Use of homoeriodictyol (HED) for reducing the gastric acid secretion-stimulating effect of n-acetyl-4-aminophenol (Paracetamol)

Abstract

The present invention primarily relates to the use of Homoeriodictyol (HED) or a salt thereof or a mixture of HED and one or more salts thereof or a mixture of several salts thereof for reducing the gastric acid secretion-stimulating effect of N-Acetyl-4-aminophenol (Paracetamol).

Claims

1. A method for reducing gastric acid secretion-stimulating effects of paracetamol comprising administering to a patient in need thereof an effective amount of homoeriodictyol and/or salt(s) thereof, and reducing the gastric acid secretion-stimulating effects of the paracetamol, wherein the patient in need thereof is a patient suffering from an inflammatory condition of the gastric mucosa and/or a patient suffering from chronic pain.

2. The method of claim 1, wherein the homoeriodictyol and/or salt(s) thereof is/are administered simultaneously with paracetamol.

3. The method of claim 1, wherein the homoeriodictyol and/or salt(s) thereof is/are administered separately from paracetamol.

4. The method of claim 1, wherein the gastric acid secretion-stimulating effect is reduced by at least 10%, compared to an identical preparation without the HED and/or salt thereof.

5. The method of claim 1, wherein the gastric acid secretion-stimulating effect is reduced by at least 50%, compared to an identical preparation without the HED and/or salt thereof.

6. The method of claim 1, wherein the gastric acid secretion-stimulating effect is reduced by at least 70%, compared to an identical preparation without the HED and/or salt thereof.

7. A method for reducing gastric acid secretion-stimulating effects of paracetamol comprising administering to a patient in need thereof a pharmaceutical preparation comprising paracetamol and homoeriodictyol (HED) and/or salt(s) thereof in an amount sufficient to reduce the gastric acid secretion-stimulating effect of the paracetamol, and reducing the gastric acid secretion-stimulating effects of the paracetamol, wherein the patient in need thereof is a patient suffering from an inflammatory condition of the gastric mucosa and/or a patient suffering from chronic pain.

8. The method of claim 7, wherein the preparation further comprises one or more of eriodictyol, phloretin, hesperetin, 2,4-dihydrobenzoic acid-/V-vanillylamide, 5,7-dihydroxy-4-(4-hydroxy-phenyl)-chroman-2-one, 5,7-dihydroxy-4-(4-hydroxy-3-methoxy-phyenyl)-chroman-2-one, 5,7-dihydroxy-4-(4-pyridyl)-chroman-2-one, 7,3-dihydroxy-4′-methoxyflavan, lariciresinol, and matairesinol; and their respective stereoisomers.

9. The method of claim 7, wherein the preparation comprises one or more salts of HED, wherein the one or more salts are selected from sodium, potassium, calcium, and ammonium salts.

10. The method of claim 7, wherein the preparation comprises homoeriodictyol (HED) and/or salt(s) thereof in an amount sufficient to mask the bitter taste of paracetamol.

11. The method of claim 7, wherein the gastric acid secretion-stimulating effect is reduced by at least 10%, compared to an identical preparation without the HED and/or salt thereof.

12. The method of claim 7, wherein the gastric acid secretion-stimulating effect is reduced by at least 50%, compared to an identical preparation without the HED and/or salt thereof.

13. The method of claim 7, wherein the gastric acid secretion-stimulating effect is reduced by at least 70%, compared to an identical preparation without the HED and/or salt thereof.

14. The method of claim 7, wherein the preparation is a tablet.

Description

EXAMPLES

(1) Investigations of the Effect of HED

(2) Cellular Model

(3) Human parietal cells (human gastric tumour cell line, HGT-1) were used as cellular model. These were provided by Dr. C. Laboisse (Laboratory of Pathological Anatomy, Nantes, France). The cells are cultured at standard conditions at 37° C., 5% CO.sub.2 in DMEM (Dulbecco's Modified Eagel's Medium) with 4 g/L glucose, 10% FBS, 2% L-glutamine and 1% Penicillin/Streptomycin. For measuring the intracellular proton concentration, the cells are treated with Trypsin/EDTA, the cellular viability was determined with Trypan blue staining and 100 000 cells/well were seeded in black 96-well plates.

(4) Determination of the Intracellular pH Value in HGT-1 Cell Cultures

(5) For measuring the intracellular pH value in HGT-1 cell cultures, the dye 1,5-carboxy-seminaphtarhodafluor-acetoxymethyl ester (SNARF-1-AM) was used. The cells in the 96-well plates are washed once with Krebs-Hepes-buffer (KRHP) and incubated with the dye in a concentration of 3 μM dissolved in KRHP for 30 min at 37° C. and 5% CO.sub.2. Afterwards, the cells are washed twice with KRHP and substances stimulating gastric acid secretion, as for example 3 mM caffeine, 0.3 mM Theobromine or 3 mM Paracetamol alone or in combination with substances for reducing the gastric acid stimulating effect of the above substances, for example Homoeriodictyol (HED), in various concentrations in phenol red free DMEM in a volume of 100 μL were added; as a further control, the substances stimulating gastric acid secretion as named above were tested alone. Homoeriodictyol is dissolved in double distilled water. The final concentration of solvent, which is added to the cells, is at most 1%. The fluorescent dye is excited at the wave length of 488 nm and the emission is measured at 580 nm and 640 nm. The ratio of the fluorescence values of 580 nm and 640 nm is plotted in comparison to the calibration curve, wherefrom the pH value can be then obtained. For the calibration curve, the cells are treated with a potassium buffer in different pH values of from 7.2 to 8.2 and 2 μM Nigericin. Nigericin equilibrates the intracellular and extracellular pH value so that the intracellular pH value can be defined. The intracellular H.sup.+-concentration is obtained from the intracellular pH value. The intracellular proton index (IPX) is calculated by log 2 transformation of the ratio of treated cells and untreated cells (control) (Rubach, M.; Lang, R.; Seebach, E.; Somoza, M. M.; Hofmann, T.; Somoza, V., Mol Nutr Food Res 2012, 56, 325-335; Rubach, M.; Lang, R.; Hofmann, T.; Somoza, V., Ann N Y Acad Sci 2008, 1126, 310-4; Rubach, M.; Lang, R.; Skupin, C.; Hofmann, T.; Somoza, V., J Agric Food Chem 2010, 58, 4153-61; Weiss, C.; Rubach, M.; Lang, R.; Seebach, E.; Blumberg, S.; Frank, O.; Hofmann, T.; Somoza, V., J Agric Food Chem 2010, 58, 1976-85; Liszt, K. I.; Walker, J.; Somoza, V., J Agric Food Chem 2012, 60, 7022-7030; Walker, J.; Hell, J.; Liszt, K. I.; Dresel, M.; Pignitter, M.; Hofmann, T.; Somoza, V., J Agric Food Chem 2012, 60, 1405-12).

(6) The number of indicated replicates relates to technical replicates (tr) or the number of total replicates (n), which results from the number of technical replicates multiplied with the number of biological replicates.

(7) The following table 1 shows the measured percentage increase (positive values) or, respectively, percentage decrease (negative values) of the proton secretion in HGT-1-cells after 10 minutes stimulation by 3 mM caffeine, 0.3 mM Theobromine or 3 mM Paracetamol alone or in combination with Homoeriodictyol (HED) in varying concentrations. The data is displayed as mean and mean standard deviation, n=3-5, tr=6. Statistics: one-way Anova with Holm-Sidak post-hoc test. Significant differences (p<0.05) are indicated by letters.

(8) TABLE-US-00001 TABLE 1 Influence of the Paracetamol-induced proton secretion of HGT-1 cells by HED compared to the influence of the Theobromine- and caffeine-induced proton secretion (as described in WO 2014111436 A1): Caffeine Theobromine Paracetamol 3 mM 0.3 mM 3 mM mean SEM mean SEM Mean SEM DMEM 54.80 5.61 20.92 3.37 24.10 2.23 +0.003 mM HED 21.56 2.33 10.96 5.12 +0.03 mM HED 38.00 6.70 13.79 3.50 6.57 4.23 +0.3 mM HED 20.20 5.99 −16.92 3.27 −46.24 3.41

(9) As table 1 shows, an increase of the proton secretion of 55% is obtained in case the proton secretion of HGT-1 cells is stimulated by 3 mM caffeine, the addition of HED slightly dampens the caffeine-induced increase of the proton secretion, i.e. at simultaneous incubation of the cells with 3 mM caffeine and 0.03 mM HED, the proton secretion is only increased by 38%, whereas at simultaneous incubation of the cells with 3 mM caffeine and 0.3 mM HED, the proton secretion is only increased by 20%.

(10) In case of a stimulation of the proton secretion in HGT-1 cells by 0.3 mM Theobromine alone, an increase in the proton secretion of 21% arises. The addition of HED slightly dampens the Theobromine-induced increase of the proton secretion, i.e. at simultaneous incubation of the cells with 0.3 mM Theobromine and 0.03 mM HED, the proton secretion is only increased by 14%. At simultaneous incubation of the cells with 0.3 mM Theobromine and 0.3 mM HED, the proton secretion is even decreased by 17%.

(11) In case of a stimulation of the proton secretion in HGT-1 cells by 3 mM Paracetamol alone, the proton secretion is increased by 24%. The addition of HED clearly dampens the increase of the proton secretion after addition of Paracetamol. At simultaneous incubation of the cells with 0.3 mM Paracetamol and only 0.003 mM HED, the proton secretion is only increased by 11%. At simultaneous incubation of the cells with 3 mM Paracetamol and 0.03 mM HED, the proton secretion is only increased by 7%. At simultaneous incubation of the cells with 3 mM Paracetamol and 0.3 mM HED, the proton secretion is clearly reduced by 46%.

APPLICATION EXAMPLES

Application Example 1: HED Dosage Forms

(12) HED-1: Homoeriodictyol, purity 95% (company Interquim, Spain)

(13) HED-2: Homoeriodictyol sodium salt variant 1:

(14) 10 g Homoeriodictyol (95%) are provided in a flask and 44 g of 3% NaOH-solution are added. The firstly dark brown solution quickly becomes pappy, is then diluted with water until a homogenous suspension and stirred for 1 hour and then freeze dried or spray dried. Yield: 10.44 g; HPLC: 94.2% HED-Na against standard.

(15) HED-3: Homoeriodictyol sodium salt variant 2:

(16) 10 g Homoeriodictyol (95%) are suspended in 100 ml ethyl acetate and 44 g of 3% Na—OH solution is added. The firstly clear solution becomes cloudy and the product precipitates. After 1 hour of stirring at room temperature, the suspension is filtered via a pressure ratched and the filter residue is dried in a vacuum drying chamber. Yield: 9.63 g; HPLC 98.5% HED-Na against standard.

(17) HED-4: Homoeriodictyol sodium salt, natural, prepared according to WO 2004/041804.

(18) HED-5: 60.8 wt.-% water are provided and 6.1 wt.-% gum Arabic and 24.3 wt.-% maltodextrin (of maize starch) are dissolved. 8.8 wt.-% of the Homoeriodictyol sodium salt (HED-2, HED-3 or HED-4) are added and mixed with an ultraturrax or another homogenizer. The emulsified suspension is then spray dried in a spray tower (inlet temperature 185-195° C., outlet temperature 70-75° C.) and a spray product loaded with 18-22% of Homoeriodictyol is obtained.

(19) HED-6: 2.5 wt.-% Homoeriodictyol (HED-1) are dissolved in 1,2-propylene glycol.

(20) HED-7: 1 wt.-% of Homoeriodictyol sodium salt (HED-2, HED-3 or HED-4) are dissolved in ethanol.

(21) In the following applications, only ingredients in pharma quality are used.

Application Example 2: Sachets

(22) 3.00 g water free citric acid

(23) 2.50 g Aspartame

(24) 1.00 g ascorbic acid

(25) 82.00 g sucrose

(26) 10.00 g Paracetamol powder

(27) 1.50 g HED-5 (spray dried HED sodium salt on maltodextrin)

(28) The ingredients are mixed and subsequently packed in portions of 5 g.

(29) Dosage: 5 g of powder are added to 100 mL and are administered.

Application Example 3: Effervescent Tablet

(30) 50.00 g Paracetamol

(31) 27.00 g sorbitol

(32) 5.00 g sodium cyclamate

(33) 0.80 g saccharin

(34) 5.00 g HED-5 (spray dried HED sodium salt on maltodextrin)

(35) 2.00 g 1,2-propylene glycol

(36) The components are mixed and subsequently

(37) 100.00 g sodium hydrogen carbonate

(38) 136.00 g citric acid

(39) are added. After 2 hours, tablets are pressed thereof (2 g/tablet)

(40) Dosage: 1 tablet is added to 100 ml water and then administered.

Application Example 4: Juice

(41) 5 ml of juice contain:

(42) 200 mg Paracetamol

(43) 20 mg HED-1

(44) Further components: Purified water, sucrose, sodium citrate, tragacanth, citric acid, methyl-4-hydroxy benzoate (E218), propyl-4-hydroxy benzoate (E216).

Application Example 5: Powder for Producing a Solution

(45) 1 sachet (6 g) contains:

(46) 600 mg Paracetamol

(47) 80 mg HED-2 (Homoeriodictyol sodium salt variant 1)

(48) Further components: Citric acid, sodium citrate, sucrose 3.7 g, saccharin-sodium salt, sodium cyclamate, maize starch, highly disperse silicon dioxide, ascorbic acid, maltodextrin, butyl hydroxy anisole, modified starch, curcumin, orange flavour.

Application Example 6: Effervescent Tablet

(49) 1 effervescent tablet contains:

(50) 500 mg Paracetamol

(51) 100 mg HED-5

(52) Further components: Ascorbic acid, citric acid, lactose 1H2O, macrogol 6000, methyl cellulose, sodium cyclamate, sodium hydrogen carbonate, sodium sulphate, povidone K25, saccharin sodium salt, simeticon, sorbic acid, citrus flavour.

Application Example 7: Juice

(53) 1 bottle (100 ml) contains:

(54) 4.000 mg Paracetamol, i.e. one dose (5 ml) contains 200 mg Paracetamol. 200 mg HED-3 (Homoeriodictyol sodium salt variant 2)

(55) Further components: sodium metabisulphite (E223), propylene glycol, saccharin sodium salt, sorbitol solution 70% (non-crystallized) (E420), purified water, flavour type cherry.

Application Example 8: Tablets

(56) 1 tablet contains:

(57) 50 mg Paracetamol

(58) 10 mg HED-5

(59) Further components: Maize starch, povidone, talc, croscarmellose sodium, microcrystalline cellulose, magnesium stearate.

Application Example 9: Solution

(60) 100 ml solution contain:

(61) 4000 mg Paracetamol

(62) 8000 mg HED-6 (2.5% solution of HED-1 in 1,2-propylene glycol)

(63) Further components: Glycerol, sodium metabisulphite, citric acid, sodium hydroxide, acesulfame potassium salt, flavour type red fruit, macrogol, saccharine sodium salt, purified water.

Application Example 10: Tablets

(64) 1 tablet contains:

(65) 500 mg Paracetamol

(66) 50 mg HED-3

(67) Further components: Microcrystalline cellulose, maize starch, stearic acid, povidone

Application Example 11: Fruit Gum

(68) TABLE-US-00002 Ingredients Indication of quantity in wt.-% A B Gelatine 7.6% 7.6% Paracetamol   5%   5% Symrise citrus flavour with 0.8% 0.8% dye HED-1 0.5% — HED-5 — 2.5% Water Ad 100% Ad 100%

(69) For producing a fruit gum with 200 mg Paracetamol per dose, the ingredients listed in the above recipe are added to a boiled sugar syrup mixture (89° Brix).