10 at 20% in weight, sucrose-ester at 30% in weight and maltodextrin at 50% in weight. This specific form of embodiment is also referred to as ternary composition of CoQ10/maltodextrin/sucrose-ester 2/5/3, referring to the ratios of weight to weight (w/w).

(12) Some forms of embodiment, which can be combined with other forms of embodiment described here, can provide to use sucrose-ester or mixtures of sucrose-esters having HLB value comprised between 14 and 16.

EXAMPLES

Example 1: Composition According to the Present Invention 1/8/1

(13) Ternary Composition of Coenzyme Q10/Maltodextrin/Sucrose-Ester 1/8/1.

(14) In some non-restrictive forms of embodiment, for the preparation of the ternary composition of this example, a co-grinding technique is adopted in which 15 g of a mixture in w/w ratio 1/8/1 of coenzyme Q10 (1.5 g), maltodextrin (12 g) and sucrose-ester (1.5 g), obtained for example using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation for 30 minutes at a speed of 200 rpm. At the end of the process, the product in fine powder form is discharged and sieved. A ternary composite material coenzyme q10/maltodextrin/sucrose-ester is obtained in a w/w ratio of 1/8/1 with a coenzyme q10 titer of 10% in weight.

COMPARATIVE EXAMPLES

(15) Example A (state of the art according to WO'997): ternary compositions of ubidecarenone/copovidone/glycine 1/8/1.

(16) For the preparation of the ternary composition of this example, 15 g of a mixture in w/w ratio 1/8/1 of ubidecarenone (1.5 g), copovidone (12 g) and glycine (1.5 g), obtained using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation in the same conditions as applied in Example 1. At the end of the process, the product in fine powder form is discharged and sieved. A ternary composite material ubidecarenone/copovidone/glycine is obtained in a w/w ratio of 1/8/1 with an ubidecarenone titer of 10%.

(17) Example B (replacement of the carrier component of Example A by the carrier used in the present invention): ternary compositions of ubidecarenone/maltodextrin/glycine 1/8/1.

(18) For the preparation of the ternary composition of this example, 15 g of a mixture in w/w ratio 1/8/1 of ubidecarenone (1.5 g), maltodextrin (12 g) and glycine (1.5 g), obtained using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation in the same conditions as applied in Example 1. At the end of the process, the product in fine powder form is discharged and sieved. A ternary composite material ubidecarenone/maltodextrin/glycine is obtained in a w/w ratio of 1/8/1 with an ubidecarenone titer of 10%.

(19) Example C (replacement of the ternarizing component of Example A by the ternary component used in the present invention): ternary compositions of ubidecarenone/copovidone/glycine 1/8/1.

(20) For the preparation of the ternary composition of this example, 15 g of a mixture in w/w ratio 1/8/1 of ubidecarenone (1.5 g), copovidone (12 g) and sucrose-ester (1.5 g), obtained using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation in the same conditions as applied in Example 1. At the end of the process, the product in fine powder form is discharged and sieved. A ternary composite material ubidecarenone/copovidone/sucrose-ester is obtained in a w/w ratio of 1/8/1 with an ubidecarenone titer of 10%.

(21) Example D (effect of the emulsifier): binary compositions ubidecarenone/sucrose-ester 1/1.

(22) For the preparation of the binary composition of this example, 15 g of a mixture in w/w ratio 1/1 of ubidecarenone (7.5 g), and sucrose-ester (7.5 g), obtained using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation in the same conditions as applied in Example 1. At the end of the process, the product in fine powder form is discharged and sieved. A binary composite material ubidecarenone/sucrose-ester is obtained in a w/w ratio of 1/1.

(23) Examples E for comparison with the state of the art according to WO'689:

(24) We shall now present some examples for comparison (A1-F1-A2-G2) made according to the examples 27 and 28 in WO'689 and some variations of said examples intended to show the differences and advantages of the present invention both in terms of ternary composition consisting of ubidecarenone, sucrose-esters and maltodextrins with respect to the pentenary compositions used in WO'689, and also in terms of the techniques used, based on co-grinding, compared with the techniques with solvents and spray-drying as used in WO'689.

(25) Hereafter the following abbreviations may be used: siloid=silicon dioxide or silica, mdx=maltodextrin(s), hpmc=hydroxypropyl methylcellulose, vit E=vitamin E or DL-alpha-tocopheryl acetate.

(26) The experimental tests and comparisons carried out and discussed hereafter show that the ternary composition which consists of ubidecarenone (CoQ10), sucrose-ester(s) and maltodextrin(s) is better than the pentenary compositions of WO'689, irrespective of the technique used (co-grinding compared with spray-drying) and also that the maltodextrins class is better than hpmc as hydrophylic polymer carrier. It is clear from the following that the composition of the example 1 according to the present invention is in any case better than the other examples A1-F1, A2-G2 as proposed.

(27) Examples of Comparison to Evaluate Co-Grinding

(28) A1 Replication of Example 27 of WO'689 Using Co-Grinding

(29) Compositions of Ubidecarenone/HPMC/Sucrose-Ester/Vitamin E/Siloid 33.3/31.2/33.3/0.1/2

(30) 30 g of a mixture of ubidecarenone (9.99 g, 33.3%), HPMC (9.36 g, 31.2%) sucrose-ester (9.99 g, 33.3%), vitamin E (0.03 g, 0.1%) and siloid (0.6 g, 2%) obtained using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation in the same conditions as applied in Example 1. At the end of the process, the product is discharged and sieved. A composite material ubidecarenone/hpmc/sucrose-ester/vitamin E/siloid is obtained with an ubidecarenone titer of 33.3%.

(31) B1 Replication of Example 28 of WO'689 Using Co-Grinding

(32) Compositions of Ubidecarenone/HPMC/Sucrose-Ester/Vitamin E/Silicon Dioxide 25/47.9/25/0.1/2.

(33) 30 g of a mixture of ubidecarenone (7.5 g, 25%), HPMC (14.37 g, 47.9%) sucrose-ester (7.5 g, 25%), vitamin E (0.03 g, 0.1%) and siloid (0.6 g, 2%) obtained using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation in the same conditions as applied in Example 1. At the end of the process, the product is discharged and sieved. A composite material ubidecarenone/hpmc/sucrose-ester/vitamin E/siloid is obtained with an ubidecarenone titer of 25%.

(34) C1 Comparison with Example 27 in WO'689 but without Vitamin E and Silicon Dioxide

(35) Ternary Compositions of Ubidecarenone/HPMC/Sucrose-Ester 33.3/33.3/33.3.

(36) 30 g of a mixture in a ratio w/w 1/1/1 of ubidecarenone (10 g, 33.3%), HPMC (10 g, 33.3%) and sucrose-ester (10 g, 33.3%), obtained using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation in the same conditions as applied in Example 1. At the end of the process, the product is discharged and sieved. A composite ternary material ubidecarenone/hpmc/sucrose-ester is obtained with an ubidecarenone titer of 33%.

(37) D1 Comparison with Example 28 in WO'689 but without Vitamin E and Silicon Dioxide

(38) Ternary Compositions of Ubidecarenone/HPMC/Sucrose-Ester 25/50/75.

(39) 30 g of a mixture of ubidecarenone (7.5 g, 25%), HPMC (15 g, 50%) and sucrose-ester (7.5 g, 25%), obtained using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation in the same conditions as applied in Example 1. At the end of the process, the product is discharged and sieved. A composite ternary material ubidecarenone/HPMC/sucrose-ester is obtained with an ubidecarenone titer of 25%.

(40) E1 Comparison with Example 27 in WO'689 but without Vitamin E and Silicon Dioxide and with Maltodextrin Instead of HPMC

(41) Ternary Compositions of Ubidecarenone/Maltodextrin/Sucrose-Ester 33.3/33.3/33.3.

(42) 30 g of a mixture of ubidecarenone (10 g, 33.3%), maltodextrin (10 g, 33.3%) and sucrose-ester (10 g, 33.3%), obtained using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation in the same conditions as applied in Example 1. At the end of the process, the product is discharged and sieved. A composite ternary material ubidecarenone/maltodextrin/sucrose-ester is obtained with an ubidecarenone titer of 33%.

(43) F1 Comparison with Example 28 in WO'689 but without Vitamin E and Silicon Dioxide and with Maltodextrin Instead of HPMC

(44) Ternary Compositions of Ubidecarenone/Maltodextrin/Sucrose-Ester 25/50/25.

(45) 30 g of a mixture of ubidecarenone (7.5 g, 25%), maltodextrin (15 g, 50%) and sucrose-ester (7.5 g, 25%), obtained using a mixer with a rotary body, are loaded into a jar of a planetary mill and subjected to mechanical-chemical activation in the same conditions as applied in Example 1. At the end of the process, the product is discharged and sieved. A composite ternary material ubidecarenone/maltodextrin/sucrose-ester is obtained with an ubidecarenone titer of 25%.

(46) Examples of Comparison to Evaluate Co-Solubilization and Spray-Drying Technique

(47) A2 Replication of Example 27 in WO'689

(48) Compositions of Ubidecarenone/HPMC/Sucrose-Ester/Vitamin E/Silicon Dioxide 33.3/31.2/33.3/0.1/2

(49) HPMC (9.36 g, 31.2%) is dispersed by stirring in 250 ml of ethanol to which the sucrose-ester (9.99 g, 33.3%) is added at ambient temperature. The ubidecarenone (9.9 g, 33.3%) is dissolved in 250 ml of methylene chloride and the solution added to the previous one. Then vitamin E (0.03 g, 0.1%) and silicon dioxide (0.6 g, 2%) are added and the whole is homogenized. The solution is spray-dried.

(50) B2 Replication of Example 28 in WO'689

(51) Compositions of Ubidecarenone/HPMC/Sucrose-Ester/Vitamin E/Silicon Dioxide 25/47.9/25/0.1/2.

(52) HPMC (14.37 g, 47.9%) is dispersed by stirring in 400 ml of ethanol to which the sucrose-ester (7.5 g, 25%) is added at ambient temperature. The ubidecarenone (7.5 g, 25%) is dissolved in 400 ml of methylene chloride and the solution added to the previous one. Then vitamin E (0.03 g, 0.1%) and silicon dioxide (0.6 g, 2%) are added and the whole is homogenized. The solution is spray-dried.

(53) C2 Example of Comparison Like Example 27 in WO'689 but without Vitamin E and Silicon Dioxide

(54) Ternary Compositions of Ubidecarenone/HPMC/Sucrose-Ester 33.3/33.3/33.3.

(55) HPMC (10.0 g, 33.3%) is dispersed by stirring in 250 ml of ethanol to which the sucrose-ester (10.0 g, 33.3%) is added at ambient temperature. The ubidecarenone (10.0 g, 33.3%) is dissolved in 250 ml of methylene chloride and the solution added to the previous one. The solution is spray-dried.

(56) D2 Example of Comparison Like Example 28 in WO'689 but without Vitamin E and Silicon Dioxide

(57) Ternary Compositions of Ubidecarenone/HPMC/Sucrose-Ester 25/50/75.

(58) HPMC (15.0 g, 50%) is dispersed by stirring in 400 ml of ethanol to which the sucrose-ester (7.5 g, 25%) is added at ambient temperature. The ubidecarenone (7.5 g, 25%) is dissolved in 400 ml of methylene chloride and the solution added to the previous one. The solution is spray-dried.

(59) E2 Example of Comparison Like Example 27 in WO'689 but without Vitamin E and Silicon Dioxide and with Maltodextrin Instead of HPMC

(60) Ternary Compositions of Ubidecarenone/Maltodextrin/Sucrose-Ester 33.3/33.3/33.3.

(61) Maltodextrin (10.0 g, 33.3%) is dispersed by stirring in 250 ml of ethanol to which the sucrose-ester (10.0 g, 33.3%) is added at ambient temperature. The ubidecarenone (10.0 g, 33.3%) is dissolved in 250 ml of methylene chloride and the solution added to the previous one. The solution is spray-dried.

(62) F2 Example of Comparison Like Example 28 in WO'689 but without Vitamin E and Silicon Dioxide and with Maltodextrin Instead of HPMC

(63) Ternary Compositions of Ubidecarenone/Maltodextrin/Sucrose-Ester 25/50/75.

(64) Maltodextrin (15.0 g, 50%) is dispersed by stirring in 400 ml of ethanol to which the sucrose-ester (7.5 g, 25%) is added at ambient temperature. The ubidecarenone (7.5 g, 25%) is dissolved in 400 ml of methylene chloride and the solution added to the previous one. The solution is spray-dried.

(65) G2 Example of Comparison Like Example 1 According to the Present Invention, but Using the Spray-Drying Technique

(66) Ternary Compositions of Ubidecarenone/Maltodextrin/Sucrose-Ester 1/8/1

(67) Maltodextrin (24.0 g, 80%) is dispersed by stirring in 400 ml of ethanol to which the sucrose-ester (3 g, 10%) is added at ambient temperature. The ubidecarenone (3 g, 10%) is dissolved in 200 ml of methylene chloride and the solution added to the previous one. The solution is spray-dried.

Experimental Characterization of the Composition According to the Present Invention

(68) Solubility

(69) The solubility in water at 37 C. (physiological temperature) of the composition according to the present invention was compared with that of the compositions in the examples of comparison.

(70) To analyze the solubility, an excess of composition according to the present invention is put in powdered form into water at 37 C., so as to have a sediment.

(71) Verification is carried out after 1 hour and 24 hours (balanced) using the suitable analytical method (for example UV or HPLC spectrophotometry) of the maximum quantity of coenzyme Q10 that passes into the solution. The results are shown in the following Table 1, where the composition according to the present invention is shown as an example with that of Example 1.

(72) TABLE-US-00001 TABLE 1 Increase Solubility 1 hour Solubility 24 hrs over CoQ10 (g/ml) (g/ml) 1 hour 24 hrs CoQ10 1 4 Example 1 280 330 280 83 Example A 50 150 50 38 Example B 3 22 3 5.5 Example C 40 110 40 27.5 Example D 4 18 3.5 4.5

(73) Based on the results of Table 1, a greater solubility is found of the composition according to the present invention, in this case as in Example 1. This advantageous technical effect was not foreseeable in the state of the art based only on the fact that, for example, the carrier maltodextrin is more soluble in water than the previous carrier copovidone.

(74) In fact, if we replace copovidone by maltodextrin (cf. the results of Example A and Example B), we have solubility after 24 hours equal to 22 g/ml, therefore much lower than example A (50 g/ml).

(75) Furthermore, this advantageous technical effect in terms of the increase in solubility was also not foreseeable based on the emulsifying power of the sucrose-ester, since a binary co-ground composition, as in Example D, in the same w/w ratio as found in Example 1, has a solubility of 4 g/ml at 1 hour and 18 g/ml at 24 hours.

(76) In the same way, the same applies if we replace the sucrose-ester with glycine in the composition in Example C compared with Example A.

(77) Without being constrained by the theory, Applicant believes that the composition according to the present invention, reaching almost maximum solubility already after 1 hour (280 g/ml compared with 330 g/ml after 24 hours) compared with Example A (50 g/ml compared with 150 g/ml after 24 hours) where it is much lower, in vivo it will be more easily assimilated because it is readily available immediately, considering the mean transit time of the gastro-intestinal tract.

(78) Tables 2 and 3 show the results for solubility tests carried out as described above, between Examples E (A1-F1, A2-G2) described above and Example 1 according to the present invention.

(79) TABLE-US-00002 TABLE 2 SPRAY DRYING EXAMPLE A2 B2 C2 D2 E2 F2 G2 N. COMPONENTS 5 5 3 3 3 3 3 TITER IN Q10 33.3%, 25%, 33.3%, 25%, 33.3%, 25%, 10%, (%), CARRIER hpmc hpmc hpmc hpmc mdx mdx mdx SOLUBILITY 70 105 80 110 140 160 200 (g/ml)

(80) TABLE-US-00003 TABLE 3 CO-GRINDING EXAMPLE A1 B1 C1 D1 E1 F1 Example 1 COMPONENTS 5 5 3 3 3 3 3 TITER IN Q10 33.3%, 25%, 33.3%, 25%, 33.3%, 25%, 10%, (%), CARRIER hpmc hpmc hpmc hpmc mdx mdx mdx SOLUBILITY 60 100 100 130 170 190 280 (g/ml)

(81) Considering the data shown above, the best composition, both in terms of co-grinding and spray-drying, is composition 1/8/1 of Example 1 according to the present invention.

(82) Again according to the data shown above, the use of maltodextrin leads to better solubility results compared with hpmc. Without being constrained by the theory, the better performance of maltodextrin could be explained by a fine particle distribution of the components on the maltodextrin, which is soluble in water and therefore, when it comes into contact with it, helps the solubilization. On the contrary, hpmc not only gels but, given complete solubilization, when it is spray-dried it incorporates the CoQ10 more intimately and therefore releases it more slowly, also because of the gelled layer.

(83) It should be noted that all the ternary preparations obtained through co-grinding are also better than their homologous preparations prepared through spray-drying: see F1 against F2, E1 against E2, D1 against D2 and C1 against C2.

(84) It can be seen that, increasing the titer in CoQ10, although on the one hand the performance is slightly worse, since there is less carrier present, there is therefore less possibility for the CoQ10 to interact, the maltodextrin always behaves better than hpmc: see F2 against D2, E2 against C2 for co-grinding and F1 against D1, E1 against C1 for spray-drying. Furthermore, the results show that co-grinding is better than spray-drying: see F1 against F2, E1 against E2, D1 against D2 and C1 against C2.

(85) Moreover, the performances of compositions with 5 components (pentenary) obtained by spray-drying are worse than those of the compositions with 3 components (ternary) obtained by spray-drying: see B2 against D2, A2 against C2. It can be deduced that the other two components, beyond the sucrose-ester, the hydrophylic carrier and the CoQ10, make the performance worse, and that the ternary composition is better irrespective of the spray-drying or co-grinding technique used.

(86) From the data shown it also emerges that the performances of the compositions with 5 components obtained by spray-drying are better than the compositions with 5 components obtained by co-grinding: see A2 against A1 and B2 against B1. It can be deduced that the other two components, beyond the sucrose-ester, the hydrophylic carrier and the CoQ10, make the performance worse for co-grinding, and therefore it is important to choose the ternary compositions.

(87) Finally, the performances of the ternary compositions obtained by co-grinding are better than the pentenary compositions obtained by co-grinding: see D1 against B1 and C1 against A1. In this case too it can be deduced that the other two components, beyond the sucrose-ester, the hydrophylic carrier and the CoQ10, make the performance worse for co-grinding, and therefore it is important to choose the ternary compositions.

(88) Dissolution Rate Test (DRT)

(89) Dissolution rate tests were carried out with the powders obtained in the examples A1-F1, A2-F2, weighed so as to use always 50 mg of Coenzyme Q10.

(90) 500 ml of pH buffer 1.2 were used, with an added 1% of Tween80 kept at T=37 C. and stirred at a speed of 50 rpm. Aliquots were taken at 15, 30, 60 and 120 minutes. For the DR test in Example 1, an aliquot was also taken at 5 minutes. Each aliquot was analyzed in HPLC under the following conditions:

(91) Detector: UV at 275 nm

(92) Column: Zorbax Extend 300 C18 or equivalent 4.6150, 5 m

(93) T column: 25 C.

(94) Mobile phase: ethanol/methanol 20/80

(95) Flow: 2.0 ml/mn

(96) TR: 8 min.

(97) FIG. 1 shows the graphs for the DR tests carried out. The y-axis shows the percentage releasability of CoQ10 and the x-axis shows the time in minutes. The dotted lines show the examples made with spray-drying and the solid lines show the examples made with co-grinding.

(98) The data shown above for the DR tests confirm what was said regarding the fact that the best composition, both in terms of co-grinding and spray-drying, is composition 1/8/1 of Example 1 according to the present invention. It is also clear that the compositions with hpmc are not able to release 100% of CoQ10 within the 120 minutes of the test, probably because hpmc tends to gel.

(99) Absorption in Cardiomyoblasts

(100) Despite the large number of clinical studies on the effects of Coenzyme Q10 supplementation, there are very few experimental data concerning the content of coenzyme Q10 in mitochondria and the cell energy condition after supplementation. Probably, controversial results of clinical and in vitro studies are mainly due to the bio-availability of the form of coenzyme Q10 used. For this reason, with reference to FIG. 2 attached, the capacity of the composition according to the present invention as per Example 1 (column III) and the composition in Example A (column II) for intracell penetration and the penetration of the mitochondrial membrane in cardiomyoblasts was also assessed, compared with Coenzyme Q10 as such (column I).

(101) For this analysis, T67 cells of human astrocytoma and H9C2 cells from rats, deriving from the heart, were cultured in a Dulbecco's modified Eagle medium (DMEM), supplemented with 10% of fetal bovine serum (FBS), 100 Ul/ml of penicillin, 100 g/ml of streptomycin and 40 g/ml of gentamicin, in an atmosphere of 5% CO.sub.2 at 37 C., with saturation humidity. The vitality of the cells and their number were measured using the exclusion method with Tripan blue (Lowry et al.). The mitochondria were isolated using known procedures (Chomyn, 1996). The cells treated were washed carefully with PBS before the extraction procedures. The extraction of the coenzyme Q10 from the isolated cells and mitochondria was performed as described by Takada et al. (Takada et al., 1984). The quantification of the coenzyme Q10 was done by means of HPLC analysis. From 50 to 100 l of ethanol extract were subjected to chromatography on a C18 column (Kinetex, Phenomex, 2.6 m, 1004.6 mm), using a mobile phase that consists of an ethanol:water ratio of 97:3 v/v with a flow rate of 0.6 ml/min. The concentrations of coenzyme Q10 were obtained by comparing the peak areas with those of the standard solutions in at least three independent experiments.

(102) FIG. 2 is a graph showing on the y-axis the increase in CoQ10 compared to the base value after a specific administration, and on the x-axis two groups of column graphs, in particular group A, relating to the increase tested on cells, and group B relating to the increase tested on mitochondria. In particular, FIG. 2 shows an increase in CoQ10 with respect to the base value after the administration of: CoQ10 as such (column I), composition according to the present invention as in Example 1 (column III) and composition as in Example A (column II):

(103) In particular, an improvement can be seen in the passage of CoQ10 from whole cells to the mitochondria with the composition according to the present invention. In particular, with CoQ10 as such and with the composition as in Example A, 4% of CoQ10 passes from the whole cells to the mitochondria, whereas in Example 1 according to the present invention 5.3% of CoQ10 passes from the whole cells to the mitochondria, with a percentage increase of 32.5%.

(104) The results also show that the intracellular content of Coenzyme Q10 correlates positively with the mitochondrial function and resistance to oxidative stress. The determination of the content of Coenzyme Q10 in lines of cultured cells after the administration of different compositions of Coenzyme Q10 has also shown that the most efficient formulation is the one according to the present invention as in Example 1 (column III). These results show that an adequate channeling of the coenzyme Q10 is important to ensure an appropriate cell uptake. The increased bio-availability also allows treatments with low doses of Coenzyme Q10 which allow to prevent an a-specific accumulation.