Patent Description:
The wall of the human gastrointestinal tract constitutes one of the major areas of interaction between our body and the external environment and plays a fundamental role in the regulation of the immune system, and therefore in the maintenance of the state of health of a body. The intestinal barrier is semi-permeable and is responsible for the absorption of nutrients, immune detection, and defence against potentially harmful antigens as well as pathogenic microorganisms [<NUM>]. In particular, the intestinal epithelium contains: absorbent cells, secretory cells, and intestinal stem cells. The absorbent cells are distinguished in: enterocytes, in the small intestine tract, and colonocytes, in the colon tract. The secretory cells include goblet cells (or GC), Paneth cells, and enteroendocrine cells. The enterocytes and colonocytes are the most-present cell types in the intestinal epithelium, with a relative abundance exceeding <NUM>%. In addition to the physical barrier function between the immune system and the commensal microbiota, the enterocytes and epithelial colonocytes are of key importance for the absorption of water as well as nutrients (mainly the role of the enterocytes) and electrolytes (essentially the function of the colonocytes) [<NUM>] [<NUM>].

The first line of physical defence which prevents bacteria in the intestinal lumen from coming into direct contact with intestinal epithelial cells is the mucous layer of the intestinal tract. The intestinal mucous layer consists mainly of mucins, i.e., highly glycosylated proteins, which form a gelatinous structure (mucus). The mucus covers and protects the intestinal epithelium. For example, the most abundant mucin in the intestine is MUC-<NUM> (Mucin <NUM>, Oligomeric Mucus/Gel-Forming), which is secreted by the goblet cells [<NUM>] [<NUM>] [<NUM>].

Under physiological conditions, the intestinal wall is characterized by an oxygen gradient with values around <NUM>% in the lamina propria, which are reduced to <NUM>% at the intestinal villi, and less than <NUM>% in the lumen. Therefore, considering the oxygen gradient and the daily fluctuations that make the microenvironment extremely complex, it can be said that the intestinal mucosa is in a state of physiological hypoxia [<NUM>] [<NUM>].

Adaptation to oxygen fluctuations depends on the activation of the HIF-<NUM> (Hypoxia Inducible Factor) cascade. Under hypoxic conditions, the HIF-<NUM> factor is activated at the nuclear level, which triggers a reprogramming of gene expression which controls the fate of the cell, generally activating alternative energy generation mechanisms and increasing oxygen uptake [<NUM>].

In particular, the activation of HIF-<NUM> promotes:.

It is commonly assumed that a dysfunction of the intestinal barrier and an uncontrolled flow of antigens through the intestinal epithelium can constitute a threat to the immune system of susceptible individuals, and affect the balance between host and microbiota, triggering inflammatory mechanisms in the intestine and/or other districts of the body [<NUM>]. Environmental factors such as stress, diet, lifestyle, smoking habits, and changes in the intestinal bacterial component may be further associated with an altered immune response [<NUM>].

The alteration of intestinal barrier integrity favours the development of sometimes exaggerated immune responses which are associated with the development of inflammatory diseases of the gastrointestinal tract. Under normal physiological conditions, increased permeability is usually not sufficient to cause intestinal disease, since the epithelial barrier can regenerate and regain its functions once the injurious stimulus disappears. However, under certain pathological conditions, this ability to self-regulate may be lost, and may contribute to increased permeability, thereby facilitating chronicization of intestinal inflammation. The presence of certain environmental factors (infections, stress) increases intestinal permeability and the passage of substances which under normal conditions would never penetrate the epithelial barrier [<NUM>] [<NUM>].

IBD (Inflammatory Bowel Diseases, or ulcerative colitis and Crohn's disease) and IBS (Irritable Bowel Syndrome) are gastrointestinal disorders which affect <NUM>% and <NUM>% of the world population, respectively. The two pathological conditions have similar symptoms with the difference that IBD is characterized by an inflammatory state generally found with endoscopic examination; while IBS is considered a functional disease without objectively measurable structural or physiological abnormalities, thus making its diagnosis more difficult [<NUM>].

Numerous studies in the literature on dysbiosis and IBD have shown the role of the intestinal microbiota as an important cause of intestinal inflammation. In addition to genetic and environmental factors, altered barrier function also leads to an abnormal immune response and increased susceptibility to intestinal inflammation. In fact, although the aetiology of IBD and IBS remains unknown, all patients share an increased intestinal permeability with respect to healthy subjects. Increased intestinal permeability is the result of structural changes in the proteins that make up tight junctions (or TJ), and would result from an exaggerated inflammatory response. An increase in IFN-γ and TNF-α levels was indeed detected in patients with IBD or IBS [<NUM>] [<NUM>] [<NUM>] [<NUM>] [<NUM>] [<NUM>].

Even in celiac disease, an autoimmune condition of unknown etiology which derives from gluten intolerance, structural changes of the TJ are found, which favour the entry of gliadin (gluten constituent protein) into the intestinal mucosa [<NUM>] [<NUM>]. This results in a sustained immune response which contributes to increasing intestinal permeability. In general, the altered function of the intestinal barrier is also involved in the patho-physiology of food allergies. Increased intestinal permeability may promote the passage of food antigens and increase the risk of an allergic response in sensitive subjects. In fact, in patients with food allergies there is greater intestinal permeability, as measured by the lactulose and mannitol permeability test [<NUM>].

Butyrate is a short-chain saturated fatty acid (Fig. <NUM>) produced by the fermentation of indigestible carbohydrates by some types of bacteria resident in the colon; as a product of intestinal microbiota it is considered a postbiotic. Butyrate is the primary energy substrate for colonocytes and is critical for the maintenance of intestinal epithelial homoeostasis [<NUM>] [<NUM>].

Butyrate is transported from the intestinal lumen to the cytosol of the colonocyte largely actively, to then diffuse into the mitochondria and be oxidized with CO<NUM> release, ATP regeneration and H<NUM>O production from O<NUM> (aerobic respiration) [<NUM>]. The consumption of O<NUM> within the colonocyte results in a state of physiological hypoxia which acts as an activation signal for HIF-<NUM> (Hypoxia Inducible Factor) [<NUM>] [<NUM>] [<NUM>] [<NUM>]. As described above, the HIF-<NUM> factor promotes the expression of MUC-<NUM> and ITF, favouring the protection of the intestinal wall [<NUM>] [<NUM>] [<NUM>]. In addition, butyrate:.

The daily dose of sodium butyrate used in clinical trials varies by application, from <NUM> to <NUM>. The main competitor recommends up to <NUM> of sodium butyrate for intestinal well-being [<NUM>] [<NUM>] [<NUM>] [<NUM>] [<NUM>].

Palmitoylethanolamide (or PEA) is a lipid mediator (Fig. <NUM>) belonging to the family of N-acylethanolamines to which neuroprotective, anti-inflammatory and analgesic properties are attributed. PEA is found in many foods in nature, such as legumes, egg yolks and peanuts, but is also present in the tissues of animals and humans [<NUM>] [<NUM>] [<NUM>]. PEA represents a PPAR-α agonist with affinity comparable to Wy-<NUM> (synthetic receptor agonist) which produces a strong anti-inflammatory effect. The binding of PEA to PPAR-α induces heterodimerization between PPAR-α and RXR (Retinoid X Receptor) and the formation of the activated receptor complex. Thereby, the receptor is formed in the active form which translocates into the nucleus where it binds the peroxisome proliferation response elements. Accordingly, it thereby reduces the transcription of pro-inflammatory genes. In addition, as an entourage effect, PEA could activate target endocannabinoid receptors, i.e., CB1 and CB2 receptors and TRPV1 (transient receptor potential vanilloid receptor type <NUM>) channels, exerting a pain-relieving effect. Theoretically, the analgesic action of endocannabinoids, arachidonoylglycerol and anandamide, depends on the levels of PEA production. In pain conditions, the endogenous expression of PEA is usually reduced and that of arachidonoylglycerol and anandamide increases. However, the increase in the latter is not such as to exert an analgesic action due to the low PEA production levels. Therefore, the supplementation of PEA could represent an effective alternative to enhance endogenous anti-nociceptive mechanisms exerted by endocannabinoids[<NUM>] [<NUM>] [<NUM>].

In vitro, in vivo, and ex vivo studies demonstrate the utility of PEA in combating IBD-associated chronic intestinal inflammation. In fact, the PEA seems to be able to:.

A recent clinical study of <NUM> patients with IBS showed that the administration of <NUM> PEA and <NUM> polydatin markedly improved abdominal pain severity but did not impact its frequency [<NUM>].

The daily dose of PEA used in clinical trials to control pain ranges from <NUM> to <NUM>. The daily dose most commonly found in the works is <NUM>. There is no statistically significant association between PEA dose and efficacy in counteracting pain. Similarly, the duration of treatment, ranging from <NUM> to <NUM> days, does not appear to significantly impact the desired outcome. In addition, on the whole, PEA treatment is well tolerated [<NUM>].

Generally the condition of intestinal inflammatory state can be treated therapeutically, for example, with the use of NSAID anti-inflammatory drugs or of those of a steroidal nature. However, such drugs have no effect in reducing the permeability of the intestinal wall and are often the cause of undesirable adverse reactions, especially when the underlying administration is long-term.

Bioactive lipids such as palmitoylethanolamide (PEA) and short chain fatty acids (SCFAs), such as butyrate belong to a large group of bioactive lipids with anti-inflammatory effects in the gastrointestinal tract. PEA reduces inflammation markers in a murine model of inflammatory bowel disease (IBD), and butyrate, produced by gut microbiota, is effective in reducing inflammation and pain in irritable bowel syndrome and IBD animal models [<NUM>].

The Applicant has advantageously developed a combination for use in treating and preventing disorders and pathologies of the intestinal tract associated with or caused by inflammatory processes comprising the following active ingredients: a) sodium butyrate and b) palmitoylethanolamide (or PEA). Sodium butyrate is administered at daily dosages between <NUM> and <NUM> daily and PEA is administered at daily dosages between <NUM> and <NUM> daily wherein the weight ratio between sodium butyrate and PEA administered daily is comprised between <NUM>:<NUM> and <NUM>:<NUM>.

The combination developed by the Applicant is based on the restoration of barrier integrity by acting in a targeted and parallel manner on two levels:.

The action on two distinct levels in parallel allows to stop the vicious cycle of inflammation, allowing the intestinal tissues to find a homoeostasis condition.

Such a combination is advantageous because its administration in patients with disorders or pathologies associated with or caused by inflammatory processes allows to reduce the healing times compared to the administration of the individual active ingredients sodium butyrate a) and PEA b).

Finally, said combination is well tolerated and does not present any adverse effect.

For the purposes of the present invention, the verbs "comprise" or "contain" are intended to define a set of elements, expressly indicating some, without excluding the presence of others not expressly indicated; while the term "substantial" or "constituted" is intended to define a set of elements, expressly indicating them all, and thus excluding the presence of components not expressly listed.

The combination developed by the Applicant is for use in treating disorders or pathologies of the intestinal tract associated with or caused by inflammatory processes. Said disorders or pathologies of the intestinal tract associated with or caused by inflammatory processes are preferably selected from: irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), leaky bowel syndrome, celiac disease and food allergies.

For the claimed combination, the weight ratio of sodium butyrate to PEA is preferably comprised between <NUM>:<NUM> and <NUM>:<NUM>, more preferably is equal to <NUM>:<NUM>.

Said combination is preferably administered in the form of two oral formulations, of which the first oral formulation contains sodium butyrate as active ingredient while the second oral formulation contains PEA as active ingredient, or said combination is preferably administered in the form of a single oral formulation containing both the active ingredients sodium butyrate a) and PEA b).

When said combination is preferably administered in the form of two oral formulations, of which the first oral formulation contains sodium butyrate as active ingredient and of which the second oral formulation contains PEA as active ingredient, preferably said second oral formulation containing PEA also contains vitamin B2.

When said combination is preferably administered in the form of two oral formulations, of which the first oral formulation contains sodium butyrate as active ingredient and of which the second oral formulation contains PEA as active ingredient, said first oral formulation more preferably contains sodium butyrate in an amount of <NUM> and more preferably said second oral formulation contains PEA in an amount of <NUM>, and both are preferably administered twice daily.

When said combination is preferably administered in the form of oral formulations, said oral formulations are preferably food supplements or nutraceutical formulations.

Food supplements are intended as a food product designed to supplement the common diet as they constitute a concentrated source of nutrients, such as vitamins and minerals, or of other substances having a nutritional or physiological effect, in particular, but not exclusively, amino acids, essential fatty acids, fibres and extracts of plant origin, both mono- and multi-compound, in predominant forms.

When said combination is preferably administered in the form of oral formulations, said oral formulations are preferably adjuvants.

Adjuvant is intended as a pharmacological substance which, added to a real drug, modifies its effect in order to enhance or maximize its effectiveness.

When said combination is preferably administered in the form of oral formulations, they contain the aforementioned active ingredients in combination with conventional excipients and which may possibly control and/or delay their release.

Claim 1:
Combination for use in treating and preventing disorders and pathologies of the intestinal tract associated with or caused by inflammatory processes comprising the following active ingredients:
a) sodium butyrate;
b) palmitoylethanolamide (or PEA)
in which butyrate is administered at daily dosages between <NUM> and <NUM> daily and PEA is administered at daily dosages between <NUM> and <NUM> daily wherein the weight ratio between sodium butyrate and PEA administered daily is between <NUM>:<NUM> and <NUM>:<NUM>.