TECHNICAL FIELD

The present invention relates to a new medication for the treatment (prophylaxis and/or therapy) of dyslipoproteinaemia, also called dyslipidemia, as well as of diseases or medical complications being related thereto, such as treatment of a lipoprotein imbalance present in the same serum of humans, more specifically an abnormally high low density lipoprotein (LDL) fraction.

BACKGROUND ART

Dyslipoproteinaemia is a general diseases indication characterized by abnormal changes in lipoprotein transport, lipid metabolism, and lipoprotein concentrations in serum. Depending on the respective aetiology, pathologic conditions are generally classified into primary dyslipoproteinaemia as a result of genetic determinations, and secondary dyslipoproteinaemiasas a result and consequence of various basic originating diseases, such as adipositas, diabetes mellitus, alcoholism, liver diseases, or as a result of prior medications such as corticosteroids, diuretics, beta-blocker, etc.

According to the present invention, dyslipoproteinaemia also means abnormal cholesterol and/or triglyceride levels in the blood, notably human blood and serum, in particular hypercholesterinaemia, hypertriglyceridemia, or imbalance of lipoprotein profile and concentrations in the blood, especially human blood or serum. Notably, increased levels of cholesterol, triglycerides and/or lipoproteins, in particular the low density lipoproteins (LDL) are typically correlated with a risk of cardiovascular diseases.

Conventionally, the management of dyslipoproteinaemia/lipidaemia, or abnormal or imbalanced lipid/lipoprotein levels are commonly based on treatments with statins (HMG-CoA reductase inhibitors), or in the case of hypertriglyceridemia treatment with fibrates (peroxisome proliferator- activated receptor-a agonists). Statins are optionally combined with ezetimib, niacin, and bile acid sequestrants. Before medication, dietary modification or dietary supplementation, bodyweight reduction, and treatment of the basic originating disease in the event of secondary

hyperlipoproteinaemia are usually the first approaches.

SU M MARY OF THE INVENTION

There is a need, and hence it is an object, to provide, especially by broadening and improving, the therapeutic possibilities of dyslipoproteinaemia and related disorders, including in particular pathologic and risk conditions associated with dyslipoproteinaemia and lipoprotein imbalance.

This object is solved by the therapeutic uses as defined in the claims 1 and 2. Preferred embodiments are defined in subclaims.

With the present invention, a completely novel therapeutic concept is established, offering broader and improved therapeutic possibilities for the treatment (prophylaxis and/or therapy) of

dyslipoproteinaemia, and of diseases correlated or associated with dyslipoproteinaemia or lipoprotein imbalance, in particular imbalanced lipoprotein profiles and specifically abnormal high lipoprotein levels, especially increased LDL. Accordingly, the present invention establishes a new therapeutic approach to reduce the risk of cardiovascular diseases, in particular arteriosclerosis, myocardial infa rction, stroke, peripheral artery disease, and vascular stenosis or restenosis, when respectively correlated or associated with dyslipoproteinaemia or lipoprotein imbalance.

As a basis for the afore-mentioned new therapeutic concepts, it was surprisingly found that inhibitors of phosphodiesterase 4 (PDE 4) are capable of decreasing lipoprotein levels present in blood, notably in human blood and serum. More surprisingly, and particularly valuable for the above specified therapeutic applications, critical lipoprotein fractions and their lipid components cholesterol and/or triglyceride, and in particular LDL and LDL-cholesterol, have been found to be selectively and remarkably decreased upon administration of PDE4 inhibitors.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 shows changes in the blood serum levels (in absolute amounts mg/dl) of cholesterol depending on specific lipoprotein fractions upon administration of a PDE4 inhibitor, respectively after 4 weeks (Tl) and after 8 weeks (T2), compared to the initial values at start (TO); Fig. 2 shows changes in the blood serum levels (in absolute amounts mg/dl) of triglycerides depending on specific lipoprotein fractions upon administration of a PDE4 inhibitor, respectively after 4 weeks (Tl) and after 8 weeks (T2), compared to the initial values at start (TO);

Figs. 3 and 4 respectively shows the course of individual changes of LDL-C and LDL-TG from start (TO) until T2 in the blood serum levels (in mg/dl) of LDL-C (Fig. 3) and LDL-TG (Fig. 4).

DESCRIPTION OF PREFERRED EMBODIM ENTS

As known, phosphodiesterase 4 (PDE 4) is a member of a large, divergent family of phosphodiesterase (PDE) enzymes that catalyse the catabolism of cAM P and cGMP to AMP and GMP, respectively. PDE4 itself encompasses four types, PDE4A, PDE4B, PDE4C and PDE4D, respectively sub-divided into various isoforms. According to the present invention, all PDE4 types and isoforms are meant, and inhibitors specific for each PDE4 type can be used according to the present invention. Literature and reviews about PDE4, its subtype isoforms and the inhibitory class effect of PDE4 inhibitors can be found, for example, in H. Wang et al. in Biochem.J. 408, 193-201 (2007); K.F. Rabe in Br J Pharmacol. 163(1), 53- 67 (2011); Hatzelmann and Schudt in JPET 297, 267-279 (2001).

The new therapeutic effect found for PDE4 inhibitors according to the present invention credibly can be achieved as the function of the class of PDE4 inhibitors.

Prior therapeutic uses of PDE4 inhibitors are known and widespread. However, since the PDE4 family of enzymes are most prevalent in immune cells and cells in the central nervous system, their therapeutic utility hitherto has been limited essentially in diseases and pathologic disorders associated with inflammation and nervous system disorders, based on neuroprotective and anti-inflammatory effects. Consequently, PDE4 inhibitors have been investigated as treatments for a diverse group of different diseases, including central nervous system disorders such as major depressive disorder (clinical depression), anxiety disorders, schizophrenia, Parkinson's disease, Alzheimer's disease, multiple sclerosis, attention deficit-hyperactivity disorder, Huntington's disease, autism and inflammatory conditions such as chronic obstructive pulmonary disease (COPD), asthma and rheumatoid arthritis.

PDE4 inhibitors have also been described in relationship with cardiac and cardiovascular diseases, however only within the classical PDE4-correlated and -associated inflammatory-related conditions and circumstances. Examples of such conventional, inflammation-related therapies of PDE4 inhibitors are e.g. WO2O09/067600A and WO2004/050624A. In the context of such classical therapeutic approach, WO2004/105751A further describes the treatment of cardiac hypertrophy by using a PDF4 inhibitor.

PDE4 inhibitors inhibit the degradation of cAMP into AMP. Since cAMP is a second messenger important in many biological processes and acts by intracellular signal transduction in many different organisms, conveying cAMP-dependent pathways, typically through activation of protein kinases, and are thereby involved in several biochemical processes, including the regulation of glycogen, sugar, and lipid metabolism (e.g. activation of lipases). Among a vast number of different reports and

investigations about possible metabolic effects of PDE 4 inhibitors, Lynn and Bornfeldt in Biochemical and Biophysical Research Communications 290, 663-669 (2002) report about PDE4 inhibitors to promote the expression of ATP cassette binding protein 1 (ABCA1) and cholesterol efflux from THP-1 cells and macrophages.

By the newly found capability of PDE4 inhibitors to beneficially influence lipoprotein profiles and in particular by their capability to counter-act lipoprotein imbalance in blood and serum of humans, the present invention now provides the possibility of treating endothelial dysfunction and cardiovascular diseases when correspondingly correlated or associated with dyslipoproteinaemia or lipoprotein imbalance, in particular abnormally high low density lipoprotein (LDL) in blood or serum. Accordingly, particularly useful treatments include for example arteriosclerosis, myocardial infarction, stroke, peripheral artery diseases, vascular stenosis, respectively independent from, or not correlated with or associated with inflammatory diseases or conditions such as inflammatory-associated cardiovascular pathology and cardiac hypertrophy. For distinction between dyslipoproteinaemia or lipoprotein imbalance associated circumstances and inflammatory-associated circumstances (especially in terms of cardiovascular events), reference can be made to Marz et al., Circulation 110:3068-3074 (2004), demonstrating the different underlying biochemical mechanisms and situations with respect to cardiovascular risks depending on whether lipoprotein metabolism or inflammatory processes are involved. That is, distinct from such inflammatory-originating pathways and associated diseases or conditions and inflammatory-associated cardiovascular pathology, the present invention beneficially allows prophylactic and/or therapeutic treatments of endothelial dysfunction and cardiovascular diseases characterized by different clinical settings, especially in circumstances of abnormal high LDL levels and especially LDL-cholesterol and LDL-triglyceride levels. Accordingly, the phosphodiesterase 4 inhibitors for use according to the present invention are particularly useful in the prophylaxis and/or therapy for reducing cardiovascular risk, and for reducing the risk of re-infarction. As standard references for such reduced risks, published practical guidelines of the American College of Cardiology (ACC) and the American Heart Association (AHA) on levels of blood cholesterol and in particular of LDL-C can be referred to, see e.g. K.K. Ray et al. in European Heart Journal advance access published March 17, 2014: " The ACC/ AHA 2013 guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular disease risk in adults: the good the bad and the uncertain: a comparison with ESC/EAS guidelines for the management of dyslipidaemias 2011 ". Regarding blood TG and LDL-TG, reference can be made to Marz et al., supra, in Circulation 110:3068- 3074 (2004).

Lipoproteins, cholesterol and triglycerides are usually analysed from serum samples but can also be assessed from plasma. Normal values for TG are below 150 mg/dl. Normal values for cholesterol are below 200mg/dl. In current guidelines, total cholesterol is seen as a risk factor for cardiovascular events. Indications for treatment of hypercholesterinaemia may depend on LDL-C value and other cardiovascular risk factors.

Current guidelines advice that therapy may already be initiated in patients with LDL-C values as low as 70 mg/dl when they have a high cardiovascular risk. On the other side of the spectrum patients with a low cardiovascular risk are considered for treatment when LDL-C levels exceed 190 mg/dl.

Target values for LDL-C in lipid lowering therapy are between 70mg/dl and 115 mg/dl depending on the cardiovascular risk.

Regarding LDL-TG as risk factor of cardiovascular diseases, Marz et al. {supra) discussed increased Odd's ratios for cardiovascular disease in depending on the quantile of LDL-TG distribution pointing towards a continuous association between LDL-TG levels and cardiovascular risk.

Furthermore, and based on the finding of a new effectivity of PDE4 inhibitors, these can also be utilized in the prophylactic and/or therapeutic treatment of diseases which are associated with a decrease in low density lipoprotein (LDL), in particular a decrease in LDL-associated cholesterol (LDL-C), and/or within the prophylaxis and/or therapy is associated with a decrease of a ratio of low density lipoprotein to high density lipoprotein (i.e. decreasing the LDL/HDL-ratio in the blood, notably the serum of humans).

Similarly and again based on the findings of the present invention, the treatment of prophylaxis and/or therapy can also be utilized when it is associated with a decrease in low density lipoprotein- triglycerides (LDL-TG). It is preferred that during the treatment (prophylaxis and/or therapy) the term "said decrease" means that the respective decrease is at least 10%, more preferably at least 20%. Our observations indicate that PDE4 inhibitors lower LDL-C by up to 45mg/dl (30%). The effect on LDL-TG is more pronounced as LDL-TG was reduced by up to 29 mg/dl (60%) in our observations. Optimisations may achieve more.

Active compounds from the class of PDE4 inhibitors (i.e. having the function of inhibiting

phosphodiesterase 4) can be used according to the present invention. These include, without being limited to, PDE4 inhibitors selected from the group consisting of: 3-cyclopropylmethoxy-4- difluoromethoxy-N- (3, 5-dichloropyrid-4-yl)-benzamide (Roflumilast) as well as Roflumilast-N-Oxide, Apremilast, Piclamilast, cis-4-cyano-4- [3-cyclopentyloxy-4-methoxyphenyl] cyclohexane-l-carboxylic acid (Cilomilast), Ibudilast, AN2728, Diazepam, 3, 7-dihydro-3- (4-chlorophenyl)-l-propyl-lH-purine-2, 6-dione (Arofylline), Atizoram, Catramilast, CC-1088, CDP-840, CGH-2466, Cipamfylline, CP-80633, Denbutylline, Drotaverine, Etazolate, Filaminast, Glaucine, HT-0712, ICI-63197, Indimilast, Irsogladine, Lavamilast, Methanesulfonic acid 2- (2, 4-dichlorophenylcarbonyl)-3-ureidobenzo-furan-6-yl ester (Lirimilast), Luteolin, Mesembrenone, Mesembrine, Mesopram, Oglemilast, Revamilast, Ro 20-1724, E6005, Ronomilast, RPL-554, RS-25344, and YM-976, Rolipram, 3- [3- (cyclopentyloxy)-4- methoxybenzyl]-6- (ethylamino)-8-isopropyl-3H-purine (V-11294A), N- [9-methyl-4-oxo-l-phenyl-3, 4,6, 7-tetrahydropyrrolo [3,2, 1-jk] [1,4] benzo-diazepin-3 (R)-yl] pyridine-4-carboxamide (Cl-1018), N- (3, 5-dichloro-4-pyridinyl)-2- [1- (4-fluorobenzyl)-5-hydroxy-lH-indol-3-yl]-2-oxoacetamide (AWD-12- 281), N- (3, 5-dichloropyridin-4-yl)-2- [5-fluoro-l- (4-fluorobenzyl)-l H-indol-3-yl]-2-oxoacetamide (AWD-12-343), tetrahydro-5- [4-methoxy-3- [(I S, 2S, 4R)-2-norbornyloxy] phenyl]-2 (1 H)-pyrimidone (ATIZORAM), 3-[3-(cyclopentyloxy)-4-methoxyphenyl]-l, 3-dihydro-l, 3-dioxo-2H-isoindole-2- propanamide (CDC-801), 3, 5-dichloro-4- [8-methoxy-2- (trifluoromethyl)quinolin-5-ylcarbox-amido] pyridine-l-oxide (SCH-351591, CDC-998, 1C-485, CC-1088 and KW4490; and pharmaceutical acceptable salts of the aforementioned compounds. Roflumilast is preferred as it has the advantage of being approved by health authorities.

Examples of the above listed PDE4 inhibitors and their synthesis are for example described in

WO2004/103407A, W002/064584, W095/01338, W092/12961, W093/19749, W095/00516,

W096/11690, W098/09946, W098/09946, W097/23457, WO00/26208, EP0435811, EP0731099, and EP0389282, and in representative literature references including e.g. those of Wang et al. (2007) and Rabe (2011), supra. The present invention further provides a pharmaceutical composition for the above-described therapeutic uses, wherein in the composition the PDE4 inhibitor is contained together with a pharmaceutically acceptable excipient, carrier, diluent, and/or additive (inactive ingredients). The composition can be formulated in a suitable administration and dosage form, for example as a solid, a semi-solid, gelatinous, a liquid, a suspension, a powder, a tablet, a pill, a capsule, a sustain-released dosage form, and the like. Furthermore, the administration and dosage form may for example be oral, parenteral, transdermal, nasal, for injection or for inhalation. Examples of suitable inactive ingredients and suitable administration forms are given in the above mentioned PDE 4 inhibitor related WO and EP publications.

The PDE4 inhibitor, either alone or when present in the afore-mentioned pharmaceutical composition, is used in a suitable amount effective for the prophylaxis and/or therapy of the afore-mentioned diseases or diseases conditions. For example, a suitable amount or dose of a PDE4 inhibitor is at least 0.0001 mg/kg, for example in a range of from 0.0001 to 1000 mg/kg, preferably from 0.01 to 100 mg/kg and particularly from 0.1 to 20 mg/kg by weight of the treated patient. Doses can be administered singly or repeatedly, locally or systemically, once or several times daily or weekly, or - especially of depot or slow-release formulations - optionally also at a lower frequency. It may also be useful to apply time-dependent dosage regimen, starting with a first unit dose, for example a daily dose of 50 mg-100 mg, preferably 250 mg-750 mg, and after some weeks of for example a second unit dose different from the first unit dose, for example being twice as high as the first unit dose. The dose and the time-dependent dosage regimen can be followed and controlled by, and associated with the measurement of, lipoprotein profiles and lipoprotein levels, in particular LDL and HDL levels, respectively their fractions, in pa rticular LDL-cholesterol and LDL-triglyceride, respectively alone or in combination.

A further aspect of the present invention is a medication comprising a PDE4 inhibitor with another compound active against, or reducing the risk of, dyslipidaemia, lipoprotein imbalance, endothelial dysfunction, cardiovascular diseases, arteriosclerosis, hypertension and stroke. Examples of such other active compounds can be selected, for instance, from the group of statins. Since, as described above, the mechanism and approach of treating endothelial dysfunction and cardiovascular diseases according to the present invention of using PDE4 inhibitors - by being correlated or associated with dyslipoproteinaemia, lipoprotein imbalance and in particular hypercholesterinaemia or

hypertriglyceridemia - is distinct from other risk factors and other aetiologies and causes of cardiovascular disease and endothelial dysfunction, in particular distinct from inflammatory-associated conditions, it is evident that combinatory effects can be achieved to thereby provide overall improved therapies of endothelial dysfunctions and cardiovascular diseases. Accordingly, the present invention for example further provides a useful combined medication, wherein a phosphodiesterase 4 inhibitor and a HMGCoA inhibitor, preferably selected from the group of statins, are combined for use in a treatment of cardiovascular diseases, arteriosclerosis, hypertension, dyslipoproteinaemia, or for reducing the risk of any one the these diseases. The combination can be embodied in separate or in common dosage forms, and can be administered concurrently or temporarily shifted according to a desirable dose and time regimen.

The present invention is further described by reference to non-limiting and representative examples in the following.

Examples

In a clinical trial, five patients were treated by daily doses of a PDE4 inhibitor. As an exemplifying and representative compound, roflumilast was used as the PDE4 inhibitor. Lipoprotein profiles were measured at start of the therapy (time TO), four weeks after start (Tl) and eight weeks after start (T2). Treatment was further monitored thereafter. In an embodiment, a first unit dose (500 mg) was administered orally every second day for a period of four weeks, and subsequently the dose was increased to a unit of 500 mg per day until a time of 8 weeks.

For analysing and measuring the lipoprotein profiles and thus the relative concentrations and levels of lipoprotein fractions and the cholesterol and triglyceride amounts per lipoprotein, 10 ml venous blood was drawn at the indicated time points. Lipoprotein profiles and fractions were determined using density separation by means of ultra-centrifugation and subsequent determination of the lipoprotein components (Baumstark et al., Biochem. Biophys. Acta - Protein Structure and Molecular Enzymology 1037, 48-57 (1990); Frey et al., Eur. J. Appl. Physiol. 60, 441-444 (1990)). The components and parameters determined include: cholesterol and triglyceride proportions of HDL and LDL, and of sub- fractions LDL-1 to LDL-6, and HDL-2a-2b and -3. Statistical analysis of the results was carried out by SPSS 23.0. The changes, in absolute amounts of mg/dl, of the cholesterol proportions of the lipoprotein fractions at time Tl and T2 compared to TO are shown in Fig. 1.

Furthermore, the absolute changes, in mg/dl, of the triglyceride proportions of the lipoprotein fractions at time Tl and T2 in comparison with TO is shown in Fig. 2.

The individual changes of LDL-T and LDL-TG in the patient's blood are shown in Figs. 4 and 5.

As shown in the Figures, a substantial reduction of certain lipoprotein sub-fractions, in particular a remarkable reduction of LDL-C by approximately 30 mg/dl was observed. LDL-TG was also substantially reduced. The reduction of LDL-C and LDL-TG are maintained over longer time periods, notably after 12 weeks (not shown).

Accordingly, it is demonstrated that, independent from effects on systemic inflammation, the PDE4 inhibitor can effectively improve therapeutic concepts, such as reduction of cardiovascular risks, by a reduction of independent cardiovascular risk factors, in particular by reduction by LDL-C. Further independently from LDL-C, also a reduction of LDL-TG is possible, as further independent beneficial influence on the cardiovascular risk and as indications of further therapeutic beneficial situations. Slight reductions of HDL-C and HDL-TG were also observed.

Accordingly, it has been shown that PDE4 inhibitors have a beneficial influence on the systemic lipoprotein profile in human blood and serum, in particular by improving the LDL to HDL ratio, and in particular by remarkably reducing the LDL-C and HDL-C, but independently also the LDL-TG.

Accordingly, the use of PDE4 inhibitors can help to reduce cardiovascular risks, and furthermore can establish effective treatments in systemic lipoprotein-related disorders, such as dyslipoproteinaemia, and diseases correlated/associated with dyslipoproteinaemia, lipoprotein imbalance or related disorders.