ITMI20102380A1 - PREVENTION AND THERAPY METHODS FOR PARTS AND BIRTHS PRETERMINE. - Google Patents

Description

"METHODS OF PREVENTION AND THERAPY FOR PRETERMINE PARTS AND BIRTHS"

DESCRIPTION

The present invention relates to the therapy and / or prevention of pathological conditions which induce preterm births and births.

Preterm deliveries are defined as deliveries that occur before the 37th week of gestation. Among preterm births (PPs), those of greater clinical importance occur before the thirty-second week of gestation. The incidence of PP represents about 10% of births, but this percentage is increasing due, in large part, to assisted reproduction and multiple pregnancies.

Preterm birth generally does not pose a health risk to the mother. In contrast, preterm births (NPT) are the cause of the main neonatal diseases and are correlated with 70% of neonatal deaths. This percentage is destined to decrease due to progress in the hospital management of premature babies even if it is always necessary to evaluate in the long term, among the survivors, the consequences of an NPT especially at the neurological level.

Spontaneous or iatrogenic PP can be induced by infectious processes, nutritional deficiencies, alcoholism, smoking, shortening of the cervix, uterine malformations, polyamnios, multiple pregnancies, placenta previa, placental abruption, premature rupture of membranes, iron deficiency, anemia, diabetes , pre-eclamsia, hypertension and hereditary thrombophilia. Although the causes of PP are multiple and multifactorial, the inflammatory state at the maternal-fetal interface is currently considered to be the crossroads of factors known to be associated with NPT. The inflammatory state is mediated by pro-inflammatory cytokines.

Although it is clear from the international bibliography that there is a strong correlation between NPT and inflammation (Ref. Christiaens Inge, Dean B. Zaragoza, Larry Guilbert, Sarah A. Robertson, Bryan F. Mitchell, David M. Olson "Inflammatory processes in preterm and term parturition ", Journal of Reproductive Immunology 79, 50-57, 2008; Debra Lyon, Ching-Yu Cheng, Lois Howland, Debra Rattican, Nancy dal, Rita Pickler, Lisa Brown and Jacqueline McGrath" Integrated Review of Cytokines in Maternal, Cord, and Newborn Blood: Part Iâ € ”Associations With Preterm Birth" Biological Research for Nursing 11 (4), 371-376, 2010), the clinical trials, conducted to determine the level and type of cytokines, designed testing the concentration of pro-inflammatory cytokines in different biological samples such as maternal, fetal or cord blood, placenta, amniotic fluid, cervicovaginal secretion, gave contradictory results. In fact, it seems unclear whether the predictability of a PP and its induction can be linked to the pro-inflammatory cytokines found in the circulation or in the cervicovaginal secretion.

The generic term of cytokine describes small proteins which, once secreted, mediate and regulate immunity, inflammation and hematopoiesis. They are divided into "prò" and "anti" inflammatory. Pro-inflammatory cytokines generally include interleukins (IL) -lbeta, IL-6, IL-11, IL-12, IL-17, IL-18, IL-8, tumor necrosis factor alpha (TNF-alpha) , and interferon gamma (IFN-gamma). Anti-inflammatory cytokines include IL-4, IL-10, IL-13, IL-16, IFN-Î ±, and tumor growth factor b (TGF-b). Another class of cytokines, hematopoietic growth factors, also known as "colony-stimulating factors (CSFs)", regulate the production of red blood cells, leukocytes and platelets and are important markers of the maternal-fetal state.

The relationship between prò- and anti-inflammatory cytokines is an important element in the immune response of the mother and fetus. The different ratio of cytokines is also necessary in conception, pregnancy, childbirth and in the first phase of fetal life.

In fact, during pregnancy there is a shift between the levels of T helper 1 (Thl) and T helper 2 (Th2) with an increase in the production of Th2 cells. Thl cells synthesize ì'IL-2, IFN-gamma, and TNF-beta and induce cellular immunity. Th2 cells synthesize IL-4, IL-5, IL-6, and IL-10 and induce the production of antibodies.

Although any classification of cytokines cannot be exhaustive, given the complexity of their multiple roles, cytokines are certainly associated with labor and delivery.

The onset of labor appears as an inflammatory reaction, involving not only an influx of leukocytes into the cervical and endometrial tissues but also an increase in the synthesis of cytokines and prostaglandins. In fact, at the moment of labor, the synthesized cytokines induce prostaglandin synthetase-2 which increases the production of prostaglandins (PGs), with consequent activation of cervical ripening and uterine contractions. In particular, IL-Î ™ β increases the production of matrix metal proteases (MMPs), which induce cervical remodeling and possible rupture of membranes, while IL-6 and TNF-Î ± activate leukocytes to invade the uterine tissue, thus stimulating the expression of oxytocin receptors and increasing prostaglandins. IL-8, produced by the placenta, also cooperates in the modification of the cervix by inducing the influx of leukocytes. In addition, these cytokines influence each other: IL-Î ™ β and TNF-Î ± activate IL-8, which then promotes the synthesis of IL-Î ™ β and IL-6.

The inflammatory process in pregnancy can be triggered by both infectious and non-infectious events.

Infections of the ascending genital tract or systemic infections, causing the release of pro-inflammatory cytokines such as IL-Î ™ β, IL-6, and TNF-Î ±, must be promptly treated with antimicrobial drugs in order to avoid or to decrease the likelihood of triggering uterine contractions and preterm labor.

On the other hand, the treatment of inflammation appears more complex if contractions and preterm labor arise as a result of inflammatory processes that involve high levels of pro-inflammatory cytokines in the circulation as in the case of infections localized in districts other than the cervical vaginal or as in the case inflammation independent of infections

It has already been reported as the increase in body mass, lifestyle (smoking, alcohol and drugs), nutritional status, heart rhythm, age, hypertension, iron deficiency, anemia, diabetes, pre-eclamsia and hereditary thrombophilia cause inflammatory phenomena, especially in pregnancy. Recently, relaxin, a collagenolytic hormone that increases matrix metalloproteinase production, has also been associated with PP. The expression of relaxin, in fact, regardless of infections, induces the synthesis of IL-6 and IL-8.

Despite the importance of an inflammatory event in pregnancy, there is a lack of reliable clinical studies on the relationship between infection-independent inflammation and NPT.

From what has been summarized, a better understanding of the relationship between cytokines at the systemic level and at the level of cervico-vaginal secretions and NPT can certainly be fundamental in the search for interventions that, in a non-invasive way and without adverse effects, can decrease inflammation and synthesis of prostaglandins, lengthening the gestational period and thus preventing both the onset of contractions and PP.

There are numerous subjective and objective diagnostic methods capable of predicting a PP, but none of those listed below is considered to be totally reliable.

Due to the lack of a reliable method to predict and define the changes in the uterus that could lead to PP, the subjective assessment of the woman is taken seriously: i) the appearance of uterine contractile activity, perceived by the woman as painful or not painful; ii) appearance of fixed or intermittent menstrual cramps in the supra-pubic region; iii) low back pain; iv) pelvic pressure.

However, the subjective evaluation of the woman must necessarily be accompanied by an objective diagnosis, bearing in mind that in a large percentage of PP the symptoms that precede it can be very scarce.

Assuming that the execution of a digital cervical examination for the identification of the PP threat has shown its limits as a minimal cervical dilation (<3 centimeters) normally takes place in pluriparous and is not necessarily an indicator of an imminent PP, other diagnostic methods must be performed.

The objective evaluation of a PP can be performed by ultrasound, cardiotocography, electrohysterogram, vaginal pH measurement, change in vaginal secretions, possible vaginal bleeding, Nugent scores, haematological parameters such as red blood cells, hemoglobin, total serum iron, serum ferritin, the level of proinflammatory cytokines such as IL-Î ™ β, IL-6, IL-8 and TNF-a, that of fetal fibronectin and PGs in the cervico-vaginal fluid.

Ultrasound, cardiotocography and electrohysterogram are particularly useful methods of evaluating uterine activity.

The ultrasound, in fact, provides information on the length of the neck and on the presence of the Funneling, that is, a funnel-shaped flaring of the internal uterine orifice. A cervix <30 mm, and the presence of funneling is a high risk index of PP, while a cervix> 30 mm and the absence of funneling is a low risk index of PP.

Cardiotocography, very common in obstetrics, is useful for evaluating both fetal well-being (heartbeat and movements) and the presence, frequency and extent of uterine contractions.

The electrohysterogram distinguishes, more specifically than tocography, the physiological uterine activity from the increased contractile activity that leads to PP, but it is not easy and economical to apply.

Measurement of vaginal pH, change in vaginal secretions, any signs of bleeding, past Nugent, haematological parameters are inexpensive diagnostic tests that are always performed. Much more rarely are specific and expensive assays carried out to define the level of proinflammatory cytokines such as IL-1b, IL-1 and IL-6 and TNF-Î ± in the circulation and in the cervico-vaginal fluid as well as that of PGs such as PGE2 and PGF2an in cervico-vaginal fluid.

The same statement can also be made for a biochemical test approved by the Food and Drug Administration: fetal fibronectin.

Fetal fibronectin (fFN) is a glycoprotein that acts as a glue between the maternal decay and the amniotic membranes. In physiological pregnancies it is elevated in cervico-vaginal secretions during the first 24 weeks of pregnancy, but decreases between the twenty-fourth and thirty-fourth weeks. The presence of high concentrations of fFN in cervico-vaginal secretions between the twenty-fourth and thirty-fourth week of gestation is therefore referred to as a parameter associated with a detachment of the amniotic membranes from the uterus wall and a PP threat in pregnant women. symptomatic and asymptomatic. However, the presence of infectious processes, traces of blood or lubricants used for the visit can alter the result of this examination.

From what has been summarized, it is clear that no analysis alone can be predictive of a PP and that to avoid uncertain and ambiguous diagnoses, several analyzes must be performed in parallel.

In short, a close relationship between the levels, type and localization of pro-inflammatory cytokines and PP has never been ascertained so far, in order to decrease the incidence of NPT through more targeted prevention and therapy.

The medical treatment of PP is still a problem that is not easy to solve. There are different types of drugs such as oxytocin receptor antagonists, beta agonists and calcium antagonists. In fact, none of these drugs have been shown to be effective in stopping labor. Conversely, the purpose of these drugs is primarily to prolong pregnancy in order to have time to induce fetal lung maturity through betamethasone. Tocolytic drugs are the most used in an attempt to inhibit uterine contractile activity, although in numerous studies they have not shown significant effects. Still debated is the use of non-steroidal anti-inflammatory drugs (NSAIDs, prostaglandin inhibitors), and their use appears questionable.

The only improvement on the perinatal outcomes is linked to the administration of cortisone drugs with the sole purpose of inducing fetal lung maturation between the twenty-fourth and thirty-fourth week of gestation. These therapies together with the great progress in neonatal care have greatly improved the prognosis of infants born before term but have not brought positive results in the prevention of births and PP, or in reducing the incidence of premature births and births.

The aim of the present invention is to provide a method for preventing and / or treating multifactorial pathological conditions that induce preterm births and births.

Prevention means the use of the present invention in pregnant women who, despite the absence of uterine contractions and other objective data, are at risk of PP due to previous or contextual non-infectious pathologies.

By therapy is meant the use of the present invention in pregnant women in whom, in the preterm period, a threat of premature birth is subjectively and objectively diagnosed.

This task and other purposes which will be evident below have been achieved by means of lactoferrin, or at least a fragment thereof, for use in the therapy and / or in the prevention of pathological conditions that induce preterm births and births, where said use includes the administration of lactoferrin, or at least a fragment thereof, to a pregnant woman in the preterm period.

Within the scope of the present invention, the therapy and / or prevention of pathological conditions, in particular the multifactorial ones, which induce preterm deliveries and births is correlated to the decrease, disappearance and / or absence of the onset of the typical symptoms of childbirth and of the preterm birth reported by the woman, confirmed by objective diagnostic methods by comparing the results obtained before and after therapy with particular reference to the decrease or absence of contractions, the decrease in the concentration of fetal fibronectin, of proinflammatory cytokines in the circulation and in the cervicovaginal secretion and prostaglandins PGE2e PGF2Î ± in the cervico-vaginal secretion as well as the restoration of physiological haematological values such as the number of red blood cells, and the concentration of hemoglobin, total serum iron and serum ferritin.

By "topical administration" or "topical" is meant a local administration of lactoferrin, preferably in the mucous membranes of the vaginal cavity.

By "oral administration", "per os" or "orally" is meant an oral administration which allows for the gastrointestinal absorption of lactoferrin.

Within the scope of the invention, human, bovine or recombinant human and bovine lactoferrin or its fragments can be administered for oral use by means of capsules, tablets or chewable tablets formulated with the excipients such as corn starch, microcrystalline cellulose, vegetable magnesium stearate or others. excipients well known to a person skilled in the art including preservatives, antioxidants, or dyes. Optionally, said formulations for oral use can be in gastro-protected and / or slow-release form.

Gastroprotection can be obtained by means of a superficial covering of the aforementioned capsules, tablets or chewable tablets which remains stable at a pH lower than 3 typical of the stomach and dissolves at a pH higher than 5 typical of the small intestine. Materials for surface coating include fatty acids, polymers and other materials known to a person skilled in the art.

The slow release can be obtained by microencapsulation or compounds that form a cavity where human, bovine or recombinant human and bovine lactoferrin or its fragments are placed in order to obtain administrations of larger quantities that are released over time at concentrations lower than the total content .

In the context of the present invention, human, bovine or recombinant human and bovine lactoferrin or its fragments can be administered for topical use including intravaginal one by means of creams, lyophilisate, lyophilisate conveyed by liposomes, nano particles, microcapsules, polysaccharides or other vehicles free of cytotoxicity, fast or slow release tablets or ovules, non-aqueous solutions, anhydrous gels, suspensions, emulsions, or ointments formulated with suitable excipients and known to a person skilled in the art.

In the context of the present invention, by lactoferrin "fragment" is meant specific sequences of lactoferrin obtained after digestion with proteolytic enzymes. The specific sequences can be the N lobe, the C lobe, and large peptide fragments belonging to the N lobe and the C lobe. The N lobe indicates the molecular weight (PM) fraction of about 33 KDa corresponding to the fragment from the amino acid 1 at 280 and part of the same lobe with a MW of about 20 KDa corresponding to the fragment from the amino acid 86 to 258 of the enzymatically digested whole lactoferrin and subsequently purified (Siciliano R, Rega B, Marchetti M, Seganti L, Antonini G, Valenti P. " Bovine lactoferrin peptidic fragments ìnvolved in inhibition of herpes simplex virus type 1 infection "Biochem Biophys Res Commun. 1999 Oct 14, 264 (1), 19-23).

The lobe C indicates the molecular weight fraction of about 38 KDa corresponding to the 345-692 fragment and another PM 47 KDa fraction corresponding to the 285 to 692 fragment of the enzymatically digested and subsequently purified integral lactoferrin fragment. By large peptide fragments we mean the fragments aa 86-258 PM 20 KDa and aa 285-692 PM 47 kDa (Siciliano R, Rega B, Marchetti M, Seganti L, Antonini G, Valenti P.

"Bovine lactoferrin peptidìc fragments involved in inhibition of herpes simplex virus type 1 infection" Biochem Biophys Res Commun. 1999, 264 (1), 19-23).

Lactoferrin, which can have various origins including recombinant human or bovine, is a highly cationic glycoprotein capable of chelating two ferric ions per molecule with high affinity. Lactoferrin is important in iron homeostasis being able to prevent and treat disorders of iron metabolism (patent IT1367078). The oral administration of lactoferrin also influences the homeostasis of inflammation, through the inhibition of the synthesis of some pro-inflammatory cytokines (patent IT1367078).

The unproven correlation between the induction of PP and the pro-inflammatory cytokines found in the circulation or in the cervicovaginal secretion strongly supports the novelty of the present invention which claims that the prevention and treatment of a PP cannot be based on the exclusive reduction of cytokines at the cervico-vaginal level but involves a simultaneous decrease in the inflammatory state at the systemic level. To support this statement, it is sufficient to mention the association between a premature birth and a pathology of the oral cavity such as periodontal disease. In fact, in pregnant women suffering from periodontal disease and more at risk of PP, the only intravaginal topical therapy aimed at the decrease of pro-inflammatory cytokines in the cervico-vaginal secretion was found to be ineffective, and the PP is induced equally as the levels blood levels of pro-inflammatory cytokines remain elevated.

In one aspect, the present invention relates to lactoferrin, or at least a fragment thereof, for use in the therapy and / or prevention of pathological conditions inducing preterm deliveries and births, where said use comprises the administration of lactoferrin, or at least a its fragment, to a pregnant woman in the preterm period.

In another aspect, the present invention relates to a method for the therapy and / or prevention of pathological conditions that induce preterm births and births, where said method comprises the administration of lactoferrin, or at least a fragment thereof, to a pregnant woman in preterm period.

Infectious events sustained by fungi such as Candida spp, or by Gram-positive or Gram-negative bacteria are, if left untreated, frequent causes of preterm births and births. Non-infectious events due to preterm births and births include nutritional deficiencies, alcoholism, smoking, shortening of the cervix, uterine malformations, polyamnios, multiple pregnancies, placenta previa, placental abruption, premature rupture of membranes, iron deficiency, anemia, diabetes, pre-eclamsia, hypertension and hereditary thrombophilia.

It has surprisingly been found that, in infectious events or in severe infections localized in other districts than the cervical vaginal one, the simultaneous administration of oral and topical intravaginal lactoferrin prevents and counteracts uterine contractions and the consequent start of labor and of childbirth and preterm birth. In other non-infectious events listed above that increase the risk of PP, including iron deficiency and anemia, lactoferrin should be administered preventively by mouth in the prevention of pathological conditions that induce preterm births and births and, at the onset of symptoms of PP, should be administered both topically and orally at the same time. In the latter cases, intravaginal and oral lactoferrin are prepared and formulated separately in two different preparations.

The simultaneous intravaginal and oral administration of lactoferrin is particularly advantageous for women in whom, associated with infectious episodes including periodontal or non-infectious diseases, a series of events predisposing to PP are observed such as the onset of uterine contractility ascertained subjectively and objectively, the change in vaginal pH, iron deficiency, anemia, an increase in the concentration of proinflammatory cytokines in the circulation and in the vaginal cervical secretion, prostaglandins and fetal fibronectin.

The efficacy of the simultaneous intravaginal and oral administration of lactoferrin may be indicated by a subjectively and objectively demonstrated decrease in uterine contractility. Furthermore, in infectious events an inhibition of the number of microorganisms in the cervicovaginal area, a decrease in pro-inflammatory cytokines in the cervico-vaginal secretion and in the circulation, prostaglandins and fetal fibronectin and in non-infectious or infectious ones can be observed. distal from the cervix, a decrease in circulating pro-inflammatory cytokines, prostaglandins and fetal fibronectin.

Surprisingly, a new function of lactoferrin (of any human or bovine origin or recombinant) administered intravaginally, in combination with oral administration, has been found, which manifests itself with a rapid decrease in the frequency and intensity of uterine contractions, as well as prostaglandins with particular reference to PGE2 and PGF2 in the cervicovaginal secretion. This new function is also manifested by the decrease in the concentration of fetal fibronectin which indicates how the therapy and / or prevention of PP by simultaneous oral and intravaginal administration of lactoferrin are also attributable to the stabilization of fetal or maternal-fetal membranes, as shown by the decrease in fetal fibronectin. , glue between the maternal decidua and the amniotic membranes.

The efficacy of intravaginal lactoferrin associated with oral lactoferrin is also associated with a decrease in pro-inflammatory cytokines in the cervico-vaginal secretion and in the circulation.

If therapy with intravaginal lactoferrin associated with oral in all pathologies, preferably in multifactorial ones, which induce PP including hypoferremia and anemia, is performed daily, preterm deliveries and births are significantly reduced in their incidence and delivery occurs at term or in 40% of subjects between the thirty-sixth and thirty-seventh week of pregnancy with an average birth weight ranging between 2,800 grams for the newborn and 2,500 g for the newborn.

Preferably, for the use of the present invention, lactoferrin, or at least a fragment thereof, is administered within 8 hours from the recording of the uterine contractile activity.

Preferably, for the use of the present invention, lactoferrin, or at least a fragment thereof, is administered within 8 hours from the recording of the increase of prostaglandine in the cervico-vaginal secretion.

Preferably, for the use of the present invention, lactoferrin, or at least a fragment thereof, is administered within 8 hours from the recording of the increase of fetal fibronectin in the cervico-vaginal secretion in order to stabilize the maternal and fetal membranes.

Preferably, for the use of the present invention, lactoferrin, or at least a fragment thereof, is administered within 8 hours from the recording of the increase of proinflammatory cytokines in the cervico-vaginal secretion and in the circulation.

Preferably, for the use of the present invention, lactoferrin, or at least a fragment thereof, is administered within 8 hours from the recording of the onset of iron deficiency and anemia.

Preferably, for the use of the present invention, lactoferrin, or at least a fragment thereof, is administered simultaneously by topical intravaginal and oral route.

Preferably, for the use of the present invention, lactoferrin, or at least a fragment thereof, is administered topically intravaginally or, for the prevention of pathological conditions that induce preterm births and births, orally.

Preferably, for the use of the present invention, the PP is induced by at least one infectious process.

Preferably, for the use of the present invention, PP is induced by a non-infectious process selected from nutritional deficiencies, alcoholism, smoking, shortening of the cervix, uterine malformations, polyamnis, multiple pregnancies, placenta previa, placental abruption, premature rupture of membranes, iron deficiency, anemia, diabetes, pre-eclamsia, hypertension and hereditary thrombophilia.

Preferably, for the use of the present invention, the preterm period is before the thirty-second or between the thirty-second and thirty-seventh week of gestation.

Preferably, for the use of the present invention, the daily intravaginal dose of lactoferrin, or at least a fragment thereof, is between 200 and 600 mg, the daily oral dose of lactoferrin, or at least a fragment thereof, is It is between 200 and 400 mg and the total daily dose is between 400 and 1000 mg, preferably 600 mg.

Preferably, for the use of the present invention, lactoferrin is of human, bovine or recombinant human or bovine origin.

More preferably, for the use of the present invention lactoferrin is of bovine origin.

Preferably, for the use of the present invention, at least one lactoferrin fragment is a large fragment obtained by enzymatic digestion.

Preferably, for the use of the invention, at least one lactoferrin fragment is selected from N lobe (1 to 280 aa), C lobe (345 to 692 aa) or peptide fragments including those 86 to 258 aa and da 285 to 692 aa.

Preferably, for the use of the invention, lactoferrin is saturated in iron, zinc, copper and / or manganese in a degree ranging from 1 to 100%.

In particular, in the use according to the present invention, the first appearance of subjective signs reported by the pregnant woman (subjective diagnosis) is immediately confirmed by objective diagnostic methods.

In the event of a diagnosis of threat of delivery and preterm birth, topical intravaginal therapy is started within 8 hours with a lactoferrin, preferably bovine, in combination with a lactoferrin, preferably bovine, for oral use assisted by antibiotic therapy in the case of a diagnosis of a ' microbial infection. After 24 h and daily, the subjective data reported by the woman in therapy are recorded, while the data obtained with objective diagnostic methods are recorded on a weekly basis. If the PP threat is also associated with a maternal anemic state, oral therapy with lactoferrin, preferably bovine, must be started within 8 hours and obligatorily. The efficacy of lactoferrìna is assessed by the disappearance of the symptoms reported by the woman, confirmed by objective diagnostic methods by comparing the results obtained before and after therapy with particular reference to the decrease or absence of contractions, to the decrease in the concentration of fetal fibronectin , of proinflammatory cytokines and prostaglandins PGE2 and PGF2 in the cervico-vaginal secretion and in the circulation as well as the restoration of haematological values such as the number of red blood cells, hemoglobin, total serum iron, serum ferritin and hematocrit.

Efficacy studies have been conducted with respect to lactoferrìna in use according to the present invention including pregnant women who at the twentieth week showed no indication of a PP, together with pregnant women who, although showing no symptoms at the twentieth week, had pathologies that could cause and increase the risk of PP such as those affected by periodontal disease, iron deficiency, anemia, diabetes, hypertension and thrombophilia. Some of these, during pregnancy, have contracted infections in the urinary tract, genitourinary and amniotic fluid and have had febrile episodes.

On the other hand, pregnant women who had undergone cerclage due to shortening of the cervix and the presence of funneling were excluded.

In the absence or in the presence of infectious phenomena, Tables 1-6 summarize the mean values of both the subjective data reported by the woman and the objective data checked with different diagnostic methods in women with PP threat at different weeks of pregnancy, before and after therapy with intravaginal application of 100 mg of lactoferrin, preferably bovine, every 4, 6, 8 or 12 hours depending on the frequency of uterine contractions and the gestational period always associated with oral therapy of 100 mg of lactoferrin, preferably bovine , twice a day.

In the most severe cases, the same intervals of administration were performed with an intravaginal and oral application of 200 mg of lactoferrin, preferably bovine.

Table _ 1: _ Intravaginal application of lactoferrin (100 mg every 6 hours) in women with PP threat between the 22nd and 27th week of pregnancy not induced by infectious phenomena in association with 100 mg of lactoferrin orally twice a day ¬.

Subjective symptoms Before After 7 days of After 30 days and objective of PP between 22 therapy and 27 weeks of therapy

pregnancy

Onset of activity 1 to 5 <1-5 Not all uterine contractile * contractions / contractions / days per day

Cramps type Yes in 100% Decreased intermittent menstrual absences of the subjects frequency

in the supra-pubic region

Low back pain Yes in 100% Absent decrease in intensity subjects

Pelvic pressure Yes in 80% Absent Absent of the subjects

IL-6 (pg / ml) 450 280 120 Average values

Fetal Fibronectin> 50> 50 <50 (ng / ml)

Average values

PGE2 (pg / ml) 432 250 25 Average values

PGF2a (pg / ml) 920 740 345 Average values

Note :

* The criterion "painful or not painful contraction" is excluded because it is too subjective.

Table 2: Intravaginal application of lactoferrin (100 mg every 6 hours) in women with PP threat between the 28th and 34th week of pregnancy not induced by infectious phenomena in association with 100 mg of lactoferrin orally twice a day.

Subjective symptoms Before the After 7 days of After 30 days and objective of PP between 28 therapy therapy therapy and 34 weeks of

pregnancy

Onset of activity> 4/20 min or <4/20 min l / 2h uterine contractile *> 8 / h

Cramps type Yes in 100% Decreased intermittent menstrual absences of the subjects frequency

in the supra-pubic region

Low back pain Yes in 100% Absent decrease in intensity subjects

Pelvic pressure Yes in 80% Absent Absent of the subjects

IL-6 (pg / ml) 480 260 130 Average values

Fetal Fibronectin> 50> 50 <50 (ng / ml)

Average values

PGE2 (pg / ml) 480 278 100 Average values

PGF2c ((pg / ml) 840 670 373 Average values

Note :

* The criterion "painful or not painful contraction" is excluded because it is too subjective.

Table _ Intravaginal application of lactoferrin (100 mg every 6 hours) in women with PP threat between 35th and 37th week of pregnancy not induced by infectious phenomena in association with 100 mg of lactoferrin orally twice a day.

Subjective symptoms and Before After 7 days After 15-objective PP between 35 and 37 therapy therapy 30 days weeks of pregnancy therapy Appearance of activity 1 / 1-3 min 1/10 min 8 / day uterine contractile *

Menstrual-type cramps Yes in 100% Decrease in intermittent absences in the frequency subject's region

supra-pubic

Low back pain Yes in 100% Decrease Absent in subjects intensity Pelvic pressure Yes in 80% Absent Absent in subjects

IL-6 (pg / ml) 450 280 120 Average values

Fetal fibronectin (ng / ml)> 50> 50 <50 Mean values

PGEZ {pg / ml) 600 420 210 Average values

PGF2n (pg / ml) 1,400 873 450 Average values

Note :

* The criterion "painful or not painful contraction" is excluded because it is too subjective.

Table _ 4: Intravaginal application of lactoferrin (100 mg every 6 hours) in women with PP threat between the 22nd and 28th week of pregnancy induced by infectious phenomena in association with 100 mg of lactoferrin orally twice a day.

Subjective symptoms Before After 7 days of After 30 and objective of PP between 22 days of therapy and 27 weeks of pregnancy therapy

Onset of activity 1 to 5 <1-5 Not all uterine contractile * contractions / contractions / days per day

Menstrual-type cramps Yes in 100% Decrease in intermittent absences in the frequency subject's region

supra-pubic

Low back pain Yes in 100% Absent decrease in intensity subjects

Pelvic pressure Yes in 80% Absent Absent of the subjects

IL-6 (pg / ml) 2,000 1,250 220 Average values

Fetal Fibronectin> 50> 50 <50 (ng / ml)

Average values

PGE2 (pg / ml) 480 292 84 Average values

PGF2cl (pg / ml) 1,050 810 374 Average values

Table 5: Intravaginal application of lactoferrin (100 mg every 6 hours) in women with PP threat between the 28th and 34th week of pregnancy induced by infectious phenomena in association with 100 mg of lactoferrin orally twice a day.

Subjective symptomatology Before After 7 days After 30 days and objective PP between 28 therapy therapy therapy and 34 weeks of

pregnancy

Onset of activity> 4/20 min or <4/20 min l / 2h uterine contractile *> 8 / h

Cramps like Yes in 100% Decreased intermittent menstrual absences of the subjects frequency

in the supra-pubic region

Low back pain Yes in 100% Absent decrease in intensity subjects

Pelvic pressure Yes in. 80% Absent Absent of the subjects

IL-6 (pg / ml) 1,850 1170 180 Average values

Fetal Fibronectin> 50> 50 <50 (ng / ml)

Average values

PGE2 (pg / ml) 560 298 120 Average values

PGF2â € ž (pg / ml) 970 640 425 Average values

Table 6: Intr avaginal application of lactoferrin (100 mg every 6 hours) in women with PP threat between 35th and 37th week of pregnancy induced by infectious phenomena in association with 100 mg of lactoferrin orally twice a day.

Subjective symptoms Before After 7 days of After 15-ed objective of PP between 35 and therapy therapy 30 days of 37 weeks of pregnancy therapy Appearance of activity 1 / 1-3 min 1/10 min 8 / day uterine contractile *

Menstrual-type cramps Yes in 100% Decrease in intermittent absences in the frequency subject's region

supra-pubic

Low back pain Yes in 100% Absent decrease in intensity subjects

Pelvic pressure Yes in 80% Absent Absent of the subjects

IL-6 (pg / ml) 2250 1480 320 Average values

Fetal Fìbronectin> 50> 50 <50 (ng / ml)

Average values

PGEZ (pg / ml) 670 450 220 Average values

PGF2U (pg / ml) 1240 970 650 Average values

From the results of the study, following the use of lactoferrin both intravaginally and orally according to the invention, a significant decrease in subjective and objective symptoms was found in all women with the threat of premature birth between the twenty-second and thirty-seventh week of pregnancy, in the absence and in the presence of infectious phenomena.

Claims (17)

R I V E N D I C A Z I O N I 1. Lactoferrin, or at least a fragment thereof, for use in the therapy and / or prevention of pathological conditions that induce preterm births and births, where said use includes the administration of lactoferrin, or at least a fragment thereof, to a woman in preterm pregnancy. 2. Lactoferrin, or at least a fragment thereof, for use according to claim 1 wherein lactoferrin, or at least a fragment thereof, is administered within 8 hours of recording uterine contractile activity. 3. Lactoferrin or at least a fragment thereof for use according to claim 1 or 2 wherein lactoferrin, or at least a fragment thereof, is administered within 8 hours of recording the increase of prostaglandins in the cervicovaginal secretion. 4. Lactoferrin, or at least a fragment thereof, for use according to one of the preceding claims in which lactoferrin, or at least a fragment thereof, is administered within 8 hours from the recording of the increase of fetal fibronectin in the cervico-vaginal secretion to the in order to stabilize the maternal and fetal membranes. 5. Lactoferrin, or at least a fragment thereof, for use according to one of the preceding claims wherein lactoferrin, or at least a fragment thereof, is administered within 8 hours of recording the increase of proinflammatory cytokines in the cervico-vaginal secretion and in the circle. 6. Lactoferrin, or at least a fragment thereof, for use according to one of the preceding claims wherein lactoferrin, or at least a fragment thereof, is administered within 8 hours from the recording of the onset of iron deficiency and anemia. 7. Lactoferrin, or at least a fragment thereof, for use according to one of the preceding claims wherein lactoferrin, or at least a fragment thereof, is administered simultaneously topically, intravaginally and orally. 8. Lactoferrin, or at least a fragment thereof, for use according to one of the preceding claims wherein lactoferrin, or at least a fragment thereof, is administered orally in the prevention of pathological conditions that induce preterm births and births. 9. Lactoferrin, or at least a fragment thereof, for use according to one of claims 1 to 6 wherein lactoferrin, or at least a fragment thereof, is administered topically intravaginally. 10. Lactoferrin, or at least a fragment thereof, for use according to one of the preceding claims in which preterm birth is induced by at least one infectious process. 11. Lactoferrin, or at least a fragment thereof, for use according to one of the preceding claims in which preterm birth is induced by a non-infectious process selected from nutritional deficiencies, alcoholism, smoking, shortening of the cervix, uterine malformations, polyamnios, multiple pregnancies, placenta previa, placental abruption, premature rupture of membranes, iron deficiency, anemia, diabetes, pre-eclamsia, hypertension and hereditary thrombophilia. 12. Lattoferrìna, or at least a fragment thereof, for use according to one of the preceding claims in which the preterm period is before the thirty-second or between the thirty-second and thirty-seventh week of gestation. 13. Lactoferrìna, or at least a fragment thereof, for use according to one of the preceding claims in which the daily intravaginal dose of lactoferrìna, or at least a fragment thereof, is between 200 mg and 600 mg, the oral daily dose of lactoferrìna, or at least a fragment thereof, is between 200 and 400 mg and the total daily dose of lactoferrìna, or at least a fragment thereof, is between 400 and 1000 mg. 14. Lactoferrìna, or at least a fragment thereof, for use according to one of the preceding claims in which lactoferrìna is of human, bovine or recombinant human or bovine origin. 15. Lactoferrin, or at least a fragment thereof, for use according to claim 14 wherein lactoferrin is of bovine origin. 16. At least one lactoferrin fragment for use according to claim 14 wherein the fragment is selected from N lobe (1 to 280 aa), C lobe (345 to 692 aa) or peptide fragments including those 86 to 258 aa and 285 to 692 aa. 17. Lactoferrin, or at least a fragment thereof, for use according to one of the preceding claims in which the lactoferrin is saturated in iron, zinc, copper and / or manganese in a degree ranging from 1 to 100%.