Use of TGF-β for the prevention and treatment of neuralgic manifestations

Abstract

The present invention relates to a novel use of TGF-β for the prevention and / or treatment of manifestations of the neuralgic component of disorders such as pain and itching.

Description

Title of the invention: Use of TGF-p for the prevention and treatment of neuralgic manifestations

[0001] The present invention relates to a novel use of TGF-[3 for the prevention and / or treatment of manifestations of the neuralgic component of disorders such as pain and itching.

[0002] TGF-[3 is known for its anti-inflammatory properties. However, the Applicant has demonstrated that this compound surprisingly allows pathologies to be treated more effectively than conventionally used anti-inflammatory drugs (steroids and non-steroidal anti-inflammatory drugs, NSAIDs); this increased effectiveness is manifested by (i) a broader spectrum of action, TGF-[3 does not only act on inflammatory manifestations but also on symptoms which do not necessarily result from inflammatory phenomena such as neuralgia and itching; in addition and very advantageously, this effectiveness is also manifested (ii) without producing any adverse effects, unlike anti-inflammatory drugs.

[0003] The Applicant has in particular demonstrated this effectiveness for the alleviation of symptoms in people suffering from dysmenorrhea, premenstrual syndrome, premenstrual dysphoric disorder, as well as for the relief of itching, particularly in people suffering from chronic kidney disease, in particular in hemodialysis patients.

[0004] Patients with chronic kidney disease are particularly susceptible to drug side effects due to impaired drug clearance and impaired kidney function.

[0005] However, when treating such patients with anti-inflammatory drugs, whether steroids (such as corticosteroids) or non-steroids (such as NSAIDs), it is important to take into account the following side effects:

[0006] - Corticosteroids are often associated with:

[0007] *fluid retention which can lead to edema and exacerbate hypertension, which is a common complication of chronic kidney disease;

[0008] *an electrolyte imbalance: in particular, hyperkalemia and hypocalcemia, which can respectively aggravate the cardiac and bone fragility of patients;

[0009] *glucose intolerance which can exacerbate or precipitate diabetes, another major risk factor for the progression of chronic kidney disease;

[0010] *an increased risk of infection: this results from immunosuppression, which poses a significant problem in patients with chronic renal failure who are already vulnerable;

[0011] *a loss of bone density which can lead to osteoporosis and an increased risk of fractures;

[0012] *gastrointestinal ulcers which are more likely to occur with concomitant use of NSAIDs.

[0013] - nonsteroidal anti-inflammatory drugs (NSAIDs) are often associated with:

[0014] *nephrotoxicity: NSAIDs can cause acute kidney injury, reduce glomerular filtration rate (GFR) or even exacerbate chronic kidney disease due to hemodynamic changes in the kidneys and direct toxicity;

[0015] *fluid retention which, as with corticosteroids, can lead to edema and exacerbation of hypertension;

[0016] *hyperkalemia, or high potassium levels pose a cardiac risk;

[0017] *gastrointestinal problems such as ulcers, hemorrhages or perforations, particularly with prolonged use;

[0018] Have cardiovascular risks such as heart attack or stroke, especially with prolonged use.

[0019] Furthermore, anti-inflammatory drugs, whether steroids or non-steroids (nonsteroidal anti-inflammatory drugs, NSAIDs), are commonly used to treat conditions such as dysmenorrhea and, to a lesser extent, premenstrual syndrome and premenstrual dysphoric disorder. Although they can be effective, they have side effects that are quite harmful for this type of patient:

[0020] - as for corticosteroids, they can induce:

[0021] *weight gain: chronic steroid use can lead to increased appetite and water retention, resulting in weight gain;

[0022] *mood changes: patients may experience mood swings, anxiety, or even depression;

[0023] *thinning of bones (osteoporosis): long-term use may decrease the bone density, which increases the risk of fracture;

[0024] *skin alteration, in particular, thinning of the skin, bruises and acne development is common;

[0025] Have increased risk of infections: Steroids suppress the immune system, making the body more vulnerable to infections;

[0026] *increased blood sugar levels: this situation can be worrying for diabetic patients or those at risk of developing it;

[0027] *the occurrence of cataracts or glaucoma: prolonged use can affect eye health.

[0028] - as for NSAIDs, they can be associated with:

[0029] Have gastrointestinal problems: These problems can range from mild indigestion to severe ulcers and bleeding. NSAIDs reduce the stomach's protective mucus, making it more sensitive to acid.

[0030] *an increase in blood pressure: NSAIDs can cause water retention, affect kidney function and increase blood pressure;

[0031] Kidney damage: Prolonged use may impair kidney function;

[0032] *liver damage: although less common than kidney problems, NSAIDs may also affect liver function;

[0033] Have allergic reactions: skin rash, wheezing and throat swelling may occur;

[0034] *an increased risk of heart attack or stroke: this is particularly the case with prolonged use or in people suffering from pre-existing heart problems;

[0035] Have bleeding problems: NSAIDs can interfere with platelet function, leading to increased bleeding.

[0036] It therefore appears critical to be able to propose therapeutic alternatives that are both more effective than those currently available and produce few or no adverse effects.

[0037] This is what the Applicant has achieved by highlighting in the context of clinical trials the very advantageous and unexpected effects of TGF-[3.

[0038] Indeed, due to its anti-inflammatory activity, TGF-[3 acts as existing anti-inflammatory drugs would without causing the undesirable effects of these drugs; in addition, its spectrum of action is more advantageous since it is also capable of acting on neuronal manifestations.

[0039] Transforming growth factor beta (TGF-|3) is a family of cytokines involved in various biological processes. The three main isoforms in mammals are TGF-[31, TGF-[32 and TGF-[33. Each of them has specific roles, although they share a similar structure and common functions. TGF-[31 is the most abundant isoform, involved in the regulation of inflammation and tissue repair. TGF-[32 is mainly expressed in epithelial and endothelial cells and participates in embryonic morphogenesis. TGF-[32 is the dominant isoform in breast milk, playing a key role in the maturation of the infant's immune system. TGF-[33 is expressed in various tissues and is involved in wound healing.

[0040] TGF-[3s are produced in a latent form, consisting of the mature TGF-[3 protein and the propeptide called latency-associated peptide (LAP). This complex is secreted and stored in the extracellular matrix in a latent form, awaiting activation. Activation of latent TGF-[3 requires dissociation of LAP by various mechanisms, allowing the mature protein to interact with cellular receptors and exert its biological functions. TGF-[3 is highly conserved between mammalian species. The mature proteins of human and bovine TGF-[31 and those of human and bovine TGF-[32 are 100% identical, respectively.

[0041] TGF-[3 present in bovine milk can be concentrated in the different fractions such as a certain fraction of caseins (EP0527283), certain fractions of whey protein concentrate, for example whey protein concentrate mainly consisting of α-lactalbumin (EP1409539) and whey protein concentrate mainly consisting of β-lactoglobulin (EP2144623), and certain cationic milk protein isolates (EP1912513). Certain fractions such as XP828-L or Vitalarmor® GF-100 are commercially available.

[0042] TGF-[3s, in particular TGF-[32, can also be synthesized using biotechnological techniques, for example, recombinant human TGF-[32 by mammalian cell lines (e.g. by CHO cells or Chinese Hamster Ovary), but also by precision fermentation (for example by certain genetically modified yeasts).

[0043] The present invention thus relates to TGF-[3 for its use in the prevention and / or treatment of neuralgic symptoms.

[0044] In particular, the neuralgic symptoms are pain and itching.

[0045] The subject to be treated may be a human or an animal, preferably a mammal.

[0046] According to a particular embodiment, the present invention relates to TGF-[3 for its use for the prevention and / or treatment of neuralgic symptoms in people suffering from dysmenorrhea; preferably, the dysmenorrhea is secondary dysmenorrhea.

[0047] In particular, the neuralgic symptoms of dysmenorrhea include pain in the pelvis or lower abdomen, back pain, muscle pain, diarrhea.

[0048] Dysmenorrhea, or painful periods, is classified into two categories: primary dysmenorrhea and secondary dysmenorrhea. Primary dysmenorrhea is characterized by menstrual pain without identifiable pelvic pathology, while secondary dysmenorrhea is associated with underlying pelvic disorders such as endometriosis or uterine fibroids (Proctor & Farquhar, 2006, BMJ, 332(7550), 1134-1138). Dysmenorrhea typically causes pain in the pelvis or lower abdomen, and other symptoms may include backache, diarrhea, and nausea. Dysmenorrhea is one of the most common symptoms in the gynecology department. The highest frequency of dysmenorrhea occurs at the age of 20 and gradually decreases with age. Dysmenorrhea also has an impact on women's daily lives and quality of work.

[0049] The menstrual cycle, particularly menstruation, can be considered an inflammatory process in itself. This process is governed by a set of immune cells and cytokines that facilitate the shedding and regeneration of the endometrium (Maybin & Critchley, 2015, Human Reproduction Update, 21(6), 748-761). During menstruation, the drop in progesterone levels triggers an inflammatory response, activating immune cells such as macrophages and neutrophils, and leading to the production of pro-inflammatory cytokines such as IL-1, IL-6, and TNF-a. It is likely that these cytokines promote the production of prostaglandins, particularly PGE2 and PGF2a, by endometrial cells, which stimulate uterine contractions and induce pain.

[0050] In conditions such as endometriosis, a common cause of secondary dysmenorrhea, immune dysregulation plays a central role. Ectopic endometrial tissue from endometriosis can recruit macrophages and other immune cells, resulting in a chronic inflammatory state. These immune cells produce excess growth factors, cytokines, and angiogenic factors, contributing to the persistence and proliferation of ectopic endometrial tissue and associated pain (Symons et al., 2018, Trends in Molecular Medicine, 24(9), 748-762).

[0051] While inflammation and immune cells contribute to dysmenorrhea, non-inflammatory mechanisms are also involved. The pain of dysmenorrhea may be related to increased uterine activity and ischemia caused by uterine contractions. Elevated levels of vasopressin, a potent vasoconstrictor, have been found in women with primary dysmenorrhea, suggesting a role in causing uterine hypoxia and pain (Dawood, 2008, Global Women's Medicine).

[0052] Secondary dysmenorrhea is often associated with underlying pathological conditions, including endometriosis and uterine fibroids. Although these conditions can induce a chronic inflammatory state, the pain associated with them can occur in the absence of active inflammation (Proctor & Farquhar, 2006).

[0053] Several non-inflammatory mechanisms contribute to dysmenorrhea:

[0054] * Uterine hyperactivity and ischemia: uterine contractions are part of menstruation normal, but in cases of dysmenorrhea, these contractions may be stronger, last longer, and occur more frequently. This can increase intrauterine pressure and reduce blood flow, causing ischemia and pain (Dawood, 2008).

[0055] * Nerve compression and sensitization: expanding cysts or fibroids can compress nerves and cause pain. In addition, nerve fibers in the ectopic endometrial tissue of patients with endometriosis can become sensitized, resulting in chronic pelvic pain.

[0056] * Psychological stress: Stress and other psychological factors can exacerbate pain perception and physiological responses to pain in dysmenorrhea (Gagua et al., 2013, Gynecology, 26(6), 350-354.).

[0057] Nonsteroidal anti-inflammatory drugs (NSAIDs) or oral contraceptives (birth control pills) are often used in the treatment of dysmenorrhea. However, the effectiveness of these drugs is not satisfactory in the long term and they have undesirable toxic side effects on metabolism.

[0058] According to another particular embodiment, the present invention relates to TGF-[3 for its use for the prevention and / or treatment of neuralgic symptoms in people suffering from symptoms of premenstrual syndrome (PMS).

[0059] Symptoms of premenstrual syndrome include, in particular, headaches, breast tenderness, muscle pain, diarrhea, colic, changes in appetite, anxiety, feelings of stress, feelings of weakness, mood disturbances and sleep disturbances.

[0060] According to yet another particular embodiment, the present invention relates to TGF-[3 for its use for the prevention and / or treatment of neuralgic symptoms in people suffering from premenstrual dysphoric disorder (PMDD).

[0061] Among PMS, PMDD is suspected when psychological or even psychiatric symptoms such as irritability, depressed mood, anxiety and anger are predominant and intense, and are now considered part of the overall depressive symptoms. PMDD is estimated to affect between 2 and 8% of women during the menstrual period, which corresponds to the proportion of women considered to be suffering from severe PMS.

[0062] PMS and PMDD are conditions that manifest with physical, emotional, and behavioral symptoms during the luteal phase (second half) of the menstrual cycle, and which abate with the onset of menstruation (O'Brien et al., 2011, Archives of Women's Mental Health, 14(1), 13-21). While While symptoms of premenstrual syndrome are common and often manageable, premenstrual dysphoric disorder represents a severe and disabling form of the condition that has a significant impact on quality of life.

[0063] Recent research has suggested that immune cells and inflammatory processes may play a role in premenstrual syndrome and premenstrual dysphoric disorder. Both conditions are thought to be related to changes in hormone levels throughout the menstrual cycle, particularly progesterone and its neuroactive metabolites (Schmidt et al., 2017, American Journal of Psychiatry, 174(10), 980-989). These hormonal fluctuations may affect immune cell function, possibly leading to inflammatory responses.

[0064] Non-inflammatory aspects of PMS and PMDD include hormonal, genetic, and environmental factors, as well as neurotransmitter dysregulation. Studies have shown that hormonal fluctuations, particularly those of progesterone and estradiol, can influence mood and behavior (Schmidt et al., 2017).

[0065] Altered sensitivity to these hormonal changes, rather than the changes themselves, may underlie the symptoms of premenstrual syndrome and premenstrual dysphoric disorder. Genetic factors may play a role in this altered sensitivity, although the specific genes involved are not yet clear (Huo et al., 2007, Biological psychiatry, 82(10), 814-824).

[0066] Environmental factors such as stress may also exacerbate symptoms of PMS and PMDD, possibly by influencing hormone levels and immune function. Dietary factors, including alcohol and caffeine consumption, as well as lack of exercise, have been associated with more severe symptoms (Dickerson et al., 2003, American Family Physician, 67(8), 1743-1752).

[0067] In conclusion, PMS and PMDD are complex diseases that result from the interaction of hormonal, genetic, inflammatory and environmental factors; which complicates the identification of effective treatment.

[0068] The present invention also relates to TGF-[3 for its use for the prevention and / or treatment of skin itching; in particular, in patients suffering from chronic renal failure (CRF or CKD), in particular after having received hemodialysis.

[0069] CKD is a disease in which kidney function declines to less than 60% of that of a healthy individual or in which kidney abnormalities such as proteinuria persist. Kidney function declines with age, and CKD is more common in older people. It is closely associated with metabolic syndrome, which includes hypertension, diabetes, hypercholesterolemia, hypertriglyceridemia (abnormal lipid metabolism) and obesity.

[0070] CKD is irreversible. In the final stage, the nephron is damaged and unable to function properly; this is called end-stage renal disease. With decreased kidney function, the kidneys are no longer able to properly remove waste and excess water from the body, allowing waste to accumulate in the blood and body. In mild cases, patients may experience fluid and electrolyte imbalances. In severe cases, it may lead to uremia, which can be fatal.

[0071] In clinical applications, a person suffering from CKD and uremia must receive dialysis or a kidney transplant to improve their condition. However, complications such as inflammation, itchy skin, and decreased iron metabolism caused by dialysis can negatively impact the quality of life. Therefore, preventing or improving complications in a patient suffering from CKD after receiving hemodialysis is of real importance. The manifestation of chronic pruritus, also known as itching, in patients with CKD, and especially in those who are on hemodialysis, is an important medical problem. Pruritus in CKD, known as uremic pruritus or CKD-associated pruritus (CKD-aP), is a common and debilitating symptom that significantly reduces the quality of life.The pathogenesis of the symptom is not yet fully understood, but is thought to be multifactorial, involving both inflammatory and non-inflammatory pathways.

[0072] The non-inflammatory aspects of CKD-aP are primarily related to the accumulation of uremic toxins and imbalances in metabolic homeostasis, which are hallmarks of CKD. Elevated levels of divalent ions such as phosphate and calcium have been associated with chronic kidney disease. These ions are poorly excreted by diseased kidneys, causing an imbalance in mineral metabolism and contributing to pruritus (Narita et al., Kidney Int. 2006;69(9): 1626-32).

[0073] In addition, patients with CKD-aP have impaired skin barrier function, likely due to the accumulation of uremic toxins. This leads to xerosis (dry skin), which is common in patients with CKD and exacerbates pruritus (Mettang et al. Kidney Int. 2015;87(4):685-91).

[0074] In recent years, kappa-opioid receptors (KORs) and mu-opioid receptors (MORs) have been identified as being involved in the suppression and promotion of pruritus, respectively, by acting on both the peripheral and central nervous systems (Kim et al., Exp. Dermatol. 2022;31(12), 1900-1907). Alterations in the opioid system, i.e., an imbalance of stimulation between the KOR and MOR also appear to be involved in the pathogenesis of CKD-aP. Indeed, nalfurafine, a KOR agonist, nalbuphine, a MOR antagonist and KOR agonist, and difelikaline, a peripheral KOR agonist, have all shown positive results in clinical trials in patients with CKD-aP (Verduzco & Shirazian, Kidney Int. Rep. 2020;5(9): 1387-1402).

[0075] Recent studies also suggest that inflammation and immune responses may play a crucial role in the development of CKD-aP.

[0076] In conclusion, CKD-aP is a multifactorial disease involving both non-inflammatory and inflammatory pathways. Further research is needed to better delineate these pathways and identify novel therapeutic targets.

[0077] It is important to note that a very significant advantage of using TGF-[3 for the above disorders is that at the same time as its beneficial action on neuralgic manifestations, TGF-[3 also produces an anti-inflammatory effect which also contributes to the treatment of these disorders which, as we have seen above, also have an inflammatory component.

[0078] According to the present invention, the TGF-[3 used is preferably predominantly TGF-[32, that is to say, the proportion of TGF-[32 on total TGF-[3 is greater than 50%, by weight, preferably greater than 80% by weight, even more preferably greater than 90% by weight.

[0079] The TGF-[3] according to the invention can be synthesized by biological techniques such as precision fermentation.

[0080] Alternatively, the TGF-[3] according to the invention may originate from mammalian milk. According to this embodiment, the TGF-[3] is present in a milk protein fraction; or the TGF-[3] is present in a soluble milk protein fraction; or the TGF-[3] is present in a soluble cationic milk protein fraction; or the TGF-[3] is present in a soluble cationic milk protein fraction composed mainly of lactoferrin.

[0081] Preferably, the TGF-[3 used according to the invention is mainly in active form (or mature, i.e. dissociated from the LAP propeptide), preferably greater than 60%, more preferably 80% by weight in active form relative to the total content of TGF-[3.

[0082] According to the invention, the daily dose of TGF-[3 is between 1 and 100 pg, preferably between 2 and 50 pg, more preferably between 4 and 12 pg.

[0083] According to a particular embodiment, TGF-[3 is administered with lactoferrin at a dose of between 3 and 500 mg, preferably between 13 and 167 mg.

[0084] The administration of TGF-[3 according to the invention can be carried out orally, in powder form, in liquid form (drink for example) or in solid form (capsule or tablet); the administration can also be carried out topically (cutaneous, nasal, inhaled).

[0085] The TGF-[3] according to the invention is preferably formulated in a composition which may for example be a food supplement or a medicament.

[0086] More particularly, this composition may be a liquid, a suspended liquid, an emulsion, a powder, a tablet, a pill, a syrup, lozenges or pastilles, a chewing gum and a capsule, and the composition may be a drug composition, a beverage composition, a nutrient composition, a food composition and a supplement composition.

[0087] Furthermore, the composition according to the present invention may comprise a suitable carrier and additional ingredients such as, but not limited to, a solvent, a dispersion medium, a coating, an antibacterial reagent, an antifungal reagent, an absorption delaying reagent and other methods compatible with medicine.

[0088] The present invention also relates to a method of prevention and / or treatment:

[0089] - neuralgic symptoms; in particular, pain and itching;

[0090] - neuralgic symptoms in people with dysmenorrhea; in In particular, the neuralgic symptoms of dysmenorrhea include pain in the pelvis or lower abdomen, back pain, muscle pain, diarrhea;

[0091] - neuralgic symptoms in people with symptoms of premenstrual syndrome (PMS); in particular, headaches, breast tenderness, muscle pain, diarrhea, colic, changes in appetite, anxiety, feelings of stress, feelings of weakness, mood swings, and sleep disturbances;

[0092] - neuralgic symptoms in people with dysphoric disorder premenstrual dysphoria disorder (PMDD);

[0093] - itchy skin; particularly in patients with renal insufficiency chronic kidney disease (CKD) especially after receiving hemodialysis;

[0094] said method comprising a step of administering TGF-[3 to a subject requiring such prevention or treatment.

[0095] The subject to be treated may be a human or an animal, preferably a mammal.

[0096] EXAMPLES

[0097] Example 1: Effect of TGF-[3 in women with dysmenorrhea

[0098] In order to verify whether TGF- [Best is capable of relieving premenstrual syndrome and premenstrual dysphoria, several patients suffering from these symptoms have been selected for the clinical trial program, and they were followed for two to six months.

[0099] Subjects: 5 subjects, aged 31 to 41 years, participated in this clinical trial. All subjects had menstrual pain that had a moderate to severe impact on their lives and even prevented them from leading a normal life. In addition, the subjects in this clinical trial also had symptoms of premenstrual syndrome or premenstrual dysphoric disorder, and suffered from reproductive disorders such as adenomyosis, uterine adnexal mass, endometrioma, ovarian vesicles, myoma, endometriosis, and other diseases of the reproductive organs.

[0100] The basic clinical characteristics of the patients are shown in Table 1 below:

[0101] [Tables 1] Subjects 1 2 3 4 5 Age (years) 33 41 31 32 38 Length of menstrual cycle (days) 30 28 28-30 26 30 Length of menstrual period (days) 4-9 5 7 5-7 7 Level of quality of life affected by dysmenorrhea Severe impact Severe impact and inability to have a regular lifestyle Severe impact Severe impact and inability to have a regular lifestyle Moderate impact Symptoms of PMS and PMDD Muscle pain, exhaustion, insomnia Anxiety, diarrhea, insomnia Headache, anxiety, drowsiness Headache, breast swelling, muscle pain, diarrhea, insomnia Change in appetite, anxiety, pressure, exhaustion, loss of mood Anxiety, pressure, exhaustion, loss of mood, insomnia, headaches, breast swelling, muscle pain, diarrhea, intestinal colic, change in appetite. History of illness and / or gynecological surgery Adenomyosis Hernia Myoma Right-sided ovarian cystadenoma Three times surgery, ovarian cyst, adenomyosis, ovarian tumor, myoma Diagnosis before treatment Adenomyosis Dysmenorrhea Dysmenorrhea, endometriosis Dysmenorrhea, ovarian cysts, myoma Endometriosis Symptoms detected by ultrasound Adenomyosis Adenomyosis, adnexal mass, endometrioma Myoma Ovarian cysts, myomas Myoma, adnexal mass Baseline clinical characteristics of the subjects

[0102] Open pilot clinical study protocol:

[0103] The TGF-[3] used in this trial was derived from bovine whey protein concentrate with a TGF-[3]2 content of 10 pg / g ("TGF-[3 / WPC") as prepared in Example 3. During the trial period of 2 to 5 months, subjects orally took the following doses of TGF-[3]:

[0104] From Day 1 to Day 10, 12 μg of TGF-[32 once daily;

[0105] From Day 11 until the end of the trial, 8 μg of TGF-[32 once daily;

[0106] After 3 months of treatment, in case of improvement of dysmenorrhea, 4 pg of TGF- [32 once a day.

[0107] Before, in the middle and after the period of taking TGF-[32, the dysmenorrheal pain scale was assessed using a questionnaire. A score of 100 corresponded to the most severe pain imaginable and a score of 0 corresponded to no pain. Tables 2 to 6 below present the pain scores of subjects 1 to 5 during the trial period respectively. The improvement rate was calculated using the following formula:

[0108] [Improvement Rate (%)] = ([Pre-Test Pain Score]-[Post-Test Pain Score]) / [Total Pre-Test Pain Score]

[0109] The degrees of premenstrual syndrome and premenstrual dysphoric disorder in the subjects were also assessed by a questionnaire, before, during and after taking TGF-|3.

[0110] Results E ffectiveness of oral administration of TGF-[3 on the relief of menstrual pain

[0111] - Subject 1 achieved a pain improvement rate of 56.9% after 2 months of taking TGF-[3 (Table 2).

[0112] - Subject 2 achieved a 98.7% improvement in pain after 2 months of taking of TGF-[3, and 99.4% after 3 months (Table 3).

[0113] - Subject 3 showed a 64.1% improvement in pain after 5 months of TGF- [3 (table 4).

[0114] - Subject 4 achieved a 54.3% improvement in pain after 1 month of TGF-[3 and a 51.4% improvement in pain after 2 months (Table 5).

[0115] - Subject 5 achieved a 59.0% pain improvement after 3 months of taking of TGF-[3 and 100% improvement after 4 months and maintained a pain-free state during the menstrual period after 5 months of taking TGF-[32 (Table 6).

[0116] All the results show that a daily intake of TGF-[32 of 4 to 12 pg has an excellent effect on the relief of menstrual pain in subjects suffering from dysmenorrhea.

[0117] [Tables2] Days: 1 2 3 4 5 6 Total Pain Score Improvement Rate (%) Before taking 43 55 44 22 12 12 188 - 1 month of taking 49 40 17 11 3 0 120 36.2 2 months of taking 24 23 15 10 9 0 81 56.9 Pain scores for subject 1

[0118] [Tables3] Days: 1 2 3 4 5 Total Pain Score Improvement Rate (%) Before taking 59 96 6 0 20 181 - 1 month of taking 25 66 37 0 0 128 29.3 2 months of taking 0.7 1.2 0.5 0 0 2.4 98.7 3 months of taking 0.5 0.5 0 0 0 1 99.4 Pain scores for subject 2

[0119] [T ables 4] Days: 1 2 3 4 5 6 7 Total Pain Score Improvement Rate (%) Before taking 65 63 45 20 20 7 0 220 - 1 month of taking 43 48 24 6 0 0 0 121 45.0 2 months of taking 56 54 15 0 0 0 0 125 43.2 3 months of taking 66 54 40 21 0 0 0 181 17.7 4 months of taking 29 61 53 0 0 0 0 143 35.0 5 months of taking 46 29 4 0 0 0 0 79 64.1 Pain scores for subject 3

[0120] [Tables5] Days: 1 2 3 4 5 6 7 Total Pain Score Improvement Rate (%) Before taking 58 90 72 19 6 0 0 245 - 1 month of taking 48 41 23 0 0 0 0 112 54.3 2 months of taking 42 47 30 0 0 0 0 119 51.4 Pain scores for subject 4

[0121] [Tableauxô] Days: 1 2 3 4 5 6 7 Total Pain Score Improvement Rate (%) Before taking 60 81.6 80.6 74.8 41.3 18.2 - 356.5 - 1 month of taking 69 78 67 33 - - - 247 30.7 2 months of taking 65 74 65 59 54 42 36 395 -10.8 3 months of taking 35 51 33 15 12 0 0 146 59.0 4 months of taking 0 0 0 0 0 0 - 0 100 5 months of taking 0 0 0 0 0 0 - 0 100 Pain scores for subject 5

[0122] Results _ Efficacy of oral administration of TGF-[3 _ on the improvement of premenstrual syndrome (PMSI) and premenstrual dysphoric disorder (PMDD)

[0123] Tables 7 to 11 provide an overview of the improvement scale of the manifestations of premenstrual syndrome and premenstrual dysphoric disorder in patients during the period of dysmenorrhea. The score 0 corresponds to asymptomatic, the score 1 very mild, score 2 mild, score 3 moderate, score 4 severe and score 5 very severe.

[0124] Subject 1 saw improvement in muscle pain, exhaustion, and insomnia after two months of taking TGF-[3 (Table 7).

[0125] Subject 2 showed improvement in PMS and PMDD manifestations such as anxiety, insomnia, and diarrhea after 1 month of TGF-[3], and this improvement further progressed after three months (Table 8).

[0126] Subject 3 showed improvement in premenstrual headache symptoms after 1 month of taking TGF-[3, and improvement in premenstrual headache and drowsiness symptoms and premenstrual dysphoric disorder after 2 to 5 months (Table 9).

[0127] Subject 4 showed after 2 months of taking TGF-[3, an improvement in symptoms such as breast swelling, diarrhea, anxiety, pressure, exhaustion and loss of mood (Table 10).

[0128] Subject 5 showed improvement in PMS symptoms such as muscle pain and appetite changes after 3 months of taking TGF-[3 and after 4 months a marked improvement in symptoms such as anxiety, stress, weakness, loss of control, insomnia, headache, breast tenderness, muscle pain, diarrhea, colic and appetite changes. PMS symptoms almost completely disappeared after 5 months of taking TGF-[3 (Table 11).

[0129] In summary, the results in Tables 7 to 11 indicate that oral administration of TGF-[3 was found to be highly effective in improving the manifestations of premenstrual syndrome and premenstrual dysphoric disorder.

[0130] [Tables7] Muscle pain Exhaustion Insomnia Before consumption 4 3 4 After 1 month of consumption 2 3 4 After 2 months of consumption 2 2 3 Symptom scores for subject 1

[0131] [Tables8] Anxiety Insomnia Diarrhea Before consumption 5 3 3 After 1 month of consumption 4 3 2 After 2 months of consumption 3 0 0 After 3 months of consumption 1 0 1 Symptom scores for subject 2

[0132] [Tables9] Headaches Anxiety Drowsiness Before consumption 3 2 4 After 1 month of consumption 1 2 4 After 2 months of consumption 0 2 3 After 3 months of consumption 2 3 2 After 4 months of consumption 2 2 2 After 5 months of consumption 1 4 2 Symptom scores for subject 3

[0133] [TableauxlO] Headaches Breast swelling Muscle pain Diarrhea Anxiety Before consumption 3 3 2 2 4 After 1 month of consumption 2.5 3 1 2 3 After 2 months of consumption 3 1 2 1 1 Symptom scores for subject 4

[0134] [Tables 11] Anxiety Pressure Exhaustion Loss of composure Insomnia Headaches Breast swelling Sculinary pain Before consumption 3 3 3 3 4 5 5 4 After 1 month of consumption 3 3 2 3 3 4 4 3 After 2 months of consumption 3 4 4 3 2 4 3 2 After 3 months of consumption 3 3 2 2 2 3 2 0 After 4 months of consumption 0 0 0 0 0 3 0 0 After 5 months of consumption 0 0 0 0 0 1 0 0 Symptom scores for subject 5

[0135] Example 2: The clinical effect of TGF-[3 in people with chronic renal failure

[0136] In order to verify whether TGF-[3, especially TGF-[32, is able to prevent or alleviate complications in people with chronic kidney failure after receiving hemodialysis, several patients were selected for the clinical trial program and followed for three months for their states of inflammation, anemia and other relevant factors.

[0137] Subjects: 40 individuals with chronic kidney disease, all of whom underwent dialysis treatment. Only individuals who met all the conditions indicated in Table 12 were included for the efficacy analyses to avoid imprecision. The baseline characteristics of the subjects included for the analyses are presented in Table 13.

[0138] [Tablesl2] 1 Not in a state of infection 2 Not cancer patients 3 Not having undergone chemotherapy or radiotherapy within 6 months 4 No bleeding disease 5 No blood test within 3 months prior to the trial 6 No change in prescription within 3 months of the trial 7 No alcohol consumption habit 8 No milk allergy 9 No more than 500 mL of blood donation within 3 months prior to the trial

[0139] Inclusion conditions for efficacy analyses

[0140] [Tablesl3] Percentage of all subjects Total number of subjects 40 - Age 65.3+1.6 - Male 24 60% Female 16 40% Without itchy skin symptoms 11 27.5% With itchy skin symptoms 29 72.5%

[0141] Baseline characteristics of subjects included for analyses

[0142] Open pilot clinical study protocol:

[0143] The TGF-[3] used in this trial was derived from bovine whey protein concentrate with a TGF-[3]2 content of 12 pg / g ("TGF-[3 / WPC bis") as prepared in Example 5. During the 3-month trial period, subjects orally took a daily dose of 800 mg of this whey protein concentrate, corresponding to a daily dose of TGF-[3]2 of approximately 10 pg.

[0144] In the trial, before and after 3 months of consumption of the whey protein concentrate containing TGF-[3, blood concentrations of TGF-[32 were measured from blood collection in 40 patients.

[0145] Before and after 3 months of consumption, 40 patients completed the 5-D itch scale survey and the visual analog scale (VAS) to analyze the improvement of skin itching.

[0146] The 5-D Itch Scale is a multidimensional scale that assesses itching based on 5 dimensions, namely duration, degree, direction, the discreteness and distribution of itching. Each dimension is rated on a scale of 1 to 5, resulting in a total score between 5 and 25. A higher score indicates more severe itching.

[0147] The visual analog scale (VAS) is a simple and commonly used tool for measuring itch intensity. It is a straight line, usually 10 cm long, with ends marked as "no itch" (0) and "unbearable itch" (10). Patients are asked to indicate on the line the intensity of their itch. The closer the mark is to 10, the more intense the itch.

[0148] Results j. Eff et of oral administration of TGF-[3 on blood anti-inflammatory factor

[0149] As shown in Table 14, there is a significant increase in the blood concentration of TGF-[32 in people who consumed bovine whey protein concentrate containing TGF-[3 for 3 months. This shows that oral administration of TGF-[3 from bovine whey protein can significantly increase the systemic blood concentration of TGF-[32, which may help prevent or improve inflammation caused by chronic kidney disease.

[0150] [Tablesl4] Before consumption After 3 months of consumption ion p value TGF-[32 (pg / mL) 48.7 ± 4.2 61.2 + 6.7 0.0238

[0151] Blood concentration of TGF-&2 before and after 3 months of consumption of bovine whey protein concentrate containing TGF-fi.

[0152] According to the results of the trial, oral administration of TGF-[3 can increase the anti-inflammatory factor of people with chronic renal failure after hemodialysis in the blood. Therefore, the use of the bovine whey protein extract composition according to the invention has high medical potential in terms of preventing or improving complications of people with chronic renal failure after receiving hemodialysis.

[0153] Results: Impact of oral administration of TGF-[3 on the improvement of skin itching in people with chronic renal failure

[0154] Table 15 shows the evolution of itchy skin symptoms in patients before and after taking bovine whey protein concentrate containing TGF-[3.

[0155] [Tablesl5] Before consumption After 3 months of consumption p values ​​Visual analogue scale (VAS) 4.8 + 0.5 4.0 + 0.5 0.0065 5-D itch scale 13.1+0.7 11.6 + 0.7 0.0094

[0156] Skin itching in people with chronic renal failure before and after 3 months of consumption of bovine whey protein concentrate containing TGF-[3

[0157] According to the results of the trial, the bovine whey protein concentrate containing TGF-[3] can improve the itching symptoms of people with chronic renal failure after hemodialysis. Therefore, the use of the composition containing TGF-[3] has high medical potential in terms of preventing or improving the complications of people with chronic renal failure after receiving hemodialysis.

[0158] Example 3: Preparation of the whey protein concentrate enriched with TGF-[3] according to the invention

[0159] A fraction of a TGF-[3]-enriched whey protein concentrate was prepared using a selective precipitation technique for soluble milk proteins.

[0160] 1000 L of skimmed cow's milk was thermized at 70°C for 20 seconds.

[0161] Casein in skim milk was precipitated by adjusting the pH to 4.6 with acid hydrochloric acid, then the precipitated casein was separated by centrifugation to obtain the soluble phase (whey).

[0162] The pH of the whey thus obtained was adjusted to pH 5.2 with sodium hydroxide, then heated to 50°C.

[0163] The proteins precipitated by this treatment were concentrated by centrifugation, then resolubilized by neutralizing with sodium hydroxide.

[0164] The solubilized proteins were reconcentrated by an ultrafiltration membrane (MWCO 10 kDa), then spray-dried.

[0165] Thus 2 kg of whey protein concentrate powder are obtained with a protein content of 80% on MS (this concentrate is designated "TGF-[3 / WPC"). The major protein constituents of this WPC are [3-lactoglobulin (45% / protein), α-lactalbumin (30% / protein) and other acidic proteins such as serum albumin (BSA), immunoglobulin G (IgG) and lactoferrin. The measured TGF-

[32] content by ELISA (Quantikine human TGF-[32, R&D Systems) is 10 pg / g of powder, representing >90% by weight of total TGF-[3.

[0166] Example 4: Preparation of the cationic whey protein isolate enriched in TGF-[3 according to the invention

[0167] A fraction of a cationic whey protein isolate enriched in TGF-[3] was prepared using an ion exchange chromatography technique.

[0168] 10000 L of skimmed bovine milk thermized at 65°C for 30 seconds is passed to a flow rate of 2 m / h through a 50 L column containing SPEC 70 SLS resins (Sartorius Stedim Biotech GmbH) previously prepared in Na+ form to bind cationic proteins including TGF-[3, then the resins were rinsed with 50 L of water.

[0169] The cationic proteins bound to the resins were eluted with 30 L of sodium chloride solution at 50 mS / cm, then concentrated by an ultrafiltration membrane (MWCO of 10 kDa) to a concentration factor of approximately 30, followed by diafiltration to increase the protein concentration on dry matter (DM).

[0170] The retentate obtained was microfiltered with a ceramic membrane at 0.8 pm, then pasteurized at 73°C for 20 seconds, finally spray-dried.

[0171] Thus, 200 g of cationic whey protein isolate powder with a protein content of 98% on MS (isolate designated "TGF-[3 / cWP") are obtained. The main proteins of this TGF-[3 / cWPI are lactoferrin (70% / MS), lactoperoxidase (19% / MS) and other cationic proteins such as lactophorin (milk Glycam-1) and ribonuclease 4. The contents of TGF-[31, TGF-[32 and TGF-[33 were measured by the R&D Systems Quantikine ELISA kits for human TGF-[31, human TGF-[32 and human TGF-[33, respectively). The TGF-

[32] content is 150 pg / g of powder, of which 90 pg / g is in mature form (assessed without activation by acidification pretreatment). The TGF-

[31] content is 6.9 pg / g of powder, of which 3.7 pg / g is in mature form. TGF-

[33] is undetectable.

[0172] Example 5: Preparation of the whey protein concentrate enriched with TGF-[3] according to the invention

[0173] A fraction of a TGF-[3]-enriched cationic whey protein concentrate was prepared using a dry blend of the TGF-[3] / cWPI prepared in Example 4 with a standard powdered whey protein concentrate whose protein content is 82% on MS (WPC). The main proteins in this WPC are [3]-lactoglobulin (about 50% / protein), α-lactalbumin (about 20% / protein) and other acidic proteins such as serum albumin (BSA) and immunoglobulin G (IgG). The TGF-

[32] content measured by ELISA (Quantikine human TGF-[32, R&D Systems) is 0.013 pg / g of powder.

[0174] 1841 g of WPC powder and 159 g of TGF-[3 / cWPI] powder were placed in a sealed stainless steel container with a capacity of 5 liters.

[0175] The homogeneous mixture was prepared using a 3D type agitator-mixer using a TURBULA® mixer.

[0176] Thus obtained 2 kg of whey protein concentrate powder with a protein content of 83% on MS (designated "TGF-[3 / WPC bis"). The TGF-[32 content measured by ELISA (Quantikine human TGF-[32, R&D Systems) is 12 pg / g of powder. Example 6#: Food supplements

[0177] Below is an example of preparing the capsules with the other ingredients:

[0178] [Tableauxlô] Composition Per capsule For 6 capsules (Daily Intake) “TGF-[3 / WPC” prepared in Example 5 133 mg 800 mg Fish oil powder containing at least 25% omega 3 FA PI of which EPA (15%) of which DHA (10%) 100 mg 15 mg 10 mg 600 mg 90 mg 60 mg = 50% ANC* Microcrystalline cellulose, 1.3 mg Naturally occurring vitamin E 1.66 mg 10 mg = 100% RI** Vitamin B9 (folic acid) 0.033 mg 0.2 mg = 100% RI**

[0179] *ANC: recommended nutritional intake

[0180] **RDA: recommended daily intake

Claims

Claims

1. TGF-[3 for its use for the prevention and / or treatment of neuralgic symptoms.

2. TGF-[3 for its use according to claim 1, characterized in that the neuralgic symptoms are pain and itching.

3. TGF-[3 for its use according to claim 1 for the prevention and / or treatment of neuralgic symptoms in persons suffering from dysmenorrhea; preferably, secondary dysmenorrhea.

4. TGF-[3 for its use according to claim 3, characterized in that the neuralgic symptoms are pain in the pelvis or lower abdomen, back pain, muscle pain, diarrhea.

5. TGF-[3 for its use according to claim 1 for the prevention and / or treatment of neuralgic symptoms in persons suffering from symptoms of premenstrual syndrome.

6. TGF-[3 for its use according to claim 5, characterized in that the symptoms of premenstrual syndrome include headaches, breast tenderness, muscle pain, diarrhea, colic, changes in appetite, anxiety, a feeling of stress, a feeling of weakness, mood disturbances and sleep disturbances.

7. TGF-[3 for its use according to claim 1 for the prevention and / or treatment of neuralgic symptoms in persons suffering from premenstrual dysphoric disorder.

8. TGF-[3 for its use according to claim 1 for the prevention and / or treatment of itchy skin in patients with chronic renal failure.

9. TGF-[3 for its use according to claim 1 for the prevention and / or treatment of complications of persons suffering from chronic renal failure after receiving hemodialysis.

10. TGF-[3 for its use according to claim 9, characterized in that said complications include skin itching.

11. TGF-[3 for its use according to any one of the preceding claims, characterized in that the TGF-[3 is TGF-[3 from bovine milk.

12. TGF-[3 for its use according to any one of the preceding claims, characterized in that the proportion of TGF-[32 on total TGF-[3 is greater than 50% by weight, preferably greater than 80% by weight, even more preferably greater than 90% by weight.

13. TGF-[3 for its use according to any one of the preceding claims, characterized in that the TGF-[3 is predominantly in active form, preferably greater than 60% in active form.