WO2020208404A1

WO2020208404A1 - A highly available powder composition containing red rice

Info

Publication number: WO2020208404A1
Application number: PCT/IB2019/052998
Authority: WO
Inventors: Mauro Rissa
Original assignee: Pharmanutrition R&D S.r.l.
Priority date: 2019-04-11
Filing date: 2019-04-11
Publication date: 2020-10-15
Also published as: EP3952670A1

Abstract

It is described: A) a powder composition comprising: • Monascus Purpureus titrated in Monacolin K, · possibly associated with coenzyme Q10, • at least one hydrophilic surfactant, and • at least one mucolytic agent; B) a method for preparing said powder compositions A), and C) oral formulations containing the aforementioned powder compositions A) in the form of food supplements, foods for special medical purposes or cholesterol-lowering medicinal products for use in the treatment of cardiovascular disorders.

Description

A HIGHLY AVAILABLE POWDER COMPOSITION CONTAINING RED RICE

FIELD OF THE INVENTION

The present invention comprises powder compositions containing fermented red yeast rice, a method for preparing said powder compositions and related oral formulations, in the form of a food supplement, of food for special medical purposes and as a cholesterol-lowering medicinal product for use in the prevention and/or the treatment of cardiovascular disorders.

STATE OF THE ART

Hypercholesterolemia has now been definitely identified as a determining factor in the perspective of an increased cardiovascular risk. This evidence is the result of a long series of epidemiological studies that have clearly correlated cholesterolaemia (plasma cholesterol concentrations) and cardiovascular ischemic events, with specific reference to myocardial infarction and to the relative cardiovascular mortality, of which myocardial infarction and cerebral stroke represent the most frequent aetiology.

In particular, The Seven Countries Study (1999) is one of the most famous and authoritative of these epidemiological investigations.

In this study, 12,770 men aged between 40 and 59 were enrolled in seven countries: the Netherlands, Finland, Greece, Yugoslavia, Italy, Japan and the USA and monitored for 25 years. Finland was the nation with the highest mortality due to ischemic heart disease as opposed to Japan, which showed the lowest mortality. The two main risk factors that emerged from the study were arterial hypertension and hypercholesterolemia, in turn closely related to the intake of saturated fat and cholesterol in the diet. On the other hand, a study published in 1975 (Ni-Hon-San Study) showed that the incidence of ischemic heart disease in 12,000 men of Japanese origin divided into two groups, one group of which resided in Japan, was half if compared to the group composed of subjects living in Hawaii and California. The data indicate the importance of environmental factors in the development of ischemic heart disease, but also the unequivocal role of genetic imprinting.

Longitudinal studies have shown that the risk of death from ischemic heart disease is strictly proportional to cholesterol levels: the higher the plasma concentrations of cholesterol, the higher the cardiovascular mortality. This correlation indicates that as cholesterolaemia increases, also the mortality due to ischemic heart disease increases more rapidly, so that an increase in plasma cholesterol of one percentage point corresponds to an increase in mortality of almost two percentage points: compared to 200 mg/dl of cholesterolaemia, the mortality rate doubled at 250 mg/dl and quadrupled at 300 mg/dl.

A further important fact that clearly emerged from the studies indicates that the various risk factors examined (mainly hypertension, smoking, diabetes, obesity, family history of ischemic heart disease and low levels of HDL) create an enhancement synergy, so that several risk factors associated in an individual determine an increase in the probability of death from cardiovascular causes.

In conclusion, epidemiological studies have demonstrated that total cholesterolaemia and, above all, LDL values and the LDL/HDL ratio are important risk factors for ischemic cardiovascular diseases. However, epidemiological studies only allow the formulation of probability hypotheses, but are unable to demonstrate a cause-effect relationship, in this case between cholesterolaemia and cardiovascular diseases. Experimental studies (albeit with the limitations of experimental character) and clinical studies have emerged in support of the causal role of cholesterolaemia in cardiovascular pathology. Experimental studies have ascertained the importance of LDL in the pathogenesis of atherosclerosis, while clinical studies have shown that a reduction in cholesterolaemia is actually accompanied by a reduction in ischemic cardiovascular events.

The use of nutraceuticals able to restore plasma physiological concentrations of plasma cholesterol in subjects affected by mild to moderate cardiovascular risk, not yet candidates for drug therapy, is becoming increasingly consolidated, also in clinical practice. Particularly, substances such as red yeast rice (RYR) titrated in Monacolin K have shown a clinically documented efficacy in reducing LDL cholesterol in hypercholesterolemic subjects [1, 2, 3, 4] Monacolin K is a substance with a statin-like structure that acts by modulating the endogenous synthesis of LDL cholesterol by inhibiting the HMG-Co-A reductase enzyme, a key step in the synthesis of the mevalonate precursor [5, 6]. In the light of the well-established pharmacodynamic mechanism and of the equally solid clinical evidence to confirm its clinical efficacy, RYR extracts titrated in Monacolin now represent a solid certainty in adjuvant nutraceutical treatment in patients with mild to moderate cardiovascular risk, not yet candidates for drug therapy with statins and other cholesterol-lowering drugs such as fibrates and ezetimibe.

Against this solid clinical evidence of efficacy, however, there is the strong individual variability of response to RYR extracts and the often uneven standardization of the extracts titrated in Monacolin K. It is likely to hypothesize that the strong variability of the clinical response in different persons and often also in the same subject treated over time is linked to the variable bioavailability of Monacolin K from the RYR phytocomplex. Since it is a composite vegetable extract in which Monacolin represents no more than 1-5% of the total extract weight, there are interactions between the vegetable components of flavonoid, having tannin and polyphenolic nature that can interfere with the enterohepatic physiology and even significantly modify the bioavailability of Monacolin. Surprisingly, Chen et al. have shown that Monacolin K in the form of RYR extracts titrated in Monacolin K results, both in vitro and in vivo, more bioavailable than the isolated molecule [7]. This result, apparently surprising, has been explained by the probable inhibitory interaction of the RYR vegetable components, which are mainly flavonoids, polyphenols, fatty acids and sterols, with the P-gP pump of the enterocyte and with the enterohepatic microsomal systems (CYT P450) that increase the bioavailability of the active molecule on HMG-CoA inhibition. Monacolin K (Lovastatin) is classified by the FDA as BCS Class II (Biopharmaceutics Classification System), i.e. as a molecule of poor solubility and high intestinal permeability. Moreover, the increased bioavailability of Monacolin from RYR compared to the isolated molecule or its conjugated acid seems to be linked to an increased solubility of the same in enteric secretions, promoted by some RYR components in the form of salified fatty acids and sterols with surfactant action. In fact, the studies conducted seem to show that the isolated Monacolin K is more bioavailable when associated with surfactants such as sodium lauryl sulphate. There is therefore a particularly interesting possibility of further increasing the hydro-dispersion and enteric assimilation of the RYR extracts in order to maximize the bioavailability of Monacolin K and above all reduce the poor uniformity of inter-individual and intra-individual efficacy in long term treatment. SUMMARY OF THE INVENTION

The Applicant has now found that the association with hydrophilic surfactant agents and mucolytic agents capable of fluidifying the mucopolysaccharide layer that coats the enteric epithelium (UWL, Unstirred Water Layer) has turned out to be an effective strategy to improve the dissolution of the RYR extract and of the Monacolin K extract, at the same time facilitating and accelerating trans-epithelial permeation. This strategy also contributes to reducing the individual variability of the response to cholesterol-lowering therapy with RYR extracts and therefore the number of non-responders.

In fact, as better discussed in more detail later in the present description, comparative tests conducted in a liquid that simulates intestinal fluid (SIF) have shown that the presence of hydrophilic surfactants and mucolytic agents considerably increases the dissolution of Monascus Purpureus variously titrated in Monacolin K (1, 3, 5%) in intestinal fluid.

Object of the present invention are therefore powder compositions containing Monascus Purpureus variously titrated in Monacolin K and at least one hydrophilic surfactant agent, preferably at least two hydrophilic surfactants and at least one mucolytic agent.

A further object of the present invention are oral formulations, in particular in the form of food supplements, foods for special medical purposes having cholesterol lowering activity for use in the prevention and treatment of cardiovascular disorders. BRIEF DESCRIPTION OF THE FIGURES

Figure 1 is a photo of a beaker containing the simulated enteric liquid (SIF), whose label shows the different components of said SIF. Figure 2 is a picture of two beakers: the one on the left contains the powder compositions of the present invention completely dissolved in the SIF, which is compared to the one on the right that contains the pure extract of Monascus Purpureus with the same titration in Monacolin K and in the same amounts as the one of the left beaker, although this extract is not completely dissolved in the SIF. DETAILED DESCRIPTION OF THE INVENTION

For the purposes of the present invention, the definitions "comprising" and "containing" referring to the powder composition according to the present invention mean that said composition can contemplate the presence of further components in addition to those explicitly reported and described.

The at least one surfactant agent is preferably selected from polyoxyethylenated sorbitan esters (polysorbates), sucrose esters with fatty acids (sucroesters), alkyl ethers of glucose (alkyl glucosides).

Preferably, the powder composition of the present invention comprises two hydrophilic surfactants, of which at least one is a polyoxyethylenated sorbitan ester, more preferably selected from polysorbate 20, polysorbate 40, polysorbate 60 and polysorbate 80, and even more preferably is polysorbate 80.

The mucolytic agents are preferably selected from N-acetylcysteine, carboxycysteine and their mixture, since they are able to increase the hydro-dispersion of the extract in the intestine and its epithelial permeability. Preferably, the powder compositions according to the present invention contain only the N-acetylcysteine as the mucolytic agent. From the tests conducted with RYR extracts variously titrated in Monacolin (1, 3 and 5%), the dissolution process in the simulated enteric liquid (SIF) (Fig. 1), at 37°C and subjected to magnetic stirring at 200 rpm, of the powders obtained by wet granulation of the aforesaid extract with N-acetylcysteine, polysorbate 80 and sucrose ester, showed a more complete and rapid dissolution with respect to the isolated RYR extract dissolved under the same conditions (Fig. 2).

Another characteristic of the present invention is the possibility to vehiculate together with the RYR the coenzyme Q10 (inhibition of the HMG-CoA reductase enzyme, involved in the synthesis of mevalonate, a biochemical intermediate necessary for the synthesis of cholesterol as well as of coenzyme Q10) in a highly bioavailable form, notwithstanding the known poor bioavailability of the substance per oral route [8, 9], and exploiting the same mechanisms of increase of enteric hydro-dispersion of the molecule. Chronic statin intake is notoriously associated with adverse muscular effects such as dystrophy, even severe pain (myalgia) and in severe cases rhabdomyo lysis [10, 11]. These events have been attributed, by various published works, to an altered presence of coenzyme Q10 in the intracellular mitochondrial compartment of the muscle fibre with consequent altered homeostasis and cellular suffering [12, 13]. However, there is no solid clinical evidence that supplementing with CoQlO at very high dosages can reduce, other than in a very variable and not always statistically significant manner, the symptoms in subjects suffering from statin- induced muscle pain [14, 15, 16]. These data, in contrast to what might be expected, given the congruity of the biochemical hypothesis, could be explained by the poor bioavailability of CoQlO in the enteric tract. The wet granulation of RYR and CoQlO with the above absorption enhancers and mucolytic agents leads to a significant increase in the bioavailability of both substances, mediated by increased enteric hydro-dispersion, actually increasing the efficacy and therapeutic uniformity profile of RYR as well as the one of its medium-long term tolerability. Red yeast rice (RYR) or Monascus Purpureus preferably has a titration in Monacolin K variable between 1 and 20% weight/weight of the RYR.

Preferably, the red yeast rice is contained in amounts comprised between 30 and 70% by weight, more preferably between 40 and 60% by weight on the total weight of the powder composition.

The hydrophilic surfactants are contained in a total amount preferably comprised between 10 and 50% by weight, preferably between 20 and 40% by weight on the total weight of the powder composition object of the present invention.

The mucolytic agents are preferably contained in amounts comprised between 5 and 25% by weight, more preferably between 10 and 20% by weight on the total weight of the powder composition.

When the powder composition also contains the coenzyme Q10, this is present in Monascus Purpureus /coenzyme Q10 weight ratios preferably comprised between 1:3 and 1:0.5.

Finally, the object of the present invention is also the preparation method of the powder composition according to the present invention, which comprises the following steps:

a) dry-mixing Monascus Purpureus, coenzyme Q10, when present, with at least one hydrophilic surfactant when this is not the polyoxyethylenated sorbitan ester, and at least one mucolytic agent,

b) moistening the mixture obtained in step a) with water or an aqueous solution of the polyoxyethylenated sorbitan ester, when this hydrophilic humectant is present, and obtaining a granulate,

c) drying the granulate obtained in step b) until a powder is obtained,

d) finally sieving the powder obtained in step c). The oral formulations object of the present invention are preferably in the form of water-dispersible tablets, capsules or granulates administrable per oral route.

Some examples of preparations of powder compositions of the present invention and of the related final oral formulations are given below for illustrative but not limitative purposes.

Example 1

Red yeast rice (RYR)

Monacolin K titration 3% 168 g (equivalent to 5 g of Monacolin K)

Polysorbate 80 20.0 g

Sucrose ester 90.0 g

N-acetylcysteine 40.0 g

Tot 318 g

Preparation method

Mix the red yeast rice, the N-acetylcysteine and the sucrose ester with a V-shaker for 10 minutes. Check the uniform granulometry of the powder mixture and if necessary proceed with the Vascuali mechanical sieve finishing. Wet the powder mixture with a solution of demineralized water containing polysorbate 80 in a ratio of 2.5 to 1 by weight (20 grams of polysorbate 80 dissolved in 50 ml of demineralized water) under constant stirring (mechanical planetary mixer) until obtaining a wet powder. Place the dough in a baking tray in a thin layer and place in a convection oven at 40°C, checking the weight loss from time to time. Once the weight uniformity has been reached (complete loss of the wetting water), sift the granulate with a Vascuali mechanical sieve. The powder obtained is added with the excipients necessary to obtain a water-soluble sachet:

Granulate 318 g (equal to 5 g of Monacolin K) Sorbitol 1100 g

Sucralose 15.0 g

Orange aroma 30.0 g

Amorphous silica 37.0 g

Tot 1500 g

Dosage: 1 sachet of 1.5 g in the evening equal to 5.0 mg of Monacolin K

Example 2

Red yeast rice (RYR)

Monacolin K titration 3% 168 g (equivalent to 5 mg of Monacolin K)

Coenzyme Q10 100 g

Polysorbate 80 20.0 g

Sucrose ester 90.0 g

N-acetylcysteine 50.0 g

Tot 428 g

Preparation method

Mix the red yeast rice, the coenzyme Q10, the N-acetylcysteine and the sucrose ester with a V-shaker for 10 minutes. Check the uniform granulometry of the powder mixture and if necessary proceed with the Vascuali mechanical sieve finishing. Wet the powder mixture with a solution of demineralized water containing polysorbate 80 in a ratio of 2.5 to 1 by weight (20 grams of polysorbate 80 dissolved in 50 ml of demineralized water) under constant stirring (mechanical planetary mixer) until obtaining a wet powder. Place the dough in a baking tray in a thin layer and place in a convection oven at 40°C, checking the weight loss from time to time. Once the weight uniformity has been reached (complete loss of the wetting water), sift the granulate with a Vascuali mechanical sieve. The powder obtained is added with the excipients necessary to obtain a water-soluble sachet:

Granulate 428 g

Sorbitol 1000 g

Sucralose 15.0 g

Orange aroma 30.0 g

Amorphous silica 27.0 g

Tot 1500 g

Dosage: 1 sachet of 1.5 g in the evening equal to 5 mg of Monacolin K and 100 mg of coenzyme Q 10.

Bibliography

Cicero AF et al. Middle-Term Dietary Supplementation with Red Yeast Rice Plus Coenzyme Q10 Improves Lipid Pattern, Endothelial Reactivity and Arterial Stiffness in Moderately Hypercholesterolemic Subjects. Ann Nutr Metab. 2016;68(3):213-9; Burke FM. Red yeast rice for the treatment of dyslipidemia. Curr Atheroscler Ren. 2015 Apr: 17(41:495:

Liu J. Chinese red yeast rice ( Monascus Purpureus) for primary hyperlipidemia: a meta-analysis of randomized controlled trials. Chin Med. 2006 Nov 23; 1:4;

Davidson MH. A multicenter, randomized, double-blind clinical trial comparing the low-density lipoprotein cholesterol-lowering ability of lovastatin 10, 20, and 40 mg/d with fluvastatin 20 and 40 mg/d. Clin Ther 2003 Nov;25(l l):2738-53;

Sirtori CR· The pharmacology of statins. Pharmacol Res. 2014 Oct;88:3-l 1;

Egom EE, Hafeez IT Biochemistry of Statins. Adv Clin Chcm. 2016;73:127-68; 7. Chen CH et al. Improved dissolution rate and oral bioavailability of lovastatin in red yeast rice products. I at i Pharm 2013 Feb 28;444( 1 -2): 18-24;

8. Balakrishnan P et al. Enhanced oral bioavailability of Coenzyme QlO by self- emulsifying drug delivery systems. Int J Pharm. 2009 Jun 5;374(l-2):66-72;

59. Sato Y et al. Emulsification using highly hydrophilic surfactants improves the absorption of orally administered coenzyme Q10. Biol Pharm Bull. 2013:36021:2012-7;

10. Tomaszewski M. Stepien KM. Tomaszewska I. Czuczwar SI . Statin-induced myopathies. Pharmacoj Rep. 2011;63(4):859-66;

1011. Blaier O. Idshner M. Elis A. Managing statin-induced muscle toxicity in a lipid clinic. J Clin Pharm Thor. 2011 Jun;36(3):336-41;

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15 Rev. 2005 Mar-Apr; 13(2):76-9;

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Claims

1. Powder composition containing: fermented Monascus Purpureus extract titrated in Monacolin K, at least one hydrophilic surfactant and at least one mucolytic agent.

2. Powder composition according to claim 1, containing at least two of said hydrophilic surfactants selected from: polyoxyethylenated sorbitan esters

(polysorbates), sucrose esters (sucroesters) and glucose alkyl ethers (alkyl glucosides).

3. Powder composition according to claim 1 or 2, comprising at least two hydrophilic surfactant agents, at least one of them being a polyoxyethylenated sorbitan ester.

4. Powder composition according to any one of claims 1-3, wherein the polyoxyethylenated sorbitan ester is selected among polysorbate 20, 40, 60, more preferably polysorbate 80.

5. Powder composition according to any one of claims 1-4, wherein the mucolytic agents are selected from: N-acetylcysteine, carboxycysteine and related mixtures of said mucolytic agents, more preferably it is N-acetylcysteine.

6. Powder composition according to any one of claims 1-5, wherein the Monascus Purpureus extract is titrated in Monacolin K in a percentage ranging from 1% to 20% by weight on the total weight of said composition.

7. Powder composition according to any one of claims 1-6, comprising the coenzyme Q10 in the form of ubiquinone or ubiquinol.

8. Powder composition according to any one of claims 1-7, wherein the total amount of hydrophilic surfactants is comprised in a percentage ranging from 10 to 50%, preferably from 20 to 40% by weight on the total weight of the powder composition.

9. Powder composition according to any one of claims 1-8, wherein the total amount of mucolytic agents is comprised in a percentage ranging from 5 to 25%, preferably from 10 to 20% by weight on the total weight of said powder composition.

10. Powder composition according to claim 7, wherein the weight ratio Monascus Purpureus : coenzyme Q10 is respectively variable between 1:3 and 1:0.5.

11. Oral formulation comprising the powder composition according to any one of claims 1-10 in combination with suitable excipients and/or diluents.

12. Oral formulation according to claim 11, in the form of a food supplement, a food for special medical purposes or a medicinal product.

13. Oral formulation according to any one of claims 11 or 12 having cholesterol lowering activity for use in the prevention and treatment of cardiovascular disorders.

14. Method for preparing the powder composition according to any one of claims 1- 10, comprising the following steps:

(a) dry-mixing Monascus Purpureus, the coenzyme Q10 when present, with at least one hydrophilic surfactant, when it is not polyoxyethylenated sorbitan ester, and at least one mucolytic agent;

b) moistening the mixture obtained in step a) with water or an aqueous solution of polyoxyethylenated sorbitan ester, when this hydrophilic surfactant is present, and obtaining a granulate;

c) drying the granulate obtained in step b) until a powder is obtained;

d) finally sieving the powder obtained in step c).