WO2020234770A1

WO2020234770A1 - Composition for use in the prevention and/or treatment of pathologies associated to the prostate

Info

Publication number: WO2020234770A1
Application number: PCT/IB2020/054740
Authority: WO
Inventors: Umberto DI MAIO
Original assignee: Neilos S.r.l.
Priority date: 2019-05-21
Filing date: 2020-05-19
Publication date: 2020-11-26
Also published as: EP3972577A1; IT201900007044A1

Abstract

A composition is described comprising an association between escin and capsaicin. The composition can further contain an extract of Curcuma longa and/or an extract of a plant belonging to the genus Cucurbita and/or an extract of Serenoa Repens effective in the prevention and/or treatment of prostate-associated pathologies.

Description

COMPOSITION FOR USE IN THE PREVENTION AND/OR TREATMENT OF PATHOLOGIES

ASSOCIATED TO THE PROSTATE

5

10 DESCRIPTION

The present invention relates to a composition of substances, preferably obtained from natural sources, effective in the prevention and/or treatment of prostate pathologies. The prostate can incur in a huge number of pathologies, from the inflammatory or 15 infectious ones to the degenerative, benign or malignant ones. In the composition of the present invention the synergic action takes place between escin and capsaicin. The composition can further contain an extract of Curcuma longa and/or an extract of a plant belonging to the genus Cucurbita and/or an extract of Serenoa Repens.

Background of the invention

20 The pelvic pain

The pelvic pain represents one of the most disabling disorders highly impacting on the quality of life and requiring a multidisciplinary diagnostic and therapeutic approach. The pelvic pain in man represents an important symptom which can be secondary to physiological or pathological conditions.

25 The “pelvic” adjective discriminates the presence of pain at the level of lower abdomen, immediately below the abdominal region, comprising the pelvis area, the external genitalia and perineum.

The possible causes of the male pelvic pain are several and correlated to disorders affecting several apparatuses:

30 · gastrointestinal,

• urinary,

• genital, • skeletal muscle and nervous.

Based upon the onset mode, the pelvic pain can be classified as:

• Acute: it appears suddenly but, once removed the triggering cause, the pain disappears;

• Chronic: it occurs gradually, without being possible to go back to a specific cause and without the possibility of removing it; in this field a particular nosological entity may emerge, that is the Syndrome from chronic pelvic pain (English CPPS acronym: Chronic Pelvic Pain Syndrome).

In the field of genitourinary causes, the non-specific chronic prostatitis represents the most frequent cause of syndrome from chronic pelvic pain. It is characterized by the inflammation of the prostate gland, but without signs of infection. All performed surveys such as culture of urine or prostate liquid result to be negative, and at the ultrasound or CT (Computed Tomography) the prostate results to be normal, notwithstanding the presence of pain and urinary symptoms suggesting its involvement. This clinical picture could be caused by an altered perception of pain with presence of allodynia or hyperalgesia.

The prostate diseases can be of benign neoplastic, malign neoplastic or inflammatory origin.

Benign prostatic hyperplasia (BPH) is a benign disease characterized by a proliferation of the prostate tissue resulting in an increase in the gland volume. This disease is very frequent, above all in men after the age of 40/50 years. With increasing age, in fact, the prostate tends to increase its own volume; to explain such phenomenon, hormonal changes and the action of several growth factors during ageing get involved. The result of a significative prostate enlargement is the appearance of urinary disorders. The prostate tumour includes a variety of forms, from those having a very slow growth, which may not cause disorders throughout life, to other more aggressive forms, which on the contrary grow quickly. Exactly the latter, the so-called malignant ones, are the most dangerous neoplasms. First of all, they are dangerous since they can spread outside the gland, by exploiting the blood and the lymphatic system. The malignant tumours then have the power of spreading in other parts of organism, that is they may determine the onset of metastases. The prostate tumour is a pathology which sometimes is difficult to be detected, since in the initial phases tends to be asymptomatic. Moreover, when the symptoms are present, they can be confused with those deriving from the benign prostatic hyperplasia or from prostatitis. The prostate inflammation, better known in medical field with the term of prostatitis, can depend upon infectious (to be precise, bacterial) or non-infectious causes.

The prostate inflammation types are 4 and, to be precise, correspond to:

• acute prostatitis of bacterial origin (type I),

• chronic prostatitis of bacterial origin (type II),

• non-bacterial chronic prostatitis (type III),

• asymptomatic prostatitis (type IV).

The typical symptoms and signs of prostatitis of type I are: high fever with chills, pelvic pain, urinary disorders of various type. The chronic prostate inflammation of bacterial type can be: the result of a failed treatment of an acute prostatitis of bacterial origin, the consequence of an infection of urinary tracts or, at last, the complication of an epididymitis (infection whose symptoms appear in a testicular manner). The acute prostate inflammation, of bacterial origin, is a form of prostatitis caused by bacteria. The non-bacterial chronic prostate inflammation is the prostatitis appearing gradually and having persistent character, not depending from the presence of bacteria at the level of the prostate gland. Also known as prostatitis of type III or chronic pelvic painful syndrome, it occurs due to reasons that physicians and researchers, notwithstanding the several studies, have not yet cleared completely. The asymptomatic inflammation is also called prostatitis of type IV or inflammatory asymptomatic prostatitis. Notwithstanding the several studies about prostatitis of type IV, the physicians have not yet understood the triggering causes thereof and the reasons of the lack of symptoms.

The Chronic Pelvic Pain (CPP) is characterized by the pain persistence, continuous or intermittent pain, with pelvic involvement, lasting more than six months. It involves progressively different pelvic organs, by involving several functions.

Mastocyte (MC) exerts a key role in the chronic pelvic pain and in the comorbilities underlying and feeding it. In fact, it plays a key role in the acute and chronic inflammation, the pain thereof is the main symptom. MC is ubiquitous in all organs and vascularized tissues, wherein it works as immune sentinel and organizes the inflammatory response. MC includes several vasoactive, proinflammatory and neurotrophic substances, included in vesicles which are released in the surrounding tissues in differentiated way in response to the action of different“antagonist” factors (dangerous or however which cause the release thereof), by coordinating the different phases of the physiological and pathological inflammatory response. These factors modulate in time the inflammatory response of the organism to endogenous and hexogen harmful heterogeneous factors. The treatment of pelvic pain provides a therapy of aetiological type based upon the removal of the underlying cause.

Upon need, it is possible to perform a symptomatic therapy aiming at removing pain only, with traditional painkilling drugs such as FANS (ibuprofen, aspirin, ketoprofen) and paracetamol (Tachipirina). However, these drugs require several daily administrations to reach adequate therapeutic doses and they often involve even serious side effects. Therefore, there is the need to make available treatments, alternative to the already existing ones, which are effective in preventing and/or treating the above-mentioned pathologies, but which have not the side effects and/or disadvantages of the treatments existing in the state of art. These and other needs are met by the present invention, which makes available a composition characterized in that it includes a synergic combination of active substances, obtained by natural sources.

Detailed Description of the Invention

In the composition of the present invention the synergic action takes place between escin and capsaicin in admixture with one or more pharmaceutically acceptable carriers. The composition can further include an extract of Curcuma longa and/or an extract of a plant belonging to the genus Cucurbita and/or an extract of Serenoa Repens.

Escin

Escin is a vegetable compound obtained from the seeds, peels and leaves of horse- chestnut. The horse-chestnut ( Aesculus hippocastanum) is a plant of the family Ippocastanacee. The seeds and the bark of the horse-chestnut include saponins, the mixture thereof is called escin. The escin exists in two forms, alfa and beta, which can be distinguished for their melting points, haemolytic activities and solubility in water. The beta form of escin is the active component of the extract of chestnut of India. The complex mixture of triterpenoid saponins has revealed to be very effective in the treatment of the microcirculatory disorders. It acts by increasing the resistance of capillaries (phlebotonic action) and by reducing the permeability (anti-exudative action) and the inflammation thereof. Three types of pharmacodynamic actions were attributed to escin: anti-oedematous properties, anti-inflammatory activities and venotonic properties.

Escin is traditionally used in the symptomatic treatment of the functional disorders of the skin capillary fragility, the venous insufficiency (heavy legs), but even in the symptoms of haemorrhoidal crisis. Escin acts upon the factors starting inflammation/oedema, that is hypoxia of endothelial cells, followed by the reduction in ATP. The inflammatory reaction is triggered by the release of prostaglandin (signals of the inflammatory response) through phospholipase A2 and PAF (platelet activating factor). It is at the origin of oedema and of the adherence of the neutrophils to endothelium. These neutrophils in turn release enzymes of elastase type, degrading the veins’ wall and stimulating the production of FGF (fibroblast growth factor), by causing a vein enlargement. At last, the adherence of neutrophils to endothelium reduces the blood flow and stimulates hypoxia. Escin acts at the anti-inflammatory level through the reduction in the activity of the lysosomal enzymes which, since they are hyper-activated by the inflammatory processes, produce a degradation of the capillary wall; in particular, escin reduces the activity of elastase, of collagenase, of hyaluronidase and beta-glucuronidase, proteolytic enzymes which - during the inflammatory processes - degrade the internal structure of vessels and weaken endothelium of capillaries by determining the oedema formation.

Escin prevents and reduces oedema. The anti-oedematous effect is linked to a selective vascular permeability, which causes an increase in the venous and arterial tone. Escin reduces the fragility of capillaries and thus allows to avoid liquid infiltration in the tissue interstitium (first cause of oedema). The therapeutic benefit is well supported by a series of experimental surveys on different animal models, indicative of anti-oedematic, anti-inflammatory and venotonic properties. Its properties are mainly correlated to the agent molecular mechanism, by allowing a better inlet of ions into the channels, it causes a raise in the venous tension both under in-vitro and in-vivo conditions. Other mechanisms, that is the release of PGF (prostaglandin) from veins, the reduced catabolism of mucopolysaccharides of tissue, further underline the wide- ranging mechanisms of escin therapeutic activity. In particular, it was demonstrated that escin exerts a venotonic effect, mediated by the stimulation of prostaglandin F2 a in the human veins. In-vivo studies have demonstrated escin effectiveness in topical application in treating the hematoma and the chronic venous insufficiency. In fact, after 6 weeks of 2% escin-based topical treatment, the patients affected by chronic venous insufficiency noted a significant reduction in the symptom of heavy legs, pain and itch, apart from a considerable decrease in the ankles’ circumference. Escin differs from other vasoprotectors of polyphenolic type (such as rutin) for its action mode and its profile. The amphiphilic character of their structure confers an indisputable advantage to cross the double membrane layers and the topical use is supported by many drugs.

The improvement of the tonic effect (vasomotor) is more significant with respect to the flavonoids (like diosmin, for example) which can be considered more“protective”. The anti-oedematous and anti-inflammatory effect (in the resolutive sense of the tissue infiltration) is relevant and demonstrated. An interesting clinical study has demonstrated the anti-inflammatory properties against patients with negative biopsy at the prostate and with high value of PSA (used in the early diagnosis of the prostate cancer). The use of a blend of vegetable substances, escin (30 mg), bromelain (50 mg) and baicalin (190 mg) detected a significative reduction in the PSA levels. In particular, the combination of the active principles in the oral administration reduced by 30% the PSA levels.

Other in-vitro studies demonstrate the capability of escin to stop the growth of cancerous prostate cells through the stop of the cell cycle in G2/M phase by means of induction of p21 and pro-apoptotic factors.

The present invention has underlined the role of escin with its vasoconstriction action and it has detected an innovative application thereof in the treatment and in the prevention of pathologies associated to the prostate. In fact, such saponine has a peripheral vasoconstricting action and it increases diuresis by means of an increased excretion of Sodium and Chlorides. Even if it determines vasoconstriction, escin does not cause hypertension. Moreover, such substance acts onto the microcirculation by decreasing both then number and the diameter of the pores of the membranes of the arterial capillaries and by reducing the passage of the liquids from the capillaries to the tissues. At last, escin is converted in the adrenal gland into a substance having corticoid-like activity performing a strong reparatory action of oedema and of hematomas of traumatic and/or allergic origin.

Capsaicin

Capsaicin (8-methyl-N-vanillyl-6-noneamide) is the active principle of chili pepper, belonging to the genus Capsicum (among the most important species we remind Capsicum annum) and responsible for its spiced flavour and spicy feeling.

“Capsaicin”, main component of chili pepper, was used in past to control various pain forms; the topical treatment of capsaicin both in the inflammatory pain, and in the neuropathic pain is of particular interest.

Capsaicin is a powerful agonist of TRPV1 (transient receptor potential vanilloid) receptor. Such receptor is a not selective cationic channel, mainly expressed on the sensorial neurons, but it was localized even in other non-neuronal tissues: it is activated by several stimuli, both endogenous and exogenous ones (resiniferatoxin, anandamide, inflammation mediators, high temperature > 43°C, acid pH < 5.3). The exposition to capsaicin determines a biphasic response. The first phase is excitative, due to the bond of capsaicin with TRPB1 with consequent opening of the channel allowing the passage of Na⁺ and Ca²⁺ ions by depolarizing the membrane; the resulting electric pulses reach the brain and are responsible for the typical feeling of local burning. An analgesia phase follows, lasting in time, in which the pain-producing fibres become insensitive to the nociceptive stimuli of any nature. The refractoriness to the pain-producing stimuli is not only due to receptor desensitisation, but to a more complex condition called “de-functionalization” consisting in functional and structural changes in the nervous fibre. The chili pepper includes more than 20 capsaicinoids. Each molecule is formed by three regions: A aromatic ring, B amidic bond, C hydrophobic lateral chain. The capsaicinoids differ by the lateral chain (Nr. of C atoms, presence/absence of double bonds); the different structure is responsible for the different biological effects, related to the bond affinity to TRPV1 , to the agonist or antagonist action, to the spicy power. All capsaicinoids have analgesic proprieties, the mechanisms thereof are very different among the various molecules. Capsaicin and dihydrocapsaicin are the most powerful agonists of TRPV1 with excitative effect, responsible for an intense spicy and irritating effect. Other forms such as homocapsaicin have weak activity on TRPV1 with more inhibiting effects, by determining a minimum irritating effect, but however an adequate analgesic effect.

The mechanisms are several and can be summed up in the following points:

I. Receptor inactivation.

II. Increase in the intracellular inflow of Ca²⁺; as this is a second messenger, it brings to activate Ca-depending protease with consequent activation of additional biological routes such as the depolymerization of the microtubules locking the axonal transportation by determining a dysfunction of the nervous fibre. III. Mitochondrial dysfunction with synthesis defect of neurotransmitters.

The first commercial products of capsaicin for topical use containing active low concentrations (0.025/0.1) showed, apart from the slight analgesic effect, the annoying local irritation, the need for several applications, the contamination of the personal environment (mucosae, clothes, instruments, contact lenses, etc.) and then a poor therapeutic compliance.

In order to face such problems a preparation was implemented containing high-dose (8%) capsaicin. In all studies, the results were positive. One single administration determined a reduction in pain, quantified by NPRS (Numeric Pain Rating Scale), scale for evaluating pain, significatively wider with respect to control, and the analgesic effect lasted until the end of the observation period (12 weeks).

Capsaicin is commonly used in the treatment of pelvic pain. In some studies capsaicin demonstrated to have a positive effect on the increase in the bladder functionality. Whereas capsaicin is widely studied and known for its capability in reducing pain, several studies have even shown that it can help to kill tumour cells, including those of the prostate cancer.

In fact, it has been demonstrated that capsaicin makes the tumour cells to “kill themselves” (a process called apoptosis). This substance succeeds in doing so by attacking the portion producing energy of cells, called mitochondrion. Moreover, it does this without affecting healthy cells surrounding the diseased cells. Capsaicin promotes the suicide of tumour cells (apoptosis) and improves even other factors inhibiting the development of the prostate cancer.

Capsaicin, in fact, has demonstrated to have anti-cancer activities in different tumour cells, thereamong the prostate cancer. Several molecular mechanisms were proposed on its chemo-preventive action, thereamong the accumulation of ceramide, the stress induction of the endoplasmic reticulum and NFkB inhibition. However, the precise mechanisms therewith capsaicin exerts its anti-proliferative effect in the prostate cancer cells remain questionable. Then, the involvement of autophagia on the action mechanism of capsaicin on LNCaP and PC-3 cells of the prostate carcinoma was tested. The results showed that capsaicin induces the death of the prostate cancer cells independently from time and concentration, by increasing the levels of light-chain protein associated to the microtubules (LC3-II, an autophagia marker) and the accumulation of p62 protein. The normal prostate cells were more resistant to cytotoxicity induced by capsaicin and did not accumulate p62 protein. These results suggest that the autophagia locking induced by ROS-mediated capsaicin, contributes to the non-proliferation of the prostate cancer cells, thanks to the antitumour molecular mechanism of capsaicin.

Moreover, the molecule has pain-relieving activity on the urine flow. Through its action mechanism it acts by locking selectively the unmyelinated C fibres, usually silent, which activate under conditions of hypersensitivity and bladder hyperactivity, by involving increased frequency and urine urgency, incontinence and reduced bladder capacity. The interaction between capsaicin and vanilloid receptors induces desensitization of C fibres, both by interrupting the responses to the nociceptive stimuli and by inhibiting the hyperactivity of the detrusor muscle. In this way it carries out both a pain-relieving action and an action for adjusting a correct urine flow.

Curcuma lonqa

Curcuma longa is a plant belonging to the family Zingiberaceae comprising a total of about 80 species. Commonly also called saffron from Indies, or simply designated as curcuma, it represents the most used species. Its name derives from the Sanskrit “Kum-kuma” and it is the main ingredient of the Indian curry. Most properties attributed to curcuma indeed depend upon curcumin, which is its most famous active principle ally of well-being.

This molecule - which from the chemical point of view can be classified among polyphenols - is responsible for the typical golden yellow colour of curcuma and of the curries containing it, but not only this.

In fact, antioxidant, anti-inflammatory, anti-infectious, anti-microbial, hepatoprotective, thrombus suppressive, cardioprotective, antiarthritic, proapoptotic, antitumour and chemo-preventive (that is preventing tumours) activities would have been attributed to curcuma. Its protective action against tumours (for example colon or breast tumour) would depend upon the negative adjustment of molecules involved in the inflammation (inflammatory cytokines), of transcription factors, of some enzymes (protein kinase), of some genes involved in cancer and reactive species of oxygen. For example, in case of breast cancer, curcumin seems to exert its anti-tumour effect through a complicated mechanism involving the mechanisms of cellular proliferation and apoptosis, the receptors of estrogens and the growth factor HER2.

The anti-inflammatory action of curcumin, on the contrary, seems to depend upon its capability of controlling different molecules promoting the inflammation (such as interleukin-6, interleukin-1 beta and the Tumour necrosis factor-alpha), some growth factors and their receptors, the protein kinases and other enzymes, the molecules involved in the mechanisms of cellular adhesion and NF-kB, fundamental transcription factor in inflammation.

Curcumin is a product of natural origin mainly endowed with anti-inflammatory activity, which might be useful in the treatment of prostatitis and benign prostatic hyperplasia. Apart from being one of the most powerful anti-inflammatory drugs of natural origin, it has chemical-physical properties suitable to its distribution at prostate level (since it is a lipophile molecule and relatively small, it has no problems in overcoming the blood- prostate barrier). The action mechanism of curcumin consists in modulating the expression of several proteins, thereamong pro-inflammatory cytokines (TNF-a, IL-8,

I L- 1 b , IL-6), apoptotic proteins, NF-kB, COX-2, STAT3, MDA, etc.

The main markers of prostate inflammation are TNF-a, IL-8, I L- 1 b , IL-6, as demonstrated by a study of Penna and collaborators, which have found high concentrations of these pro-inflammatory cytokines and above all of chemokine IL-8 .in the sperm of patients with prostatitis. One of curcumin activities is exactly that of inhibiting TNF-a, I L- 1 b , IL-6 and IL-8 (the latter to a lesser extent) and this could make it a valid active principle to be used in case of prostatitis, since it reduces its main markers. The same activity was demonstrated in prostatitis models (induced by castration and administration of 17b-ob3ΐ^ίoI): the administration of curcumin, in this case, is capable of reducing the expression of TNF-a, IL-6, IL-8.

A clinical, randomized study with long-term follow up of patients evaluated the effect of combination of Serenoa repens (160 MG), Urtica dioica (120 mg), curcumin (200 mg), quercetin (100 mg) and prulifloxacin (600 mg) with respect to prulifloxacin only for the treatment of the bacterial chronic prostatitis. After one month of treatment, in 89% of patients of the first group the disappearance of the prostatitis symptoms was observed, whereas in the second group the disappearance of symptoms was observed in 27% of patients only. This spice carries out a definitely beneficial action in the treatment of the chronic prostatitis (or syndrome of pelvic pain) as well as of the bacterial prostatitis.

Curcuma combines well with other integrators and it is used often as ingredient in the formulations. It can also be used in combination with traditional therapies and it could even help to improve the effectiveness of antibiotics when they are taken together for the bacterial prostatitis. It was studied that curcuma: reduces the pro-inflammatory interleukin-8 cytokines and alfa tumour necrosis factor in the blood tissues and it is effective on the patients with prostate problems.

Several studies underlined the specific role in fighting the bacterial prostatitis, especially in combination with other integrators and antibiotics. A combination of curcumin, quercetin, saw palmetto and nettle, together with the moderate use of an antibiotic, is optimum for the bacterial chronic prostatitis.

In the effort of finding, then, a non-toxic alternative to the therapy for the prostate cancer, the attention of researchers was put on curcuma. The results of the researches showed that curcumin is a powerful inductor of apoptosis (form of programmed cell death) of the cells of both androgen-dependant and independent prostate tumour.

Plant belonging to the family Cucurbitaceae

Cucurbita is a plant genus belonging to the family Cucurbitaceae originating from Centre-South America. The general term Cucurbita comes from the Latin name of pumpkin and, in turn, from the Sanskrit c’arbata curved, rounded. In botany, pumpkins divide into four species: Cucurbita maxima - Cucurbita moschata - Cucurbita pepo - Cucurbita melanosperma. The most studied varieties are represented by the Cucurbita maxima Duchesne (yellow or sweet pumpkin) and by Cucurbita pepo (commonly called zucchini). The pumpkin seed oil is obtained by squeezing the homonymous plant and it varies in relation to the varieties and the botanic species therefrom the seeds are obtained. The pumpkin is a natural antidote against the free radicals, dangerous substances for our organism and which speed-up the cell aging process. Betacarotene contained in the pumpkin has several anti-inflammatory and anti-tumour properties as the National Cancer Institute discovered; on the contrary, lycopene is more effective in case of prostate, lung and stomach tumour. The studies showing an important effect in reducing blood sugar in the animal model, by using extracts both of pulp and of the seeds, are interesting. The effect could be due both to the presence in the fruit pulp of D-chiro-inositol and of other particular polysaccharides, substances promoting the action of insulin, and to substances such as trigonelline and nicotinic acid present in the seeds. The presence of vitamin E and other substances with antioxidant action, particularly some polysaccharides which are bound to proteins in the fruit pulp, can stimulate the activity of enzymes such as superoxide dismutase and glutathione peroxidase, fundamental enzymes in reducing the oxidative damage in cell.

Several works showed that diets rich in carotenoids, abundant in pumpkin may have some role in reducing the incidence of prostate tumour. Moreover, different substances isolated from the pumpkin pulp and seeds, moschatine and cucurmosine demonstrated to have the capability of inhibiting the growth of specific types of tumour cells, particularly some types of melanoma. The pumpkin seeds include substances such as cucurbitin, phytosterine and delta sterols able to inhibit the aromatase enzyme influencing the tonicity of the pelvic floor.

Moreover, the pumpkin seed oil (extracted both from Cucurbita maxima and from Cucurbita pepo) is widely used as phyto-therapeutic remedy useful in the treatment of benign prostatic hypertrophy. Even carotenoids and zinc (both useful for the prostate’s health), manganese, magnesium, phosphorous and iron are abundant, even if the main constituent however remains oleic acid (24-38%) linoleic acid (43-56%), tocopherols and carotenoids (lutein and b-carotene). Some own substances of pumpkin seeds are useful in fighting benign prostatic hyperplasia, a condition which determines an increase in the prostate volume in turn caused by an increase in the number of (not tumour) cells of the gland. This pathology usually appears with frequent urinations, recurrent infections and inflammations of the urinary tracts. It was shown that the pumpkin seeds can help in alleviating the symptoms of this pathology, thanks to their content of zinc and, above all, phytosterols such as beta sitosterol, sitostanol, avenasterol. Such phytosterols have a chemical structure very similar to that of the androgenic and estrogenic hormones. These would be capable of inhibiting the conversion of testosterone into dihydrotestosterone, responsible for the cell growth at the basis of the prostatic hypertrophy. Even zinc is important for the good operation of the prostate and the pumpkin seeds, since they are rich therein, even under this aspect contribute to explicate a protective action against this gland.

It has long been known that the seeds contain betasterols structurally similar to androgens and estrogens. These substances have demonstrated to be useful to lower the cholesterol levels and to improve the symptoms of prostatic hypertrophy, effect which seems to be partially linked to the capability of inhibiting the conversion of testosterone into dihydrotestosterone. This latter substance is a hormone derived from testosterone thanks to 5-alfa-reductase enzyme, involved - among other things - in the hyperproliferation of the prostatic cells.

In a randomized clinical study, double-blind with placebo, performed on 47 Korean patients, being 53.3 years old on the average, affected by benign prostatic hyperplasia, the effectiveness of pumpkin seed oil was tested, administered for 12 months alone (320 mg/per day) or in association with Serenoa repens oil (320 mg/per day + 320 mg/ per day of the other one); the results were compared to those of the group treated with placebo (320 mg of sweet potato starch) or with serenoa oil only (320 mg/per day). None of the proposed treatments guaranteed a significative decrease in the prostate volume; the PSA serum values reduced in the group ingesting the two oils together, whereas the life quality improved after 6 months. The two phytotherapeutic drugs ingested separately guaranteed an improvement of the prostatic symptoms expressed through IPSS (International Prostatic Symptom Score) within three months, whereas the maximum urinary flow improved after six months in the group treated with pumpkin oil seed, and after 12 months in the group treated with serenoa oil. The combination of Serenoa with another variety still belonging to the genus Cucurbita demonstrated a reduction in the symptoms of BPH with a very reduced amount of the active principles of interest ( Cucurbita pepo 80 mg and Serenoa repens 80 mg).

Serenoa repens

Serenoa repens, in English called Saw Palmetto, is a dwarf palm of the family Arecaceae growing mainly in South-East of United States along the coast of the Atlantic Ocean and in the South tropical areas. Its fruits, having colour from dark brown to black and rich in active principles such as free fatty acids and sterols (in particular beta-sitosterol), are recognized by the undisputed healthy properties reported previously to the advantage of prostate and hair.

The sterol lipid extract of Serenoa repens, highly titrated in fatty acids, not only helps in keeping healthy the prostate, but as reported by a lot of scientific evidence, thanks to the inhibition of 5-alfa reductase, it contributes even to the well-being and the trophism of hair of men affected by androgenetic alopecia. The lipido-sterolic extract of Serenoa repens, thanks to its composition and content in fatty acids thereamong the lauric acid, the myristic acid and a vegetable sterol called beta-sitosterol, has an important selective antiandrogenic action which mainly explicates through the inhibition of both isoforms I and II of 5-alfa reductase (5AR), key enzyme in the process for transforming testosterone into dihydrotestosterone. Dihydrotestosterone is the hormone mainly responsible for the prostate growth and the appearance of male androgenetic alopecia. Scientific evidence has shown that the lauric acid and the myristic acid are also capable of interacting directly with testosterone by forming esters of this hormone which then cannot be converted into DHT. On the contrary, studies on animal models have highlighted that beta-sitosterol limits the bioavailability of cholesterol and then the synthesis of testosterone, substrate of 5-alfa reductase, in the prostate and capillary follicle. An in-vitro study demonstrates that both the ethanolic and the hexane extract, two types of lipido-sterolic extract of Serenoa repense are capable of inhibiting 5-alfa reductase in co-cultures of epithelial cells and prostate fibroblasts. Another in-vitro study demonstrates that the lipido-sterolic extract of Serenoa repens is useful in treating BPH with inflammation features, by modulating presumably the equilibrium between apoptotic factors and proliferative factors. An in-vivo study compares the effectiveness of the lipido-sterolic extract of Serenoa repens (LSESR) to finasteride (inhibitor of 5-alfa-reductase) on prostatic hyperplasia induced by hyperprolactinaemia in rats. Hyperprolactinaemia was induced by 30 daily injections of sulpiride. Rats received daily gavage of lipido-sterolic extract of Serenoa repens or finasteride. They were divided into the following groups: control, castrated rats, castrated with a replacement testosterone (T) or with an implant of 5alfa-dihydrotestosterone (DHT). Hyperprolactinemia increases the humid weight and induces hyperplasia in the lateral prostate. Differently from finasteride, LSESR reduced significantly the growth and hyperplasia of the lateral prostate in the castrated rats, treated with DHT and treated with sulpiride. Finasteride was only capable of inhibiting the effect of androgens on the enlargement of the rat prostate. LSESR inhibited not only the androgenic effect but even the trophic effect of prolactin in the lateral hyperplasia of rat. Several clinical studies then confirmed the effectiveness of this vegetable sterol in improving the well being of the prostate in man. At last, additional studies highlighted that the extract of Serenoa repens has an inhibitory action against the growth factors responsible for the cell proliferation and of the inflammatory process at the base of benign prostatic hypertrophy. It is scientifically proven that the fruit of Serenoa repens contributes to the normal functionality of prostate. Clinical studies, in fact, demonstrate that the lipido- sterolic extract from fruits of Serenoa repens is effective in improving the symptoms of benign prostate hypertrophy. Several clinical studies in double-blind, randomized and controlled by placebo and performed on patients affected by benign prostatic hypertrophy (BPH) confirmed the effectiveness of Serenoa extracts. One of these studies, performed on 176 patients affected by BPH, and resulted to be no-responders to the treatment with placebo in preceding clinical studies, highlighted that the daily intake of 320 mg of extract of Serenoa repens fruit already after 30 days contributes to the normal prostate functionality. In fact, it was possible to note an evident improvement of dysuria, a reduction in polyuria and nycturia in the treated patients with respect to placebo (32.5% vs. 17.7%). Moreover, the patients thereto the lipido-sterolic extract of Serenoa was administered noticed a maximum increase in the urinary flow (28.9%) with respect to the patients who had received placebo (8.5%). There is evidence that the hexane extract of Serenoa repens (HESr) can antagonize metabolites of lipoxygenase, by determining an anti-inflammatory effect. This feedback has recently been confirmed by in-vivo and in-vitro studies, with the evidence that Serenoa can modulate the expression of multiple genes correlated to inflammation. Moreover, randomized clinical studies evaluated the Serenoa effect on biomarkers of chronic inflammation persisting in patients with LUTS/BPH, by using a not invasive method (urine test after prostatic massage), by correlating the effectiveness of the hexane extract of Serenoa to the inflammatory state of the patient. Recent acquisitions, moreover, have pointed out that the fact of using inflammation as therapeutic target is useful also in favouring the therapeutic response of other treatments. In fact, it was demonstrated that during a chronic inflammation state the effectiveness of the inhibitors of 5-alfa-reductase (5-ARI) decreases. Moreover, this inflammatory chronic state, as said, is capable of stimulating the production of interleukins and growth factors which act independently from the presence of 5-ARI. Therefore, it appears interesting for the present invention for its anti-inflammatory and anti-proliferative capabilities.

The present invention allows to obtain contemporarily:

• anti-inflammatory effect

• analgesic effect

• pro-apoptotic effect on cancerous cells

In particular, the effectiveness of the above-mentioned composition derives from the precise properties of its main components: the diuretic, anti-oedematous and anti inflammatory activities of escin and the analgesic and pain-killing action of capsaicin.

Moreover, both compounds are capable of inducing apoptosis in prostatic cancerous cells by means of different activities. The above-mentioned invention can further include the following components: curcuma, powerful anti-inflammatory of natural origin, has chemical-physical properties suitable to its distribution and the prostatic level; an extract from the plant belonging to the genus Cucurbita which thanks to its variety of constituents (sitosterols, vitamin E, zinc, carotenoids, etc.) contributes to a complete well-being of the prostate. Al last, the extract of Serenoa repens, apart from its anti-inflammatory and anti-androgenic activity, has a complete regulating action on the prostate functionality.

As previously mentioned, in the present invention the synergic action takes place between escin and capsaicin in admixture with one or more pharmaceutically acceptable carriers.

Suitable pharmaceutically acceptable carriers are those commonly known to the expert in the art for the preparation of pharmaceutical compositions for oral administration such as solutions, suspensions, powders or granulates, tablets, capsules, pellet. By way of not-limiting example, such pharmaceutically acceptable carriers can consist of binders, diluents, lubricants, glidants, disaggregating agents, solubilizing (wetting) agents, stabilizing agents, colouring agents, anti-agglomerating agents, emulsifying agent, thickening and gelling agents, coating agents, wetting agents, sequestrants, sweeteners. In the specific example of diluents, they can be: magnesium carbonate, microcrystalline cellulose, starch, lactose, sucrose; the mainly used lubricants are magnesium stearate, stearic acid, sodium stearyl fumarate. As glidants, colloidal silica, magnesium silicate can be mentioned, as disaggregating agents the crosslinked polyvinylpyrrolidones, sodium starch glycolate can be mentioned, as solubilizing agents the surfactants such as TWEEN or sodium lauryl sulphate can be mentioned and as stabilizing agents all class of preservatives (sorbic acid and derivatives, benzoic acid and derivative, parabens), antioxidants (ascorbic acid and derivatives, tocopherol), acidifying agents (phosphoric acid, tartaric acid) can be mentioned. The thickening and gelling agents can be carrageenan, pectin, starches, the coating agents include for example waxes and derivatives, the anti-agglomerating agents include calcium or magnesium carbonate, the wetting agents include sorbitol, mannitol, the EDTA sequestrants and derivatives, the colouring agents include aspartame, potassium acesulfame. The composition, the present invention relates to, and preferably a liquid or solid composition for oral use, still more preferably an aqueous solution, a suspension, a powder or granulate, a tablet, a capsule, pellet. The composition can further include an extract of Curcuma longa and/or an extract of a plant belonging to the genus Cucurbita and/or an extract of Serenoa Repens.

There is synergy when capsaicin is present in an amount ranging between 0.1 mg and 5000 mg, preferably between 1 mg and 3000 mg; escin is present between 0.1 mg and 5000 mg, preferably between 1 mg and 3000 mg. The extract of Curcuma longa, if present in the composition, is contained in an amount ranging between 10 mg and 5000 preferably between 20 mg and 3000 mg. The extract of the plant belonging to the genus Cucurbita, if present in the composition, is contained in an amount ranging between 1 mg and 5000 mg, preferably between 10 mg and 3000 mg. The extract of Serenoa repens, if present in the composition, is contained in an amount ranging between 10 mg and 5000 mg, preferably between 20 mg and 3000 mg. The following examples are provided by way of example only and not for limitative purposes of the scope of the invention, as defined by the enclosed claims.

EXAMPLE 1

Pharmaceutical form: soft gel capsules

EXAMPLE 2

Pharmaceutical form: soft gel capsules EXAMPLE 3

Pharmaceutical form: sachets EXAMPLE 4

Pharmaceutical form: sachets

The dosage form can be a food supplement or a medical device, a food complement, a cosmetic, a nutraceutical, dietetic and nutritional composition, a food product, a beverage, a nutraceutical agent, a medicament, medicated food, a pharmaceutical composition or food for special medical purposes including the above-mentioned active ingredients in admixture therebetween. The preferred administration route is the oral or topical one.

EXPERIMENTAL PART

The synergic action of the composition with respect to the single active ingredients is evaluated by means of in-vitro and/or in-vivo experimental methods.

The susceptibility of the cell system to the pro-inflammatory stimuli can be evaluated with immuno-enzymatic essays (ELISA), by detecting the amount of cytokines involved in inflammatory processes and in particular the activity of bioactive compounds on the response, inhibition/activation, of cytokines (TNF-a, IL-1 , IL-4, IL-6, IL-8, IL-10, IL-17, INF-Y, COMP), proinflammatory enzymes such as lipoxygenase (LOX) and cyclooxidases (COX2) involved during inflammatory processes induced by LPS through the“Whole Blood Assay (WBA)” or the test with“mononucleated cells of the peripheral blood” (PBMC) or other similar tests.

The pro-apoptotic activity of the active principles, the present invention relates to, can be evaluated by means of in-vitro assays capable of demonstrating the way in which the synergy of the two components inhibits significantly the cell viability with respect to the single active principles.

Cell models known to the person skilled in the art can be used, such as for example PC-3 cells, CRPC (castration-resistant prostate cancer), DU-145, LNCap and/or other analogous ones.

As in-vivo test a simple model of general inflammation is the model of oedema from carrageenan induced on mice or rats. In this test, the injection of carrageenan in the animal determines an acute inflammatory reaction with appearance of the typical signs of inflammation. With this test the response of the active principles to reduce inflammation in rats can be evaluated.

The anti-inflammatory activity can be evaluated by demonstrating the effectiveness of the compounds, the present invention relates to, in favouring apoptosis in mouse with BPH induced experimentally. The apoptosis system is a promising system as far as the medical treatment of benign prostate hypertrophy is concerned. Within the apoptotic cascade, the apoptosis inhibiting proteins (lAPs) modulate apoptosis through the direct inhibition of the cascade of caspases. The study can evaluate the effectiveness of the active principles, alone or in combination, on the capability of inducing the expression for the proteins lAPs in mice with BPH induced experimentally by injection with testosterone.

A more complex in-vivo model provides to test the active principles of the present composition to demonstrate their anti-inflammatory and analgesic effect on rats in which an experimental autoimmune prostatitis (EAP) is induced, through intradermic injection of prostatic antigens of rat and of “Freund’s adjuvant”, a mixture of antigens emulsified in mineral oils used as immuno-potentiating agents.

This takes place on pre-established days (for example 0-28 and 42) and after injection both the level of inflammatory cytokines and chemokines and the chronic pelvic pain is evaluated. In particular, the recruited rats are tested in terms of allodynia and hyperalgesia on the pre-established day (for example 42) after immunization. Allodynia and hyperalgesia are tested by using filaments of von Frey with forces of 0.4 g. The filament is applied for a total of 5 times for 1-2 seconds with an inter-stimulus interval of 2 minutes. The behaviours considered positive upon the filament stimulation are: net retraction of the abdomen, licking or scratching immediately in the stimulation area of the filament or jumping. The control, each substance singularly and then even the mixture of substances of interest for the present composition, are administered orally 2 hours before the mechanical stimulation, in this way the role of the active principles in reducing the frequency of reactions to the mechanical stimulation is evaluated, with respect to the control and to the substances administered singularly.

Claims

1) A composition comprising an association of escin and capsaicin, in admixture with one or more pharmaceutically acceptable carriers.

2) The composition according to claim 1 , wherein escin is present in the composition in form of an extract of Aesculus hippocastanum.

3) The composition according to claim 1 , wherein capsaicin is present in the composition in form of an extract of a plant of the genus Capsicum preferably Capsicum annum and/or Capsicum pubescens and/or Capsicum baccatum and/or Capsicum chinense and/or Capsicum frutescents.

4) The composition according to any one of the preceding claims, wherein an extract of Curcuma longa and/or an extract of a plant belonging to the genus Cucurbita and/or an extract of Serenoa repens is present as additional active component.

5) The composition according to any one of the preceding claims, wherein the extract of a plant belonging to the genus Cucurbita is an extract of Cucurbita maxima and/or Cucurbita pepo.

6) The composition according to any one of the preceding claims, wherein escin is present in an amount between 0.1 mg and 5000 mg, preferably between 1 mg and 3000 mg; capsaicin is present in an amount between 0.1 mg and 5000 mg, preferably between 1 mg and 3000 mg.

7) The composition according to any one of the preceding claims, wherein, if present, the extract of Curcuma longa is contained in an amount between 10 mg and 5000 mg, preferably between 20 and 3000 mg; the extract of a plant belonging to the genus Cucurbita is contained in an amount between 1 mg and 5000 mg, preferably between 10 mg and 3000 mg; the extract of Serenoa repens is contained in an amount between 10 mg and 5000 mg preferably between 20 mg and 3000 mg.

8) The composition according to any one of the preceding claims, wherein the composition is a food supplement, a medical device, a food complement, a cosmetic, a nutraceutical, dietetic and nutritional composition, a food product, a beverage, a nutraceutical, a medicament, medicated food, a pharmaceutical composition or food for special medical purposes.

9) The composition according to any one of the preceding claims in the form of liquid, solid or semi-solid composition for oral or topical use. 10) The composition according to any one of the preceding claims for the use in the prevention and/or treatment of prostate pathologies, wherein the prostate- associated pathologies are pelvic pain, chronic pelvic pain, acute prostatitis of bacterial origin, chronic prostatitis of bacterial origin, non-bacterial chronic prostatitis, asymptomatic prostatitis, overactive bladder, benign prostatic hypertrophy, prostate cancer.