EP3897677A1

EP3897677A1 - Composition for use in a method for lowering of ldl-cholesterol in plasma

Info

Publication number: EP3897677A1
Application number: EP19829701.2A
Authority: EP
Inventors: Ruddy Wattiez; Felice MASTROLEO; Robertus Christiaan Josephus Suters
Original assignee: Ezcol BV
Current assignee: Ezcol BV
Priority date: 2018-12-18
Filing date: 2019-12-17
Publication date: 2021-10-27
Also published as: WO2020130813A1

Abstract

The present invention relates to a (pharmaceutical) composition comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for use in a method for the lowering of LDL-cholesterol in subjects in need thereof. Furthermore, the invention relates to a foodstuff comprising a food supplement, wherein the food supplement has cholesterol-lowering properties and comprises compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. For the pharmaceutical composition and the composition and the food supplement the proviso is that the (pharmaceutical) composition is not, and the compounds are not provided as, a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1:10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. Furthermore, the (pharmaceutical) composition and compounds are not comprising or provided as whole R.rubrum cells or as a membrane fraction of R.rubrum.

Description

COMPOSITION FOR USE IN A METHOD FOR LOWERING OF LDL-CHOLESTEROL IN PLASMA

TECHNOLOGICAL FIELD

The present invention relates to a composition comprising bacteriopheophytins, carotenoids and quinones, in particular compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and a pharmaceutical composition comprising a bacteriopheophytin, carotenoid and quinone in particular a composition comprising said compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in a method for the lowering of LDL-cholesterol in the plasma of a subject in need thereof, such as a human subject. In particular, the present invention relates to compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and a pharmaceutical composition comprising compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in a method for the treatment or the prophylaxis of cardiovascular disease or a risk factor for developing cardiovascular disease or atherosclerosis. The present invention also relates to bacteriopheophytins, carotenoids and quinones such as compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and to a pharmaceutical composition comprising a bacteriopheophytin, carotenoid and quinone in particular said compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for use in a method for the treatment or prophylaxis of a cardiovascular risk factor and/or a risk factor for developing atherosclerosis, in particular a plasma LDL-cholesterol level of 70 mg/dL or higher, or 100 mg/dL or higher, or 130 mg/dL or higher. The invention also relates to a food supplement with cholesterol-lowering properties, comprising compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. Furthermore, the invention relates to a foodstuff comprising a food supplement, wherein the food supplement has cholesterol-lowering properties and comprises compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. For the pharmaceutical composition and the composition and the food supplement the proviso is that the (pharmaceutical) composition is not, and the compounds are not provided as, a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. Furthermore, for the composition according to the invention and for the composition for use according to the invention, the proviso is that the composition and the composition for use of the invention is not a preparation comprising the membrane fraction of Rhodospirillum spp. and/or the membrane fraction of Phaeospirillum spp., and not a preparation or pharmaceutical preparation of Rhodospirillum spp. for use in lowering plasma cholesterol, said preparation or pharmaceutical preparation comprising dead Rhodospirillum spp. , or freeze-dried Rhodospirillum spp.

BACKGROUND

Cardiovascular diseases (CVDs) are responsible for over 17.3 million deaths per year and are the leading causes of death in the world, according to the World Health Organization. CVDs include diseases of the heart, vascular diseases of the brain and diseases of blood vessels. The different types of CVDs are: CVDs due to atherosclerosis, which are ischaemic heart disease or coronary artery disease (e.g. heart attack); cerebrovascular disease (e.g. stroke); diseases of the aorta and arteries, including hypertension and peripheral vascular disease, and other CVDs, i.e. congenital heart disease; rheumatic heart disease; cardiomyopathies; and cardiac arrhythmias.

CVD is caused by a number of synergistic factors, the most important being a too high blood cholesterol level. Cholesterol is an essential building block for animal and human cells, since it is a component of cell membranes. Human cells can synthesize their own cholesterol, but cholesterol is also assimilated from food. Both processes play an important part in cholesterol metabolism.

Apart from its essential biological role as a building block for cellular membranes, cholesterol also has negative effects on human health, as a cause of cardiovascular disease (such as, for instance, myocardial infarction, stroke, and peripheral vascular disease), more specifically in relation to the occurrence of atherosclerotic lesions in the blood vessel wall. An elevated plasma cholesterol level is the most important predictive risk factor for the occurrence of cardiovascular disease and atherosclerosis.

In blood plasma, cholesterol is transported in lipoproteins, which can be subdivided into a number of different classes, based on their diameter and specific density. The very-low-density lipoproteins (VLDL), the intermediate-density lipoproteins (IDL), the low-density lipoproteins (LDL), and the high-density lipoproteins (HDL) constitute the most important classes of lipoproteins.

Experimental and clinical studies have shown that the amount of cholesterol transported in the VLDL, IDL and LDL classes of lipoproteins (the pro-atherogenic cholesterol) is a risk factor for the occurrence of cardiovascular disease. Cholesterol transported in HDL particles, in contrast, protects against the development of cardiovascular disease (anti-atherogenic cholesterol). Randomized, placebo-controlled, prospective clinical studies have demonstrated that lowering plasma cholesterol has a favorable effect on the incidence of cardiovascular disease and on mortality. A prerequisite is, though, that the reduction in cholesterol should be predominantly or substantially due to a reduction in the pro-atherogenic cholesterol present in LDL, leaving the level of anti-atherogenic cholesterol (HDL-cholesterol) preferably essentially unaltered.

For the treatment and prevention of cardiovascular disease it is therefore imperative to reduce the pro-atherogenic cholesterol such as the level of LDL-cholesterol, and to increase, in absolute or relative proportion, the anti-atherogenic cholesterol, the HDL-cholesterol.

A number of approaches are available to reduce plasma cholesterol. The most important are: to inhibit cholesterol biosynthesis;

to increase the removal of cholesterol (and/or its metabolites, specifically bile acids) from tissues into the intestinal lumen;

to reduce the absorption of cholesterol and bile acids from the gastrointestinal tract.

Drugs that are used to inhibit cholesterol synthesis are often inhibitors of the enzyme hydroxymethyl- glutaryl-coenzyme A reductase (HMGCoA reductase), the rate-limiting enzyme in the cholesterol synthesis pathway. These“statins” are molecules that inhibit enzyme action. Examples are simvastatin, pravastatin and atorvastatin. Statins are generally chemically-synthetized derivatives of naturally- occurring fungal metabolites.

Extended release niacin has been reported to lower LDL-cholesterol with 17%. Fenofibrate has been reported to lower LDL-cholesterol levels with about 20%. Ezetimibe is an intestinal cholesterol absorption inhibitor which reduces LDL-cholesterol with 18%. Colesevelam is a bile acid sequestrant which reduces LDL-cholesterol with 18%. Mipomersen is an inhibitor of apolipoprotein B-100 synthesis and was shown to reduce LDL-cholesterol levels with about 25% in patients with homozygous familial hypercholesterolemia. Lomitapide is an inhibitor of microsomal triglyceride transfer protein for example for the treatment of patients with homozygous familial hypercholesterolemia. The lomitapide reduced LDL-cholesterol levels with 50% in those patients. Proprotein convertase subtilisin / kexin type 9 inhibitor (PCSK9 inhibitor) molecules and gene-silencing approaches are under development. Inhibition of PCSK9 in a subject may enhance the LDL-cholesterol lowering activity of statins. Combined treatment of subjects with an antibody against PCSK9 (REGN727 / SAR236553) and statin atorvastatin resulted in a reduction in LDL-cholesterol levels of about 39% to 61 %. The small molecule ETC-1002 modulates adenosine triphosphate-citrate lyase as well as adenosine monophosphate-activated protein kinase. In patients suffering from hypercholesterolemia (LDL-cholesterol levels of 130-220 mg/dL), LDL- cholesterol levels were reduced with about 18% to about 27%, when treated with increasing doses of ETC-1002. Cholesteryl ester transfer protein (CETP) inhibitors raise HDL-cholesterol levels and decrease LDL-cholesterol levels. Examples of such a CETP inhibitors are anacetrapib and evacetrapib. Anacetrapib and evacetrapib have been shown in clinical trials with human subjects to increase HDL- cholesterol levels with respectively about 138% and about 129%, and to lower LDL-cholesterol levels with respectively about 40% and about 36%. WAY-252623 is an activator of the beta-isoform of the liver X receptors. In non-human primates, WAY-252623 reduced LDL-cholesterol with 70%-77%. To increase cholesterol removal, a bile acid-adsorbing resin can be used (for example cholestyramine). Because of the adsorption of bile acids to the resin, their secretion in the stool is increased, and their reabsorption from the gut into the blood is reduced, resulting in a relative loss of bile acids from the body. Consequently, the liver increases the conversion of cholesterol into bile acids, resulting in a net increase in the secretion of cholesterol (metabolites) from the body. Because bile acids (by solubilizing cholesterol) are essential for the uptake of cholesterol from the lumen into the intestinal tissue, a reduction in bile acid content in the intestinal lumen will also result in a decreased cholesterol uptake.

Maintenance of cholesterol homeostasis is vital for healthy status and achieved through a regulatory network consisting of genes involved in cholesterol synthesis, absorption, metabolism and elimination. Imbalance of cholesterol level as a results of environmental and genetic factors leads to hypercholesterolemia, a predominant risk factor for atherosclerosis ( i.e . hardening of furring of the arteries) and associated coronary and cerebrovascular diseases. Hypercholesterolemia and its associated cardiovascular diseases represent one of the greatest worldwide economic, social and medical challenges that we are facing now.

Despite the wide use of therapeutic drugs for controlling blood cholesterol, like statins inhibiting cholesterol synthesis, the fact remains that it is estimated that more than 50% of the population of the United States has cholesterol levels at the borderline levels. In addition, adverse effects associated with therapeutic drugs to control cholesterol levels, such as myopathy, liver damages and potential drug- drug interaction, have been reported. Therefore, development of additional therapies for controlling cholesterol levels is warranted, especially for those with better safety profile.

Patent EP1569667 discloses a preparation of Rhodospirillum cells for use as a medicament, for use in a food supplement, for use in a foodstuff, wherein said use is for use in a method for the lowering of plasma cholesterol in a subject.

SUMMARY

There exists a need for cholesterol-lowering compounds, compositions and pharmaceutical compositions, specifically for use in the lowering of plasma LDL-cholesterol concentration in human subjects in need thereof, in particular efficacious compounds or compositions having an acceptable and/or improved safety profile.

The present invention will be described with respect to particular embodiments but the invention is not limited thereto but only by the claims.

The embodiments of the invention described herein can operate in combination and cooperation, unless specified otherwise.

An aspect of the invention relates to a composition comprising at least one carotenoid, pheophytin such as a bacteriopheophytin or quinone for use in lowering plasma cholesterol, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a composition comprising at least one carotenoid, pheophytin such as a bacteriopheophytin or quinone for use in the lowering of LDL-cholesterol in blood plasma of a subject, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a composition comprising at least one carotenoid, pheophytin, such as a bacteriopheophytin, or quinone for use in a method for the lowering of LDL-cholesterol in blood plasma of a subject, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a composition comprising at least one carotenoid, pheophytin such as a bacteriopheophytin or quinone for use in the treatment or prophylaxis of a cardiovascular disease (CVD) such as coronary heart disease, or of risk factors associated with development of a CVD or for example atherosclerosis, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a composition comprising at least one carotenoid such as a xanthophyll for use in the treatment or prophylaxis of a cardiovascular disease (CVD) such as coronary heart disease, or of risk factors associated with development of a CVD or for example atherosclerosis, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a composition comprising at least one pheophytin such as a bacteriopheophytin for use in the treatment or prophylaxis of a cardiovascular disease (CVD) such as coronary heart disease, or of risk factors associated with development of a CVD or for example atherosclerosis, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. The pheophytin(s) is/are for example natural pheophytin(s) and are for example isolated from a natural source or are chemically synthesized. An aspect of the invention relates to a composition comprising at least one quinone or ubiquinone or ubiquinol or ubiquinone derivative for use in the treatment or prophylaxis of a cardiovascular disease (CVD) such as coronary heart disease, or of risk factors associated with development of a CVD or for example atherosclerosis. The quinone or ubiquinone or ubiquinol or ubiquinone derivative is/are for example natural quinone, ubiquinone or ubiquinol or a naturally occurring ubiquinone derivative such as rhodoquinone, and are for example isolated from a natural source or are chemically synthesized. The ubiquinol is for example any one or more of naturally occurring and/or chemically synthesized ubiquinol-5, ubiquinol-6, ubiquinol-7, ubiquinol-8, ubiquinol-9, ubiquinol-10, ubiquinol-1 1 , ubiquinol-12, ubiquinol-13, ubiquinol-14, or is for example any one or more of naturally occurring and/or chemically synthesized ubiquinol-6, ubiquinol-7, ubiquinol-8, ubiquinol-9, ubiquinol-10, ubiquinol-1 1 , ubiquinol-12, ubiquinol-13, or is for example any one or more of naturally occurring and/or chemically synthesized ubiquinol-7, ubiquinol-8, ubiquinol-9, ubiquinol-10, ubiquinol-1 1 , ubiquinol-12, or is for example any one or more of naturally occurring and/or chemically synthesized ubiquinol-8, ubiquinol-9, ubiquinol-10, ubiquinol-1 1 , or is for example any one or more of naturally occurring and/or chemically synthesized ubiquinol-9 or ubiquinol-10, such as ubiquinol-10. The ubiquinone is for example any one or more of naturally occurring and/or chemically synthesized ubiquinone-5, ubiquinone-6, ubiquinone-7, ubiquinone-8, ubiquinone-9, ubiquinone-10, ubiquinone-1 1 , ubiquinone-12, ubiquinone-13, ubiquinone-14, or is for example any one or more of naturally occurring and/or chemically synthesized ubiquinone-6, ubiquinone-7, ubiquinone-8, ubiquinone-9, ubiquinone-10, ubiquinone-1 1 , ubiquinone-12, ubiquinone-13, or is for example any one or more of naturally occurring and/or chemically synthesized ubiquinone-7, ubiquinone-8, ubiquinone- 9, ubiquinone-10, ubiquinone-1 1 , ubiquinone-12, or is for example any one or more of naturally occurring and/or chemically synthesized ubiquinone-8, ubiquinone-9, ubiquinone-10, ubiquinone-1 1 , or is for example any one or more of naturally occurring and/or chemically synthesized ubiquinone-9 or ubiquinone-10, such as ubiquinone-9 or ubiquinone-10.

The pheophytin is for example a chlorophyll derivative lacking the central Mg²⁺ ion and is for example a bacteriopheophytin a, for example a bacteriopheophytin a comprising a phytyl group or comprising a geranyl-geranyl group. The carotenoid(s) is/are for example natural carotenoid(s) and are for example isolated from a natural source or are chemically synthesized. The carotenoid(s) is/are preferably tetraterpenoids such as a xanthophyll and a carotene, though xanthophylls are preferred. The carotenoids are preferably polar carotenoids such as xanthophylls, and xanthophylls are preferred. The carotenoids are preferably oxygenated carotenoids. The carotenoids are for example xanthophylls based on the precursor phytoene. The carotenoids are for example xanthophylls based on the precursor xanthophyll rhodopin. The xanthophyll is for example an alcohol, an ether, or has both a hydroxyl group and an ether group. The xanthophyll is for example rhodovibrin hydroxyspirilloxanthin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin or 3,4-spirilloxanthin. An aspect of the invention relates to a composition comprising at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in the treatment or prophylaxis of a cardiovascular disease (CVD), under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. Particularly, the treatment or prevention of a CVD in a human subject or in the treatment or prophylaxis of risk factors associated with development of a CVD or for example with development of atherosclerosis. The geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 are for example isolated from a natural source, such as cultured R.rubrum bacteria cells, or are chemically synthesized, under the proviso that the isolated geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

An aspect of the invention relates to a composition comprising two or more, in particular three, four, five, six, seven, eight, nine, ten, eleven, twelve or thirteen, preferably thirteen compounds of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in a method for the lowering of LDL- cholesterol in blood plasma of a human subject, wherein the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

An embodiment of the invention relates to a composition for use according to the invention comprising all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

An aspect of the invention relates to a composition comprising at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use as an LDL-cholesterol level lowering medicament, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An embodiment is the composition comprising at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use as an LDL-cholesterol level lowering medicament, wherein the HDL- cholesterol level in the plasma of a human subject to whom the composition is administered, remains essentially unaltered, increases when compared to the level prior to administration of the composition, or decreases to a lesser extent than the decrease of the LDL-cholesterol level upon administering the composition to said subject.

A further aspect of the invention relates to a composition comprising at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in the treatment or prophylaxis of atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL- cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, ischemia, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. Preferred is the use of said composition in the treatment or prophylaxis of these diseases and health threats, wherein the subject to whom the composition is administered is a human subject. As a result of said use, the plasma LDL-cholesterol level of the treated human subject reduces to a level relating to lower risk for CVD and/or any of the listed diseases and health problems, such as a level of less than 200 mg/dL LDL-cholesterol, or less than 160 mg/dL, or less than 140 mg/dL, or less than 120 mg/dL, or less than 100 mg/dL, or less than 80 mg/dL, such as between 45 mg/dL and 75 mg/dL LDL-cholesterol. The level of HDL-cholesterol in plasma of a human subject to whom the composition is administered essentially remains unaltered, increases, or decreases to a limited extent such as with about 5-20%.

An aspect of the invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, preferably all thirteen of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and optionally a pharmaceutically acceptable excipient for use in lowering plasma cholesterol. Preferably the pharmaceutical composition comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, preferably all thirteen of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. More preferably, the pharmaceutical composition comprises all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, under the proviso that the pharmaceutical composition is not, or does not comprise a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of at least one, preferably at least two, more preferably all, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10and optionally a pharmaceutically acceptable excipient for use in the lowering of LDL- cholesterol in blood plasma of a subject, under the proviso that the pharmaceutical composition is not, or does not comprise a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of at least one, preferably at least two, more preferably all, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and optionally a pharmaceutically acceptable excipient for use in a method for the lowering of LDL-cholesterol in blood plasma of a subject, under the proviso that the pharmaceutical composition is not, or does not comprise a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. . A further aspect of the invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of at least one, preferably at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, more preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and optionally a pharmaceutically acceptable excipient, for use in a method for the treatment or prophylaxis of any one or more of cardiovascular disease, atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL- cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, inflammation, inflammatory disease, ischemia, infection, under the proviso that the pharmaceutical composition is not, or does not comprise a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. The inventors previously established that a petroleum ether extract of Rhodospirillum rubrum ( R.rubrum ) comprises compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and reduces the LDL-cholesterol plasma level while maintaining the plasma HDL- cholesterol level essentially unaltered when the compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 in the bacteria cell extract was administered to a subject (international patent application with application number PCT/EP2018/065878).

In the pharmaceutical composition for use in the method for the treatment or prophylaxis of any of the diseases, health problems, risk factors, the at least one, preferably at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, more preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10is/are either the sole active pharmaceutical ingredient(s) in the pharmaceutical composition, orthe pharmaceutical composition is administered to a subject to whom at least one further active pharmaceutical ingredient is administered, such as an active pharmaceutical ingredient selected from a statin, niacin, fenofibrate, ezetimibe, colesevelam, mipomersen, lomitapide, a PCSK9 inhibitor, alirocumab, evolocumab, ETC-1002, a CETP inhibitor, anacetrapib, evacetrapib, WAY-252623, a blood- pressure lowering compound, hydrochlorothiazide. For example, a statin and a PCSK9 inhibitor are administered to a subject in need thereof, and the pharmaceutical composition according to the invention is administered to said subject. The at least one further active pharmaceutical ingredient is administered to a subject in need thereof simultaneously, separately or sequentially with the pharmaceutical composition for use in the method for the treatment or prophylaxis of e.g. CVD.

An aspect of the invention relates to a method for treating or preventing a cardiovascular disease, atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL-cholesterol plasma level of at least 70 mg/dL, an LDL- cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, inflammation, inflammatory disease, ischemia, infection, the method comprising administering in a subject in need thereof an effective amount of at least one, preferably at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, more preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, under the proviso that the effective amount of the one or more compounds is not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

A further aspect of the invention relates to a method of treating a patient suffering from an LDL- cholesterol concentration in the plasma of said patient of at least 70 mg/dl_ such as above 100 mg/dL or above 150 mg/dL by administering an effective amount of at least one, preferably at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, more preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 to the patient, under the proviso that the effective amount of the one or more compounds is not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. For example, the patient is a human patient.

An aspect of the invention relates to a method of treating a patient suffering from or having an increased risk for any one or more of cardiovascular disease, atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL- cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, inflammation, inflammatory disease, ischemia, infection, by administering an effective amount of at least one, preferably at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, more preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 to the patient, underthe proviso that the effective amount of the one or more compounds is not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. In an embodiment, the effective amount of at least one, preferably at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, more preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10is administered to a patient in need thereof, said patient suffering from or having an increased risk for any one or more of said aforementioned diseases or risk factors. The occurrence or presence of for example a plasma LDL-cholesterol level of at least 140 mg/dL is a risk factor for developing atherosclerosis.

The CVDs include diseases of the heart, vascular diseases of the brain and diseases of blood vessels. The CVDs are: CVDs due to atherosclerosis, which are ischaemic heart disease or coronary artery disease (e.g. heart attack), coronary heart disease; cerebrovascular disease (e.g. stroke); diseases of the aorta and arteries, including hypertension and peripheral vascular disease, and other CVDs, i.e. congenital heart disease; rheumatic heart disease; cardiomyopathies; and cardiac arrhythmias.

The invention also relates to a food supplement with cholesterol-lowering properties, comprising at least one, preferably at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, more preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, under the proviso that these one or more compounds is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. Furthermore, the invention relates to a foodstuff comprising a food supplement, wherein the food supplement has cholesterol-lowering properties and comprises at least one, preferably at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, more preferably all thirteen, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, under the proviso that these one or more compounds is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

Furthermore, for the composition according to the invention and for the composition for use according to the invention, the proviso is that the composition and the composition for use of the invention is not a preparation comprising the membrane fraction of Rhodospirillum spp. and/or the membrane fraction of Phaeospirillum spp., and not a preparation or pharmaceutical preparation of Rhodospirillum spp. for use in lowering plasma cholesterol, said preparation or pharmaceutical preparation comprising dead Rhodospirillum spp. , or freeze-dried Rhodospirillum spp. Thus, for example for the food supplement and the food stuff of the invention, the compounds are not provided as a preparation comprising the membrane fraction of Rhodospirillum spp. and/or the membrane fraction o Phaeospirillum spp., and not as a preparation or pharmaceutical preparation of Rhodospirillum spp. for use in lowering plasma cholesterol, said preparation or pharmaceutical preparation comprising dead Rhodospirillum spp. , or freeze-dried Rhodospirillum spp.

Thus, the current invention provides a composition comprising compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, and under the proviso that the composition of the invention is not a preparation comprising the membrane fraction of Rhodospirillum spp. and/or the membrane fraction of Phaeospirillum spp., and is not a preparation or pharmaceutical preparation of Rhodospirillum spp. for use in lowering plasma cholesterol, said preparation or pharmaceutical preparation comprising dead Rhodospirillum spp. , or freeze-dried Rhodospirillum spp. , according to the invention.

SHORT DESCRIPTION OF THE DRAWINGS

Figure 1 . This figure displays the scope of Figure 2 of WO 2004/052380 A1 , displaying the lipoprotein pattern in plasma of Wistar rats that were fed a normal chow diet (‘Controls’), and from Wistar rats that were fed a chow diet containing 10% (w/w) R. rubrum (‘10% R.rub’), as explained in detail in Example 2 of WO 2004/052380 A1 . Under influence of consumed R.rubrum cells the plasma LDL-cholesterol levels in the rats decreased, whereas the plasma HDL-cholesterol levels remained essentially unaltered.

Figure 2. This figure displays the scope of Figure 5 of WO 2004/052380 A1 , showing the lipoprotein pattern in plasma of C57BI/6 mice that were fed a hypercholesterolaemic“Western-type” diet, and a hypercholesterolaemic“Western-type” diet containing 10% (w/w) R.rubrum, as outlined in detail in Example 4 of WO 2004/052380 A1 . Under influence of consumed R.rubrum cells the plasma LDL-cholesterol levels in the mice decreased, whereas the plasma HDL-cholesterol levels remained essentially unaltered.

Figure 3. Shown is the plasma cholesterol lowering effect with regard to LDL-cholesterol in an in vivo mouse model, while HDL-cholesterol levels and total cholesterol levels essentially remain unaltered upon treatment of the mice. Mice were fed control feed (‘control’) or feed enriched with an extract of bacterium Rhodospirillum rubrum comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10(‘Extract 1.1’). This extract is an example of a composition comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

DEFINITIONS

The terms first, second, third and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequential or chronological order. The terms are interchangeable under appropriate circumstances and the embodiments of the invention can operate in other sequences than described or illustrated herein.

Furthermore, the various embodiments, although referred to as“preferred” or“e.g.” or“for example” or“in particular” are to be construed as exemplary manners in which the invention may be implemented rather than as limiting the scope of the invention.

The term“comprising”, used in the claims, should not be interpreted as being restricted to the elements or steps listed thereafter; it does not exclude other elements or steps. It needs to be interpreted as specifying the presence of the stated features, integers, steps or components as referred to, but does not preclude the presence or addition of one or more other features, integers, steps or components, or groups thereof. Thus, the scope of the expression“a pharmaceutical composition comprising A and B” should not be limited to a pharmaceutical composition consisting only of compounds A and B, rather with respect to the present invention, the only enumerated compounds of the pharmaceutical composition are A and B, and further the claim should be interpreted as including equivalents of those compounds.

A cholesterol-lowering property is herein defined as the capability of a compound or a composition, such as a carotenoid, bacteriopheophytin or quinone, such as compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, a composition comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, a (pharmaceutical) composition comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, a preparation, a food supplement or a foodstuff, when administered to the body of a subject, such as an animal subject, for example a human subject, to lower the cholesterol, or LDL- cholesterol, level of the blood or the plasma of said subject. Methods for measuring the level of cholesterol in blood and plasma are known to the skilled person.

Throughout the specification the term“petroleum ether with a boiling point of between 60°C and 80°C” has its regular scientific meaning and here refers to a petroleum fraction consisting of aliphatic hydrocarbons and boiling in the range of between 60°C and 80°C.

The term“LDL-cholesterol level” and the term“LDL-cholesterol concentration” have their regular scientific meaning throughout the description and in the claims, and here refer to the amount of cholesterol that is associated with and transported by LDL in the body of the subject, e.g. a human subject. The LDL-cholesterol level is measured directly, as outlined in the Examples section. Where indicated, the LDL-cholesterol concentration is calculated from the measured total cholesterol concentration, the measured HDL-cholesterol concentration and the measured tri-acyl glycerol concentration in a sample such as a plasma sample of a human subject, and based on these measured data, the LDL-cholesterol level is calculated.

DETAILED DESCRIPTION

It is a goal of at least certain embodiments and aspects of the present invention to provide a substance or composition comprising an active pharmaceutical ingredient with plasma LDL-cholesterol lowering activity in a subject to whom the substance or the composition is administered.

It is an objective of at least some aspects and embodiments of the current invention to provide a compound or a composition comprising said compound, which is one or more of i) safe at a dose suitable for administering to a subject in need thereof, wherein‘safe’ refers to an acceptable extent, if occurring at all, of adverse side events in a subject to whom the compound or composition is administered, such as a human subject with a plasma LDL-cholesterol level of 70 mg/dL or higher, ii) active when lowering plasma LDL-cholesterol level in a subject is considered, and iii) capable of maintaining plasma HDL-cholesterol essentially unaltered or increasing HDL-cholesterol concentration in plasma in an absolute manner or relative manner when compared to plasma LDL-cholesterol level, when the compound or the composition comprising the compound is administered to a (human) subject.

At least one of the above objectives is achieved by at least some embodiments of the invention, providing at least one, preferably at least two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, more preferably all, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in a method for treating or preventing a disease or a health problem such as CVD and atherosclerosis, related to a plasma LDL-cholesterol level of 70 mg/dL or higher in a subject, for example a human subject, such as for example 100 mg/dL or higher, under the proviso that the one or more compounds is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

At least one of the above objectives is also achieved by providing a composition comprising two or more compounds, such as three, four, five, six, seven, eight, nine, ten, eleven, twelve, preferably all thirteen of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in a method for the lowering of LDL-cholesterol in blood plasma of a human subject, wherein the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. Preferably, said composition for use according to the invention comprises all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

At least one of the above objectives is also achieved by at least some embodiments, providing a pharmaceutical composition comprising a pharmaceutically effective amount of at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and optionally a pharmaceutically acceptable excipient, for use in a method for the treatment or prophylaxis of any one or more of cardiovascular disease, atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL-cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, inflammation, inflammatory disease, ischemia, infection.

An aspect of the invention relates to a composition comprising one, two or more, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use as a medicament, under the proviso that the composition and the one or more compounds is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to the use of a composition comprising one, two or more, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for the manufacture of a medicament for the treatment of high LDL-cholesterol in blood plasma of a subject, such as a human subject, under the proviso that the composition and the one or more compounds is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. The term“high LDL-cholesterol in blood plasma” should be read as a plasma LDL-cholesterol concentration of at least 70 mg/dL, such as more than 160 mg/dL or more than 120 mg/dL. An aspect of the invention relates to the use of at least one carotenoid, pheophytin such as a bacteriopheophytin or quinone for the manufacture of a medicament for the lowering of LDL- cholesterol in blood plasma of a subject, such as a human subject, such as a human subject having a plasma LDL-cholesterol level of higher than 80 mg/dL, under the proviso that the one or more carotenoids is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a composition comprising one, two or more, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in lowering plasma cholesterol, under the proviso that the composition and the one or more compounds is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a composition comprising one, two or more, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for use in the lowering of LDL-cholesterol in blood plasma of a subject, under the proviso that the composition and the one or more compounds is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a composition comprising one, two or more, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for use in a method for the lowering of LDL-cholesterol in blood plasma of a subject, under the proviso that the composition and the one or more compounds is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

An aspect of the invention relates to a composition comprising one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, or more, preferably all thirteen of compouns geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for use in the treatment or prophylaxis of a cardiovascular disease such as coronary heart disease, under the proviso that the composition and the one or more compounds is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. An aspect of the invention relates to a composition comprising compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in the treatment or prophylaxis of a cardiovascular disease, under the proviso that the composition is not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. The CVDs include diseases of the heart, vascular diseases of the brain and diseases of blood vessels. The CVDs include: CVDs due to atherosclerosis, which are ischaemic heart disease or coronary artery disease (e.g. heart attack); cerebrovascular disease (e.g. stroke); diseases of the aorta and arteries, including hypertension and peripheral vascular disease, and other CVDs, i.e. congenital heart disease; rheumatic heart disease; cardiomyopathies; and cardiac arrhythmias.

A further aspect of the invention relates to a composition comprising one, two or more, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for use in the treatment or prophylaxis of atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL-cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, ischemia, under the proviso that the composition and the one or more compounds is/are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. A further aspect of the invention relates to a composition comprising compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in the treatment or prophylaxis of atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL plasma level of at least 70 mg/dL, an LDL plasma level of at least 100 mg/dL, an LDL plasma level of at least 140 mg/dL, an LDL plasma level of at least 200 mg/dL, a total cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, ischemia, under the proviso that the composition is not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

An aspect of the invention relates to a composition comprising two, three, four, five, six-twelve or more, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in a method for the lowering of LDL-cholesterol in blood plasma of a human subject, wherein the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

An embodiment is the composition for use according to the invention comprising all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

An aspect of the current invention relates to a composition comprising at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for use in a method for the treatment or prophylaxis of a cardiovascular risk factor and/or a risk factor for developing atherosclerosis, wherein the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. A risk factor for developing cardiovascular disease, or a cardiovascular risk factor, and/or a risk factor for developing atherosclerosis is for example a total blood cholesterol level of 200 mg/dL - 239 mg/dL or 240 mg/dL and higher. A further risk factor is for example a plasma LDL-cholesterol level of 100 mg/dL or higher, such as 100 mg/dL - 129 mg/dL, 130 mg/dL - 159 mg/dL, 160 mg/dL - 189 mg/dL, or 190 mg/dL and higher. A further risk factor is for example a plasma HDL-cholesterol level in a human subject of lower than 60 mg/dL, or less than 40 mg/dL.

Without wishing to be bound by any theory, lowering the LDL-cholesterol level in plasma / serum of a subject such as a human subject can be the consequence of inhibition of the absorption of cholesterol and/or cholesterol esters from the gastrointestinal tract, and/or the consequence of inhibition of the synthesis of cholesterol in e.g. the liver, and/or the consequence of an upregulated density and number of LDL-receptors on the plasma membranes of organ cells such as the liver cells, facilitating clearance of circulating LDL-cholesterol. A portion of cholesterol in the body and in the circulation originates from dietary intake, and in addition a portion of the cholesterol present in a subject originates from de novo synthesis in amongst others and predominantly the liver. Cholesterol biosynthesized in the liver is in part transported and stored in the gall bladder, in the bile. The bile with the cholesterol is excreted to the gastrointestinal tract when required, i.e. when the subject consumes fat-rich food products, etc. It has been established that a large portion of bile cholesterol is excreted with the feces, with the remainder of the bile cholesterol being taken up from the intestine together with a portion of dietary cholesterol. Amongst other lipophilic compounds, triacyl glycerides and cholesterol from the food intake, together with the cholesterol transferred from the gall bladder to the intestine with the bile, are solubilized first as emulsion particles and then in bile salt micelles. Subsequently, intestinal mucosal cells (enterocytes) absorp the mixed bile salt micelles comprising the cholesterol. Then, the cholesterol is transferred into the lymphatic circulation as part of chylomicrons and also as part of very low density lipoproteins (VLDLs), which are transferred to the blood circulation. Relocation and transfer of cholesterol comprising particles is based on passive simple diffusion and/or based on scavenger receptor mediated transfer, for example with involvement of scavenger receptor B-l and/or CD36. In the bloodstream the cholesterol-loaded chylomicrons transfer (in part) into chylomicron remnants carrying cholesterol, which are taken up from the circulation by the liver. The VLDL-cholesterol particles are degraded into intermediate density lipoproteins (IDLs) comprising the cholesterol, and further into low- density lipoprotein-cholesterol upon discarding triacyl glycerol molecules from the IDL particles. The LDL-cholesterol particles are bound by LDL-receptor molecules on the surface of organ cells such as the liver. Upon binding, the LDL-cholesterol is taken up by these LDL-receptor carrying cells.

Cholesterol taken up by liver cells is in part relocated to the gall bladder and becomes part of the stored bile, to be secreted into the intestine after food intake. Cholesterol taken up by liver cells is also in part transferred to the endoplasmic reticulum during assembly of VLDL particles comprising cholesterol. The cholesterol in the VLDL-cholesterol particles originate in part from de novo cholesterol synthesis by the liver cells and in part from dietary cholesterol if present in the diet of the subject. The liver cells excrete the VLDL-cholesterol into the blood circulation. In the blood circulation, the VLDL- cholesterol are transformed into IDL-cholesterol, which (partly) further transforms into LDL-cholesterol.

Carotenoids and (bacterio)chlorophyll analogues such as a bacteriopheophytin are not biosynthesized by humans. Carotenoids and bacteriopheophytins present in the body thus originate from e.g. dietary intake, food supplements, etc. It has been established that uptake of polar carotenoids such as xanthophylls, and bacteriopheophtyins such as bacteriopheophytin a, from the ingested food into the blood circulation and into e.g. liver cells, essentially is established via the same transport route and mechanism applied by the body for cholesterol intake from food. That is to say, the polar carotenoids and the bacteriopheophytins are dissolved in emulsion particles and then in mixed bile salt micelles, facilitating transferto the lymphatic circulation as part of chylomicrons, and further into the blood stream. Chylomicrons, once transferred into chylomicron remnants, are taken up by the liver. Quinones are both biosynthesized by humans and by other organisms that are at least in part subject to human consumption and/or a fraction is part of the human diet, for example when prepared as part of a food product or food supplement. Quinones present in the body can thus originate from e.g. dietary intake, food supplements, etc. Uptake of quinones such as ubiquinones and ubiquinols, from the ingested food into the blood circulation and into e.g. liver cells, essentially is established via the same transport route and mechanism applied by the body for cholesterol intake from food. That is to say, the quinones are dissolved in emulsion particles and then in mixed bile salt micelles, facilitating transfer to the lymphatic circulation as part of chylomicrons, and further into the blood stream. Chylomicrons, once transferred into chylomicron remnants, are taken up by the liver.

Again, without being bound by any theory, the inventors now propose that the LDL-lowering effect of the carotenoids, bacteriopheophytins and quinones is based on the following principle of inhibition of cholesterol uptake into the circulation and inhibition of VLDL-cholesterol release from (liver) cells into the blood circulation. In the intestine, dietary polar carotenoids, bacteriopheophytins as well as quinones compete with cholesterol during the process of formation of the lipid emulsion, such that uptake of cholesterol by the lipid emulsion is inhibited at the expense of increased uptake of the polar carotenoid, bacteriopheophytin or quinone by the lipid emulsion. As a consequence, less cholesterol, which originates from the bile and, if part of the diet, from food, etc., is transferrable to bile salt micelles, required for ultimate release into the lymph system and blood circulation. In contrast, the carotenoids, bacteriopheophytins and quinones are delivered to the circulation as part of the chylomicrons and VLDL particles. The liver takes up the chylomicron remnants comprising the carotenoids, bacteriopheophytins and quinones originating from the chylomicrons, and therewith the carotenoids, bacteriopheophytins and quinones become part of the pool of fat soluble compounds present and stored in liver cells, together with de novo synthesized cholesterol by the liver cells and cholesterol taken up by liver cells. In liver cells assembly of VLDL-cholesterol particles occurs. Since the polar carotenoids, bacteriopheophytins and quinones are also present in the pool of fat-soluble compounds to be incorporated in VLDL, the carotenoid, bacteriopheophtyin or quinone again can compete with the cholesterol for incorporation in the VLDL particle. Less cholesterol is secreted into the blood circulation with the VLDL, as a consequence. Further, also less IDL-cholesterol can be formed from VLDL, and then also less LDL- cholesterol. Indeed, polar carotenoids and quinones are determined in the blood plasma of human subjects, as part of LDL-particles. It is presumed that the bacteriopheophtyins are also in the blood plasma of human subjects, as part of LDL-particles. Furthermore, a lower chylomicron-cholesterol level in the blood due to inhibition of cholesterol uptake from the intestine by the presence of polar carotenoids, bacteriopheophtyins and quinones in the intestine, induces upregulation of LDL-receptor on hepatocytes, which as a result increases uptake of circulating LDL-cholesterol by the liver cells. Increased uptake of LDL-cholesterol by the hepatocytes additionally contributes to a lower level of blood LDL-cholesterol. The mode of action may in part resemble the mode of action of ezetimibe, for the cholesterol-absorption inhibitory activity. In addition, or alternatively, as part of the inhibitory activity of polar carotenoids, bacteriopheophytins and quinoneswhen lowering plasma level of LDL-cholesterol is considered, also (or solely) inhibition of enzymes and proteins involved in cholesterol synthesis and transport such as Niemann-Pick C1 Like 1 , ATP-binding cassette transporters such as ABCG5 and ABCG8, and 3-hydroxy-3-methylglutaryl-CoA reductase, by the geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and/or rhodoquinone-10 may occur.

In an embodiment, a composition comprising at least one of, preferably at least two of, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 or a pharmaceutical composition comprising at least one, preferably at least two, more preferably all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in a method for the lowering of LDL- cholesterol in blood of a (human) subject or for use in a method for the treatment or prophylaxis of a CVD or atherosclerosis, comprises the administering of the compound or the pharmaceutical composition to the subject during the intake of food comprising cholesterol. That is to say, the carotenoid, bacteriopheophytin and/or quinone is taken orally at the start of e.g. a cholesterol-comprising meal, snack, etc., during consumption thereof, and/or shortly thereafter. Furthermore, in an embodiment, a composition comprising at least one, preferably at least two, more preferably all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 or a pharmaceutical composition comprising at least one, preferably at least two, more preferably all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in a method for the lowering of LDL-cholesterol in blood of a (human) subject or for use in a method for the treatment or prophylaxis of a CVD or atherosclerosis, comprises the administering of the composition or the pharmaceutical composition to the subject during each intake of food comprising cholesterol, during the day. That is to say, the subject is administered the composition or pharmaceutical composition at any time during the day at which a cholesterol-rich product is consumed. Therefore, for example, the geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and/or rhodoquinone-10 is administered to a human subject during breakfast, and/or during lunch, and/or during dinner, and/or during any intake of a cholesterol-comprising food product such as a snack, during any moment of the day in between meals.

In an embodiment, the (human) subject to whom the composition comprising at least one, preferably at least two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, more preferably all thirteen of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 or the pharmaceutical composition comprising at least one, preferably at least two, more preferably all of compounds geranyl- geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is administered, has a LDL-cholesterol level in mmol/L of 3, 6-5, 7, or 3, 3-5, 3, or 3, 1 -5,0, or 2, 9-4, 7, or 2,77-4,44, or 2, 6-4, 2, or 2, 5-4,0, or 2, 4-3, 8, or 2,27-3,64, or 2,17-3,48. For example, at the start of the administration of the compound, composition or the pharmaceutical composition, the human subject has an LDL-cholesterol blood level of 2,94-4,71 mmol/L or 2,50-4,00 mmol/L. Typically, the LDL-cholesterol plasma level of a human subject is at least 95 mg/dL such as at least 100 mg/dL, and typically lower than 240 mg/dL, such as lower than 200 mg/dL, typically lower than 190 mg/dL or lower than 159 mg/dL.

In particular, the administration to a human subject of a composition comprising at least one, preferably at least two, most preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, or the pharmaceutical composition comprising at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in a method for the lowering of LDL-cholesterol in blood of a (human) subject or for use in a method for the treatment or prophylaxis of a CVD or atherosclerosis, results in the lowering of the LDL- cholesterol level in the plasma or serum of the subject, for example with 10-50% or 4-35%, and in addition results in the total cholesterol level remaining essentially unaltered or becoming lower as well, and/or in addition results in the HDL-cholesterol level remaining essentially unaltered or becoming higher, with for example 4-20% or 2-30%, under the proviso that the composition or pharmaceutical composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. For example, subjects such as human subjects to whom composition comprising at least one, preferably at least two, most preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, or the pharmaceutical composition comprising at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is administered, do not suffer from any disease or risk factor related to an increased risk for developing a CVD, diabetes, atherosclerosis, or are any one or more of a CVD patient, an atherosclerosis patient, a diabetic patient, and/or suffer from hypercholesterolemia. Typically the geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is/are the sole active pharmaceutical ingredients) administered to the human subject, or the geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is/are administered to a human subject to whom also a further active pharmaceutical ingredient is administered such as a statin, ezetimibe and/or a PCSK9 inhibitor. Oral administration is preferred. Once, twice, or thrice daily administration is preferred.

The lUPAC name of the carotenoid rhodovibrin is (6E,8E,10E,12E,14E,16E,18E,20E,22E,24E,26E,28E)-31 -methoxy-2,6,10,14,19,23,27,31 - octamethyldotriaconta-6,8,10,12,14,16,18,20,22,24,26,28-dodecaen-2-ol; the molecular weight of rhodovibrin is 584.929 g/mol. Rhodovibrin is isolated from for example R.rubmm bacterium cells (See also Example 2-4, here below), and rhodovibrin is synthesized (See for example Example 5, here below). The carotenoid is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the rhodovibrin can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the carotenoid to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dl_.

The lUPAC name of the carotenoid 1 -hydroxy-spirilloxanthin is (4E,6E,8E,10E,12E,14E,16E,18E,20E,22E,24E,26E,28E)-31 -methoxy-2,6,10,14,19,23,27,31 - octamethyldotriaconta-4,6,8,10,12,14,16,18,20,22,24,26,28-tridecaen-2-ol; the molecular weight of 1 - hydroxy-spirilloxanthin is 582.913 g/mol. 1 -hydroxy-spirilloxanthin is isolated from for example R.rubmm bacterium cells (See also the Examples 2-4, here below). The carotenoid is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the 1 -hydroxy-spirilloxanthin can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the carotenoid to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

The lUPAC name of the carotenoid 3,4-didehydro-rhodopin (also referred to as 3,4-dehydro- rhodopin) is (4E,6E,8E,10E,12E,14E,16E,18E,20E,22E,24E,26E)-2,6,10,14,19,23,27,31 - octamethyldotriaconta-4,6,8,10,12,14,16,18,20,22,24,26,30-tridecaen-2-ol; the molecular weight of 3,4- dehydro-rhodopin is 552.887 g/mol. 3,4-dehydro-rhodopin is isolated from for example R.rubmm bacterium cells (See also Example 2-4, here below), and 3,4-dehydro-rhodopin is synthesized (See for example Example 5, here below). The carotenoid is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the 3,4- dehydro-rhodopin can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the carotenoid to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

The lUPAC name of the carotenoid chloroxanthin is (6E,8E,10E,12E,14E,16E,18E,20E,22E,26E)-2,6,10,14,19,23,27,31 -octamethyldotriaconta-

6.8.10.12.14.16.18.20.22.26.30-undecaen-2-ol; the molecular weight of chloroxanthin is 556,919 g/mol. Chloroxanthin is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below), and chloroxanthin is chemically synthesized (See for example Example 5, here below). Chloroxanthin is also referred to as hydroxyneurosporene. The carotenoid is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the chloroxanthin can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the carotenoid to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

administration is preferred.

The lUPAC name of the carotenoid rhodopin is (6E,8E,10E,12E,14E,16E,18E,20E,22E,24E,26E)-2,6,10,14,19,23,27,31 -octamethyldotriaconta-

6.8.10.12.14.16.18.20.22.24.26.30-dodecaen-2-ol; the molecular weight of rhodopin is 554.903 g/mol. Rhodopin is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below), and rhodopin is synthesized (See for example Example 5, here below). The carotenoid is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the rhodopin can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the carotenoid to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

The lUPAC name of the carotenoid spirilloxanthin is (4E,6E,8E,10E,12E,14E,16E,18E,20E,22E,24E,26E,28E)-2,31 -dimethoxy-2,6,10,14,19,23,27,31 - octamethyldotriaconta-4,6,8,10,12,14,16,18,20,22,24,26,28-tridecaene; the molecular weight of spirilloxanthin is 596.94 g/mol. Spirilloxanthin, also referred to as rhodoviolascin, is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below), and spirilloxanthin is chemically synthesized (See for example Example 5, here below). The carotenoid is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the spirilloxanthin can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the carotenoid to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

The lUPAC name of the carotenoid 3,4-dihyd ro-spirilloxanthin is (4E,6E,8E,10E,12E,14E,16E,18E,20E,22E,24E,26E)-2,31 -dimethoxy-2,6,10,14,19,23,27,31 - octamethyldotriaconta-4,6,8,10,12,14,16,18,20,22,24,26-dodecaene; the molecular weight of rhodopin is 598.956 g/mol. 3,4-dihydro-spirilloxanthin is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below). The carotenoid is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the 3,4- dihyd ro-spirilloxanthin can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the carotenoid to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

Geranyl-geranyl bacteriopheophytin a is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below). The pheophytin, a bacteriochlorophyll derivative lacking the Mg²⁺ ion, is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the geranyl-geranyl bacteriopheophytin a can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the pheophytin to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

Phytyl bacteriopheophytin a is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below). The pheophytin, a bacteriochlorophyll derivative lacking the Mg²⁺ ion, is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the phytyl bacteriopheophytin a can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the pheophytin to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

The lUPAC name of the quinone ubiquinol-10 is 2-[(2E,6E,10E,14E,18E,22E,26E,30E,34E)- 3,7,1 1 ,15,19,23,27,31 ,35,39-decamethyltetraconta-2,6,10,14,18,22,26,30,34,38-decaenyl]-5,6- dimethoxy-3-methyl benzene-1 ,4-diol; the molecular weight of ubiquinol-10 is 865.381 g/mol. Ubiquinol- 10 is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below), or ubiquinol-10 is synthesized. The ubiquinol is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the ubiquinol-10 can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the ubiquinol to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

The lUPAC name of the quinone ubiquinone-9 is 2,3-diniethoxy-5-niethyl-6- [(2E,6E,1 QE,14E,1 SE,22E,26E,3GE)-3,7,1 1 ,15,19,23,27,31 ,35-nonaniethylhe.xatriaconta- 2,6,10,14,1 S,22,28,3Q,34-nonaenyi]cyclobexa-2,5-diene-1 ,4-dione; the molecular weight of ubiquinone- 9 is 795.246 g/mol. Ubiquinone-9 is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below), or ubiquinone-9 is synthesized (See Examples section, here below). The ubiquinone is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the ubiquinone-9 can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the ubiquinone to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL. The lUPAC name of the quinone ubiquinone-9 is 2,3-dimethoxy-5-methyl-6- [(2E,6E,1 GE,14E,18E,22E,26E,3QE)-3,7,1 1 ,15,19,23,27,31 ,35-nonamethyihexatriaconta- 2,6,10 14,18,22,26 30 34-nonaenyi]eyciohexa-2_:5-diene-1 ,4-dione; the molecular weight of ubiquinone- 9 is 795 246 g/mol. Ubiquinone-9 is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below), or ubiquinone-9 is synthesized (See Examples section, here below). The ubiquinone is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the ubiquinone-9 can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the ubiquinone to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

The lUPAC name of the ubiquinone ubiquinone-10 is 2- [(2E,6E,10E,14E,18E,22E,26E,30E,34E)-3,7,1 1 ,15,19,23,27,31 ,35,39-Decamethyltetraconta-

2.6.10.14.18.22.26.30.34.38-decaenyl]-5,6-dimethoxy-3-methylcyclohexa-2,5-diene-1 ,4-dione; the molecular weight of ubiquinone-10 is 863.365 g/mol. Ubiquinone-10 is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below), or ubiquinone-10 is synthesized (See Examples section, here below). The ubiquinone is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the ubiquinone-10 can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the ubiquinone to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

The lUPAC name of the ubiquinone derivative rhodoquinone-10 is 2-amino-5- [(2E,6E,10E,14E,18E,22E,26E,30E,34E)-3,7,1 1 ,15,19,23,27,31 ,35,39-decamethyltetraconta-

2.6.10.14.18.22.26.30.34.38-decaenyl]-3-methoxy-6-methylcyclohexa-2,5-diene-1 ,4-dione; the molecular weight of rhodoquinone-10 is 848.354 g/mol. Rhodoquinone-10 is isolated from for example R.rubrum bacterium cells (See also Example 2-4, here below), or rhodoquinone-10 is synthesized (See Examples section, here below). The ubiquinone derivative is lipophilic and is suitably dissolved in a solvent for dissolving lipophilic agents, such as a fat or an oil or a lipid or lipid composition. That is to say, the rhodoquinone-10 can be mixed with for example an oil or a lipid composition. The oil or lipids then act as a carrier, supporting the ease of (orally) administering the ubiquinone derivative to a subject such as a human subject, such as a patient in need thereof, such as a human subject suffering from a CVD or a plasma LDL-cholesterol level of at least 90 mg/dL.

The at least one carotenoid, pheophytin such as a bacteriopheophytin or quinone, e.g. rhodovibrin, hydroxyspirilloxanthin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, preferably a combination of odovibrin, hydroxyspirilloxanthin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for use according to the invention, is in particular administered to a subject having an LDL-cholesterol level in plasma of at least 1 ,8 mmol/L (70 mg/dL), or at least 2,59 mmol/L (100 mg/dL), or at least 3,34 mmol/L (129 mg/dL), or at least 4,0 mmol/L, such as at least 5,2 mmol/L (200 mg/dL). The subject is for example a human subject, such as a patient in need of treatment of a CVD or a plasma LDL-cholesterol level of over e.g. 1 10 mg/dL. A plasma LDL- cholesterol level of higher than 100 mg/dL is associated with an increased risk for developing cardiovascular disease. Furthermore, a subject, for example a human subject, having a plasma LDL- cholesterol level of higher than 100 mg/dL has an increased risk for developing for example atherosclerosis. The subject to whom the carotenoid, pheophytin such as a bacteriopheophtyin or quinone is administered is for example a subject suffering from a CVD or being at risk for developing a CVD or atherosclerosis. The human subject has for example a plasma LDL-cholesterol level of on average 75 mg/dL, or 90 mg/dL or higher, or 100-250 mg/dL such as 120-220 mg/dL or 140-200 mg/dL, 160-180 mg/dL, related to an increased risk for developing a disease or health problem such as a CVD, atherosclerosis, arteriosclerosis, a total cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, ischemia.

A composition comprising at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 or pharmaceutical composition comprising at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for use according to the invention is in certain embodiments administered to a subject such as a human subject to whom at least one further active pharmaceutical ingredient is (co- )administered, such as at least one active pharmaceutical ingredient selected from a statin, niacin, fenofibrate, ezetimibe, colesevelam, mipomersen, lomitapide, a PCSK9 inhibitor, alirocumab, evolocumab, ETC-1002, a CETP inhibitor, anacetrapib, evacetrapib, WAY-252623, a blood-pressure lowering compound, hydrochlorothiazide, or any combination thereof such as a statin combined with a CETP inhibitor. Co-administering a statin and a PCSK9 inhibitor to a human subject together with a carotenoid, pheophytin such as a bacteriopheophtyin and/or quinone, e.g. geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, is an example of said use. The at least one further active pharmaceutical ingredient and the carotenoid, pheophytin such as a bacteriopheophytin and/or quinone are administered simultaneously, separately, or sequentially. For example, (the combination of) the geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is/are co-administered, preferably separately, with a statin. For example, the composition comprising the thirteen compounds is administered sequentially with a PCSK9 inhibitor. For example, the composition comprising rodovibrin, hydroxyspirilloxanthin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, ubiquinol-10, ubiquinone-9, ubiquinone- 10 and rhodoquinone-10 is co-administered with a statin and a PCSK9 inhibitor. Inhibition of PCSK9 enhances the lipid-lowering effect of a statin. Preferably, the rodovibrin, hydroxyspirilloxanthin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, ubiquinol-10, ubiquinone-9, ubiquinone- 10 and rhodoquinone-10 are the sole active pharmaceutical active ingredients in a composition comprising rodovibrin, hydroxyspirilloxanthin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

The at least one further active pharmaceutical ingredient is administered to a subject in need thereof simultaneously, separately or sequentially with the (pharmaceutical) composition comprising carotenoid(s), bacteriopheophytin(s) and/or quinone(s), such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for use in the treatment or prophylaxis of e.g. CVD, atherosclerosis, etc. It is preferred that the at least one carotenoid, bacteriopheophytin and/or quinone is geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and/or rhodoquinone-10. Preferably, at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 are used. More preferably, all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 are used. For example, the at least one further active pharmaceutical ingredient such as a statin or anacetrapib are combined with a composition comprising at least one, preferably at least two, preferably all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 as a single dosage form, e.g. combined in a capsule, tablet, granulate, sachet comprising powdered compounds, etc. The composition or pharmaceutical composition according to the invention and further active pharmaceutical ingredient(s) are administered orally or are administered using a different route, although oral administration is preferred.

The composition or pharmaceutical composition is also used in a method for the treatment or the prophylaxis of cardiovascular disease, wherein administering at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 to a (human) subject replaces previous administration of an active pharmaceutical ingredient that was administered for the same purpose of lowering the plasma LDL-cholesterol level in said subject. That is to say, the composition or pharmaceutical composition for use according to the invention replaces for example any pharmaceutical composition comprising a statin, niacin, fenofibrate, ezetimibe, colesevelam, mipomersen, lomitapide, a PCSK9 inhibitor, alirocumab, evolocumab, ETC-1002, a CETP inhibitor, anacetrapib, evacetrapib, WAY-252623, a blood-pressure lowering compound, hydrochlorothiazide.

Preferred is the composition or pharmaceutical composition comprising at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use according to the invention, wherein the composition or pharmaceutical composition is administered orally to a subject, preferably a human subject. Of course, alternative routes of administration are equally preferred if applicable for administering a sufficient amount of the compound(s), composition or pharmaceutical composition to a subject in need thereof, such as administration using a parenteral route, e.g. by injection intradermally or intravenously, or by infusion such as intravenously, wherein the administered amount of the compound(s), composition or pharmaceutical composition is sufficient to induce a plasma LDL-cholesterol level lowering effect in the (human) subject to whom the compound(s), composition or pharmaceutical composition is/are administered. Typically, the compound(s), composition or pharmaceutical composition is/are administered to a human subject in need thereof with the aim of lowering the plasma LDL-cholesterol level to a level of 100 mg/dL or lower such as 70 mg/dL, or lower, e.g. 60 mg/dL or lower. The human subject in need of being administered a pharmaceutically effective dose of e.g. geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and in some embodiments at least one further active pharmaceutical ingredient, is for example a human patient suffering from ischemia, a CVD, a high plasma LDL-cholesterol level of at least 150 mg/dL.

The composition or pharmaceutical composition according to the invention preferably comprises administering the carotenoids, bacteriopheophtyins and quinones to a subject, said administering resulting in lowering of the plasma LDL-cholesterol concentration in the subject, preferably to a plasma concentration of less than 3,34 mmol/L, preferably less than 2,59 mmol/L, more preferably to a plasma LDL-cholesterol concentration of less than 1 ,8 mmol/L.

Also preferred is the composition or pharmaceutical composition for use according to the invention, wherein administering (a pharmaceutically effective dose of) the at least one, preferably at least two, preferably all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 to a subject (in need thereof) results in a decrease of the LDL-cholesterol concentration in the plasma of said subject, wherein the plasma HDL-cholesterol concentration remains essentially unaltered or decreases to a smaller extent than the decrease in the plasma LDL-cholesterol concentration (HDL-cholesterol level increases in a relative sense), or wherein the plasma HDL-cholesterol concentration increases. The plasma HDL- cholesterol level is in an embodiment at least 40 mg/dL, or at least 60 mg/dL, in the plasma of a (human) subject to whom the composition or pharmaceutical composition is administered. Desired is a plasma HDL-cholesterol level of 60 mg/dL or higher. The decrease of the LDL-cholesterol concentration is expressed as the percentage of the plasma LDL-cholesterol concentration prior to the administration of (an effective dose of) (a composition comprising) the carotenoid(s), bacteriopheophtin(s) and/or quinone(s), such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, to said (human) subject (in need of cholesterol-lowering treatment), and the change of the HDL-cholesterol concentration is expressed as a percentage of the plasma HDL-cholesterol concentration prior to the administration of the composition or pharmaceutical composition for use according to the invention to said subject. The subject is for example a human subject, and the human subject is preferably a human subject in need of a reduction of the plasma LDL-cholesterol concentration in view of treating or prophylaxis of a CVD or one or more of the further diseases or risk factors for developing a disease, as here above listed. The human subject in need thereof is for example a human subject suffering from any one or more of CVD, atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL-cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, ischemia. The human subject in need of treatment with an effective amount of at least one carotenoid, bacteriopheophytin or quinone, e.g. a composition comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, is for example a human subject having an increased cardiovascular risk factor and/or an increased atherosclerosis risk factor, such as a plasma LDL- cholesterol level of 100 mg/dL or higher and/or a total plasma cholesterol level of 200 mg/dL or higher, such as 200 mg/dL - 239 mg/dL or 240 mg/dL and higher. Preferably, the human subject is administered at least two, preferably all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. More preferably the human subject is administered all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

An embodiment is the composition or pharmaceutical composition for use according to the invention, wherein administering the at least one carotenoid, bacteriopheophytin or quinone, e.g. all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, to a subject results in a decrease of the plasma LDL-cholesterol concentration in said subject with at least 5%, preferably at least 10%, more preferably at least 20%, most preferably at least 30%, based on the plasma LDL-cholesterol concentration prior to the administration of the carotenoid, bacteriopheophytin and/or quinone to said subject. The administered amount of carotenoid (s), bacteriopheophytin(s) and/or quinone(s), such as geranyl- geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, to a human subject in need thereof is an amount sufficient to lower the plasma LDL-cholesterol concentration in said human subject to a level of lower than 200 mg/dL such as lower than 160 mg/dL, or lower than 145 mg/dL, lower than 120 mg/dL, lower than 100 mg/dL, preferably 70 mg/dL or lower. The composition or pharmaceutical composition is for example administered to a human subject suffering from a risk factor for developing cardiovascular disease or atherosclerosis, such as a plasma LDL-cholesterol level of about 100 mg/dL or about 200 mg/dL, wherein the administering of the carotenoid results in a decrease of the plasma LDL-cholesterol level to about 70 mg/dL or lower, or to about 140 mg/dL or lower, respectively. Typically, administering the carotenoid(s), bacteriopheophytin(s) and/or quinone(s), such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, to a subject results in a decrease of the plasma LDL-cholesterol level of 30%- 40%.

Particularly, administering the composition or pharmaceutical composition comprising at least one carotenoid, bacteripheophytin and/or ubiquinone, e.g. in a pharmaceutically effective amount, to a subject such as a human subject in need thereof results in a decrease of the plasma LDL-cholesterol concentration to, or maintenance of the plasma LDL-cholesterol concentration at, a plasma LDL- cholesterol concentration of less than 200 mg/dL, preferably less than 159 mg/dL, more preferably less than 129 mg/dL, most preferably less than 100 mg/dL, such as less than 70 mg/dL, or even lower such as less than 60 mg/dL. Lowering the plasma LDL-cholesterol level in the human subject in need thereof by administering a therapeutically effective amount of the carotenoid(s), bacteriopheophytin(s) and/or quinone(s), such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, results in a decreased risk for developing a CVD, atherosclerosis, ischemia, etc. , and/or results in reduction and/or relief from symptoms related to e.g. a CVD, atherosclerosis, ischemia.

Preferably, the composition or pharmaceutical composition comprising at least one, preferably at least two, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is formulated as an oral dosage form, preferably a solid oral dosage form or a liquid oral dosage form, preferably a liquid oral dosage form comprising an oil and/or at least one lipid such as a phospholipid and/or a fat, preferably comprising omega-3 fatty acids and/or omega-6 fatty acids. An oral dosage form is for example self-administered conveniently by the patient in need of LDL-cholesterol lowering treatment. The oral dosage form is for example provided as a single capsule or tablet, or as multiple capsules or tablets for (self-)administration once daily or during more than one occasion during a day, such as twice or thrice daily or before or after a meal. The composition or pharmaceutical composition for use according to the invention is in an embodiment formulated as a tablet or a capsule or a powder or a granulate.

An embodiment is the composition or pharmaceutical composition comprising at least one, preferably at least two, more preferably all compounds of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use according to the invention, wherein the daily dose of the at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 to be administered to a subject is 10 microgram-150 mg per day, preferably 100 microgram-100 mg per day, more preferably 500 microgram-50 mg per day, most preferably 1 mg-25 mg per day. Of course, also part of the invention are a daily dose of the active pharmaceutical ingredient of less than 10 microgram per day or more than 150 mg per day. An embodiment is the composition or pharmaceutical composition for use according to the invention, wherein the daily dose of the at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 to be administered to a subject is 10 microgram-30 g per day, preferably 100 microgram-20 g per day, more preferably 500 microgram-10 g per day, most preferably 1 mg-5 g per day. Of course, also part of the invention are a daily dose of the active pharmaceutical ingredient of less than 10 microgram per day or more than 30 g per day. For example, a daily dose of the composition or pharmaceutical composition comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is 50 mg, 100 mg, 150 mg, 250 mg, 500 mg, 600 mg, 750 mg, 1 g, 2 g, 2,5 g, 3 g, 4 g, 40 g. For example, the daily dose of rhodovibrin is 10 mg/kg - 500 mg/kg, such as 25 mg/kg - 250 mg/kg, or 50 mg/kg - 200 mg/kg, based on the weight of the human subject. The effective daily dose is the dose administered to the patient in need thereof, that results in the desired lowering of plasma LDL-cholesterol level aimed for. For example, the carotenoid, such as rhodovibrin is administered to a human subject resulting in the LDL- cholesterol level in the blood plasma lowering from any initial value before treatment of 120 mg/dL - 159 mg/dL, to a value lower than said initial value, such as a LDL-cholesterol level of between 60 mg/dL and 140 mg/dL, or lower than 120 mg/dL. For example, the at least one carotenoid, bacteriopheophytin and/or quinone, such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, is administered to a human subject resulting in the LDL-cholesterol level in the blood plasma lowering from any initial value before treatment of 95 mg/dL - 190 mg/dL, or 95 mg/dL - 155 mg/dL, to a value lower than said initial value, such as a LDL-cholesterol level of between 60 mg/dL and 140 mg/dL, or lower than 95 mg/dL. In an embodiment the administering to a subject of the composition or pharmaceutical composition comprising at least one, preferably at least two preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use according to the invention, maintains the LDL-cholesterol concentration in the plasma of the subject at a level of less than 159 mg/dL, preferably less than 129 mg/dl_, more preferably less than 100 mg/dL, most preferably less than 70 mg/dL, such as between 50 mg/dL and 159 mg/dL. It is desired that plasma LDL-cholesterol levels in a human subject are lower than 100 mg/dL, such as lower than 70 mg/dL, and that the plasma LDL-cholesterol levels are maintained at such low values, therewith diminishing the risk for developing a cardiovascular disease and/or any of atherosclerosis, dyslipidemia, arteriosclerosis, inflammation, inflammatory disease, ischemia, infection.

An aspect of the invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of at least one carotenoid, pheophtyin such as a bacteriopheophytin and/or a quinone, such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, and optionally a pharmaceutically acceptable excipient for use in lowering plasma cholesterol, wherein the pharmaceutical composition is not, or does not comprise, a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. In a preferred embodiment, the pharmaceutical composition comprises at least two, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. In another preferred embodiment the pharmaceutical composition comprises all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

An aspect of the invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of at least one carotenoid, pheophytin such as a bacteriopheophytin and/or quinone, such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, and optionally a pharmaceutically acceptable excipient for use in the lowering of LDL-cholesterol in blood plasma of a subject, wherein the pharmaceutical composition is not, or does not comprise, a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. In a preferred embodiment, the pharmaceutical composition comprises at least two, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. In another preferred embodiment the pharmaceutical composition comprises all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

An aspect of the invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of at least one carotenoid, pheophytin such as a bacteriopheophytin and/or quinone, such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, and optionally a pharmaceutically acceptable excipient for use in a method for the lowering of LDL-cholesterol in blood plasma of a subject, wherein the pharmaceutical composition is not, or does not comprise, a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. In a preferred embodiment, the pharmaceutical composition comprises at least two, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. In another preferred embodiment the pharmaceutical composition comprises all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

An aspect of the invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of at least one carotenoid, pheophytin such as a bacteriopheophytin and/or quinone, such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, and optionally a pharmaceutically acceptable excipient, for use in a method for the treatment or the prophylaxis of any one or more of cardiovascular disease, such as coronary heart disease, atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL- cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, inflammation, inflammatory disease, ischemia, infection, wherein the pharmaceutical composition is not, or does not comprise, a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. In a preferred embodiment, the pharmaceutical composition comprises at least two of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. In another preferred embodiment the pharmaceutical composition comprises all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. Furthermore, an embodiment is the pharmaceutical composition comprising a pharmaceutically effective amount of at least one carotenoid, pheophytin such as a bacteriopheophytin and/or quinone and optionally a pharmaceutically acceptable excipient, for use in a method for the treatment or the prophylaxis of a symptom or a risk factor of any one or more of a cardiovascular disease, atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, inflammation, inflammatory disease, ischemia, infection.

In the pharmaceutical composition for use in the method for the treatment or the prophylaxis of any of the diseases, health problems, risk factors, symptoms, the at least one carotenoid, pheophytin such as a bacteriopheophytin and/or quinone, e.g. geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, is/are either the sole active pharmaceutical ingredient(s) in the pharmaceutical composition, or the pharmaceutical composition is administered to a subject to whom at least one further active pharmaceutical ingredient is administered, such as an active pharmaceutical ingredient selected from a statin, niacin, fenofibrate, ezetimibe, colesevelam, mipomersen, lomitapide, a PCSK9 inhibitor, alirocumab, evolocumab, ETC-1002, a CETP inhibitor, anacetrapib, evacetrapib, WAY-252623, a blood- pressure lowering compound, hydrochlorothiazide, or any combination thereof of two or three active pharmaceutical ingredients.

An aspect of the invention relates to a method for treating or preventing a cardiovascular disease, atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL-cholesterol plasma level of at least 70 mg/dL, an LDL- cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, inflammation, inflammatory disease, ischemia, infection, comprising administering in a subject in need thereof an effective amount of a composition comprising at least one carotenoid, pheophytin such as a bacteriopheophytin and/or quinone, such as geranyl- geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, wherein the composition is not, or does not comprise, a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. In a preferred embodiment, the composition comprises at least two of compounds geranyl- geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. In another preferred embodiment the composition comprises all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. The invention also relates to a method for treating any symptom or risk factor related to any one or more of these health problems and diseases.

Preferred is the pharmaceutical composition for use, wherein (an effective dose of) the pharmaceutical composition is administered to a (human) subject (in need thereof) suffering from an LDL-cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a CVD. Equally preferred is the pharmaceutical composition for use, wherein the pharmaceutical composition is administered to a subject suffering from a total plasma cholesterol level of 200 mg/dL or higher and/or a plasma LDL-cholesterol level of higher than 100 mg/dL.

An aspect of the invention relates to a method of treating a patient suffering from an LDL- cholesterol concentration in the plasma of said patient of above 100 mg/dL by administering a composition comprising an effective amount of at least one carotenoid, pheophytin such as a bacteriopheophytin and/or quinone, such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 to the patient, wherein the composition is not, or does not comprise, a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. In a preferred embodiment, the composition comprises at least two of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. In another preferred embodiment the composition comprises all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

An aspect of the invention relates to a method of treating a patient suffering from or having an increased risk for any one or more of cardiovascular disease, atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL- cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, inflammation, inflammatory disease, ischemia, infection, by administering a composition comprising an effective amount of at least one carotenoid, pheophytin such as a bacteriopheophytin and/or quinone, such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 to the patient, wherein the composition is not, or does not comprise, a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride. In a preferred embodiment, the composition comprises at least two, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. In another preferred embodiment the composition comprises all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

The subject is suffering from a CVD and/or the subject is at risk for developing a CVD. The CVD being due to atherosclerosis, relating to any of ischaemic heart disease or coronary artery disease (e.g. heart attack), cerebrovascular disease (e.g. stroke), diseases of the aorta and arteries, including hypertension and peripheral vascular disease, and/or the CVD is congenital heart disease, rheumatic heart disease, cardiomyopathies, or cardiac arrhythmias. An aspect of the current invention relates to a food supplement or a feed supplement with LDL- cholesterol lowering properties, comprising at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, preferably thirtheen of a carotenoid, pheophytin such as a bacteriopheophytin and/or quinone, such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. In the present invention a food supplement and a feed supplement is defined as a formulation that is consumed in addition to a normal diet and that contains compounds or components that do not occur in a normal diet, or that occur in low amounts or in insufficient amounts, while sufficient or increased consumption of these components is desired. Preferably, a food supplement according to the invention is composed such that it is suitable for human consumption. Consequently, a food supplement as defined in the present invention should preferably have a texture, taste and smell, but also a nutritional value, that makes the supplement suitable for human consumption.

Preferably, a feed supplement according to the invention is composed such that it is suitable for animal consumption, such as consumption by poultry such as laying hens, chicken, cow, pig, goat, horse, sheep, dog, cat, rabbit, etc. Consequently, a feed supplement as defined in the present invention should preferably have a texture, taste and smell, but also a nutritional value, that makes the supplement suitable for animal consumption.

In a preferred embodiment, the food supplement or feed supplement comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve or at least thirteen, preferably all thirteen of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

In another preferred embodiment the food or feed supplement comprises all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

A food supplement or a feed supplement according to the invention preferably contains between 0.01 % and 99.9% (w/w) of at least one carotenoid, pheophytin such as a bacteriopheophtyin and/or quinone, such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, for example 0.05%- 50% by weight based on the total weight of the food supplement or the feed supplement. Preferably, a food supplement or a feed supplement contains between 10% and 90% (w/w), or between 30% and 75% (w/w), of a preparation of at least one carotenoid, pheophytin such as a bacteriopheophytin and/or quinone, such as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. To make a food supplement or a feed supplement according to the invention suitable for consumption, components are preferably added to improve, for instance, texture, taste or smell.

Consequently, a food supplement or a feed supplement according to the invention preferably comprises (additional) sources of protein, carbohydrate and fat, and vitamins, minerals, electrolytes, trace elements, and other suitable components, so that the food supplement or the feed supplement itself is suitable for use as a nourishing food.

As a source of protein each and every protein that is suitable for use in nutritional formulations, and mixtures of these, are preferably used in a food supplement or a feed supplement according to the invention. This type of proteins encompasses for instance animal proteins such as whey proteins, whey protein concentrates, whey powder, egg protein, egg albumin, casein, or milk albumin, and plant proteins such as soy protein, soy meal, or proteins from soy milk. For choosing the source of proteins to be used, the biological value of a protein may constitute an important criterion. Caseinate, including calcium caseinate, but also whey, milk albumin, egg albumin, and total egg proteins, for instance, are proteins with a very high biological value, because they contain a large proportion of essential amino acids.

Suitable carbohydrates to be used in a food supplement or a feed supplement according to the invention are, for instance, preferably simple short-chain carbohydrates such as mono- and disaccharides, but also polysaccharides, or a combination of both. A carbohydrate is preferably selected because of its suitable organoleptic properties, according to the invention. Preferably, a complex carbohydrate is suitably used as a food fiber, according to the invention.

A food supplement or a feed supplement according to the invention preferably contains, in some embodiments, combinations of both simple and complex carbohydrates. A food supplement or a feed supplement according to the invention preferably contains, in some embodiments, a fat selected from all edible oils and edible fats.

Vitamins and minerals are preferably added to a preparation according to the invention, in conformity with the rules of the regulatory health authorities, and preferably encompasses all vitamins and minerals endorsed by the above authorities, for instance vitamin A, B1 , B2, B12, C, D, B, and K, and folic acid, niacin, pantothenic acid, and biotin. As minerals for instance iron, zinc, iodine, calcium, magnesium, chromium, and selenium are preferably added to a preparation according to the invention.

Electrolytes such as the ions of sodium, potassium, and chloride, and trace elements and other additives do preferably also form part of a food supplement or a feed supplement according to the invention. Such components are, if present, preferably used in the recommended concentrations. Additionally, a food supplement or a feed supplement according to the invention preferably contains components improving its texture, colorings and flavorings, aromatic substances, spices, fillers, emulsifiers, stabilizing compounds, preservatives, antioxidants, fibers, and other supplements such as amino acids, choline, lecithin, fatty acids, etc. The choice of such components depends upon formulation, design, and preferences. The amounts of such components that are added are known to the skilled person, while the choice of the amounts to be added are preferably guided by for example considering the recommended daily amounts (RDA) for children and adults when a food supplement of the invention is considered. Emulsifiers are preferably added to stabilize the final product of the invention. Examples of acceptable emulsifiers are lecithin ( e.g . , derived from soy or from egg), and/or mono- and di-glycerides, according to the invention. As stabilizers, carob, guar or carrageenan are, for instance, preferably used, according to the invention.

Preservatives are preferably added to increase the shelf life of the product of the invention.

Preferably, preservatives such as sodium sorbate, potassium sorbate, potassium benzoate, sodium benzoate, or calcium disodium EDTA are used in a preparation of the invention.

In addition to the carbohydrates mentioned above, natural or synthetic sweeteners, such as saccharides, cyclamates, aspartame, acesulfame potassium, and/or sorbitol, are preferably added to the food supplement or to the feed supplement, according to the invention.

The amounts of food supplement or of feed supplement of the invention to be consumed are varying in size, and are not necessarily restricted to the dosages mentioned in the dosages advised. The term“food supplement” is not meant to be restricted to a specified weight, or to a specified dose of the food supplement. The term“feed supplement” is not meant to be restricted to a specified weight, or to a specified dose of the feed supplement.

The composition of a food supplement or a feed supplement according to the invention takes in principle any form that is suitable for human or animal consumption, according to the invention.

In a preferred embodiment of the invention, the food supplement or the feed supplement is a dry powder that is suitable to be suspended, dispersed or emulsified in an aqueous solution such as water, milk, coffee, tea, broth, and fruit juice. To that end, the powder is preferably supplied in a dispenser according to the invention.

In an alternative preferred embodiment of the invention, the food supplement or the feed supplement is formulated, starting from dry powder, as a tablet or as a granulate. To this end, preferably the composition of a food supplement or a feed supplement according to the invention is suitably supplied with fillers such as microcrystalline cellulose (MCC) and mannitol, binders such as hydroxylpropyl-cellulose (HPC), lubricants such as stearic acid, and other excipients.

A food supplement or a feed supplement according to the invention is in one embodiment preferably supplied as a fluid, in which the solid components have been suspended, dispersed or emulsified. Such a composition of the invention is preferably directly mixed into a foodstuff or a feedstuff, or is preferably for instance extruded and formatted into granules or other forms.

In an alternative embodiment of the invention, a food supplement or a feed supplement is preferably formulated in a solid form, such as a bar, a biscuit, or a roll.

A food supplement or a feed supplement of the invention is preferably formulated for oral consumption, preferably in combination with an acceptable carrier such as a capsule, a tablet, a granulate, a water-miscible powder, or another form acceptable for administration. Alternatively, a food supplement of the invention is preferably processed into a foodstuff, according to the invention. Alternatively, a feed supplement of the invention is preferably processed into a feedstuff, according to the invention.

One aspect of the present invention relates to a foodstuff comprising a food supplement according to the invention. The food supplement comprising at least one carotenoid, pheophytin such as a bacteriopheophytin and/or quinone, such as one or more, preferably all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. One aspect of the present invention relates to a feedstuff comprising a feed supplement according to the invention. The feed supplement comprising at least one, preferably at least two, more preferably all of compoundsgeranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

In a preferred embodiment, the food supplement comprised in the foodstuff comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve or at least thirteen, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

In another preferred embodiment the food supplement comprised in the the foodstuff comprises all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

A food supplement or a feed supplement may suitably be used to reduce intestinal cholesterol absorption, thus reducing the cholesterol level of blood plasma, in particular the plasma LDL-cholesterol concentration, while preferably leaving the HDL-cholesterol concentration essentially unaltered or raising the plasma HDL-cholesterol concentration in a subject, such that the plasma levels of HDL- cholesterol increase in absolute sense or relative to the plasma LDL-cholesterol concentration. The subject being a human subject or an animal subject such as a chicken or laying hen. The invention relates to a food supplement with cholesterol-lowering properties, comprising at least one, preferably at least two, more preferably all compounds of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

Feeding an animal such as a broiler, hen, pig, cow, duck, goat, goose, turkey, bovine calf, sheep, in particular a pig, a broiler and a laying hen, with the feed supplement or the feed stuff comprising the feed supplement, results in the lowering of the cholesterol level in the blood serum (plasma) of said animal and/or the lowering of the cholesterol content in the meat of said animal and/or in other parts or products derived from said animal and/or in the lowering of the cholesterol content in the eggs laid by the e.g. chicken, duck, goose, turkey, etc. This lowering of the cholesterol content, in particular the LDL- cholesterol content provides for e.g. meat, eggs, etc., suitable for human consumption, and comprising a lower content of cholesterol, in particular LDL-cholesterol. Therewith, the human diet comprises less cholesterol, in particular less LDL-cholesterol, when the consumed meat and/or eggs by the human subject are derived from animals fed with the feed supplement or the feedstuff of the invention. As a consequence, less cholesterol is transported from the intestine to the blood circulation, the liver and further organs of the human subject.

In an embodiment, a food supplement of the invention is applied in a foodstuff with cholesterollowering properties. In an embodiment, a feed supplement of the invention is applied in a feedstuff with cholesterol-lowering properties.

A method to prepare a cholesterol-lowering foodstuff or feedstuff of the invention involves the production of a foodstuff or feedstuff, respectively, incorporating a food supplement or a feed supplement according to the invention. Such a method preferably involves a step in which a foodstuff or a feedstuff is first prepared in the normal way, followed by the addition of at least one carotenoid, pheophtyin such as a bacteriopheophytin and/or quinone, such as preferably all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, to the prepared foodstuff or feedstuff. Preferably, at least two of compounds as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is added. More preferably, all of compounds as geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is added.

Also, it is possible to add at least one, preferably at least two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 to the foodstuff or to the feedstuff during its production.

A foodstuff with cholesterol-lowering properties according to the invention or a feedstuff with cholesterol-lowering properties according to the invention contains typically between 0.1 % and 20% (w/w), preferably between 1 % and 10% (w/w), of the food supplement or feed supplement according to the invention and described above. The feedstuff is for example chicken feed or feed for laying hen. The invention relates to a foodstuff comprising a food supplement, wherein the food supplement has cholesterol-lowering properties and comprises at least one, preferably at least two, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

With regard to the aspects of the invention and the embodiments described, the carotenoid(s) is/are for example natural carotenoid(s) and is/are for example isolated from a natural source or is/are chemically synthesized. The pheophytin(s) is/are for example natural pheophytin(s) and are for example isolated from a natural source or are chemically synthesized. The quinone, ubiquinone or ubiquinol or ubiquinone derivative such as rhodoquinone is/are for example natural quinone, ubiquinone or ubiquinol or a naturally occurring ubiquinone derivative such as rhodoquinone, and are for example isolated from a natural source or are chemically synthesized. The pheophytin is for example a chlorophyll derivative lacking the central Mg²⁺ ion and is for example a bacteriopheophytin a, for example a bacteriopheophytin a comprising a phytyl group or comprising a geranyl-geranyl group. The carotenoid(s) is/are preferably tetraterpenoids such as a xanthophyll and a carotene, though xanthophylls are preferred. The carotenoids are preferably polar carotenoids such as xanthophylls, and xanthophylls are preferred. The carotenoids are for example xanthophylls based on the precursor phytoene. The carotenoids are for example xanthophylls based on the precursor xanthophyll rhodopin. The xanthophyll is for example an alcohol, an ether, or has both a hydroxyl group and an ether group.

For all described, exemplified, and/or claimed embodiments of the invention and aspects according to the invention in the description, claims, abstract, examples, drawings: the proviso is that for the composition according to the invention and for the composition for use according to the invention and for the compounds in e.g. the foodstuff of the invention and the food supplement of the invention or the feed of the invention, the composition and the composition for use of the invention and and the compounds in e.g. the foodstuff of the invention and the food supplement of the invention or the feed of the invention, is/are not a preparation comprising the membrane fraction of Rhodospirillum spp. and/or the membrane fraction of Phaeospirillum spp., and is/are not a preparation or pharmaceutical preparation of Rhodospirillum spp. for use in lowering plasma cholesterol, said preparation or pharmaceutical preparation comprising dead Rhodospirillum spp. , or freeze-dried Rhodospirillum spp. Thus, for example for the food supplement and the food stuff of the invention, the compounds are not provided as a preparation comprising the membrane fraction of Rhodospirillum spp. and/or the membrane fraction of Phaeospirillum spp., and not as a preparation or pharmaceutical preparation of Rhodospirillum spp. for use in lowering plasma cholesterol, said preparation or pharmaceutical preparation comprising dead Rhodospirillum spp. , or freeze-dried Rhodospirillum spp. Furthermore, for example the (pharmaceutical) composition of the invention does not comprise or consist of whole R.rubrum cells and is not or does not comprise a preparation comprising the membrane fraction of Rhodospirillum spp. and/or Phaeospirillum spp., and is not or does not comprise a preparation of Rhodospirillum spp. for use in lowering plasma cholesterol, said preparation comprising dead Rhodospirillum spp. , or freeze-dried Rhodospirillum spp. , according to the invention. Thus, the current invention provides a composition comprising compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, and under the proviso that the composition of the invention is not a preparation comprising the membrane fraction of Rhodospirillum spp. and/or the membrane fraction of Phaeospirillum spp., and is not a preparation or pharmaceutical preparation of Rhodospirillum spp. for use in lowering plasma cholesterol, said preparation or pharmaceutical preparation comprising dead Rhodospirillum spp., or freeze-dried Rhodospirillum spp., according to the invention.

While the invention has been described in terms of several embodiments, it is contemplated that alternatives, modifications, permutations and equivalents thereof will become apparent to one having ordinary skill in the art upon reading the specification. The invention is not limited in any way to the illustrated embodiments. Changes can be made without departing from the scope which is defined by the appended claims.

EXAMPLES

Measurement of cholesterol levels

Total cholesterol assay - mouse plasma

The measurement of total plasma cholesterol in the mouse plasma was conducted using the in vitro kit “cholesterol” from Beckman Coulter Nederland B.V. (Woerden, Netherlands, Datasheet from 09-2011 , coded BAOSR6x16.02; catalogue No. OSR6116) intended for the quantitative determination of Cholesterol concentrations in serum, EDTA plasma, heparinized plasma on Beckman Coulter AU analyzers. Reference is made to the protocol used as provided by Beckman Coulter reagent kit.

Measurements of cholesterol are used primarily in the diagnosis and treatment of disorders involving excess cholesterol in the blood, and lipid and lipoprotein metabolism disorders.

Total serum or plasma cholesterol analysis has proven useful in the diagnosis of hyperlipoproteinemia, atherosclerosis, hepatic and thyroid diseases. Total and HDL cholesterols, in conjunction with a triglyceride determination, provide valuable information for the prediction of coronary heart disease.

Method & materials

Mouse plasma was used. See Example 1 and 2, here below.

The system reagent of the kit (OSR6116 / OSR6216 /OSR6516) was obtained from Beckman Coulter Nederland B.V. and consists of:

Phosphate buffer (pH 6.5) - 103 mmol/L

Cholesterol Esterase (Candida/Pancreatic) - > 0.2 kli/L (3.3 pkat/L)

4-Aminoantipyrine - 0.31 mmol/L

Cholesterol Oxidase (Brevibacterium) - > 0.2 kU/L (3.3 pkat/L)

Phenol - 5.2 mmol/L

Peroxidase (Horseradish) - > 10.0 kU/L (166.7 pkat/L)

Preservatives The Cholesterol reagents are ready for use. No preparation was required.

Methodology

The reagent is brought into contact with the plasma in aBeckman Coulter analyzer, which causes the cholesterol esters in the serum to be hydrolyzed by the cholesterol esterase (CHE). The free cholesterol produced in this reaction is oxidized by cholesterol oxidase (CHO) to cholest-4-en-3-one with the simultaneous production of hydrogen peroxide (H2O2), which oxidatively couples with the 4- aminoantipyrine and phenol in the presence of the peroxidase to yield a chromophore. The red quinoneimine dye formed through this reaction can be measured spectrophotometrically at 540/600 nm as an increase in absorbance.

Measurements obtained typically fall within these risk categories, when a human subject is considered:

< 200 mg/dl_ - normal total cholesterol level

200 - 239 mg/dL - bordering on high total cholesterol level

> 240 mg/dL - high total cholesterol level

Triglycerides assay - mouse plasma

The measurement of plasma triglyceride levels in the mouse plasma was conducted using the in vitro kit“tryglycerides” from Beckman Coulter Nederland B.V. (Woerden, Netherlands, Datasheet from 1 1 - 2010, coded BAOSR6x1 18.02; catalogue No. OSR601 18) intended for the quantitative determination of Triglyceride concentrations in human serum, EDTA, or heparinized plasma samples on Beckman Coulter All analyzers. Reference is made to the protocol used as provided by Beckman Coulter reagent kit.

Triglycerides are the major form of fat found in nature and their primary function is to provide energy for the cell. Measurements of triglyceride are used in the diagnosis and treatment of patients with diabetes mellitus, nephrosis, liver obstruction, other diseases involving lipid metabolism, or various endocrine disorders. Clinically, triglyceride assays are used to help classify the various genetic and metabolic lipoprotein disorders and in the assessment of risk factors for atherosclerosis and coronary artery disease.

Method & materials

Mouse plasma was used. See Example 1 and 2, here below.

The system reagent of the kit (OSR601 18 / OSR61 1 18 / OSR661 18) was obtained from Beckman Coulter Nederland B.V. and consists of:

PIPES buffer (pH 7.5) - 50 mmol/L

Lipase ( Pseudomonas ) - > 1 .5 kU/L (25 pkat/L)

Glycerol kinase ( Bacillus stearothermophiius) - ³ 0.5 kU/L (8.3 pkat/L)

Glycerol phosphate oxidase ( Pseudomonas ) - > 1 .5 kU/L (25 pkat/L) Ascorbate oxidase ( Curcubita species ) - > 1 .5 kll/L (25 pkat/L)

Peroxidase (Horseradish) - > 0.98 kU/L (16.3 mkat/L)

ATP - 1 .4mmol/L

4-Aminoantipyrine - 0.50mmol/L

Magnesium acetate - 4.6 mmol/L

MADB - 0.25mmol/L

Preservatives

For OSR601 18 and OSR61 1 18, the Triglyceride Reagents are ready for use. No preparation was required. For OSR661 18, the pipe supplied had to be inserted into the 180 ml_ reagent vial before use on the analyzer. The pipe was for single use only.

Methodology

This triglyceride procedure is based on a series of coupled enzymatic reactions. The triglycerides in the sample are hydrolyzed by a combination of microbial lipases to give glycerol and fatty acids. The glycerol is phosphorylated by adenosine triphosphate (ATP) in the presence of glycerol kinase (GK) to produce glycerol-3-phosphate. The glycerol-3-phosphate is oxidized by molecular oxygen in the presence of GPO (glycerol phosphate oxidase) to produce hydrogen peroxide (H2O2) and dihydroxyacetone phosphate. The formed H2O2 reacts with 4-aminophenazone and N,N-bis(4-sulfobutyl)-3,5- dimethylaniline, disodium salt (MADB) in the presence of peroxidase (POD) to produce a chromophore, which is read at 660/800nm. The increase in absorbance at 660/800 nm is proportional to the triglyceride content of the sample.

<150 mg/dl_ - Normal

150-199 mg/dL - Borderline High

200-499 mg/dL - High

>500 mg/dL - Very High

LDL Cholesterol assay - mouse plasma

The measurement of plasma LDL cholesterol in the mouse plasma was conducted using the in vitro kit “LDL-cholesterol” from Beckman Coulter Nederland B.V. (Woerden, Netherlands, Datasheet from 08- 2009, coded BAOSR6x96.01 ; catalogue No. OSR6196) intended for the quantitative determination of LDL-Cholesterol concentrations in human serum, EDTA, or heparinized plasma samples on Beckman Coulter AU analyzers. Reference is made to the protocol used as provided by Beckman Coulter reagent kit.

Measurements of cholesterol are used primarily in the diagnosis and treatment of disorders involving excess cholesterol in the blood, and lipid and lipoprotein metabolism disorders. LDL-Cholesterol plays a causal role in the development of coronary heart disease (CHD). In 1988 the National Cholesterol Education Program Adult Treatment Panel (NCEP-ATP) developed recommendations for the diagnosis and treatment of patients with hypercholesterolemia. These recommendations defined LDL-Cholesterol as the primary target of therapy. The 2001 update of these guidelines (NCEP-ATP III) put further emphasis on better risk identification and more aggressive cholesterol-lowering treatment.

Method & materials

Mouse plasma was used. See Example 1 and 2, here below.

The system reagent of the kit (OSR6196 / OSR6296) was obtained from Beckman Coulter Nederland B.V. and consists of:

MES Buffer (pH 6.3)

Cholesterol esterase ( Pseudomonas ) - 1875 U/L

Cholesterol oxidase ( Nocardia ) - 1 125 U/L

Peroxidase (Horseradish) - 975 U/L

Detergent 1 - 0.75 %

Detergent 2 - 0.25 %

DSBmT - 0.25mmol/L

4-aminoantipyrine - 0.375mmol/L

Ascorbate Oxidase - 2250 U/L

Preservative

The Cholesterol reagents are ready for use. No preparation was required.

Methodology

The LDL-Cholesterol test is a two reagent homogenous system. The assay is comprised of two distinct phases. In phase one a unique detergent solubilizes cholesterol from non-LDL- lipoprotein particles. This cholesterol is consumed by cholesterol esterase, cholesterol oxidase, peroxidase and 4- aminoantipyrine to generate a colorless end product.

In phase two a second detergent in the reagent releases cholesterol from the LDL - lipoproteins. This cholesterol reacts with cholesterol esterase, cholesterol oxidase and a chromogen system to yield a blue color complex which can be measured bichromatically at 540/660nm. The resulting increase in absorbance is directly proportional to the LDL-C concentration in the sample.

The guidelines (NCEP-ATP III) classify LDL - Cholesterol levels as follows, when a human subject is considered:

1 . < 100 mg/dL - Optimal

2. 100 - 129 mg/dL - Near optimal/above optimal

3. 131 - 159 mg/dL - Borderline high

4. 160 - 189 mg/dL - High 5. > 190 mg/dL - Very high

HDL Cholesterol assay - mouse plasma

The measurement of plasma HDL cholesterol in the mouse plasma was conducted using the in vitro kit “HDL-cholesterol” from Beckman Coulter Nederland B.V. (Woerden, Netherlands, Datasheet from 08- 2009, coded BAOSR6x95.01 ; catalogue No. OSR6195) intended for the quantitative determination of HDL-Cholesterol concentrations in human serum, EDTA, or heparinized plasma samples on Beckman Coulter AU analyzers. Reference is made to the protocol used as provided by Beckman Coulter reagent kit.

Many epidemiological investigations have demonstrated the strong and independent inverse association between HDL-Cholesterol and the risk of coronary artery disease. It has been proposed that HDL particles, through the uptake and transport of Cholesterol from peripheral tissue to the liver (reverse Cholesterol transport), protects against the development of atheromatous plaques. Under the guidelines issued by The National Cholesterol Education Program Adult Treatment Panel 2 (NCEP ATP 2) it is recommended that both HDL-Cholesterol and Total Cholesterol should be measured in the initial screening for hypercholesterolemia.

Method & materials

Mouse plasma was used. See Example 1 and 2, here below.

The system reagent of the kit (OSR6195 / OSR6295) was obtained from Beckman Coulter Nederland B.V. and consists of:

Goods Buffer (pH 6.0)

Cholesterol esterase ( Pseudomonas ) - 375 U/L

Cholesterol oxidase ( E.coli ) - 750 U/L

Peroxidase (Horseradish) - 975 U/L

Ascorbate oxidase ( Curcubita sp.) - 2250 U/L

DSBmT - 0.75mmol/L

4-aminoantipyrine - 0.25mmol/L

Detergent - 0.375%

Preservative - 0.05%

The Cholesterol reagents are ready for use. No preparation was required.

Methodology

The HDL-Cholesterol test is a two reagent homogenous system for the selective measurement of serum or plasma HDL-Cholesterol in the presence of other lipoprotein particles. The assay is comprised of two distinct phases. In phase one, free cholesterol in non-HDL-lipoproteins is solubilized and consumed by cholesterol oxidase, peroxidase, and DSBmT to generate a colorless end product.

In phase two a unique detergent selectively solubilizes HDL- lipoproteins. The HDL cholesterol is released for reaction with cholesterol esterase, cholesterol oxidase and a chromogen system to yield a blue color complex which can be measured bichromatically at 600/700nm. The resulting increase in absorbance is directly proportional to the HDL-C concentration in the sample.

The guidelines (NCEP ATP 2) classify HDL- C levels as follows, when a human subject is considered:

1 . < 40 mg/dL as indicative of a major risk factor for Coronary Heart Disease.

2. > 60 mg/dL as a negative risk factor for Coronary Heart Disease.

Example 1

Plasma LDL-Cholesterol lowering activity of R. rubrum cells

Dried Rhodospirillum rubrum bacteria, 25 g, were purchased from Algosource Technologies (Saint- Nazaire, France) and stored at 4°C until use.

Eight-week old male mice (21 -27 g) were of the C57BL/6J strain that is well known for their high cholesterol level upon exposure to a high fat (a so-called Western) diet. The mice were obtained from Charles River Laboratories (France).

The feed for the mice was obtained from Altromin Spezialfutter GmbH (Lage, Germany). Hydrogels were obtained from ClearH20 (Westbrook, USA) and contained approximately 65 gram of 97% water in a gel.

The 65 g hydrogel was heated for 30 minutes at 70°C and then 1 .5 g control diet (in the pretest diet and in the diet for the control group), or 1 .5 g dried bacteria (the diet for the treated group) was gently mixed into the liquefied hydrogel with a spatula. Subsequently, 13.5 g high fat diet was also mixed into the hydrogel. The mixing was continued until a smooth homogenous product was obtained. The hydrogel was weighed before and after mixing with the diets and also just prior to placement into the animal cages. The hydrogel-diets were stored in the dark at 4°C for a maximum of 3 days.

The mice were placed in cages with litter underneath a mesh. Enrichment was provided in the form of a polymer shelter and a piece of paper towel. The mice were provided the hydrogel-diet cups as their only source of food and water.

Upon reception from the supplier the mice were quarantined together for 7 days. After individually housing, the mice were provided the feed-hydrogel mix in cups containing 60 gr hydrogel, 13.5 gr high fat diet and 1 .5 g control diet for 7 days. The feed-hydrogel product was refreshed 3 times over that period.

After 7 days two groups were formed: a treatment group (n=4) receiving the feed-hydrogel mix in cups containing 13.5 g high fat diet and 1 .5 g dried R.rubrum bacteria for 7 days and a control group (n=4) receiving the feed-hydrogel mix in cups containing 13.5 g high fat diet and 1 .5 g control diet for 7 days. Again, hydrogel-diet product was refreshed 3 times over that period.

Every second day the mice and the remaining hydrogel-diet cup were weighed to assess the effect of the two feeds on the mice weight development and their food consumption.

At the end of the second week the mice were weighed and anaesthetized with 0.05 ml of 200 mg/ml Nembutal (Kela, Lot 26344A13). After sufficient sedation the chest cavity was opened, and blood was taken with a 24 G needle on a 1 ml syringe (both heparin-treated) and stored in a 2 ml Vacutainer EDTA tube on ice.

The blood was centrifuged for 10 minutes at 1500 rpm at 4°C to separate the cells from the plasma. The plasma was stored at 4°C overnight.

The cholesterol was measured in the plasma by company SynLab (Mons, Belgium). In this study subcontractor SynLab was chosen to perform the cholesterol analysis using an ELISA-based method that required less plasma for their measurements.

Statistical significance was tested in Microsoft Excel with Student’s t-test (unpaired, equal size, equal variance).

Results

The goal was to examine the effect of administering Rhodospirillum rubrum (GEPEA/Algosolis) on the blood cholesterol level in mice. The feed was mixed into hydrogel and was offered ad libitum. All mice are comprised in the pre-test (n=8). After one week the mice were divided into the control group (same feed as pretest, n=4) and the treated group (feed with R.rubrum bacteria cells, n=4) and kept on these respective feeds for one week.

For one mouse (nr 6) not enough blood was obtained, and subsequently not enough plasma resulted.

After the second week the treated group (n=4) showed a slight weight gain of 1 .75 g (±0.35 g) whereas the control group (n=4) -1 .5 g (± 1 .37 g) showed a weight loss (p=0.0018). This difference occurred after the second week (the treatment week), but was not present after the first week (pre-test: all 8 mice).

The weekly food consumption by the mice showed no difference between the consumption of the groups in the second week or as compared to the first week (pre-test: all 8 mice).

Total plasma cholesterol levels for treated mice that were administered R.rubrum cells were 6% lower than those of the control group with a p value of 0.047 (statistically significant; p-values below 0.05 when compared to the corresponding control parameter).

LDL cholesterol levels of the treated mice that were administered R.rubrum cells, were 39% lower than those of the control group with a p value of 0.044 (statistically significant; p-values below 0.05 when compared to the corresponding control parameter).

Non-HDL cholesterol levels (mostly comprised of LDL and VLDL) of the treated mice were 32% lower than those of the control group with a p value of 0.053. In conclusion, feeding mice with feed comprising Rhodospirillum rubrum bacteria cells produced by GEPEA/Algosolis had a statistically significant reductive effect on the total plasma cholesterol levels (-6%) and on the LDL cholesterol levels (-39%) of C57BL/6J mice placed on a high fat diet.

The species of bacterium is from the genus Rhodospirillum, or is a mixture of different Rhodospirillum spp. selected for example from Rhodospirillum rubrum, Rhodospirillum centenum, Rhodospirillum photometricum, Rhodospirillum oryzae, Rhodospirillum sulfurexigens, Rhodospirillum salexigens, Rhodospirillum salinarum, Rhodospirillum sodomense, and Rhodospirillum tenue. For example, the Rhodospirillum is Rhodospirillum rubrum.

Example 2

Effect of composition comprising carotenoids, bacteriopheophytins and quinones on plasma LDL-cholesterol levels

Experimental procedures

The Rhodospirillum rubrum strain S1 H was stored in liquid nitrogen in a 10% w/w sucrose- 0.85% w/w saline solution. To regrow the strain, the cells were taken out the liquid nitrogen and thawed for 30 minutes at room temperature. Cells were streaked on a sistrom succinate agar plate and a rich Luria Bertani (LB) medium to grow colony forming units. The agar plates were incubated at 30°C in dark and aerobic conditions for up to 4 days.

After 4 days, 10 single colonies were picked up and transferred to 10 tubes with 2 mL of Sistrom succinate liquid medium and incubated at 30°C in dark, aerobic and orbital shaking at 150 rpm. After 4- 5 days, the cells were grown and reached an OD6so = 0.5-0.6. To check the axenicity of the cultures, the cells were streaked on a sistrom succinate agar plate and a rich LB medium agar plate and incubated up to 1 week to look for heterotrophic contaminants. When the axenicity check was approved, the 2 mL cultures were transferred to 15 mL of Sistrom succinate liquid medium and incubated at 30°C in dark, aerobic and orbital shaking at 150 rpm. After 4-5 days the cells were grown and reached an ODeao = 0.5-0.6. Then, the 15 mL cultures were transferred to 100 mL of Sistrom succinate liquid medium and incubated at 30°C in dark, aerobic and orbital shaking at 100 rpm. After 4-5 days, the cells were grown and reached an OD6so = 0.5-0.6. Once the axenicity was checked on Sistrom succinate and LB medium, these cells constituted the inoculum cultures.

Cell culturing

Culture conditions

Bioreactor

Light anaerobic conditions were applied for culturing the Rhodospirillum rubrum strain S1 H cells. Ten axenic inoculum cultures were pelleted by centrifugation at 5000*g for 10 minutes. The supernatant was discarded and the pellets were pooled in 25 mL of Melissa liquid medium with acetate as carbon source to constitute a concentrated inoculum. The bioreactor was sterilized by wet-heat sterilization, 20 minutes of water vapor exposure at 121 °C and 1 .2 bar in an autoclave. After sterilization the bioreactor was closed. Bottles with Melissa medium, 1 M H2SO4, and for the effluent were coupled aseptically in a laminar flow cabinet (LAF).

Extraction protocols

30 g of bacterial pellet of Rhodospirillum rubrum strain S1 H cells, are mixed using 440 ml of a biphasic mixture of petroleum ether (boiling point 60-80°C) and methanolic saline during 2 h at room temperature (biphasic mixture: 220 ml containing 20 ml of NaCI 0.3% by mass and 200 ml of methanol + 220 ml of the petroleum ether (boiling point 60-80°C)). After centrifugation at 5000 RPM during 20 minutes at room temperature, the upper phase (the petroleum ether phase) was removed and stored at room temperature. Lower phase (the methanolic saline solution) was optionally submitted to a second extraction with an additional 220 ml of petroleum ether for 2 h at room temperature. The second upper phase was mixed with the first one and dried using a rotavapor system. The resulting viscous liquid was named“FRACTION 1.1”, or“fl .1 or“Extract 1.1” in Figure 3. This fl .1 is an example of a composition comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

Materials and methods for mass spectrometry analysis

MALDI-ToF

Matrix-assisted laser desorption ionization time-of-flight (MALDI-ToF) mass spectrum was recorded using a QToF Premier mass spectrometer equipped with a Nd:YAG laser, operating at 355 nm with a output frequency of 50 Hz. Time-of-flight mass analyses were performed in reflection mode at a resolution of about 10.000. Samples of fl .1 were analyzed using (DCTB)trans-2-[3-(4-tertbutylphenyl)- 2-methylprop-2-enylidene] malononitrile. This matrix was prepared as a 40 mg/mL solution in CHCI3. The matrix solution (1 mg/mL) was applied to a stainless steel target and air dried. The samples were dissolved in THF and I microliter aliquots of this solution were applied onto the target area already bearing the matrix crystals and air dried. For the recording of the single-stage MS spectra, the quadrupole (rf-only mode) was set to pass ions from 200 to 2500 Th, and all ions were transmitted into the pusher region of the time-of-flight analyzer where they were mass analyzed with 1 s integration time.

Q-tOF (5600 ABSCIEX) - nanoESI-MS

Samples of fl.1 were diluted in 0.1 % formic acid in acetonitrile, centrifuged at room temperature during 5 min at 13.000 RPM and the supernatants were infused directly in the Mass spectrometer (flow rate : 89 microliter/hour) using nano-esi source. The acquisition parameters were: ion source gas1 : 4; Curtain gas 15; ionspray Voltage floating 2.300, heater temperature 150°C; Polarity : positive; ToF mass range : 100-2.000.

Ion Trap (HCT Ultra Brucker) - nanoESI-MS Samples of f 1.1 were diluted in 0.1 % formic acid in acetonitrile, centrifuged at room temperature during 5 minutes at 13.000 RPM and the supernatants were infused directly in the Mass spectrometer (flow rate : 89 microliter/hour) using nano-esi source. The acquisition parameters were: capillary 1 .900 Volt; Dry gas : 6 l/min; Dry temp : 250°C; Polarity : positive; scan mode : Standard - enhanced ; scan range : 100-2.000; Smart target 20.000; Max accu time : 200ms.

Results

With MALDI-ToF analyses of f 1.1 , the presence of the carotenoids rhodovibrin (molecular ion indicated as M+H+ value was 585,5 for rhodovibrin), 1 -hydroxyspirilloxanthin (molecular ion indicated as M+H+ value was 583,5 for 1 -hydroxy-spirilloxanthin), 3,4-dehdyro-rhodopin (molecular ion indicated as M+H+ value was 587,5 for 3,4-dehydro-rhodopin), chloroxanthin (molecular ion indicated as M+H+ value was 557,5 for chloroxanthin), rhodopin (molecular ion indicated as M+H+ value was 555,4 for rhodopin), spirilloxanthin (molecular ion indicated as M+H+ value was 597,4 for spirilloxanthin) and 3,4-dihydro- spirilloxanthin (molecular ion indicated as M+H+ value was 599,5 for 3,4-dihyd ro-spirilloxanthin) in fl .1 was determined .

The carotenoids rhodovibrin, 1 -hydroxyspirilloxanthin, 3,4-dehdyro-rhodopin, chloroxanthin, rhodopin, spirilloxanthin and 3,4-dihyd ro-spirilloxanthin were also identified in the fl.1 extract when applying nano-ESI Q-ToF analysis. Ten mI of the fraction 1.1 was dried in presence of matrix.

With MALDI-ToF analyses of fl.1 , the presence of the pheophytins geranyl-geranyl bacteriopheophytin a (molecular ion indicated as M+H+ value was 883,5 for geranyl-geranyl bacteriopheophytin a) and phytyl bacteriopheophytin a (BPha (phytyl)) (molecular ion indicated as M+H+ value was 889,5 for phytyl bacteriopheophytin a) in fl.1 was determined.

The pheophytins geranyl-geranyl bacteriopheophytin a and phytyl bacteriopheophytin a were also identified in the fl.1 extract when applying nano-ESI Q-ToF analysis. Ten mI of the fraction 1.1 was dried in presence of matrix.

With MALDI-ToF analyses of fl.1 , the presence of the quinones ubiquinol-10 (molecular ion indicated as M+H+ value was 865,7 for ubiquinol-10), ubiquinone-9 (molecular ion indicated as M+H+ value was 795,6 for ubiquinone-9), ubiquinone-10 (molecular ion indicated as M+H+ value was 848,7 for ubiquinone-10) and rhodoquinone-10 (molecular ion indicated as M+H+ value was 863,7 for rhodoquinone-10) in fl.1 was determined.

The quinones ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 were also identified in the fl.1 extract when applying nano-ESI Q-ToF analysis. Ten mI of the fraction 1.1 was dried in presence of matrix.

Analysis ofthe fl.1 extract using Q-tOF (5600 ABSCIEX) - nanoESI-MS confirmed the presence of rhodovibrin, 1 -hydroxyspirilloxanthin, 3,4-dehdyro-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-dihyd ro-spirilloxanthin, geranyl-geranyl bacteriopheophytin a, phytyl bacteriopheophytin a, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 present in fl.1 . NanoESI MS spectra were obtained for the fraction 1.1 with a triple tof mass spectrometer (ABSCIEX) using acetonitrile 99%, 1 % HCOOC as organic solvent. In the Q-tOF (5600 ABSCIEX) - nanoESI-MS spectrum for fl .1 with carotenoid rhodovibrin amongst others, peaks were revealed at the following m/z values (approximate relative intensity in brackets): 647.6 (2.5); 651 .6 (3.3); 881 .5 (2); 927.5 (1).

Maldi-ToF data analysis with the fl .1 extract with carotenoid 3,4-dehydro-rhodopin revealed, amongst others, peaks at the following m/z values (approximate relative intensity in brackets): 500.3 (1 ); 882.5 (1); 898.5 (4).NanoESI MS spectra are obtained for the fraction 1.1 with an lonTrap mass spectrometer (Bruker) using 1 % HCOOC in acetonitrile.

From the above data it is clear that the 1.1 fraction comprises all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10. Thus, the inventors have herewith found an example of a composition comprising all of the above-mentioned carotenoids, bacteriopheophytins and quinones.

Mice test - effect of administering fl.1 with 13 carotenoids, bacteriopheophytins and quinones on plasma cholesterol level

The mice test for testing the influence of a diet comprising fl .1 with carotenoids, bacteriopheophytins and quinoneson plasma cholesterol level was performed at SCKOEN animalarium (BE) following 2 weeks acclimation of 40 C57BL/6 male mice. After initial weighing of the food and the mice, they were placed in individual ventilated cage. Food consumption was checked every day and hydrogel weighed every 2 days. Based on previously performed preliminary palatability tests, the bacterial extracts were resuspended in sunflower oil, and 5% regular sugar was added to the chow (Cafetaria-diet) to ensure high palatability.

The first week, the first group of mice received the Cafetaria Diet + sunflower oil replacing R. rubrum extract fl.1 with carotenoids, bacteriopheophytins and quinones ad libitum while the second group of mice received the Cafetaria Diet + the control diet replacing R. rubrum extract fl.1 ad libitum.

The second week, the control group continued on the same diet while the three experimental groups received the Cafetaria Diet + 10% of either the fl.1 extract comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 in sunflower oil. Thus, one group of mice was fed with feed comprising the fl.1 extract of Rhodospirillum rubrum, a second group of mice, the control group, was fed feed without extract of Rhodospirillum rubrum.

Effects of feeding control feed or feed comprising fl.1 as a composition comprising geranyl- geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 on cholesterol levels in plasma is detailed below.

End of mice test After 2 weeks of testing, the mice where weighted and euthanized using intraperitoneal pentobarbital injection prior to dissection. Whole-blood was removed in EDTA-tubes, centrifuged to obtain plasma and placed at 4°C for further analysis.

Mice weight

After week 1 and week 2, no differences between the groups were detected.

Blood analysis

Total cholesterol, HDL and LDL fractions

Figure 3 shows the results of the cholesterol-, HDL- and LDL analysis in the blood of the treated mice.

Extract 1.1 (which is Fraction 1.1 , fl.1 ) as an example of a composition comprising geranyl- geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 has a significant effect on the plasma LDL- cholesterol concentration since the Extract 1.1 , as a composition comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, decreased plasma LDL-cholesterol levels for more than 40%, i.e. about 46%, compared to the control group (p<0.001) while the total plasma cholesterol levels stayed essentially unchanged. See Figure 3. Furthermore, also the HDL-cholesterol level in the mice group that were fed the fl .1 fraction with the carotenoids, bacteriopheophytins and quines, stayed essentially unaltered after the experimental period. Total cholesterol in plasma was 4,39 pg/mI for the fl .1 group of mice compared to 4,46 pg/mI for the control group, HDL-cholesterol was 2,93 mg/ml for the fl.1 group and 2,72 mr/mI for the control group, and LDL-cholesterol was 0,36 mg/ml for the fl .1 group (p < 0.001) and 0,67 mg/ml for the control group, respectively.

Conclusions

From this mice test using R. rubrum S1 H extracts fl.1 comprising carotenoids, bacteriopheophytins and quinones, it is determined that the extract fl.1 , as an example of a composition with geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10has a beneficial effect on lowering LDL-cholesterol level to a large extent, while at the same time keeping the HDL-cholesterol level essentially unaltered when the extract comprising geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 is administered orally.

Thus this test has shown that a composition comprising the geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, has an LDL-cholesterol lowering effect.

Example 3

Preparative isolation of carotenoid rhodovibrin from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl.1 of R. rubrum, with rhodovibrin, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the rhodovibrin present in the fraction Fl.1 . The choice of the chromatography support and approach ( e.g . a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to rhodovibrin isolation (See Example 4, here below, referring to Bcma-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated rhodovibrin will be analyzed by mass spectrometry. As a next step, rhodovibrin is isolated from the fraction fl.1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the carotenoid. The carotenoid is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated rhodovibrin obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. See also Example 1 and 2 for the LDL-cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated carotenoid rhodovibrin with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

Preparative isolation of carotenoid 1-hydroxy-spirilloxanthin from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl.1 of R.rubrum, with 1 -hydroxy-spirilloxanthin, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the 1 -hydroxy-spirilloxanthin present in the fraction Fl.1 . The choice of the chromatography support and approach (e.g. a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to 1 -hydroxy-spirilloxanthin isolation (See Example 4, here below, referring to Bcma-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated 1 -hydroxy- spirilloxanthin will be analyzed by mass spectrometry. As a next step, 1 -hydroxy-spirilloxanthin is isolated from the fraction fl.1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the carotenoid. The carotenoid is separated from the remainder of fraction fl .1 to an extent as close as possible to homogeneity. The isolated 1 -hydroxy-spirilloxanthin obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. See also Example 1 and 2 for the LDL-cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated carotenoid 1 -hydroxy-spirilloxanthin with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

Preparative isolation of carotenoid 3,4-dehydro-rhodopin from the ether extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl.1 of R.rubrum, with 3,4-dehydro-rhodopin, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the 3,4-dehydro-rhodopin present in the fraction Fl.1 . The choice of the chromatography support and approach (e.g. a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to 3,4-dehydro-rhodopin isolation (See Example 4, here below, referring to Bona-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated 3,4-dehydro- rhodopin will be analyzed by mass spectrometry. As a next step, 3,4-dehydro-rhodopin is isolated from the fraction fl.1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the carotenoid. The carotenoid is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated 3,4-dehydro-rhodopin obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. See also Example 1 and 2 for the LDL- cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated carotenoid 3,4-dehydro-rhodopin with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

Preparative isolation of carotenoid chloroxanthin from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl.1 of R.rubrum, with chloroxanthin, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the chloroxanthin present in the fraction Fl.1 . The choice of the chromatography support and approach (e.g. a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to chloroxanthin isolation (See Example 4, here below, referring to Bona-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated chloroxanthin will be analyzed by mass spectrometry. As a next step, chloroxanthin is isolated from the fraction fl .1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the carotenoid. The carotenoid is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated chloroxanthin obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl .1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. See also Example 1 and 2 for the LDL-cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated carotenoid chloroxanthin with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

Preparative isolation of carotenoid rhodopin from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl .1 of R.rubrum, with rhodopin, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the rhodopin present in the fraction Fl.1 . The choice of the chromatography support and approach (e.g. a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to rhodopin isolation (See Example 4, here below, referring to Bona-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated rhodopin will be analyzed by mass spectrometry. As a next step, rhodopin is isolated from the fraction fl.1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the carotenoid. The carotenoid is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated rhodopin obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. See also Example 1 and 2 for the LDL-cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated carotenoid rhodopin with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

Preparative isolation of carotenoid spirilloxanthin from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl.1 of R.rubrum, with spirilloxanthin, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the spirilloxanthin present in the fraction FI.1 . The choice ofthe chromatography support and approach (e.g. a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to spirilloxanthin isolation (See Example 4, here below, referring to Bona-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated spirilloxanthin will be analyzed by mass spectrometry. As a next step, spirilloxanthin is isolated from the fraction fl .1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the carotenoid. The carotenoid is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated spirilloxanthin obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl .1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. See also Example 1 and 2 for the LDL-cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated carotenoid spirilloxanthin with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

Preparative isolation of carotenoid 3,4-dihydro-spirilloxanthin from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl .1 of R.rubrum, with 3,4-dihydro-spirilloxanthin, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the 3,4-dihydro-spirilloxanthin present in the fraction Fl.1 . The choice of the chromatography support and approach (e.g. a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to 3,4-dihydro-spirilloxanthin isolation (See Example 4, here below, referring to Bona-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated 3,4-dihydro- spirilloxanthin will be analyzed by mass spectrometry. As a next step, 3,4-dihydro-spirilloxanthin is isolated from the fraction fl.1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the carotenoid. The carotenoid is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated 3,4-dihydro-spirilloxanthin obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. See also Example 1 and 2 for the LDL-cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated carotenoid 3,4-dihydro-spirilloxanthin with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy. Preparative isolation of pheophytin geranyl-geranyl bacteriopheophytin a from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl.1 of R.rubrum, with geranyl-geranyl bacteriopheophytin a, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the geranyl-geranyl bacteriopheophytin a present in the fraction Fl.1 . The choice of the chromatography support and approach (e.g. a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to geranyl-geranyl bacteriopheophytin a isolation (See Example 4, here below, referring to Bcma-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated geranyl-geranyl bacteriopheophytin a will be analyzed by mass spectrometry. As a next step, geranyl-geranyl bacteriopheophytin a is isolated from the fraction fl.1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the pheophytin. The pheophytin is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated geranyl-geranyl bacteriopheophytin a obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the pheophytin compound. See also Example 1 and 2 for the LDL-cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated pheophytin geranyl-geranyl bacteriopheophytin a with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

Isolating the bacteriopheophytin phytyl bacteriopheophytin a from R. rubrum bacteria cells

A method of isolation and characterisation of phytyl bacteriopheophytin a has been described in Bcma- Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating phytyl bacteriopheophytin a from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of phytyl bacteriopheophytin a using HPLC-MS For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bcma-Lovasz et al. (2013)). The retention time and mass of phytyl bacteriopheophytin a are depicted in Table 1 of Bona-Lovasz et al. (2013), and these are 26,75 minutes and 888,5 g/mol respectively. The isolated phytyl bacteriopheophytin a obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the pheophytin compound. Negative control is a group of mice fed regular diet omitting the pheophytin and omitting the R.rubrum fraction.

Preparative isolation of ubiquinol ubiquinol-10 from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl.1 of R.rubrum, with ubiquinol-10, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the ubiquinol-10 present in the fraction Fl.1 . The choice of the chromatography support and approach ( e.g . a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to ubiquinol- 10 isolation (See Example 4, here below, referring to Bona-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated ubiquinol-10 will be analyzed by mass spectrometry. As a next step, ubiquinol-10 is isolated from the fraction fl.1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the ubiquinol. The ubiquinol is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated ubiquinol-10 obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the ubiquinol compound. See also Example 1 and 2 for the LDL-cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated ubiquinol ubiquinol-10 with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

Preparative isolation of ubiquinone ubiquinone-9 from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl.1 of R.rubrum, with ubiquinone-9, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the ubiquinone-9 present in the fraction Fl.1 . The choice of the chromatography support and approach (e.g. a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to ubiquinone- 9 isolation (See Example 4, here below, referring to Bona-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated ubiquinone-9 will be analyzed by mass spectrometry. As a next step, ubiquinone-9 is isolated from the fraction fl.1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the ubiquinone. The ubiquinone is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated ubiquinone-

9 obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the ubiquinone compound. See also Example 1 and 2 for the LDL-cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated ubiquinone ubiquinone-9 with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

Preparative isolation of ubiquinone ubiquinone-10 from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl.1 of R.rubrum, with ubiquinone-10, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the ubiquinone-10 present in the fraction Fl.1 . The choice of the chromatography support and approach (e.g. a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to ubiquinone-

10 isolation (See Example 4, here below, referring to Bona-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated ubiquinone-10 will be analyzed by mass spectrometry. As a next step, ubiquinone-10 is isolated from the fraction fl.1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the ubiquinone. The ubiquinone is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated ubiquinone-10 obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the ubiquinone compound. See also Example 1 and 2 for the LDL-cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R.rubrum. The isolated ubiquinone ubiquinone-10 with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

Preparative isolation of ubiquinone derivative rhodoquinone-10 from the extract fl.1 derived from cells of bacterium R. rubrum

The extract fraction fl.1 of R.rubrum, with rhodoquinone-10, and obtained as described here above in Example 2, is submitted to analytical chromatography for defining suitable conditions for the separation and isolation of the rhodoquinone-10 present in the fraction Fl.1 . The choice of the chromatography support and approach (e.g. a hydrophobic chromatography using silica and two organic solvents) is based on the here above described results and is based on the parts of the cited literature relating to rhodoquinone-10 isolation (See Example 4, here below, referring to Bona-Lovasz et al. (2013), page 925, last paragraph - page 926, first paragraph). The analytically separated rhodoquinone-10 will be analyzed by mass spectrometry. As a next step, rhodoquinone-10 is isolated from the fraction fl .1 by applying preparative (HPLC) chromatography based on hydrophobic column material. The HPLC is coupled to a mass spectrometer for analysis of the fractions comprising the ubiquinone derivative. The ubiquinone derivative is separated from the remainder of fraction fl.1 to an extent as close as possible to homogeneity. The isolated rhodoquinone-10 obtained with preparative chromatography is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the ubiquinone derivative compound. See also Example 1 and 2 for the LDL- cholesterol lowering effect in plasma of mice, of the fraction 1.1 of R. rubrum. The isolated ubiquinone derivative rhodoquinone-10 with cholesterol-lowering activity is further characterized by applying methods including for example mass spectrometry, UV spectroscopy, Infrared spectroscopy, NMR spectroscopy.

As shown by this Example, all of the present carotenoids, bacteriopheophytins and quinones could be extracted from the extract fraction 1.1 . This shows that a composition comprising at least one, preferably at least two, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 can be made.

Example 4

Isolating the carotenoid rhodovibrin from R. rubrum bacteria cells

A method of isolation and characterisation of rhodovibrin has been described in Bona-Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating rhodovibrin from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of rhodovibrin using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of rhodovibrin are depicted in Table 1 of Bona-Lovasz et al. (2013), and these are 21 .43 minutes and 584.5 g/mol respectively. The isolated rhodovibrin obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

The isolation of rhodovibrin is also outlined in the Main List, entry No. 167 of the Carotenoids Handbook, edited by Britton et al. (2004). Rhodovibrin isolated according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Isolating the carotenoid 1-hydroxy-spirilloxanthin from R. rubrum bacteria cells

A method of isolation and characterisation of 1 -hydroxy-spirilloxanthin has been described in Bona- Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating 1 -hydroxy-spirilloxanthin from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of 1 -hydroxy-spirilloxanthin using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of 1 -hydroxy- spirilloxanthin are depicted in Table 1 of Bona-Lovasz et al. (2013), and these are 20,85 minutes and 582.5 g/mol, respectively.

The isolated 1 -hydroxy-spirilloxanthin obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

Isolating the carotenoid 3,4-dehydro-rhodopin from R. rubrum bacteria cells

A method of isolation and characterisation of 3,4-dehydro-rhodopin has been described in Bona-Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating 3,4-dehydro-rhodopin from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of 3,4-dehydro-rhodopin using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of 3,4-dehydro- rhodopin are depicted in Table 1 of Bona-Lovasz et al. (2013), and these are 22,07 minutes and 552,5 g/mol respectively.

The isolated 3,4-dehydro-rhodopin obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl .1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

The isolation of 3,4-dehydro-rhodopin is also outlined in the Main List, entry No. 90 of the Carotenoids Handbook, edited by Britton et al. (2004). 3,4-dehydro-rhodopin isolated according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Isolating the carotenoid chloroxanthin from R. rubrum bacteria cells

A method of isolation and characterisation of chloroxanthin has been described in Bona-Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating chloroxanthin from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of chloroxanthin using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of chloroxanthin are depicted in Table 1 of Bona-Lovasz et al. (2013), and these are 24.61 minutes and 556.5 g/mol respectively.

The isolated chloroxanthin obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

The isolation of chloroxanthin is also outlined in the Main List, entry No. 100 of the Carotenoids Handbook, edited by Britton et al. (2004). Chloroxanthin isolated according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Isolating the carotenoid rhodopin from R. rubrum bacteria cells A method of isolation and characterisation of rhodopin has been described in Bona-Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating rhodopin from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of rhodopin using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of rhodopin are depicted in Table 1 of Bona- Lovasz et al. (2013), and these are 23.63 minutes and 554.4 g/mol respectively.

The isolated rhodopin obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

The isolation of rhodopin is also outlined in the Main List, entry No. 93 of the Carotenoids Handbook, edited by Britton et al. (2004). Rhodopin isolated according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Isolating the carotenoid spirilloxanthin from R. rubrum bacteria cells

A method of isolation and characterisation of spirilloxanthin has been described in Bona-Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating spirilloxanthin from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of spirilloxanthin using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of spirilloxanthin are depicted in Table 1 of Bona-Lovasz et al. (2013), and these are 23.87 minutes and 596.4 g/mol respectively.

The isolated spirilloxanthin obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

The isolation of spirilloxanthin is also outlined in the Main List, entry No. 166 of the Carotenoids Handbook, edited by Britton et al. (2004). Spirilloxanthin isolated according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Isolating the carotenoid 3,4-dihydro-spirilloxanthin from R. rubrum bacteria cells

A method of isolation and characterisation of 3,4-dihydro-spirilloxanthin has been described in Bona- Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating 3,4-dihydro-spirilloxanthin from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of 3,4-dihydro-spirilloxanthin using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of 3,4-dihydro-spirilloxanthin are depicted in Table 1 of Bona-Lovasz et al. (2013), and these are 24.46 minutes and 598.5 g/mol respectively.

The isolated 3,4-dihydro-spirilloxanthin obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl .1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

The isolation of 3,4-dihydro-spirilloxanthin is also outlined in the Main List, entry No. 168 of the Carotenoids Handbook, edited by Britton et al. (2004). 3,4-dihydro-spirilloxanthin isolated according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Isolating the pheophytin geranyl-geranyl bacteriopheophytin a from R. rubrum bacteria cells

A method of isolation and characterisation of geranyl-geranyl bacteriopheophytin a has been described in Bona-Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating geranyl-geranyl bacteriopheophytin a from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth ( R . rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of geranyl-geranyl bacteriopheophytin a using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of geranyl-geranyl bacteriopheophytin a are depicted in Table 1 of Bona- Lovasz et al. (2013), and these are 23,60 minutes and 882,5 g/mol respectively.

The isolated geranyl-geranyl bacteriopheophytin a obtained according to the method of Bona- Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the pheophytin compound. Negative control is a group of mice fed regular diet omitting the pheophytin and omitting the R.rubrum fraction.

Isolating the pheophytin geranyl-geranyl bacteriopheophytin a

An isolation protocol for geranyl-geranyl bacteriopheophytin a is described by Walter et al. (1979; starting at p. 91 1 , last paragraph - up to p.914, 1^st paragraph). Reference is therefore made to this protocol for providing geranyl-geranyl bacteriopheophytin a. In brief, first, 31 g of frozen R. rubrum cells were defrosted in a mortar and crushed with 100 ml of methanol and 30 mg of butyl hydroxyl toluol (BHT). The resulting suspension was diluted with 150 ml of ether and filtered through a slurry of 12 g of Celite in ether using a glass suction filter G3, such that a clear extract was obtained. The filtered methanol extract was saved. The filter residue was washed 3 times with 30 ml of ether and the resulting combined filtrate was diluted with 150 ml pentane. The separated water/methanol phase was shaken with 60 ml ether/pentane (1 :1) and 5 ml of NaCI solution was added. Both organic solutions were first washed with 30 ml buffer with pH 7 and then with 30 ml of NaCI solution. This crude extract contains geranyl-geranyl bacteriopheophytin a and geranyl-geranyl bacteriochlorophyll a. To separate the geranyl-geranyl bacteriopheophytin a from the geranyl-geranyl bacteriochlorophyll a, the crude extract is dried over Na₂SC>₄ after addition of 20 ml methanol in a rotary evaporator at 30°. Five ml of methanol is then poured overthe crude product four times, while the solution is pipetted off, filtered and evaporated after every round. The residue that does not dissolve in methanol was dissolved in ether. After addition of methanol, the solvent was evaporated. The remaining residue is washed with 6 ml of methanol and then the non-dissolved part discarded, the solution re-united with the first methanol extracts and evaporated. This product is washed again 4 times with 2.5 ml methanol, and solvent was evaporated. The geranyl-geranyl bacteriopheophytin a was dissolved in methylene chloride (20 ml) and 20 ml 1 N HCI was added, and the mixture was shaken. The organic phase was then washed with 20 ml buffer at pH 7. The water phase was washed with 30 ml methylene chloride, and the combined methylene chloride solutions were dried over Na2SC and dried in a rotary evaporator. The geranyl-geranyl bacteriopheophytin a was obtained. This was dissolved in 2 ml methylene chloride, then diluted with 2 ml methanol, and left for 16 hr at room temperature in a not completely closed container. The precipitate was filtered and obtained, and was washed with 8 ml methanol, and then dried (yield was 1 10 mg). The geranyl-geranyl bacteriopheophytin a was purified by two cycles of: dissolving the geranyl-geranyl bacteriopheophytin a in methylene chloride; diluting with methanol; evaporating the solvents with a rotary evaporator; washing the solid geranyl-geranyl bacteriopheophytin a with 6 ml methanol. Isolating the bacteriopheophytin phytyl bacteriopheophytin a from R. rubrum bacteria cells

A method of isolation and characterisation of phytyl bacteriopheophytin a has been described in Bona- Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating phytyl bacteriopheophytin a from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum ) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of phytyl bacteriopheophytin a using HPLC-MS For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of phytyl bacteriopheophytin a are depicted in Table 1 of Bona-Lovasz et al. (2013), and these are 26,75 minutes and 888,5 g/mol respectively.

The isolated phytyl bacteriopheophytin a obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl .1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the pheophytin compound. Negative control is a group of mice fed regular diet omitting the pheophytin and omitting the R.rubrum fraction.

Isolating the ubiquinol ubiquinol-10 from R. rubrum bacteria cells

A method of isolation and characterisation of ubiquinol-10 has been described in Bona-Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating ubiquinol-10 from a crude preparation of R rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of ubiquinol-10 using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of ubiquinol-10 are depicted in Table 1 of Bona-Lovasz et al. (2013), and these are 31 .08 minutes and 846.7 g/mol respectively.

The isolated ubiquinol-10 obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the ubiquinol compound. Negative control is a group of mice fed regular diet omitting the ubiquinol and omitting the R.rubrum fraction.

Isolating the ubiquinone ubiquinone-9 from R. rubrum bacteria cells

A method of isolation and characterisation of ubiquinone-9 has been described in Bcma-Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating ubiquinone-9 from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of ubiquinone-9 using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of ubiquinone-9 are depicted in Table 1 of Bcma-Lovasz et al. (2013), and these are 31 .43 minutes and 794.6 g/mol respectively.

The isolated ubiquinone-9 obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the ubiquinone compound. Negative control is a group of mice fed regular diet omitting the ubiquinone and omitting the R.rubrum fraction.

Isolation & purification of ubiquinone-9 from a natural source

Isolation of ubiquinone-9 (CoQg) from Agaricus campestris (edible mushrooms) is described in detail by Erickson et al. (1960; p. 315). Reference is made to Erickson et al. (1960; p. 315) for details, and in brief, 7600 g of mushrooms was blended with 7.2 I of 95% ethanol to form a paste. To this paste 260 g of pyrogallol was added and the mixture was heated to reflux temperature. A solution of 900 g potassium hydroxide in 1 .8 litre water was slowly added and the entire mixture was stirred for 30 minutes at reflux temperature. When the mixture had cooled to room temperature it was extracted successively with 5.4, 1 .8, 1 .8 and 1 .8 litre portions of hexane. The extracts were combined and evaporated to dryness under vacuum to yield 12.4 g residue. Next, the residue was combined with 200 ml of warm hexane and kept at 5 deg for 16 hrs. The precipitate was centrifuged and washed with small portions of hexane until the precipitate became colourless. Then the precipitate was discarded while the washings and supernatant were combined and evaporated to dryness under vacuum. Successive precipitations were then carried out with 25, 10, 5 and 2.5 ml of warm acetone respectively. Each time the precipitate was washed with small portions of acetone until it became colourless and the washings and supernatant were collected. The combined washings and supernatants were then evaporated to dryness under vacuum to yield 362 mg of solid. The solid was eluted in a glass column with 2% ethyl ether hexane. This yielded several fractions of Coenzyme Cb. The impure fractions were then further eluted and evaporated to dryness. This yielded 1 1 .2 mg of an orange solid, which was crystallized with 1 ml of absolute ethanol. A yield of 70% C0Q9 was achieved.

Isolating the ubiquinone ubiquinone-10 from R.rubrum bacteria cells

A method of isolation and characterisation of ubiquinone-10 has been described in Bcma-Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating ubiquinone-10 from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of ubiquinone-10 using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et al. (2013)). The retention time and mass of ubiquinone-10 are depicted in Table 1 of Bcma-Lovasz et al. (2013), and these are 32.81 minutes and 862.7 g/mol, respectively.

The isolated ubiquinone-10 obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the ubiquinone compound. Negative control is a group of mice fed regular diet omitting the ubiquinone and omitting the R.rubrum fraction.

Isolation & purification of ubiquinone-10 from a natural source

Isolation of C0Q10 from a natural source is performed using Rhodospirillum rubrum and is detailed by Parson & Rudney (1965; p 1856, left column, 2^nd paragraph). Reference is made to the protocol outlined in Parson & Rudney (1965), and in brief, first, R. rubrum is grown and the cells harvested, centrifuged, washed with water and lyophilized. The lyophilized cells were then extracted with three 70 ml portions of ether-ethanol (3:1). The extract was evaporated, redissolved in 25 ml of petroleum-ether and filtered. This petroleum-ether solution was then extracted four times with 5 ml portions of 95% methanol. Ubiquinone was eluted with 7% ether in petroleum ether and recrystallized under the influence of ethanol-methanol (1 :1).

Isolation of C0Q10 from a natural source is performed using Ustilago zea harvested cells (Erickson et al. , 1960; pp. 315, right column, 4^th paragraph - 316, end). Reference is made to Erickson et al. (1960) for the detailed protocol, and in brief, these cells are dried by slurrying with acetone, centrifugation and drying in a vacuum oven. The dried cell cake (4.56 kg) was then ground to 60 mesh and extracted by stirring with successive 10.8-, 7.2-, 3.6- and 3.6 litre portions of hexane at reflux temperature, followed by cooling and separating of the solution. Extracts were then combined and evaporated to dryness yielding 13.4 g of tarry solid. Next, the cells were extracted with 4 litre portions of boiling absolute ethanol and evaporated under vacuum to dryness yielding 102.9 g of tarry residue. The hexane extract was purified by precipitation of impurities and chromatography to give 249 mg of yellow oil containing the C0Q10 (66%). The ethanol extract was purified in the same manner and yielded 600 mg of material containing C0Q10.

Isolating the ubiquinone derivative rhodoquinone-10 from R.rubrum bacteria cells

A method of isolation and characterisation of rhodoquinone-10 has been described in Bona-Lovasz et al. (2013). Reference is made to page 925, last paragraph - page 926, first paragraph, for the detailed steps for isolating rhodoquinone-10 from a crude preparation of R. rubrum. In brief, every step of this extraction was carried out under argon and in the dark. Five to 20 ml of bacterial fermentation broth (R. rubrum) was provided and centrifuged for 15 minutes, and then the supernatant was discarded. Next, 1 ml of methanol and 2 ml of hexane was added to the pellet and the mixture was vortexed for 2 minutes. Phase separation was achieved by adding 1 ml of water to the mixture, followed by vortexing for 1 minute and centrifugation for 20 minutes. The upper supernatant phase was then used directly for separation of rhodoquinone-10 using HPLC-MS. For HPLC, a Spherisorb C18 column with a mobile phase consisting of acetone-water and a non-linear gradient of 50%-100% acetone, was applied (see: abstract on page 912 of Bona-Lovasz et at. (2013)). The retention time and mass of rhodoquinone-10 are depicted in Table 1 of Bona-Lovasz et al. (2013), and these are 32.24 minutes and 847.7 g/mol, respectively.

The isolated rhodoquinone-10 obtained according to the method of Bona-Lovasz et al. (2013) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl .1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the ubiquinone derivative compound. Negative control is a group of mice fed regular diet omitting the ubiquinone derivative and omitting the R.rubrum fraction.

Isolation & purification of rhodoquinone-10 from a natural source

Isolation of rhodoquinone-10 from a natural source is performed using Rhodospirillum rubrum and is detailed by Parson & Rudney (1965; p 1856, left column, 2^nd paragraph). Reference is made to the protocol outlined in Parson & Rudney (1965), and in brief, first, R. rubrum is grown and the cells harvested, centrifuged, washed with water and lyophilized. The lyophilized cells were then extracted with three 70 ml portions of ether-ethanol (3:1). The extract was evaporated, redissolved in 25 ml of petroleum-ether and filtered. This petroleum-ether solution was then extracted four times with 5 ml portions of 95% methanol. Rhodoquinone-10 was eluted with 16% ether in petroleum ether.

As shown by this Example, all of the present carotenoids, bacteriopheophytins and quinones can be isolated from a natural source. This shows that a composition comprising at least one, preferably at least two, preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 can be made. Example 5

Synthesizing the carotenoid rhodovibrin

Synthetic synthesis of the carotenoid rhodovibrin has been detailed by Surmatis et al. (1966). Briefly summarized, Surmatis et al. (1965) provided rhodovibrin by reacting crocetindialdehyde with (7-hydroxy- 3,7-dimethyl-2-octenyl)triphenylphosphonium bromid in a benzene-methanol solution. After purification by chromatography on grade I alumina, recrystallization was carried out with benzene to yield 42.3% of 1 -hydroxy-1 ,2-dihydroapo-3-lycopenal as a dark red crystalline solid. Condensation of this compound with (7-methoxy-3,7-dimethyl-2,4-octadien-1 -yl)triphenylphosphorium bromide in boiling methyl alcohol gave rhodovibrin in a yield of 65% as a violet crystalline solid. Reference is made to Surmatis et al. (1965), p. 188, left column, 4^th paragraph for the details of the rhodovibrin synthesis procedure.

The synthesized rhodovibrin obtained according to the method of Surmatis et al. (1966) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

The synthesis of rhodovibrin is also outlined in Carotenoids Volume 2 Synthesis (1996), page 142-143, page 297. The synthesis of rhodovibrin is also outlined in the Main List, entry No. 167 of the Carotenoids Handbook, edited by Britton et al. (2004). Rhodovibrin synthesized according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Chemical synthesis the carotenoid 3,4-dehydro-rhodopin

Synthetic synthesis of the carotenoid 3,4-dehydro-rhodopin has been detailed by Schneider et al. (1967). Briefly summarized and referring to the original paper, Schneider et al. (1967) provided 3,4- dehydro-rhodopin by using acetone in a Reformatsky-type reaction with prop-2-ynyl bromide to give the alcohol2-methylpent-4-yn-2-ol. Methylation in dimethyl sulphoxide with methyl iodide in the presence of barium oxide furnished the corresponding methyl ether 2-methoxy-2-methylpent-4-yne. Lithium derivatives of methylpent-4-yn-2-ol and 2-methoxy-2-methylpent-4-yne reacted with methyl vinyl ketone to give the 2,6-dimethyloct-7-en-4ybe-2,6 diol and 7-methoxy-3,7-dimethyloct-1 -en-4-yn-3-ol, respectively. These were reduced with lithium aluminium hybride to give 6,6-dimethylocta-4,7-diene- 2,6-diol and 7-methoxy-3,7-dimethylocta-1 ,4-dien-3-ol. Treatment with triphenylphosphine hydrobromide then furnished the required Wittig reagents (7-hydroxy-3,7-dimethylocta-2,4- dienyl)triphenylphosphonium bromide and (7-methoxy-3,7-dimethylocta-2,4- dienyl)triphenylphosphonium bromide. Condensation of (7-hydroxy-3,7-dimethylocta-2,4- dienyl)triphenylphosphonium bromide with C30 apo-8-lycopenal gave the 3,4-didehydro-rhodopin. The synthesized 3,4-dehydro-rhodopin obtained according to the synthesis method of Schneider et al. (1967) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl .1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

The synthesis of 3,4-dehydro-rhodopin is also outlined in Carotenoids Volume 2 Synthesis (1996), page 141 (compound 90). The synthesis of 3,4-dehydro-rhodopin is also referred to in the Main List, entry No. 90 of the Carotenoids Handbook, edited by Britton et al. (2004). 3,4-dehydro-rhodopin synthesized according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Chemically synthesizing the carotenoid chloroxanthin

Synthetic synthesis of the C40 carotenoid chloroxanthin has been detailed by Barber et al. (1966). Briefly summarized, Barber et al. (1966) provided chloroxanthin following the following procedure. Sodium methoxide in methanol was added dropwise to a stirred solution of 7,8-dihydro-apo-8’-lycopenal and (7- hydrocy-3,7-dimethyloct-2-enyl)triphenylphosphonium bromide in dry methanol. The solution was stirred, diluted with water and extracted with benzene. The crude product was purified by repeated chromatography on alumina using benzene as eluant. Collection, evaporation and crystallisation of the residue from light petroleum gave chloroxanthin as orange-red prisms. Reference is made to page 2173, right column, last paragraph - page 2174, left column, first paragraph, for the detailed procedure (Barber et al. , 1966).

The synthesized chloroxanthin obtained according to the method of Barber et al. (1966) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

The synthesis of chloroxanthin (compound 100) is also outlined in Carotenoids Volume 2 Synthesis (1996), page 144-145. The synthesis of chloroxanthin is also outlined in the Main List, entry No. 100 of the Carotenoids Handbook, edited by Britton et al. (2004). Chloroxanthin synthesized according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Synthesizing the carotenoid rhodopin

Synthetic synthesis of the carotenoid rhodopin has been described by Surmatis et al. (1966). Briefly summarized, Surmatis et al. (1966) condensed crocetindialdehyde with (3,7-dimethyl-2,6- octadienyl)triphenylphosphonium bromide to yield apo-3-lycopenal in a 41 .6% yield as purple plates. This was then condensed with (7-hydroxy-3,7-dimethyl-2-octenyl)triphenylphosphonium bromide by the Wittig reaction and this yielded 57% rhodopin as a dark red crystalline solid. Reference is made to p. 188, right column, 4th paragraph of Surmatis et al. (1966), for the details of the chemical synthesis procedure.

The synthesized rhodopin obtained according to the method of Surmatis et al. (1966) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubmm fraction.

The synthesis of rhodopin is also outlined in Carotenoids Volume 2 Synthesis (1996), page 144- 145. The synthesis of rhodopin is also outlined in the Main List, entry No. 93 of the Carotenoids Handbook, edited by Britton et al. (2004). Rhodopin synthesized according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Synthesizing the carotenoid spirilloxanthin

Synthetic synthesis of the carotenoid spirilloxanthin has been described by Surmatis et al. (1963). Briefly summarized, Surmatis et al. (1963) prepared 6-methoxy-6-methyl-2-heptanone by stirring 6-methylhept- 5-en-2-one in a solution of sulfuric acid and methyl alcohol for 24 hrs at room temperature. Condensation of 6-methoxy-6-methyl-2-heptanone with ethyl bromoacetate by the Reformatsky reaction, followed by dehydration with phosphorus oxychloride in pyridine yielded 7-methoxy-3,7-dimethyl-2-octenoic acid ethyl ester. Reaction of this ester with N-bromosccinimide, followed by dehydrobromination with dimethlaniline yielded the unsaturated esterethyl 7-methoxy-3,7-dimethylocta-2,4-dienoate. This ester was reduced with lithium aluminium hybride in ethyl ether to give 7-methoxy-3,7-dimethyl-2,4-octadien- 1 -ol. Stirring the 7-methoxy-3,7-dimethyl-2,4-octadien-1 -ol with triphenylphosphonium bromide in methanol yielded the Wittig salt (7-methoxy-3,7-dimethyl-2,4-octadien-1 -yl)triphenylphosphonium bromide. A Wittig condensation with crocetin dialdehyde and the Wittig salt was carried out in sodium methylate in methanol stirring at reflux temperature to give spirriloxanthin as spindle-shaped violet coloured crystals in a 30% yield. Reference is made to page 2738, left column, 6^th paragraph of Surmatis et al (1963), for the details of the chemical synthesis procedure.

Surmatis et al. (1963) describe a second method to chemically synthesize spirilloxanthin. Briefly summarized, Surmatis et al. (1963) outlined the ethynylation of 6-methoxy-6-methyl-2-heptanone with sodium acetylide in liquid ammonia to yield 7-methoxy-3,7-dimethyl-1 -octyn-3-ol. Selective hydrogenation of 7-methoxy-3,7-dimethyl-1 -octyn-3-ol resulted in 7-methoxy-3,7-dimethyl-1 -octen-3-ol. Stirring 7-methoxy-3,7-dimethyl-1 -octen-3-ol with triphenylphosphonium bromide in methanol resulted in the Wittig salt (7-methoxy-3,7-dimethyl-2-octen-1 -yl)triphenylphosphonium bromide. Condensing the Wittig salt with crocetin dialdehyde resulted in 1 ,T-dimethoxy-1 ,2,1’,2’-tetrahydrolycopene. This was then reacted with N-bromosuccinimide, which resulted in a 24% yield of spirilloxanthin. Reference is made to page 2738, right column, first paragraph, of Surmatis et al. (1963) for the details of the chemical synthesis procedure.

Another method to chemically synthesize spirilloxanthin was presented by Schneider et al. (1967). Briefly summarized, Schneider et al. (1967) used acetone in a Reformatsky-type reaction with prop-2-ynyl bromide to give the alcohol2-methylpent-4-yn-2-ol. Methylation in dimethyl sulphoxide with methyl iodide in the presence of barium oxide furnished the corresponding methyl ether 2-methoxy-2- methylpent-4-yne. Lithium derivatives of methylpent-4-yn-2-ol and 2-methoxy-2-methylpent-4-yne reacted with methyl vinyl ketone to give the 2,6-dimethyloct-7-en-4ybe-2,6 diol and 7-methoxy-3,7- dimethyloct-1 -en-4-yn-3-ol, respectively. These were reduced with lithium aluminium hybride to give 6,6- dimethylocta-4,7-diene-2,6-diol and 7-methoxy-3,7-dimethylocta-1 ,4-dien-3-ol. Treatment with triphenylphosphine hydrobromide then furnished the required Wittig reagents (7-hydroxy-3,7- dimethylocta-2,4-dienyl)triphenylphosphonium bromide and (7-methoxy-3,7-dimethylocta-2,4- dienyl)triphenylphosphonium bromide. Condensation of (7-methoxy-3,7-dimethylocta-2,4- dienyl)triphenylphosphonium bromide with crocetindial yielded spirriloxanthin. Reference is made to page 1689, left column, 3^rd paragraph, of Schneider et al. (1967) for the details of the chemical synthesis procedure.

The synthesized spirilloxanthin obtained according to the method(s) of Surmatis et al. (1963) and/or Schneider et al. (1967) is tested in a mice activity test performed substantially as described here above in Example 1 and Example 2. A minimum of 5 mice is included in each group. A positive control group of mice, consisting of mice fed with active extract fl.1 , is included in the test in order to be able to evaluate quantitatively the cholesterol lowering effect of the carotenoid compound. Negative control is a group of mice fed regular diet omitting the carotenoid and omitting the R.rubrum fraction.

The synthesis of spirilloxanthin is also referred to in Carotenoids Volume 2 Synthesis (1996), page 142- 143. The synthesis of spirilloxanthin is also outlined in the Main List, entry No. 166 of the Carotenoids Handbook, edited by Britton et al. (2004). Spirilloxanthin synthesized according to these procedures is subjected to plasma LDL-cholesterol level lowering tests, as herein described.

Chemical synthesis of ubiquinone-10

Chemical synthesis methods for ubiquinone-10 are outlined by Inoue et al. (1974). Details of the chemical synthesis of C0Q10 are outlined in Inoue et al. (1974) and in brief, employed the selective carbon-carbon bond formation between tt-allylnickel halide complexes and organic halides, specifically TT-allylnickel bromides (see p. 3098, 1 ^st paragraph of Inoue et al. (1974)). Inoue et al. (1974) reacted hydroquinone diacetate bromide with tt-decaprenylnickel bromide in HMPA at 75 deg for 7 hrs, after which the resulting diacetate molecule was then converted by removing the acetate groups with LiAIH and subsequent oxidation by aqueous FeCh to Coenzyme Qio. The yield was 69%. Reactions involving the TT-allylnickel bromides were carried out under nitrogen or argon. (See also: Inoue et al., 1974; p. 3101 , left column, 2^nd paragraph-end).

C0Q10 is chemically synthesized as detailed by Ravada et al. (2009). Reference is made to the detailed protocol in Ravada et al. (2009), and in brief, the synthesis is carried out in nine steps and makes use of isoprenol as a pre-cursor. The method starts with prenylation of 2,3-dimethoxy-5- methylhydroquinone using isoprenol in presence of a Lewis acid, followed by selective oxidation of the trans methyl group of the isoprenyl side chain and subsequent allylic bromination. Next, ptoluenesulfination of the resulting bromide followed by coupling with solanesyl bromide and de- ptoluenosulfination to yield the dimethyl derivative of CoQio-quinol. Finally, CAN oxidation of the dimethyl quinol followed by purification by column chromatography and crystallization yields C0Q10 in 72% yield and 13% overall yield (See also: Ravada et al., 2009; abstract and p. 86 last paragraph right column - 87, first paragraph left column).

Chemical synthesis of rhodoquinone-10

A chemical synthesis method for ubiquinone-10 is outlined by Moore & Folkers (1965). Details of the chemical synthesis of rhodoquinone-10 are outlined in Moore & Folkers (1965) and in brief, synthetic rhodoquinone-10 is synthesised from precursor Coenzyme Qio by ammonolysis. Treating C0Q10 with ammonium hydroxide in a solvent of diethyl ether-ethanol (1 :1) gives synthetic rhodoquinone-10 (See for further details: Moore & Folkers, 1965; p. 1410, left column, 1^st paragraph).

This Example has shown that the synthesis of several carotenoids and quinones is possible, to create a composition comprising at least one, preferably two, more preferably all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

Example 6

Isolation and purification of carotenoids, bacteriopheophytins and quinones from the R.rubrum fraction 1.1

Materials and Equipment

HPLC materials:

Analytical HPLC column ACE5 C18-AR 100x4.6 mm

Semi-preparative HPLC column ACE5 C18-AR 100x21 .2 mm

Water, methanol, isopropylalcohol, acetonitrile, n-pentane, n-hexane and tetrahydrofuran (THF) were of LC-MS quality.

HPLC system:

Waters 600E solvent manager with 100 pi analytical and 5000 pi preparative injection loops.

Waters column oven set at 30°C

Waters 486 UV detector

Waters WFCIII fraction collector set at collection a fraction per minute

Under control of Empower2 software

LC-MS system:

Shimadzu Prominence-i HPLC system Shimadzu LCMS-2020

R.rubrum fraction 1.1 was prepared as here above described for Example 2. Preparing R.rubrum fraction 1.1 from 60 g of biomass (bacterial cells) resulted in a yield of 8 ml (approx. 7 g) of oily petroleum ether extract Fraction 1.1 (after removal of the petroleum ether).

Initial method:

The HPLC method was initially based on Indriatmokoa et at. (2015) Procedia Chemistry 14, 202-210 “Separation of Photosynthetic Pigments by High-Performance Liquid Chromatography: Comparison of Column Performance, Mobile Phase, and Temperature”.

The current column material was similar to the column described in this paper: a C18-type column material, but now functionalized to be more specific for aromatic compounds.

A 10% to 90% gradient of acetonitrile in isopropanol at a very low flow rate of 0.15 ml/min on an ACE5 C18-AR 100x4.6 mm column (injections of 10 pi) was tested. The semi-prep system had an injection loop of 5 ml: 0.5 mg could be injected when a solution of 0.1 mg/ml is used.

Alternatively, THF was used as the solvent for the R.rubrum extract Fraction 1.1 and a gradient elution of THF in isopropanol was used (flow was 0,5 ml/min.), as well as, alternatively, a gradient elution of THF in methanol (flow was 0,5 ml/min.). The fraction 1.1 of R.rubrum dissolves very well in THF. Solutions of 1 mg/ml and 10 mg/ml were prepared. Samples of 50 mg fraction 1.1 are injectable in this semi-preparative HPLC approach. Elution of the compounds from the column resulted in two compound peaks (230 nm detection).

Then, applying the same column and the same solutions, elution using a gradient of THF in n-pentane (0.5 ml/min ) provided four distinct compound fractions at 230 nm, at Rt’s of 3, 3.5, 5.5 and 7.5 mins. The eluted compound peak at Rt is 3 minutes could be separated in two compound fractions. Similar results and compound fractions were obtained when n-pentane was replaced for n-hexane. Elution of compounds was established with a gradient of THF-n-hexane at 0.5 ml/min. Fractions are collected with a fraction collector and the solvent (mobile phase) from the fractions is removed under vacuum.

Eluted compound fractions, obtained from the HPLC column, were analyzed with LC-MS. Analysis of fractions was performed on a Shimadzu LCM2020 with a gradient of isopropylalcohol- acetonitrile with a low flow of 0.15 ml/min using a Shimadzu Prominence HPLC system and using the ACE5 C18-AR 100x4.6 mm column

With a solution of R.rubrum fraction 1.1 dissolved in THF at 20 mg/ml and using an injection loop of 5000 pi, semi-preparative HPLC runs of 50-100 mg of fraction 1.1 material are run. REFERENCES

Barber MS, Jackman LM, Manchand PS, Weedon BCL. 1966. Carotenoids and related compounds. Part XVI. Structural and synthetic studies on spirilloxanthin, chloroxanthin, spheroidene and spheroidenone. J Chem Soc: Org. 0: 2166-2174.

Bcma-Lovasz J, Bona A, Ederer M, Sawodny O, Ghosh R. A rapid method for the extraction and analysis of carotenoids and other hydrophobic substances suitable for systems biology studies with photosynthetic bacteria. Metabolites, 2013. V3: pp.912-930.

Carotenoids Volume 2 Synthesis, Edited by G. Britton, S. Liaaen-Jensen, H. Pfander, Birkhaeuser Verlag, 1996, ISBN 3764352973, ISBN 0817652973.

Carotenoids Handbook, Edited by G. Britton, S. Liaaen-Jensen, H. Pfander, Compiled by A.Z. Mercadante, E.S. Egeland, Springer Basel AG, 2004, ISBN 9783034895880, DOI 10.1007/978-3-0348- 7836-4.

Erickson RE, Brown KS, Wolf DE, Folkers K. 1960. Coenzyme Q XX. Isolation of coenzymes Q9 and Q10 from two basidiomycetes. Archives of biochemistry and biophysics. 90; 314-317.

Inoue S, Yamaguchi R, Saito K, Kikumasa S. 1974. The synthesis of Coenzyme Q. Bulletin of the chemical society of Japan. 47(12); 3098-3101 .

Moore HW, Folkers K. 1965. Coenzyme Q. LXII. Structure and synthesis of rhodoquinone, a natural aminoquinone of the coenzyme Q group. Journal of the American Chemical Society. 86(6): 1409- 1410.

Parson WW, Rudney H. 1965. The biosynthesis of ubiquinone and rhodoquinone from p- hydroxybenzaldehyde in Rhodospirillum rubrum. The Journal of Biological chemistry. 240(4): 1855- 1863.

Ravada SR, Emani LR, Garaga, Meka B, Golakoti T. 2009. Synthesis of Coenzyme Qio. American Journal of Infectious Diseases. 5(2): 83-89.

Schneider DF, Weedon BCL. 1967. Carotenoids and related compounds. Part XVII. Synthesis of spirilloxanthin, ΌH-spirilloxanthin’’and 3,4-dehydrorhodopin. J Chem Soc. 0: 1686-1689.

Shunk CH, Erickson RE, Wong EL, Folkers K. 1959. Coenzyme Q X. Synthesis of coenzyme Q9, 2,3-dimethyl-5-solanesylbenzoquinone (Q-254) and a vitamin K analog. J. Am. Chem. Soc. 81 (18): 5000.

Surmatis JD, Ofner A, Gibas J, Thommen R. Total synthesis of rhodovibrin (OH-P481 ), anhydrorhodovibrin (P481 ) and rhodopin. J Org Chem. 31 (1): 186-188 (January 1966).

Walter E, Schreiber J, Zass E, Eschenmoser A. 1979. 94. Bakteriochlorophyll aGg and Bakteriophaophytin ap in den photosynthetischen reaktionszentren von Rhodospirillum rubrum G-9+. Helvetica chimica acta. 62(94): 899-920.

Claims

1 . Composition comprising two or more compounds of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 for use in a method for the lowering of LDL-cholesterol in blood plasma of a human subject, under the proviso that the composition is not a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

2. Composition for use according to claim 1 comprising all of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4- rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

3. Composition according to claim 1 or 2 for use in the treatment or the prophylaxis of a cardiovascular disease.

4. Composition according to claim 1 or 2 for use in the treatment or prophylaxis of atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL-cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, a plasma Lp(a) level of at least 14 mg/dL, ischemia.

5. Composition for use according to any one of the claims 1 -4, wherein the composition is administered to a subject having an LDL-cholesterol level in plasma of at least 1 ,8 mmol/L (70 mg/dL), or at least 2,59 mmol/L (100 mg/dL), or at least 3,34 mmol/L (129 mg/dL), or at least 4,0 mmol/L, such as at least 5,2 mmol/L (200 mg/dL).

6. Composition for use according to any one of the claims 1 -5, wherein composition is administered to a subject to whom at least one active pharmaceutical ingredient is (co-)ad ministered, such as at least one active pharmaceutical ingredient selected from a statin, niacin, fenofibrate, ezetimibe, colesevelam, mipomersen, lomitapide, a PCSK9 inhibitor, alirocumab, evolocumab, ETC-1002, a CETP inhibitor, anacetrapib, evacetrapib, WAY-252623, a blood-pressure lowering compound, hydrochlorothiazide.

7. Composition for use according to any one of the claims 1 -6, wherein the composition is administered orally to a subject, preferably a human subject.

8. Composition for use according to any one of the claims 1 -7, wherein administering the composition to a subject results in lowering of the plasma LDL-cholesterol concentration, preferably to a plasma concentration of less than 3,34 mmol/L, preferably less than 2,59 mmol/L, more preferably to a plasma LDL-cholesterol concentration of less than 1 ,8 mmol/L.

9. Composition for use according to any one of the claims 1 -8, wherein administering the composition to a subject results in a decrease of the LDL-cholesterol concentration in the plasma of said subject, wherein the plasma HDL-cholesterol concentration remains essentially unaltered or decreases to a smaller extent than the decrease in the plasma LDL-cholesterol concentration, or wherein the plasma HDL-cholesterol concentration increases.

10. Composition for use according to any one of the claims 1 -9, wherein administering the composition to a subject results in a decrease of the plasma LDL-cholesterol concentration in said subject with at least 5%, preferably at least 10%, more preferably at least 20%, most preferably at least 30%, based on the plasma LDL-cholesterol concentration prior to the administration of the composition to said subject.

1 1 . Composition for use according to any one of the claims 1 -10, wherein administering the composition to a subject results in a decrease of the plasma LDL-cholesterol concentration to, or maintenance of the plasma LDL-cholesterol concentration at, a plasma LDL-cholesterol concentration of less than 200 mg/dL, or less than 159 mg/dL, or less than 129 mg/dL, preferably less than 100 mg/dL, such as less than 70 mg/dL.

12. Composition for use according to any one of the claims 1 -1 1 , wherein the composition is formulated as an oral dosage form, preferably a solid oral dosage form or a liquid oral dosage form, preferably a liquid oral dosage form comprising an oil.

13. Composition for use according to any one of the claims 1 -12, wherein the composition is formulated as a tablet or a capsule or a powder or a granulate.

14. Composition for use according to any one of the claims 1 -13, wherein the daily dose of composition to be administered to a subject is 10 microgram-150 mg per day, preferably 100 microgram-100 mg per day, more preferably 500 microgram-50 mg per day, most preferably 1 mg-25 mg per day.

15. Composition for use according to any one of the claims 1 -14, wherein administering the composition to a subject maintains the LDL-cholesterol concentration in the plasma of the subject at a level of less than 159 mg/dL, preferably less than 129 mg/dL, more preferably less than 100 mg/dL, most preferably less than 70 mg/dL, such as between 50 mg/dL and 159 mg/dL.

16. Pharmaceutical composition comprising a pharmaceutically effective amount of at least two, preferably all, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and optionally a pharmaceutically acceptable excipient, for use in a method for the lowering of LDL- cholesterol in blood plasma of a subject, under the proviso that the pharmaceutical composition does not comprise a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

17. Pharmaceutical composition comprising a pharmaceutically effective amount of at least two, preferably all, of compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 and optionally a pharmaceutically acceptable excipient, for use in a method for the treatment or prophylaxis of any one or more of cardiovascular disease, atherosclerosis, dyslipidemia, arteriosclerosis, hypercholesterolemia, familial hypercholesterolemia, hyperlipidemia, an LDL-cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a total plasma cholesterol level of at least 200 mg/dL, an Lp(a) level of at least 14 mg/dL, inflammation, inflammatory disease, ischemia, infection, under the proviso that the pharmaceutical composition does not comprise a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

18. Pharmaceutical composition for use according to claim 16 or 17, wherein the pharmaceutical composition comprises compounds geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

19. Pharmaceutical composition for use according to claim 16-18, wherein the compounds geranyl- geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10 are the sole active pharmaceutical ingredients in said pharmaceutical composition.

20. Pharmaceutical composition for use according to claim 16-18, wherein the pharmaceutical composition is administered to a subject to whom at least one further active pharmaceutical ingredient is administered, such as an active pharmaceutical ingredient selected from a statin, niacin, fenofibrate, ezetimibe, colesevelam, mipomersen, lomitapide, a PCSK9 inhibitor, alirocumab, evolocumab, ETC- 1002, a CETP inhibitor, anacetrapib, evacetrapib, WAY-252623, a blood-pressure lowering compound, hydrochlorothiazide, or any combination thereof such as a statin and a PCSK9 inhibitor.

21 . Pharmaceutical composition for use according to any one of the claims 16-20, wherein the pharmaceutical composition is administered to a subject suffering from an LDL-cholesterol plasma level of at least 70 mg/dL, an LDL-cholesterol plasma level of at least 100 mg/dL, an LDL-cholesterol plasma level of at least 140 mg/dL, an LDL-cholesterol plasma level of at least 200 mg/dL, a CVD.

22. Food supplement with cholesterol-lowering properties, comprising at least two, preferably all, compounds of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4- spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10, under the proviso that the two or more compounds are not provided as a petroleum ether extract of Rhodospirillum rubrum obtainable by extraction of Rhodospirillum rubrum cells with a mixture of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride, wherein the extract is obtained by extraction for between 10 minutes and 48 hours at between 8°C and 37°C, while mixing the cells with the mixture, and wherein the Rhodospirillum rubrum cells are extracted with the mixture in a volume ratio of between 10:1 and 1 :10 of petroleum ether with a boiling point of between 60°C and 80°C and methanol comprising sodium chloride.

23. Food supplement with cholesterol-lowering properties according to claim 22, wherein the food supplement comprises all of geranyl-geranyl bacteriopheophytin a, phytyl derivative of bacteriopheophytin a, hydroxyspirilloxanthin, rhodovibrin, 3,4-rhodopin, chloroxanthin, rhodopin, spirilloxanthin, 3,4-spirilloxanthin, ubiquinol-10, ubiquinone-9, ubiquinone-10 and rhodoquinone-10.

24. Foodstuff comprising a food supplement according to claim 22 or 23.