RU2808426C2 - Composition for treatment and/or prevention of lower urinary tract symptoms - Google Patents

Abstract

FIELD: medicine; urology; pharmacology.

SUBSTANCE: invention is intended for the treatment and/or prevention of symptoms of the lower urinary tract. A composition is used containing an aqueous extract of pollen obtained from a mixture of plants belonging to the families Pinaceae and Poaceae, an aqueous extract of seeds of plants belonging to the genus Cucurbita, and vitamin E or its esters for the treatment and/or prevention of lower urinary tract symptoms (LUTS).

EFFECT: use of the invention makes it possible to increase the effectiveness of treatment and/or prevention of genitourinary pain syndromes and clinical syndromes indicating dysfunction of the lower urinary tract.

9 cl, 18 dwg, 6 tbl, 9 ex

Description

Field of technology

The present invention relates to a composition based on plant extracts for use in the treatment and/or prevention of lower urinary tract symptoms (LUTS) (LUTS).

Lower urinary tract symptoms (LUTS) commonly occur in men and women of all ages. The prevalence and severity of LUTS increases with age and is a major concern for the aging population. It is now accepted that LUTS are a group of progressive, age-related, non-gender or organ-specific symptoms that include a combination of filling, voiding and post-void phase symptoms.

One of the main symptoms is urinary incontinence. It is known (DeMaagd et al., “Management of Urinary Incontinence,” P&T®, June 2012, Vol. 37 No. 6) that approximately 10 million patients in the United States experience urinary incontinence. For example, in 2007, it was estimated that more than 25 million people in the United States experienced episodes of urinary incontinence. The prevalence of urinary incontinence appears to be higher in women than in men aged 80 years and younger, but men and women are affected almost equally after age 80.

LUTS can have multiple causes located in the urinary tract or some distance from it. LUTS may result from infection or inflammation (eg, cystitis, prostatitis), structural abnormalities (eg, stones, tumors, prolapses, benign prostatic hypertrophy), medical problems (eg, diabetes, multiple sclerosis), or drug use (eg, anticholinergics). , opioids).

Although LUTS usually do not cause serious illness, they are a common reason for seeking medical attention, can seriously affect quality of life, and are a sign of serious urinary tract disease. These LUTS can cause complications and may, for example, be associated with sexual dysfunction, depression, hypertension and anxiety.

Due to the etiological complexity, treatment of LUTS must be tailored to each patient and must take into account the type and intensity of symptoms, their progression, the presence of comorbidities and the patients' quality of life. In addition, patients' personal opinions should be taken into account.

Not all people with LUTS require treatment.

However, it may be necessary to turn to pharmaceutical treatment or even surgery to treat LUTS.

State of the art

The preferred pharmaceutical treatment for a patient with moderate to severe symptoms that affect quality of life is alpha-blocker monotherapy. They are alpha-adrenergic receptor agonists. They work by relaxing the smooth muscle fibers of the bladder neck and prostate gland in men and/or the proximal urethra in women. Their effect is rapid and stable over several years, with significant improvement in LUTS.

However, alpha-blockers have significant side effects, such as orthostatic hypotension, headaches, dizziness, and impaired accommodation. They should also be used with caution in elderly patients, patients with coronary artery disease, and in cases of concomitant antihypertensive treatment. They are contraindicated before cataract surgery or if there is a history of orthostatic hypotension.

5-alpha reductase inhibitors (I5ARs) are second-line pharmacological agents. They are used more specifically in men and block the conversion of testosterone to dihydrotestosterone (DHT), which is the active metabolite of prostate growth. Their activity has been demonstrated in terms of symptoms, flow, reducing the risk of acute urinary retention, reducing the need for surgery and reducing prostate volume by 20%.

However, their effect is slow, between 3 and 6 months, and they cause sexual disorders (decreased libido, erectile dysfunction, gynecomastia).

Combination therapy between an alpha blocker and a 5-alpha reductase inhibitor is more effective in the long term for urinary symptoms than either single agent, but the side effects are cumulative.

Other treatment methods are known using anticholinergics (antimuscarinics) or phosphodiesterase type 5 inhibitors (IPDE5).

However, they have only limited effectiveness, for example in emergency situations for anticholinergics, causing undesirable effects such as dry mouth, constipation, confusion, and especially increased urinary dysfunction.

Collagen is also known to be used especially in the treatment of urinary incontinence in women.

However, collagen is administered by injection, which requires intervention and may reduce compliance.

Herbal medicine is also used in the treatment of LUTS. It is a therapy based on plant extracts and has excellent tolerability due to the absence of side effects. The most widely used molecules are extracted from Pygeum africanum and Serenoa repens .

However, even if the literature is inconsistent with their subject matter, their effectiveness on LUTS, although significant, remains modest. Their combination with other treatment methods is not recommended.

For this purpose, compositions for maintaining prostate health in men are known.

Known, for example, are the publications of HG Preuss et al. (“A Critical Review of Cernitin™ for Symptomatic Relief of Lower Urinary Tract Symptoms (LUTS) in Men,” RESEARCH COMMUNICATIONS IN PHARMACOLOGY AND TOXICOLOGY, 2013) or WG Chambliss et al. (“Flower Pollen Extract and its Effects on Urinary Support, Bladder and Smooth Muscle Contents”, 2013), which describe the activity of CERNITIN™ (=GRAMINEX® flower pollen extract) in men with benign prostatic hypertrophy, which can cause lower urinary tract symptoms or other prostate-related disorders CERNITIN™ is a mixture of rye (Secale cereale L.), timothy (Phleum pratense) and corn (Zea mays L.) pollen containing an aqueous extract and an oil extract of the pollen in a 20:1 ratio.

Other compositions based on GRAMINEX® containing, for example, pumpkin seed extract containing 25% fatty acids (“Prostate Support - Weil Vitamin Advisor”, 2016), or pumpkin seed extract standardized to contain 10% fatty acids and vitamin E. (PROSTACERN™) is known to help with prostate problems in men.

Such products are primarily intended for men with prostate problems.

Men may experience problems with urinary incontinence due to this, but the reasons are different from those in women. Mainly:

- disorders associated with the prostate gland, such as an increase in its volume (prostatic hypertrophy), which compresses the urethra. The male subject then suffers from retained drips or even nighttime urine leaks;

- prostatectomy: which is the removal of the prostate gland;

- health problems such as urethral tumor or urethral stricture, which is defined as a narrowing of the urethra and which prevents normal urination.

However, even though diseases associated with the prostate gland may generally lead to lower urinary tract symptoms, such compositions do not appear, at first glance, to be directly indicated for the treatment and/or prevention of LUTS and, in particular, , urinary incontinence in women.

However, the product PURE™ Control C/P (“Incontinence, frequent urination, bladder health - Women Living Naturally”, June 17, 2017) is known as a treatment for urinary incontinence in women. It consists of CERNITIN™ pollen extract (a mixture of rye pollen (Secale cereale L.), timothy grass (Phleum pratense) and corn (Zea mays L.) containing aqueous extract and pollen oil extract in a ratio of 20:1) and pumpkin seed extract , standardized to contain lignans and phenolic derivatives rather than fatty acids. This fatty acid-free pumpkin seed extract (commonly known to be high in fatty acids) appears to help support healthy urination during the day and at night after 6 weeks of treatment in postmenopausal women.

Technical problem

Taking into account the above, to address the known problems associated with the side effects of used treatments, and to improve the effectiveness against LUTS, in particular urinary incontinence, especially in women in the broad sense, for example, women aged 16 years to 75, demonstrating excellent portability, the applicant has developed a new composition based on plant extracts.

In addition, mastering the production parameters of plant extracts ensures traceability, standardization and reproducibility within the specification of the extracts.

Technical solution

The solution to the problem posed by the present invention relates to a composition comprising a pollen extract containing an aqueous extract of pollen obtained from a mixture of plants belonging to the families Pinaceae and Poaceae, an aqueous extract of seeds of plants belonging to the genus Cucurbita , and vitamin E or its esters for treatment and/or prevention of lower urinary tract symptoms (LUTS) in women.

Benefits provided

The Applicant has been able to demonstrate that the use of the composition which is the subject of the invention makes it possible, in particular, to prevent or improve lower urinary tract symptoms (LUTS) associated with the filling, voiding and/or post-void phases in women, regardless of their age, and, more preferably, urinary incontinence, in particular the treatment and/or prevention of stress urinary incontinence (SUI), urinary incontinence caused by urgency (or urge) incontinence and mixed urinary incontinence (MUI), including including, in particular, reducing the frequency of nighttime urination, which causes awakening.

Brief description of the drawings

The present invention and the advantages resulting therefrom will be better understood by reading the following description and non-limiting embodiments illustrated with reference to the accompanying drawings, in which:

Fig. 1 shows the results obtained using the SANDVIK scale in comparison with various compositions in the context of the Study Example 5 on urinary incontinence in women before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 2 shows the results obtained using the MUH scale (Measurement of Urinary Handicap) in comparison with various compositions in the context of the study according to Example 5 on urinary incontinence in women before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 3 shows the results obtained using the ICIQ scale (International Consultation on Incontinence Questionnaire) in comparison with various compositions in the context of the study according to Example 5 on urinary incontinence in women before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 4 shows the results obtained by ICIQ urinary incontinence type with Composition A in the context of the study according to Example 5 on female urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 5 shows the results obtained by ICIQ urinary incontinence type with Composition B in the context of the study according to Example 5 on female urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 6 shows the results obtained by ICIQ urinary incontinence type with Composition C in the context of the study of Example 5 on female urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 7 shows the results obtained by MUH urinary incontinence type with Composition A in the context of the study of Example 5 on female urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 8 shows the results obtained by MUH urinary incontinence type with composition B in the context of the study according to Example 5 on female urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 9 shows the results obtained by MUH urinary incontinence type with Composition C in the context of the study of Example 5 on female urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 10 shows the results obtained using the SANDVIK scale with composition C in the context of the study according to Example 5 on any type of urinary incontinence in general women before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 11 shows the results obtained using the ICIQ scale in the context of Example 6 on the effectiveness of the composition according to the invention (according to Example 1 = PSCEP; compared with placebo) in women with urgency urinary incontinence (or urge) or mixed urinary incontinence before treatment (D0 ), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 12 shows the results obtained using the MUH scale in the context of Example 6 on the effectiveness of the composition according to the invention (according to Example 1 = PSCEP; compared with placebo) in women with urgent urinary incontinence (or urge) or mixed urinary incontinence before treatment (D0 ), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 13 shows the results obtained using the SANDVIK score in the context of Example 6 on the effectiveness of the composition according to the invention (according to Example 1 = PSCEP; compared with placebo) in women with urge (or urge) or mixed urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 14 shows the results obtained using the ICIQ scale in the context of Example 7 on the effectiveness of the composition according to the invention (according to Example 1 = PSCEP; compared with placebo) in women with stress urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 15 shows the results obtained using the MUH scale in the context of Example 7 on the effectiveness of the composition according to the invention (according to Example 1 = PSCEP; compared with placebo) in women with stress urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 16 shows the results obtained using the SANDVIK scale in the context of Example 7 on the effectiveness of the composition according to the invention (according to Example 1 = PSCEP; compared with placebo) in women with stress urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 17 shows the results obtained in the context of Example 8 on the effectiveness of the composition according to the invention (according to Example 1) on the frequency of nighttime urination causing awakening in women with urinary (or urge) incontinence or mixed urinary incontinence before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90).

Fig. 18 shows the results obtained in the context of Example 9 regarding the activity of the composition according to the invention (according to Example 1) in comparison with each active ingredient of the composition, taken independently, at the M3 muscarinic receptor.

Detailed description

In the present description, unless otherwise indicated, it should be understood that when an interval is specified, it includes the upper and lower limits of the specified interval.

The invention relates to a composition used for the treatment and/or prevention of lower urinary tract symptoms (LUTS) in women, including a pollen extract containing an aqueous extract of pollen obtained from a mixture of plants belonging to the families Pinaceae and Poaceae , an aqueous extract of seeds of plants belonging to genus Cucurbita , and vitamin E or its esters.

The term "treatment" refers to the improvement, prevention or modification of a disease or disorder or at least one significant symptom thereof. It is also an improvement, prevention or change in at least one measurable physical parameter associated with the disease or disorder being treated that is not necessarily felt by the subject. In another embodiment, the term “treating” refers to inhibiting or slowing the progression of a disease or disorder, either physically, such as by stabilizing an expressed symptom, or physiologically, such as by stabilizing a physical parameter, or both. The term "treatment" also refers to delaying the onset of a disease or disorder. In some specific embodiments of the invention, the composition of interest is administered as a prophylactic measure. In this context, the term "prevention" refers to reducing the risk of acquiring a particular disease or disorder.

By pollen extract according to the invention is meant a pollen extract containing an aqueous pollen extract obtained from a mixture of plants belonging to the Pinaceae and Poaceae families.

Poaceae, also called Gramineae , is a family of monocotyledonous plants in the order Poales . The common name is “ Poaceae ” or “ Grass ”. This family of approximately 12,000 species grouped into 780 genera includes most of the species commonly known as "herbs" and "cereals". Usually these are herbaceous plants, less often woody (bamboo).

Like all anemophilous pollen, Poaceae pollen is spherical or slightly ellipsoid in shape with reduced ornamentation. The only hole (or pore) is round: this is one of the criteria for monocots. Poaceae pollen is small in size and lightweight. Size about 40 microns. For cereals, the size is from 60 to 100 microns.

The Pinaceae (pine) family or Abietaceae includes gymnosperms; has 220-250 species, divided into 11 genera. These are trees or shrubs from temperate regions, either with evergreen leaves that are needle- or scale-shaped, or deciduous, like larch. In this family, species native to France are found among the genera Abies (fir), Picea (spruce), Larix (European larch) and Pinus (pine).

Pinaceae abundantly produces large pollen grains, typically ranging in size between 40 and 100 microns. They are poreless. Pollen grains from pine, fir, spruce and cedar have two balls that make them easier to suspend in the air. Larch and Douglas fir pollen grains are more or less spherical and without a ball.

The pollen extract according to the invention may also contain an oily pollen extract obtained from the same Pinaceae and/or Poaceae plants, containing fat-soluble active ingredients obtained by extraction, for example maceration, infusion, digestion, digestion, percolation. or by leaching, preferably macerating at room temperature from 15 to 27°C, the plant material in an oily solvent such as an ether, eg diethyl ether, a ketone, eg acetone or oil.

Preferably, the pollen extract according to the invention consists of an aqueous pollen extract obtained from a mixture of plants belonging to the family Pinaceae and Poaceae , that is, it contains only an aqueous pollen extract and does not contain another pollen extract such as an oil extract.

In accordance with the present invention, an aqueous extract means an extract containing water-soluble active ingredients obtained by extraction, for example by hydrodistillation, maceration, infusion, digestion, digestion, percolation or even leaching, preferably by maceration at a low temperature of 15 to 40°C of the plant material in an aqueous solvent that is, the solvent is one containing water alone or preferably mixed with other solvent(s) such as an alcohol, a ketone and/or a non-ionic surfactant.

By way of non-limiting examples, the aqueous solvent is selected from a mixture primarily containing water in combination with an alcohol such as ethanol, a ketone such as acetone, and/or a nonionic surfactant.

The solvent can then be partially or completely removed to obtain an extract.

Preferably, the composition in accordance with the present invention, used for the treatment and/or prevention of lower urinary tract symptoms (LUTS) in women, includes a pollen extract consisting of an aqueous pollen extract obtained from a mixture of plants belonging to the family Pinaceae and Poaceae , and an aqueous extract of seeds of plants belonging to the genus Cucurbita, and vitamin E or its esters.

The pollen extracts used in the composition according to the present invention are preferably obtained from a mixture of plants of the family Poaceae , selected from the genera Secale , Zea and/or Dactylis , and the family Pinaceae , selected from the genus Pinus .

More specifically, the aqueous pollen extract according to the present invention is preferably obtained from a mixture of plants of the family Poaceae selected from the species Secale cereale L. (rye), Zea mays L. (corn), Pinus sylvestris L. (pine) and/or Dactylis glomerata L. (urchin) and Pinaceae selected from Pinus sylvestris L. (Scots pine) and/or Pinus pinaster (maritime pine) ).

The composition used in accordance with the invention may further comprise an aqueous and/or oleaginous, preferably aqueous, extract of pistils obtained from the same plants.

Preferably, the plants used, from which the pollen and possibly the pistils are obtained, are freshly harvested.

The pollen used in the present invention may be pollen collected by insects (such as bee pollen) or collected through human intervention.

Bee pollen, for example, contains pollen as well as nectar and bee saliva. Pollen collected as a result of human intervention lacks such additional ingredients. Preferably, said pollen in the present compositions is obtained only through human intervention. This once again allows us to standardize the final product.

Once collected, the pollen and preferably the pistils can be used fresh or dried, preferably dried and possibly debacteriated.

According to a preferred embodiment of the invention, the aqueous pollen extract is prepared from a mixture of:

- aqueous extract of Secale cereale L. pollen;

- aqueous extract of Zea mays L. pollen;

- aqueous extract of Pinus sylvestris L. pollen;

- aqueous pollen extract of Dactylis glomerata L.

Preferably, the composition used in accordance with the invention further comprises an aqueous extract of Zea mays L. pistil.

According to a particularly preferred embodiment of the invention, the aqueous pollen extract is obtained from a mixture of:

- from 45% to 90% of the aqueous extract of Secale cereale L. pollen by weight of the total weight of the extract;

- from 1% to 35% of aqueous extract of pollen of Zea mays L. by weight of the total weight of the extract;

- from 0.01% to 5% of aqueous extract of Pinus sylvestris L. pollen by weight of the total weight of the extract;

- from 3% to 30% of aqueous extract of pollen of Dactylis glomerata L. by weight of the total weight of the extract.

Even more preferably, the composition used in accordance with the invention further contains from 0.1% to 10% by weight of an aqueous extract of Zea mays L. pistil, based on the total weight of the extract.

As a non-limiting example of a method for producing an aqueous extract of pollen and, more advantageously, pistil obtained from plants belonging to the family Pinaceae and/or Poaceae , the method may be referred to as a method which includes the sequential steps:

a) water extraction of pollen;

b) aqueous extraction of pollen and, more advantageously, pistil;

c) spray drying the extracts obtained in steps a) and b); and

d) extracting said pollen and pistil extracts of the plant(s) obtained in step c).

The applicant was able to demonstrate that the extraction step was particularly sensitive. Therefore, the extraction temperature must be strictly below 45°C. Above this temperature, one or more of the pollen and possibly the pistils described above are no longer present. The quality of the composition and/or its effectiveness is therefore not guaranteed.

Preferably, the extraction temperature should be less than 42°C.

The duration of the extraction step for each extract is preferably at least 6 hours, preferably at least 10 hours, even more preferably at least 12 hours.

In addition, the applicant was able to demonstrate that excessive mixing resulted in degradation of the extract(s). Thus, when extraction is combined with stirring, the latter should not exceed 6000 revolutions per minute (rpm), preferably 4500 rpm, more preferably 2800 rpm.

Preferably, the method for producing an aqueous extract of pollen and, more advantageously, pistil, according to the invention includes the following steps:

a) extraction with water and/or a water-acetone mixture of the pollen of Secale cereale L., Zea mays L., Pinus sylvestris L. and Dactylis glomerata L., at a temperature below 45°C for at least 6 hours, to obtain the first extract;

b) extraction with water and/or a water-acetone mixture of pollen and preferably Zea mays L. pistil at a temperature below 45°C for at least 6 hours to obtain a second extract;

c) mixing the first and second extracts obtained in a) and b);

d) spray drying the mixture obtained in step c); And

e) recovering the mixture of pollen and preferably pistil extracts obtained in step d).

The process developed in this way ensures good traceability of the pollen and pistils used.

The composition used according to the invention also includes an aqueous extract of seeds of plants belonging to the genus Cucurbita.

Cucurbita is a genus of climbing or climbing plants belonging to the family Cucurbitaceae. It consists of 12-14 species, most of which are grown under the name pumpkin. The three main domesticated species are Cucurbita pepo, Cucurbita maxima and Cucurbita moschata .

The seed extract of plants belonging to the genus Cucurbita used in accordance with the invention is an aqueous extract containing fatty acids in an amount of less than 10%, for example between 5 and 8% by weight of the total weight of the extract.

Pumpkin seeds contain, in particular, active substances known for their diuretic effect, such as unsaturated fatty acids, sterols, vitamins and zinc.

Preferably, the composition used in accordance with the invention comprises an aqueous extract of the seeds of Cucurbita pepo, Cucurbita maxima and Cucurbita moschata, alone or as a mixture.

More preferably, the extract used in the composition of the invention is an aqueous extract of pumpkin seeds ( Cucurbita pepo).

Once collected, the seeds are preferably dried and then optionally ground.

According to a preferred embodiment of the invention, the aqueous extract of Cucurbita seeds is obtained from pumpkin seeds ( Cucurbita pepo ), dried, then crushed and extracted by maceration at room temperature in water in a ratio of between 10:1 and 50:1. for example 20:1, 30:1 or 40:1.

Preferably, the aqueous extract of Cucurbita seeds used in accordance with the invention is an extract of Cucurbita pepo containing fatty acids in an amount of less than 10%, for example 5 to 8% by weight of the total weight of the extract. More preferably, the aqueous extract of Cucurbita seeds used according to the invention is an extract containing carbohydrates, fatty acids in an amount of less than 10%, for example 5 to 8%, proteins, fiber and sodium.

According to a particularly preferred embodiment, the aqueous extract of Cucurbita pepo seeds used according to the invention contains 52% carbohydrates, 6% fatty acids, 24% proteins, 14% fibers and 0.1%/0.25% sodium/sodium chloride.

The composition used according to the invention also contains vitamin E or its esters.

Vitamin E is a fat-soluble vitamin providing eight organic molecules, four tocopherols (α-tocopherol, β-tocopherol, γ-tocopherol and δ-tocopherol) and four tocotrienols (α-tocotrienol, β-tocotrienol, γ-tocotrienol and δ-tocotrienol ). The most biologically active form is α-tocopherol. The most common form in the diet is γ-tocopherol. These molecules are present in large quantities in vegetable oils. They act, along with vitamin C and glutathione, primarily as antioxidants against reactive oxygen derivatives.

Vitamin E esters refer to vitamin E that has been chemically stabilized to prevent, for example, any oxidation problems. In this form, it does not have antioxidant capacity. Passing through the body, tocopheryls are hydrolyzed, converted into tocopherols and become active. Examples of a vitamin E ester may include the acetate or succinate form, such as D-alpha tocopheryl acetate.

Preferably the composition used in accordance with the invention includes:

- from 5 to 40% wt. from the total weight of the composition of the aqueous pollen extract;

- from 20 to 80% of the mass. from the total weight of the composition of the aqueous extract of Cucurbita seeds; And

- from 0.1 to 10% wt. from the total weight of the composition of vitamin E or its esters.

More preferably, the composition contains:

- from 10 to 30% wt. based on the total weight of the aqueous pollen extract composition, for example 10%, 15%, 20%, 25%, 30% by weight;

- from 30 to 50% by weight of the total weight of the composition of the aqueous extract of Cucurbita seeds; for example 30%, 35%, 40%, 45%, 50%; And

- from 1 to 5% wt. by weight based on the total weight of the composition of vitamin E or its esters, for example 1%, 2%, 3%, 4%, 5%.

The composition used in accordance with the invention also includes a physiologically acceptable vehicle, proportional to the acceptable benefit/risk ratio, containing known excipients commonly used in herbal medicine, such as binders, disintegrants, fillers, dispersing agents, agglomerating agents, lubricants, wetting agents, surfactants, emulsifiers, thickeners, flow agents, flavorings, sweeteners, colors, film formers, stabilizers and/or preservatives, preferably free of nanoparticles.

One skilled in the art should take care to select these optional excipients and their amounts so that they do not affect the beneficial properties of the compositions used in accordance with the invention.

Examples of excipients may include cellulose, preferably microcrystalline cellulose, and silica.

Moreover, preferably, the composition used in accordance with the invention may include another active agent selected from manganese or alternative extracts of Betula alba, Cerasus avium, Equisetum arvense, Phaseolus vulgaris, Achillea millefolium, Agropyron repens, Galium verum, Lavandula officinalis, Mentha piperita , and Urticaria dioica .

The addition of at least one of these other active agents, preferably manganese, will enhance the effect of the composition used according to the invention.

The composition advantageously contains from 0.1 to 10% by weight of the total weight of the composition of one of these active elements, taken individually or in combination, for example manganese.

The composition used in accordance with the invention is preferably in a form suitable for oral administration, in one or more identical or different formulations. It is provided in any herbal form commonly used for oral administration, such as capsule, tablet, liquid capsule, soft capsule, dragee, sachet, tube, bottle, chewing gum, pellet, emulsion, suspension, liquid, solution, ampoule, drink, syrup, powder, solid form, soft gel, semi-solid form.

Advantageously, the composition is in the form of a tablet, capsule, soft capsule, soft gel, semi-solid form, solid form, liquid form or powder.

Preferably, the daily dose of the composition used according to the present invention is in the range of 360 mg to 1000 mg, preferably 600 to 1000 mg, more preferably 600 to 800 mg. This daily dose is preferably administered in 1, 2 or 3 doses (morning, afternoon and/or evening), for example in the form of 1, 2, 3, 4 or 6 tablets, preferably in one dose, more preferably over a period of at least at least 2 months, for example 2, 3 or 6 months, preferably 3 months.

The compositions of the invention are used for the treatment and/or prevention of lower urinary tract symptoms (LUTS) associated with the filling, voiding and/or post-voiding phases in women and, in particular, urinary incontinence.

Filling phase symptoms are symptoms felt during the filling phase of the bladder, without distinction between day and night. Preferably these are the following symptoms:

- daytime pollakiuria, that is, increased frequency of urination during the day;

- nocturia, which means the need to urinate, which wakes the patient. Nocturia should be distinguished from “nocturnal voiding frequency,” which refers to the totality of nocturnal voiding from the time the patient goes to bed, but does not include the concept of awakening. The term nocturnal pollakiuria should be reserved for situations where it is desired to describe an abnormal increase in the frequency of nocturnal urination, and hence the concept of a pathological threshold is introduced, defined either by a series of events or by a possible impact in terms of quality of life or discomfort;

- Urinary urgency or urge to urinate is a sudden, overwhelming and often uncontrollable urge to urinate. This term refers to the expression of a sudden, irresistible urge to urinate that is difficult or impossible to put off. This need is abnormal in its cruelty and intensity. Often accompanied by only moderate or even small amounts of urine. It differs from the normal development of the need from the feeling of a full bladder to the feeling of wanting to pass urine when the bladder is full. Normal physiological need is a precursor to urination, which can be postponed for a certain time to satisfy social conveniences and environmental restrictions;

- Urinary incontinence is the involuntary loss of urine. The term “urinary incontinence” should be defined according to: the mechanism and circumstances surrounding the occurrence of leaks, their severity, their frequency, the presence of any contributing factors, their social, hygienic or quality of life impact, the presence of any measures taken to avoid leaks and, finally, the concept of a medical request from a patient;

- stress urinary incontinence (SUI) - involuntary leakage of urine during exercise, coughing and sneezing;

Urinary incontinence caused by urge incontinence (or urge) is the involuntary leakage of urine accompanied or immediately preceding urge;

- mixed urinary incontinence (SUI) is the involuntary leakage of urine associated with urgency, as well as involuntary leakage of urine during exercise, coughing or sneezing;

- Overflow incontinence is leakage due to a person's inability to completely empty the bladder. This leads to frequent and recurring urine leaks;

- enuresis - involuntary urination. The term nocturnal enuresis describes enuresis that occurs during sleep;

- chronic urinary incontinence - constant leakage of urine;

- bladder sensitivity:

normal: the patient describes a gradually increasing need to urinate until he feels an urgent need;

increased: the patient describes a very early and persistent need to urinate;

decreased: the patient feels an increase in the volume of the bladder, but does not feel the need to urinate;

absent: the patient does not feel any sensations.

Various symptoms relate to the voiding phase:

- weakness of the stream: the patient feels a decrease in the strength of the urine stream when urinating;

- stream like a watering can;

- intermittent stream: urination was interrupted one or more times;

- fluctuating flow: delayed onset of urination;

- urination with a push: a stream of urine obtained with a concomitant abdominal push into the stomach;

- terminal drops, pulling urination: gradual and slow completion of urination, which ends with dripping.

Post-void phase symptoms are the symptoms that the patient experiences immediately after finishing urination:

- feeling of incomplete emptying of the bladder - a subjective feeling that the bladder has not emptied completely after urination;

- Retained drip is the involuntary loss of urine that occurs immediately after urination, most often when leaving the toilet for a man or when getting up from the toilet for a woman.

Dyspareunia, vaginal dryness and urinary incontinence are the main symptoms described during sexual intercourse and can also be treated with the compositions used in accordance with the invention.

Many symptoms can be alleviated in patients due to urogenital prolapse, such as: a feeling of heaviness in the pelvis or endovaginal foreign body, the need to move the vaginal mass to be able to pass urine or have a bowel movement, and can also be treated.

The sensation of pain that can be treated by the compositions of the invention can be characterized by its frequency of occurrence, intensity, duration, the presence of any contributing factors or its location, such as:

- cystalgia, which is a general term for pain felt in the suprapubic or retropubic region. The pain usually worsens when the bladder is full and may persist after urination. This term should be preferred to the term "interstitial cystitis", which corresponds to a pathological condition more precisely defined according to a set of diagnostic criteria;

-pain in the urethra, urethralgia, in which pain is felt in the urethra and for which the patient comes in;

- pain in the vulva, which is pain that is felt strictly at the level of the vulva;

- vaginal pain, which is pain that is felt inside the vaginal cavity outside the vulvar segment;

- pain in the scrotum, the description of which does not indicate a more specific localization in the umbilical cord, testicle, epididymis or even skin;

- perineal pain, which is pain felt in women between the vulva and anus, and in men between the scrotum and anus;

- pelvic pain, which is pain that is less clearly defined in terms of its exact location, is more diffuse and is not directly related to the filling-void cycle of the bladder.

Preferably, the compositions according to the invention are used for the treatment and/or prevention of urinary incontinence in women.

In particular, they are used for the treatment and/or prevention of stress urinary incontinence (SUI), urge incontinence (or urge), mixed urinary incontinence (SUI), overflow incontinence, enuresis and/or chronic incontinence even more preferably for the treatment and/or prevention of stress urinary incontinence (SUI), urge incontinence (or urge incontinence), mixed urinary incontinence (SUI), including, in particular, reducing the frequency of nighttime urination that causes awakening.

More preferably, compositions according to the invention comprising a pollen extract consisting of an aqueous extract of pollen obtained from a mixture of plants belonging to the families Pinaceae and Poaceae, an aqueous extract of the pistil of Zea mays L. , an aqueous extract of seeds of plants belonging to the genus Cucurbita, and vitamin E or its esters are used in the treatment of urinary incontinence, particularly stress urinary incontinence (SUI), urge (or urge) incontinence, and mixed urinary incontinence (SUI), particularly reducing the frequency of nighttime awakening urination.

It is known that the functions of the lower urinary tract, which consist in the periodic accumulation and drainage of urine, depend on the activity of the smooth and striated muscles of the bladder, urethra and external urethral sphincter.

While urine is stored, the outlet is closed and the smooth muscle of the bladder is at rest. When bladder capacity reaches the voiding threshold, activation of the pontine micturition activation center causes contraction of the bladder and reciprocal relaxation of the urethra, resulting in emptying of the bladder.

During voiding, the parasympathetic sacral (pelvic) nerves have an excitatory (cholinergic and purinergic) effect on the bladder through the pelvic nerves and an inhibitory (nitrergic) effect on the urethra. These peripheral systems are united by excitatory and inhibitory regulation of the spinal cord and brain. Injury or disease of the nervous system, as well as medications and diseases of peripheral organs can cause dysfunction of the lower urinary tract.

In the case of overactive bladder (OAB), therapeutic targets promoting urine storage may be found in the urothelium, detrusor muscle, autonomic and afferent pathways, spinal cord, and brain.

The urothelium appears to have specialized sensory and signaling properties, including: (1) expression of nicotinic, muscarinic, tachykinin, adrenergic, bradykinin receptors and transient receptor potential (TRP), (2) close physical connection with afferent nerves, and (3) the ability to release chemical molecules such as adenosine triphosphate (ATP), acetylcholine and nitric oxide.

Increased expression and/or sensitivity to such urothelial sensory molecules that lead to afferent sensitivity may be responsible for OAB. Targeting afferent pathways and/or bladder smooth muscle by modulating the activity of receptors (eg, neurokinin, ATP, or β3-adrenergic) and ion channels (eg, TRPV1 or K) may be effective in suppressing OAB.

In the case of stress urinary incontinence (stress), pharmaceutical therapy targeting the neuron-mediated urethral continence reflex during stressful conditions such as sneezing or coughing may be effective in increasing output resistance. Therapeutic targets include adrenergic and serotonergic receptors in the spinal cord, as well as adrenergic receptors in the urethral sphincter, which may correspondingly enhance urethral reflex activity during stressful conditions and increase baseline urethral pressure.

Thus, the compositions of the invention used for the treatment and/or prevention of urinary incontinence may act at the level of the nervous system, in particular the afferent pathways and smooth muscles, acting, for example, on one or more of such molecular targets in the urothelium, urethra and bladder.

The compositions are also used for the treatment and/or prevention of genitourinary pain syndromes and clinical syndromes indicative of lower urinary tract dysfunction.

Syndromes are a collection of different symptoms, but they are not necessarily pathognomonic of a medical condition. The term "syndrome" can only be used when multiple symptoms are present in the same patient. The various syndromes treated correspond to functional disorders for which no specific cause has been identified, it being understood that conventional research methods have eliminated obvious local pathology associated with the life of the organism, such as, for example, infection, tumor or fluid-electrolyte disturbance.

Urogenital pain syndromes are all chronic. Pain, which is the predominant complaint, may be associated with other symptoms relating to the lower urinary tract, genital area or digestive tract:

- Bladder pain syndrome is an increase in suprapubic pain when filling the bladder, associated with daytime or nighttime pollakiuria in the absence of urinary tract infection or bladder tissue pathology;

- urethral pain syndrome is recurrent pain in the urethra, occurring predominantly during urination in the absence of a urinary tract infection or bladder tissue pathology;

- vaginal pain syndrome is constant or intermittent vaginal pain that may vary depending on the urinary cycle or may be associated with symptoms indicating lower urinary tract dysfunction or sexual dysfunction;

- vaginal pain syndrome is constant or recurrent vaginal pain associated with symptoms indicating dysfunction of the lower urinary tract or sexual dysfunction, in the absence of proven vaginal infection or any other obvious vaginal pathology;

- scrotal pain syndrome is constant or recurrent pain in the scrotum associated with symptoms indicating dysfunction of the lower urinary tract or sexual disorders, in the absence of inflammation of the ovary or any other local pathology;

- perineal pain syndrome is constant or recurrent pain in the groin associated with symptoms indicating dysfunction of the lower urinary tract or sexual dysfunction, in the absence of proven infection or any other obvious pathology of the perineum;

- Pelvic pain syndrome is persistent or recurrent pelvic pain associated with symptoms indicating dysfunction of the lower urinary tract, digestive system or sexual dysfunction, in the absence of proven infection or any other obvious pathological disease.

In clinical practice, an empirical diagnostic approach is often used to decide how to manage a patient with symptoms suggestive of lower urinary tract dysfunction:

- “clinical overactive bladder syndrome” or “urgent pollakiuria syndrome” is defined by the occurrence of urge with or without urinary incontinence, usually associated with pollakiuria or nocturia. This syndrome is indicative of detrusor overactivity, confirmed by urodynamic testing, but is not specific as it may also be associated with other types of lower urinary tract dysfunction. In addition, the term “clinical overactive bladder syndrome” presupposes the absence of urinary tract infection or obvious local pathology associated with the life of the body (tumors, etc.);

- "dysuric syndrome", indicating obstruction under the bladder, should be used in men with predominant voiding disorders and in the absence of urinary tract infection or obvious local pathology. In women, dysuria should indicate an overactive bladder and not subbladder obstruction.

Examples

The present invention will be further illustrated by the following examples.

The compositions in accordance with the present invention contain a pollen extract containing (and preferably consisting of) an aqueous extract of pollen, an aqueous extract of Cucurbita seeds containing less than 10% fatty acids and vitamin E or its esters, and preferably, in addition, an aqueous Zea mays L pistil extract is prepared in accordance with methods generally known and used by those skilled in the art.

Example 5 : Study of urinary incontinence activity in women

The study was conducted on 160 patients aged 18 to 75 years (mean age 60.35 years) suffering from LUTS, in particular urinary incontinence.

The dropout rate was 21.875% due to physician recommendation, change in treatment, side effects, lack of specific cause and loss to follow-up (9 patients).

Among the patients participating in the study, treatment compliance was very satisfactory; the tablets were taken at the correct time and in the correct form.

Results were obtained after once daily administration for at least 2 months (and up to 3 months) of composition C (formulation according to Example 1) and were compared with administration under the same conditions:

- compositions A containing 120-200 mg of aqueous extract of pollen P2 and 5-10 mg of oily extract of pollen EA5 (=GRAMINEX®), 100-300 mg of aqueous extract of pumpkin seeds and 16-30 mg of D-alpha-tocopheryl acetate;

- composition B containing 120-200 mg of aqueous extract of pollen P2 and 5-10 mg of oily extract of pollen EA5 (= GRAMINEX®); or

- placebo (tablet containing microcrystalline cellulose, isomalt, magnesium stearate, silicon dioxide and coating agent (Aqua Polished Gray 089.45 MS)).

The effectiveness of the various formulations was assessed using validated questionnaires, namely the SANDVIK, MUH (Measurement of Urinary Disorders) and ICIQ (International Consultation on Incontinence Questionnaire) scales.

The severity assessment developed by SANDVIK (Incontinence Severity Index; Sandvik H, Hundskaar S et al. Validation of a severity index in female urinary incontinence and its implementation in an epidemiological survey. J Epidemiol Community Health 1993; 47: 497-99) makes it possible classify the severity of urinary incontinence into four stages (minimal, moderate, severe, very severe).

MUH (Measurement of Urinary Handicap) scale published by Amarenco in 1992 (Amarenco G, Kerdraon J, Perrigot M. Pelvic handicap evaluation scale: measurement of urinary handicap (MUH). In: et anorectale. Edited by Pelissier J, Costa P, Lopez S, Mares P, Masson Ed, 1992, 498-504) provides quantitative measurements of various urinary symptoms. This questionnaire can be used regardless of the type of urinary incontinence. It has 7 questions with a response scale of 0 to 4. It was modified for female non-neurological urinary incontinence by removing the question “other urinary incontinence” and replacing it with a question about the frequency of stress urinary incontinence. The result is a score from 0 to 28 with four parts: urge (0 to 8), stress (0 to 8), pollakiuria (0 to 8), and dysuria (0 to 4).

MUH scale (Measurement of urinary handicap) published by Amarenco in 1992 (Amarenco G, Kerdraon J, Perrigot M. Pelvic handicap evaluation scale: measurement of urinary handicap (MUH). In: et anorectale. Edited by Pelissier J, Costa P, Lopez S, Mares P, Masson Ed, 1992, 498-504) provides quantitative measurements of various urinary symptoms. This questionnaire can be used regardless of the type of urinary incontinence. It has 7 questions with a response scale of 0 to 4. It was modified for female non-neurological urinary incontinence by removing the question “other urinary incontinence” and replacing it with a question about the frequency of stress urinary incontinence. The result is a score ranging from 0 to 28, with four intermediate scores: urge (0 to 8), stress (0 to 8), pollakiuria (0 to 8), and dysuria (0 to 4).

ICIQ scale (International Consultation on Incontinence Questionnaire; Avery K, Donovan J, Peters TJ et al. (2004) International Consultation on Incontinence Questionnaire (ICIQ): a brief and robust measure for evaluating the symptoms and impact of urinary incontinence. Neurourol Urodyn 2004 ; 23: 322-30.) is one of the most recent questionnaires developed for women to assess urinary incontinence (stress, urge, or mixed). It has only four parameters. Of these, three assess urinary incontinence as a symptom (frequency, amount and circumstances of incontinence), and the fourth item measures the impact on quality of life on a visual analogue scale.

Specifically, the results obtained before treatment (D0), after 60 days of treatment (D60) and after 90 days of treatment (D90) were measured using these different scales and are compiled in Table 5 below. The results thus obtained are also shown in FIG. 1-3.

In these figures 1 to 3, as well as in figures 4 to 9, the following references are made:

a-s. Two by two comparison. Within the same group, different lowercase letters indicate statistically significant differences between each time (Bonferroni correction).

A-B. Two by two comparison. For the same given time, different capital letters indicate statistically significant differences between groups (Bonferroni correction).

The results of the study with the three questionnaires analyzed (SANDVIK, MUH and ICIQ) show that the three groups of patients who received compositions A, B or C, respectively, showed a statistically significant improvement compared to the placebo group.

Among the three compositions used, it appears that composition C (according to Example 1 in accordance with the present invention) is the one that brought the greatest improvement in patients, followed by the treatment group with composition A, then finally with composition B.

For example, after 60 days of treatment with Composition C in accordance with the present invention, an average reduction in urinary severity and/or symptoms of about 46.55% (SANDVIK score), 68.14% (MUH score) and 59.44% (ICIQ score) was obvious in treated patients.

In comparison, the average reductions obtained with compositions A and B, respectively, were only 42.05% and 38.07% (SANDVIK estimate), 52.22% and 37.68% (MUH estimate), and 44.73% and 32.42% (ICIQ estimate).

After 90 days of treatment, the average reduction with composition C according to the invention is respectively 61.68% (SANDVIK), 74.9% (MUH) and 64.14% (ICIQ).

In comparison, the average reductions obtained with compositions A and B, respectively, are only 54.8% and 48.22% (SANDVIK estimate), 57.07% and 45.71% (MUH estimate), and 52.73% and 35.06% (ICIQ estimate).

These results are consistent with a reduction in the severity of urinary incontinence, initially classified as very severe, severe and moderate, and identified at the end of treatment with a significant percentage of mild cases and even patients who are no longer incontinent and who have fully recovered.

These results demonstrate the effectiveness of the treatment, more specifically with composition C (Example 1 in accordance with the present invention), with maximum effectiveness after 60 days, the positive effect is maintained 90 days after the start (at the time of the last visit).

More specifically, the results obtained by type of urinary incontinence for each of the tested compositions A to C, respectively, are illustrated in FIG. 4-6 with ICIQ scale, fig. 7-9 with MUH scale and FIG. 10 with the SANDVIK scale for composition C in accordance with the present invention.

According to the results obtained, according to a preferred embodiment of the invention, urinary incontinence caused by urge incontinence (or urge) and mixed urinary incontinence, in particular, are statistically significantly improved after 60 days.

For example, as illustrated in FIG. 6, after 60 days of treatment with composition C according to the invention, there was an average reduction in the severity and/or symptoms of urination with an ICIQ score of 36.87% for stress urinary incontinence, 62.68% for urge urinary incontinence and 75.69% for mixed urinary incontinence , with scores of 40.28%, 64.49% and 80.4% respectively after 90 days of treatment.

Likewise, as shown in FIG. 9, after 60 days of treatment with composition C according to the invention, there is an average reduction in urinary severity and/or symptoms with an MUH score of 70.66% for stress urinary incontinence, 64.15% for urge urinary incontinence and 72.67% for mixed urinary incontinence, the scores were 82.66%, 70.53%, and 75.47%, respectively, after 90 days of treatment.

As a result, as shown in FIG. 10, from the assessment obtained using the SANDVIK scale, it is possible to observe a significant improvement in the severity of urinary incontinence in patients treated with composition C according to the invention: for example, we go from 39.4% to very severe (2.6%) and severe (36.8%) cases to 8.4% (4.2% + 4.2%) on D60 with the appearance of 54.2% of mild cases and even 4.2% of fully recovered patients or even 45.8% ( lungs) and 20.8% (fully recovered), respectively, on D90.

Example 6 : Study of the safety and effectiveness of the composition of the present invention in women with urinary incontinence caused by urge incontinence (or urge) or mixed urinary incontinence (randomized, double-blind, placebo-controlled clinical trial)

The purpose of this study is to determine the response to the composition according to the invention in women diagnosed with overactive bladder (OAB), manifested by urinary incontinence due to urge incontinence (or urge) or mixed urinary incontinence.

In this study, 52 women were divided into two groups. Some were treated with the composition according to the invention (according to example 1 = PSCEP in Fig. 11-13) (n = 26), and others were treated with placebo (n = 26). Effectiveness was assessed using validated questionnaires, namely SANDVIK, MUH (Measurement of Urinary Disability) and ICIQ-UI SF (International Consultation on Urinary Incontinence Questionnaire-Short Form).

No side effects were observed in any of the groups.

results

There were no differences between the two groups regarding demographic characteristics and urinary incontinence.

As shown in FIG. 11, after 90 days using ICIQ-UI SF, the group receiving the composition according to the invention "PSCEP" showed a significant improvement (p < 0.001) compared to baseline and compared to the placebo group also showed a significant improvement (p = 0.023 ).

As shown in FIG. 12, MUH showed a significant improvement in the group receiving the composition according to the invention "PSCEP" (p < 0.001) compared to baseline. Compared with the placebo group, a significant improvement after 90 days (p = 0.009) was observed in women receiving the composition according to the invention "PSCEP".

As illustrated in FIG. 13, with the Sandvik index there was a significant improvement after 90 days of treatment in the group with the composition according to the invention "PSCEP" compared to the baseline (p < 0.001); however, no statistically significant differences were found compared with the placebo group (p = 0.23).

Example 7 : Study of the safety and effectiveness of the composition according to the invention in women with stress urinary incontinence (randomized, double-blind, placebo-controlled clinical trial)

The purpose of this study is to determine the response to the composition according to the invention in women diagnosed with stress urinary incontinence.

In this study, 24 women were divided into two groups. Some were treated with the composition according to the invention (according to example 1 = PSCEP in Fig. 14-16) (n = 12), and others were treated with placebo (n = 12). Effectiveness was assessed using validated questionnaires, namely SANDVIK, MUH (Measurement of Urinary Disability) and ICIQ-UI SF (International Consultation on Urinary Incontinence Questionnaire-Short Form).

No side effects were observed in any of the groups.

results

As shown in FIG. 14, with ICIQ-UI SF, a significant improvement in the group receiving the composition according to the invention "PSCEP" compared to the placebo group was observed after 3 months (p = 0.048).

As shown in FIG. 15, with MUH there was a significant improvement in the group receiving the composition according to the invention "PSCEP" (p < 0.001) compared to baseline, and a significant improvement after 3 months of treatment compared to the placebo group (p = 0.021).

As shown in FIG. 16, with Sandvik index after 3 months of treatment in the group receiving the composition according to the invention "PSCEP", there was a significant improvement compared to inclusion (p = 0.006). The placebo group showed no significant change at three months (p = 0.102). However, there are no statistically significant differences from the active group (p = 0.225), probably due to the “insufficient” number of patients.

Example 8 : Study of the effectiveness on the frequency of night urination of the composition according to the invention.

In this study, 26 women diagnosed with overactive bladder (OAB) symptoms exhibiting urinary incontinence due to urge incontinence (or urge incontinence) or mixed urinary incontinence were treated with a composition according to the invention (according to Example 1) (n = 26).

“Nocturnal urinary frequency” (or nocturia) refers to the totality of nocturnal urination from the time the patient goes to sleep, taking into account the concept of awakening. Patients included in the study were divided into the following categories:

0 or 1 urination per night (/night);

2 urinations/night;

3-4 urinations/night;

5-6 urinations/night; or

more than 6 urinations/night

results

As illustrated in FIG. 17, the composition according to the invention (according to Example 1) significantly reduces the frequency of nighttime urination in patients with urinary incontinence due to urge incontinence (or urge) or mixed urinary incontinence:

with the composition according to the invention, the percentage of patients who had more than 6 urinations per night at the beginning of the study (enrolment) disappears after 90 days of treatment: thus, no patient falls into this category with more than 6 urinations. /night;

the percentage of those who had 3-4 urinations per night was reduced by 50%;

the percentage of those who had 2 urinations per night was reduced by 55%;

therefore, we then find 3.3 times more patients in the patient group who had 0-1 voids per night.

Example 9 : Study of the activity of the composition according to the invention in relation to the M3 muscarinic receptor.

Acetylcholine (ACh) is a neurotransmitter released by nerve cells to transmit signals to other cells, that is, neurons, muscle cells and glandular cells. Acetylcholine works by binding to receptors on the surface of these cells and activating them. There are two main classes of acetylcholine receptors, nicotinic and muscarinic, which are found in different tissues. In the bladder, the action of acetylcholine depends on the presence of muscarinic receptors, which are located on the smooth muscles of the detrusor muscle. Of the 5 types of muscarinic receptors called M1, M2, M3, M4 and M5, M2 and M3 are present in the bladder. M2 predominates in quantity, and M3 is primarily the mediator of direct contractile responses to acetylcholine. When a healthy bladder contracts, acetylcholine is released from the cholinergic nerves and activates the M3 receptor in the detrusor muscle, resulting in voiding of urine accompanied by relaxation of release.

However, in overactive bladder (OAB), either more acetylcholine is released during normal bladder distension, or the acetylcholine receptors in the detrusor muscle are more sensitive. This leads to increased contractility of the bladder. For overactive bladder, pharmacological treatment is direct treatment, such as administration of muscarinic receptor antagonists that act directly on M3 receptors.

The purpose of the study is to test the activity of the composition according to the invention (according to example 1) on the muscarinic receptor M3 using HEK293 cells that express the receptor endogenously (Crittenden C. et al., 2012, poster).

The composition according to the invention was tested separately, as well as each active ingredient of the composition (according to Example 1), taken independently.

Compounds were tested using a benchmark Gq GPCR compound profiling technology: called FLIPR Tetra, using the Screen Quest™ Fluo-8® No Wash Calcium Assay Kit (Euromedex).

Carbacholine was used as a control agonist, and intracellular calcium increases were assessed using FLIPR Tetra technology as a readout for GPCR activation.

The robustness of the assay was confirmed by determining the affinity of carbachol in various assays. Dose-response curves of carbacholin (10 concentrations) were constructed during preliminary experiments, and the calculated EC50 value of carbacholin was used to test the antagonistic activity of the composition according to the invention and each active ingredient taken independently.

4-DAMP was used as a control M3 receptor antagonist. The measured IC50 value is 0.26 nM or 0.00183 μg/ml, as described previously (Dörje et al. , 1991).

Positive (carbacholin) and negative (4-DAMP or HBSS) controls were included in each trial to standardize data, assess assay reliability, and monitor performance.

HEK293 cells were plated on clear poly-D-lysine flat support, 96 black 96-well plates, incubated overnight at 37°C with 5% CO ₂ . The next day, the plates were washed, the medium was replaced with 50 μl of saline buffer (HBSS), and the cells were loaded with Fluo-8 NW calcium dye (Molecular Devices). Fluo-8 NW dye was used according to the manufacturer's instructions: dissolved in DMSO, then mixed in assay buffer (1X) consisting of 10 ml 10X Pluronic® F127 Plus (Molecular Devices) in 90 ml HBSS buffer.

The dye solution (110 μl) was added to the cell plates and incubated at room temperature for 1.5 hours. Fluorescence was then measured using a FLIPR Tetra plate analyzer. During the first 10 seconds, the baseline was read. After 10 seconds, 20 μl of different compounds were tested in 10 different doses and added to the wells, and fluorescence was monitored for 3 minutes at Ex/Em = 490/525 nm.

A second baseline was then measured for 10 seconds, then 20 μl of carbachol (agonist control) was added to the wells and fluorescence was monitored for 3 minutes at Ex/Em = 490/525 nm.

results

The dose-response curves of the composition according to the invention (according to Example 1) for each of the active ingredients tested independently and the control antagonist were normalized. The IC50 data is shown in the table below and shown in FIG. 18:

The results show that the IC50 value for the composition according to the invention is significantly lower than the IC50 value of each of the active ingredients tested independently.

Thus, according to these results, significant antagonistic activity against M3 is observed with the composition according to the invention, demonstrating synergy between the various active ingredients of the composition according to the invention, while each of the active ingredients, taken independently, has no (or very little) antagonistic activity on M3; these M3 activity results also provide a possible indication of a potential mechanism of action.

Claims (14)

1. The use of a composition comprising a pollen extract consisting of an aqueous pollen extract obtained from a mixture of plants belonging to the family Pinaceae and Poaceae , in which the aqueous pollen extract is obtained from a mixture of: - from 45% to 90% of the aqueous extract of Secale cereale L. pollen by weight of the total weight of the extract; - from 1% to 35% of aqueous extract of pollen of Zea mays L. by weight of the total weight of the extract; - from 0.01% to 5% of aqueous extract of Pinus sylvestris L. pollen by weight of the total weight of the extract; - from 3% to 30% of aqueous extract of pollen of Dactylis glomerata L. by weight of the total weight of the extract, an aqueous extract of seeds of plants belonging to the genus Cucurbita , and vitamin E or its esters for the production of medicaments for the treatment and/or prevention of lower urinary tract symptoms (LUTS) associated with the filling, voiding and/or post-micturition phases in women, wherein Lower urinary tract symptoms (LUTS) associated with the filling, voiding, and/or postvoiding phases in women constitute urinary incontinence. 2. Use according to claim 1, characterized in that the aqueous extract of Cucurbita seeds contains fatty acids in an amount of less than 10% by weight of the total weight of the extract. 3. Application according to claim 2, characterized in that the aqueous extract of Cucurbita seeds contains fatty acids in an amount of 5 to 8% by weight of the total mass of the extract. 4. Use according to any of the preceding claims, characterized in that the composition additionally contains an active agent selected from an aqueous extract of the pistil of Zea mays L. , manganese, extract of Betula alba, Cerasus avium, Equisetum arvense, Phaseolus vulgaris, Achillea millefolium, Agropyron repens, Galium verum, Lavandula officinalis, Mentha piperita, Urticaria dioica . 5. Use according to claim 4, characterized in that the composition additionally contains an aqueous extract of Zea mays L. pistil. 6. Use according to any of the preceding claims, characterized in that the composition is in a form suitable for oral administration. 7. Use according to claim 6, characterized in that the composition is in the form of a tablet, capsule, soft capsule, soft gel, soft dosage form, semi-solid, solid dosage form, liquid dosage form or powder. 8. Use according to claim 1, wherein urinary incontinence in women is stress urinary incontinence, urinary incontinence caused by urge incontinence (or urge), and mixed urinary incontinence. 9. Use according to claim 8 to reduce the frequency of night urination, which causes awakening.