COMPOSITION TO PROMOTE THE ACTIVITY OF SIRTUINS

Description

[0001] The invention relates to a composition for promoting the activity of sirtuins, in particular for the treatment and / or prophylaxis of diseases associated with reduced sirtuin activity. These include, in particular, aging phenomena and diseases associated with impaired cell differentiation.

[0002] Sirtuins are mostly type III histone deacetylases, which are highly evolutionarily conserved and occur in almost all life forms. The name is derived from "silent information regulator." Sirtuins deacetylate lysine residues, particularly those at the N-terminal end of histone proteins, which, as a component of chromatin, are responsible for DNA packaging. NAD+< is required as a cofactor for deacetylation. Thus, after cleavage of the acetyl group, nicotinamide and O-acetyl-ADP-ribose are formed in addition to the deacetylated protein. Because the lysine carries a positive charge after deacetylation, the affinity of the histone to the negatively charged DNA backbone increases. This leads to a stronger DNA block, and transcription is downregulated. Sirtuins therefore have a direct influence on cellular processes because they are able to regulate the expression of certain proteins and enzymes.In humans and other mammals, seven different sirtuins are distinguished: SIRT1 to SIRT7. In addition to histones, sirtuins also deacetylate intracellular signaling molecules.

[0003] Due to the influence of sirtuins on the expression of proteins and enzymes, some of which are linked to certain diseases, considerable research has been devoted to this enzyme class in recent years. In particular, sirtuins appear to have a positive effect on various aging symptoms and diseases that occur more frequently with age, for example, because they are associated with altered gene regulation during cell differentiation. Some media outlets have therefore already referred to them as "anti-aging enzymes." Among other things, the effects of sirtuin activators, such as resveratrol, are being investigated.

[0004] The diseases influenced by sirtuins include the aforementioned aging phenomena, particularly those of the lungs, such as the onset of senile emphysema, also known as age-related emphysema or primary atrophic emphysema. In addition to lung aging, the literature also discusses the connection between sirtuins and other diseases, such as cardiac hypertrophy, gout, atherosclerosis or arteriosclerosis, obesity, amyotrophic lateral sclerosis, or cancer, particularly in the field of hematology, such as acute or chronic leukemia or malignant lymphomas. The regulation of gene activity via histones plays a significant role in cell differentiation. A connection has been postulated between an age- or disease-related loss of sirtuin activity and the onset of aging phenomena.

[0005] Mojhgan Sheikhpour et al., J Cancer Sci Ther 2014, 6:10 (doi.org / 10.4172 / 1948-5956.) discloses a study investigating the antiproliferative effects of two compatible solvents, ectoine and 5-hydroxyectoine, on a human lung cancer cell line (QU-DB). Ectoine and hydroxyectoine were found to have inhibitory and cytotoxic effects on the cancer cell line.

[0006] The task therefore arises to provide substances or compositions that counteract the age-related loss of sirtuin activity.

[0007] Surprisingly, it was found that ectoine, hydroxyectoine, and salts, esters, and amides of ectoine / hydroxyectoine are capable of significantly reducing the loss of sirtuin activity. Sirtuin activity primarily relates to the number, structure, and / or function of the sirtuins; that is, reduced sirtuin activity can occur when sirtuins are less active or even when fewer sirtuins are present. In this way, both therapeutic treatment and preventative effects can be achieved.

[0008] Ectoine and hydroxyectoine are tetrahydropyrimidine derivatives synthesized under stress conditions by extremophilic, particularly halophilic, microorganisms. Various uses have been described for ectoine and hydroxyectoine, for example, as moisturizers, for the treatment of vascular leak syndrome (VLS) (DE 10 2006 056 766 A1), or for the treatment of atopic dermatitis (DE 103 30 243 A1).

[0009] The systematic name for ectoine is 2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid, for hydroxyectoine 5-hydroxy-2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid.

[0010] The structure of natural L-ectoine ((S)-2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid) is shown below:

[0011] The structure of natural hydroxyectoine ((4S,5S)-5-hydroxy-2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid) is given below:

[0012] The use of the indicated stereoisomers is preferred but not mandatory, i.e. the use of other stereoisomers or the racemate is also possible.

[0013] The composition according to the invention can generally be used for the treatment and / or prophylaxis of aging phenomena and for influencing self-renewal processes, cell differentiation, and cell proliferation. The composition is particularly important in the treatment of aging phenomena of the lungs, especially senile emphysema. This is characterized by the degeneration of the alveolar structures in the lungs, which leads to a reduction in the lung surface available for gas exchange. The patient therefore experiences "poor air." It has been shown that repeated intrapulmonary administration of an ectoine-containing solution in rats leads to a significant reduction in the loss of sirtuin activity in lung tissue.Although the treatment of lung diseases caused by particulate matter exposure has already been described in WO 2005 / 002556 A1, it was surprisingly discovered that general lung aging symptoms can also be treated with ectoine / hydroxyectoine. Tissue aging is based on altered gene activity during self-renewal and cell differentiation in (lung) tissue, which is regulated by histones via DNA accessibility and thus also dependent on sirtuins.

[0014] Accordingly, the present invention may exclude the treatment / prophylaxis of lung diseases caused by exposure to airborne particulate matter, as can be seen from WO 2005 / 002556 A1. Likewise, the treatment / prophylaxis of pneumonia, asthma, COPD, ARDS (Acute Respiratory Distress Syndrome), cystic fibrosis, pulmonary fibrosis, silicosis, sarcoidosis, allergies, and bronchial hyperreactivity, which can be seen from WO 2013 / 034299 A1.

[0015] Another group of lung diseases that can be treated or prevented with the composition according to the invention is idiopathic interstitial pneumonia (IIP). The cause of these diseases is unknown, which is why they are referred to as idiopathic. They primarily affect the connective tissue of the lungs. Patients suffer from shortness of breath, which in the initial stages may be limited to physical exertion, but often worsens over time and, in the final stages, progresses to respiratory failure. Oxygen deficiency in the blood can occur. These diseases are also frequently associated with impaired cell proliferation.

[0016] According to the current classification, seven different forms of the disease are distinguished: Idiopathic pulmonary fibrosis (IPF), non-specific interstitial pneumonia (NSIP), cryptogenic organizing pneumonia (COP), acute interstitial pneumonia (AIP), respiratory bronchiolitis with interstitial lung disease (RB-ILD), desquamative interstitial pneumonia (DIP), and lymphoid interstitial pneumonia (LIP).

[0017] In addition, there are unclassifiable IIPs for diseases that cannot be clearly defined.

[0018] Idiopathic pulmonary fibrosis (IPF) is particularly common. Treatment with the composition according to the invention plays a special role here because the disease is difficult to combat with anti-inflammatory drugs. The same applies to acute interstitial pneumonia (AIP). Both diseases typically have a poor prognosis, which is why the treatment of these diseases according to the invention plays a special role.

[0019] The signs of aging that can be treated or prevented with the composition according to the invention also include intrinsic signs of skin aging. These are understood to be signs of aging of the skin that are not attributable to external influences such as UV radiation or sun exposure in general. They are caused by a reduced reactivity of the skin cells, are genetically determined, and usually begin by the age of 30 at the latest. The skin loses elasticity and tone. Due to the reduced cell division of the skin in advanced age, the collagen and elastin content in the skin decreases significantly.

[0020] Signs of skin aging caused by exposure to airborne dust can also be treated or prevented by the composition according to the invention. In particular, airborne dust (also called fine dust) with an average particle size of ≤ 15 µm, especially ≤ 10 µm, can cause signs of skin aging. Such airborne dust is often produced by the combustion of fossil fuels, but also in the form of sand, spores, pollen, rock dust, in agriculture, mining, tobacco consumption, tire wear, brake wear, or forest fires.

[0021] In general, the invention particularly encompasses the treatment / prophylaxis of aging phenomena and diseases attributable to impaired cell differentiation or cell proliferation, which are influenced or caused intracellularly by endogenous factors. Strengthening the expression of sirtuins by ectoine / hydroxyectoine improves cell differentiation in the tissue.

[0022] All human epithelia and endothelia that come into contact with ectoine / hydroxyectoine are positively influenced in their self-renewal and cell differentiation. Accordingly, the invention relates to a composition for the treatment of endothelial, epithelial, and neurological diseases. In addition to the diseases already mentioned above, there are other diseases that can be described as signs of aging in the sense that they occur primarily in people of advanced age, or that are associated with impaired cell differentiation / proliferation. These include: cardiac hypertrophy, gout, arteriosclerosis / atherosclerosis, obesity, amyotrophic lateral sclerosis (ALS), and cancer. The cancers are particularly those from the field of hematology, such as acute or chronic leukemia and malignant lymphomas. Epithelial and mesenchymal tumors, i.e.Generally, carcinomas and sarcomas. Other treatable cancers include: lung cancer, skin cancer, colorectal cancer (especially colon cancer), breast cancer, prostate cancer, pancreatic cancer, esophageal cancer, laryngeal cancer, stomach cancer, cervical cancer, ovarian cancer, testicular cancer, kidney cancer, bladder cancer, thyroid cancer, oral cancer, pharyngeal cancer, and brain tumors.

[0023] Other treatable hematological diseases include anemia and MDS (myelodysplastic syndrome). Ectoine / hydroxyectoine is also suitable for the treatment and prophylaxis of these diseases. The treatment of amyotrophic lateral sclerosis is due to the fact that sirtuins have a stimulating effect on the growth of neurons and axons.

[0024] The sirtuin whose activity is increased or whose loss of activity is reduced is SIRT-1, which has cardioprotective, anticancer, anti-inflammatory, and antioxidant effects, among others. SIRT6 is also of particular importance. It has been shown that a deficiency of these sirtuins leads to premature aging in mice (Cheng et al., Proc Natl. Acad Sci USA 2003, 100, 10794-10799; Mostoslavsky et al., Cell 2006, 124, 315-329).

[0025] Pharmacologically acceptable salts of ectoine / hydroxyectoine include alkali or alkaline earth salts, in particular the salts of potassium, sodium, magnesium and calcium, but also salts with organic bases such as non-toxic aliphatic or aromatic amines.

[0026] By reacting the carboxyl group of ectoine / hydroxyectoine with alcohols or amines, corresponding esters or amides can be obtained, which can also be used according to the invention. In this case, the COOH group of ectoine / hydroxyectoine is replaced by a carboxylic acid ester function COOR or a carboxylic acid amide function CONHR' or CONR'R", where R, R', and R" independently represent saturated or unsaturated, straight-chain or branched alkyl, cycloalkyl, aryl, heteroaryl, alkylaryl, arylalkyl, alkoxyalkyl, alkylthioalkyl, aryloxyalkyl, or arylthioalkyl groups. In particular, it can be a C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, or C18 alkyl group. In hydroxyectoine, the hydroxy group can also be reacted with a carboxylic acid of different chain length to form a corresponding ester.The respective esters or amides may also be present in ionic or zwitterionic form, ie the invention includes the use of salts of the said esters or amides.

[0027] The use of ectoinamide of 2-hydroxy-5-aminobenzoic acid is also possible. The structural formula is shown below:

[0028] It is therefore the 2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid amide of 2-hydroxy-5-aminobenzoic acid. Preferably, it is the corresponding amide of L-ectoine: (S)-2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid amide. The use of the hydroxyectoine amide of 2-hydroxy-5-aminobenzoic acid is also conceivable.

[0029] The composition may contain common excipients, such as carriers, preservatives, bactericides, solubilizers, vitamins, stabilizers, anti-foaming agents, osmotically active substances, dyes, surfactants, emulsifiers, humectants, viscosity-increasing agents, etc. When administered in solid form, e.g., via powder inhalers, it is advisable to use only readily absorbable, non-irritating carriers such as micronized lactose.

[0030] Preservatives include thiomersal, organic mercury compounds such as phenylmercury, benzalkonium chloride, chlorhexidine, benzyl alcohol, glucose, ethanol and quaternary ammonium salts.

[0031] The formulations of the invention may also contain suitable buffer systems or other pH-adjusting adjuvants to establish and maintain a desired pH value. Typically, this is a pH value between 4 and 8, preferably between 5 and 7.5, more preferably approximately 7. Suitable buffer systems are citrate, phosphate, TRIS, glycine, borate, and acetate. These buffer systems can be prepared from substances such as citric acid, monosodium phosphate, disodium phosphate, glycine, boric acid, sodium tetraborate, acetic acid, or sodium acetate.

[0032] The compositions may contain other active ingredients, but it is perfectly possible and sufficient for the treatment or prevention of the signs of aging to use compositions containing only ectoine and / or hydroxyectoine or corresponding salts, esters or amides as the active ingredient.

[0033] The concentration of ectoine / hydroxyectoine and / or corresponding salts, esters, or amides can in particular be in a range from 10 to 500 mM, preferably 50 to 500 mM, particularly preferably 100 to 500 mM or 100 to 200 mM. These concentrations have proven suitable for producing the effect according to the invention. In particular, the proportion of ectoine / hydroxyectoine and / or corresponding salts, esters, or amides in the composition can be in a range from 0.05 to 20 wt. %, preferably 0.1 to 10 wt. %. A good effect has been observed, for example, in a range between 0.5 and 2 wt. %.

[0034] The composition can be in the form of a solution, preferably an aqueous solution. It can also be in the form of a suspension, emulsion, or microemulsion.

[0035] To improve the application and shelf life of the composition according to the invention, the composition containing the active ingredient can also be encapsulated in nanostructures or administered in the form of liposomes. This is particularly advantageous if the composition does not contain a preservative. Corresponding methods for encapsulation and for the production of liposomes are generally known from the prior art.

[0036] Furthermore, a spray device is disclosed with which the composition according to the invention can be inhaled. The spray device comprises means for atomizing the composition and has an outlet opening that allows the user to inhale it. A spray device is particularly important in connection with the treatment or prevention of aging symptoms of the lungs, in particular senile emphysema. Suitable spray devices can be used for inhaling an aerosol or a powder. Examples of suitable powder inhalers include Spinhaler, Diskhaler, Turbohaler, Rotahaler, or Aerolizer.

Claims

1. Composition comprising as active ingredient ectoine, hydroxyectoine and / or salts, esters or amides of these compounds for use in a method of treatment and / or prophylaxis of lung ageing symptoms, excluding COPD, and / or idiopathic interstitial pneumonia (IIP), wherein the lung ageing symptom is senile emphysema.

2. Composition for use according to claim 1, characterized in that the idiopathic interstitial pneumonia is idiopathic pulmonary fibrosis (IPF) or acute interstitial pneumonia (AIP).

3. Composition for use according to any one of claims 1 to 2, characterized in that a reduction in the loss of sirtuin activity is induced.

4. Composition for use according to any one of claims 1 to 3, characterized in that the composition is present as a solution, suspension, emulsion or microemulsion.

5. Composition for use according to claim 4, characterized in that the solution is an aqueous solution.

6. Composition for use according to any one of claims 1 to 5 for application by inhalation.

7. Composition for use according to one of claims 1 to 6, characterized in that the concentration of ectoine, hydroxyectoine and / or salts, esters or amides of these compounds in the composition is 10 to 500 mM, preferably 100 to 500 mM.

8. Composition for use according to one of claims 1 to 7, characterized in that the composition is encapsulated in nanostructures or is present in the form of liposomes.