KR102499386B1 - New use of melatonin - Google Patents

Abstract

As an invention related to a novel use of melatonin, the composition containing melatonin has the effect of alleviating intrauterine inflammation and oxidative stress and improving blood flow in the placenta, thereby reducing, improving, preventing, or reducing side effects caused by premature birth of premature babies. It can be used as an active ingredient of a composition for treatment.

Description

New use of melatonin {New use of melatonin}

It relates to a new use of melatonin, the use of which reduces, ameliorates, prevents or treats side effects caused by premature birth in premature infants.

According to the World Health Organization, a baby born less than 37 weeks of gestation or less than 259 days from the last menstrual period is called a preterm infant or premature baby. The main symptoms of side effects due to being born prematurely include lung damage due to underdevelopment of the lungs, brain damage due to underdevelopment of cerebrovascular vessels, and cardiovascular damage due to poor blood flow in the placenta.

Injecting corticosteroids into pregnant women who are expected to give birth prematurely can induce lung development by stimulating the alveoli of the fetus. However, the lung development stage of neonates born before 28 weeks of pregnancy is the tubular stage, and alveoli have not yet developed, so there is a limit to treating newborns with corticosteroid injections in the early developmental stage before birth. In addition, corticosteroids may have systemic side effects such as increased blood sugar in pregnant women and fetuses, so they are not usable in all cases, and their safety is controversial. In addition, if premature birth is expected, magnesium oxide is injected to the pregnant woman to prevent brain damage after birth, but the effect is insignificant, so premature babies born before 28 weeks of gestation often suffer from inflammatory brain damage, cerebral hemorrhage, and ischemic brain damage after birth. . In addition, many studies have shown that one of the reasons for damage to the fetal cardiovascular system in pregnancy accompanied by intrauterine inflammation is uteroplacental insufficiency.

Under such a technical background, research on a composition for preventing and treating side effects caused by premature birth in premature babies (Korean Registered Patent No. 10-1405620) has been actively conducted, but the situation is still insufficient.

One aspect is to provide a pharmaceutical composition comprising melatonin for reducing, ameliorating, preventing or treating side effects of premature birth.

Another aspect is to provide a functional health food composition containing melatonin for reducing, improving, preventing or treating side effects of premature birth.

Another aspect is to provide a method for reducing, improving, preventing or treating side effects of premature birth in premature infants, comprising administering a pharmaceutical composition containing melatonin to a subject.

One aspect provides a pharmaceutical composition containing melatonin for reducing, improving, preventing or treating side effects of premature birth.

The term "improvement" refers to all actions that improve or beneficially change symptoms due to side effects caused by premature birth or reduction of side effects caused by premature birth by administration of the pharmaceutical composition containing melatonin.

The term “treat” refers to an improvement in a condition (eg, one or more symptoms) of a subject suffering from or at risk of developing a disease, delaying disease progression, or delaying the onset of symptoms. or any form of treatment or prevention that provides an effect, including slowing the progression of symptoms, and the like. Accordingly, the term "treatment" also includes prophylactic treatment of a subject to prevent the occurrence of symptoms.

The terms “treatment” and “prevention” are not intended to mean cure or complete elimination of symptoms. They refer to any form of treatment that provides a benefit to a patient suffering from a disease, including improving the patient's condition (eg, one or more symptoms), delaying the progression of a disease, and the like.

The term "treatment-effective amount" refers to a side effect of premature birth, improvement of condition (eg, one or more symptoms), delay of disease progression, etc. means an amount sufficient to produce the desired effect, including

The terms “preterm infant,” “preterm infant,” or “preterm infant” may refer to a baby born less than 37 weeks of gestation or less than 259 days from the date of the last menstrual period. The causes of premature birth are poor socioeconomic conditions, mothers aged less than 16 years or older than 35 years old, prolonged standing or activities requiring physical stress, acute or chronic medical conditions, multiple births, If you have previously delivered a premature baby, if you have an obstetric disease such as uterine malformation, 　placenta previa, gestational hypertension or gestational diabetes, it may be a case where the fetus is in poor condition.

In one embodiment, the side effect may be lung damage of preterm infants, neonatal respiratory distress syndrome (RDS), bronchopulmonary dysplasia (BPD), and the like.

In one embodiment, the side effect may be brain damage of a premature infant, intracranial hemorrhage (ICH), and the like.

In one embodiment, the side effect may be cardiovascular damage of a premature infant, congestive heart failure (CHF), uteroplacental blood flow insufficiency, and the like.

In one embodiment, the premature infant may be less than 28 weeks of gestation.

Premature infants with a gestation period of less than 28 weeks have immature development of their body organs, so general pharmaceutical compositions often do not exhibit appropriate effects. there is.

In the case of a premature infant having a short gestational period, the cause of the premature birth may be due to intrauterine inflammation, premature labor, premature membrane rupture or cervical incompetence.

In one embodiment, the composition may be administered to a subject by intraperitoneal injection.

The term “subject” refers to all animals, including humans, ie, mothers, pregnant with fetuses that have or may have adverse effects due to premature birth, and more specifically, humans or non-human primates, mice , and mammals such as dogs, cats, horses, and cattle.

In one embodiment, the subject may have intrauterine inflammatory symptoms during pregnancy.

The symptoms of intrauterine inflammation may be accompanied by high fever, abdominal pain, odorous vaginal discharge, rapid fetal heartbeat, and the like.

In one embodiment, the composition may have an effect of alleviating intrauterine inflammation and oxidative stress.

The composition may be prepared by including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients for administration. A pharmaceutically acceptable carrier may be saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, liposome, and a mixture of one or more of these components, and, if necessary, an antioxidant , buffers, bacteriostatic agents and other conventional additives may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate formulations for injections such as aqueous solutions, suspensions, emulsions, pills, capsules, granules, or tablets, and may act specifically on target organs. A target organ-specific antibody or other ligand may be used in combination with the carrier. Furthermore, it can be preferably formulated according to each disease or component by using an appropriate method in the art or by using a method disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA). there is.

The composition may be prepared for oral, topical, parenteral, intranasal, intravenous, intramuscular, subcutaneous, intraocular, or transdermal administration. For example, it can be used in an injectable form. Accordingly, it can be mixed with any pharmaceutically acceptable vehicle for injectable compositions, particularly for direct injection into the area to be treated. The composition may include, inter alia, an isotonic sterile solution or a lyophilized composition which allows for the formulation of an injectable solution upon addition of sterile water or appropriate physiological saline. The pharmaceutical composition is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is based on the type, severity, activity of the drug, drug It may be determined according to factors including sensitivity to , time of administration, route of administration and excretion rate, duration of treatment, drugs used concurrently, and other factors well known in the medical field. The administration may be administered once a day or divided into several times.

Specifically, the effective amount of the pharmaceutical composition may vary depending on the patient's age, condition, body weight, absorption rate of the active ingredient in the body, inactivity rate, excretion rate, disease type, concomitant drugs, administration route, severity of obesity , may increase or decrease according to gender, weight, age, etc. The daily dosage is about 0.0001 to 10 mg/kg, preferably 0.001 to 1 mg/kg, and may be administered once or several times a day.

The pharmaceutical composition according to one embodiment reduces fetal IL1β, HIF-1β and CXCR2 expression levels, and increases surfactant protein B and vimentin expression levels to prevent lung damage.

It also reduces the amount of IBA1 expression in the fetus, preventing brain damage.

In addition, it normalizes the Doppler waveform change and Tei index of the blood vessels of the fetus, thereby preventing damage to the cardiovascular system.

Another aspect provides a health functional food composition containing melatonin for reducing, improving or preventing side effects of premature birth.

Among the terms or elements mentioned in the description of the composition, the same as those already mentioned are as described above. The content of the composition as an active ingredient contained in the health functional food may be appropriately selected without limitation depending on the type of food, desired use, etc., for example, 0.01 to 15% by weight of the total food weight may be added. there is. Also, for example, the health drink composition may be added at a rate of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml.

In one embodiment, the side effect may be lung damage in premature infants, brain damage in premature infants, or cardiovascular system damage in premature infants.

Another aspect provides a method for reducing, improving, preventing or treating side effects of premature birth in premature infants, comprising administering a pharmaceutical composition containing the melatonin to a subject.

The meaning of terms such as “pharmaceutical composition”, “side effects due to premature birth”, “subject”, “administration”, “prevention” or “treatment” may be within the aforementioned range.

In one embodiment, the subject may be an animal other than human.

According to the pharmaceutical composition or health functional food containing melatonin according to one aspect, it can be applied to reduce, improve, prevent or treat lung damage, brain damage or cardiovascular damage in premature infants.

In addition, since the composition has an effect of alleviating intrauterine inflammation and oxidative stress and improving blood flow in the placenta, it can be used as an active ingredient of a composition for reducing, improving, preventing, or treating side effects of premature birth.

1 is a result showing changes in the expression level of IL1β, IL6, Tnfα, HIF-1β and CXCR2 in lung tissue of a mouse fetus 24 hours after injection of LPS or PBS.
Figure 2 is a result showing the change in surfactant protein B and vimentin expression level of lung tissue of a mouse fetus 24 hours after injection of LPS or PBS.
Figure 3 is a result showing structural damage changes in lung tissue of mouse fetuses 24 hours after injection of LPS or PBS.
Figure 4 is a result showing the change in the expression level of oxidative stress markers in lung tissue of a mouse fetus 24 hours after injection of LPS or PBS.
5 is a result showing changes in Yap1/Taz expression levels in lung tissue of mouse fetuses 24 hours after injection of LPS or PBS.
6 is a result showing the change in IBA1 expression level in the brain tissue of a mouse fetus 24 hours after injection of LPS or PBS.
7 is a result showing Doppler waveform changes of blood vessels of a mouse fetus 6 hours after LPS or PBS injection.
8 is a result showing the change in the Tei index of the blood vessels of mouse fetuses 6 hours after LPS or PBS injection.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

Example. Preparation of the mouse model

Pregnant ICR mice (Koatech, Pyeongtack, Korea) or CD1 mice (Charles River Laboratories, Wilmington, MA, USA) were used. To create an intrauterine inflammation rat model, gestation day 17 (E17) rats were inhaled anesthetized with isoflurane (Baxter # NDC 10019-360-60, Deerfield, IL, USA). The abdominal area was disinfected and a 1.5 cm vertical incision was made in the center of the lower abdomen. A solution of 25 μg of LPS (Escherichia coli, 055: B5; Sigma-Aldrich, St. Louis, MO, USA) dissolved in 100 μl of Phosphate-buffered saline (PBS) was administered between the first and second amniotic cavities in the lower right part of the uterus. injected into This procedure was performed carefully so as not to puncture the amnion. Afterwards, the abdominal wall was closed and the pregnant mice were allowed to recover.

Experimental Example 1. Confirmation of composition for preventing lung damage and promoting lung maturation

In order to confirm whether the composition has an effect of preventing lung damage caused by premature birth in premature babies, the following experiment was conducted.

Pregnant rats were divided into 4 groups: control group, LPS group, melatonin alone administration group, and melatonin and LPS administration group together. LPS, one of the components of the cell wall of Gram-negative bacteria, has been used in mouse models to study complications associated with preterm birth. On day E17, control rats were intraperitoneally injected with PBS, and 30 minutes later, intrauterine PBS was injected. In the LPS group, pregnant mice were intraperitoneally injected with LPS, and 30 minutes later, intrauterine LPS was injected. In the melatonin group, mice were intraperitoneally injected with 10 mg/kg of melatonin (Sigma-Aldrich), followed by intrauterine PBS injection 30 minutes later. In addition, in the group administered with melatonin and LPS, melatonin was injected intraperitoneally, and LPS was injected intrauterinely 30 minutes later.

As shown in Figure 1, the composition according to one embodiment was found to reduce the expression levels of IL1β, HIF-1β and CXCR2 in the lungs of mouse fetuses.

In addition, as shown in Figure 2, it was found to increase the surfactant protein B and non-mentin expression of the lungs of the mouse fetus.

As shown in Figure 3, the composition according to one embodiment was shown to prevent structural damage to the lungs of mouse fetuses.

As shown in Figure 4, the composition according to one embodiment was found to reduce the expression level of oxidative stress markers in the lungs of mouse fetuses.

As shown in FIG. 5 , the composition according to one embodiment was found to prevent a decrease in Yap/Taz expression level in the lungs of mouse fetuses.

These results mean that by administering the composition according to one embodiment, lung damage, which is a side effect of premature birth of the fetus, can be prevented and lung maturation can be promoted.

Experimental Example 2. Confirmation of the brain damage prevention effect of the pharmaceutical composition

In order to determine whether the composition has an effect of preventing brain damage caused by premature birth in premature infants, the following experiment was conducted.

Pregnant rats were divided into 4 groups: control group, LPS group, melatonin alone administration group, and melatonin and LPS administration group together. LPS, one of the components of the cell wall of Gram-negative bacteria, has been used in mouse models to study complications associated with preterm birth. On day E17, control rats were intraperitoneally injected with PBS, and 30 minutes later, intrauterine PBS was injected. In the LPS group, pregnant mice were intraperitoneally injected with LPS, and 30 minutes later, intrauterine LPS was injected. In the melatonin group, mice were intraperitoneally injected with 10 mg/kg of melatonin (Sigma-Aldrich), followed by intrauterine PBS injection 30 minutes later. In addition, in the group administered with melatonin and LPS, melatonin was injected intraperitoneally, and LPS was injected intrauterinely 30 minutes later.

As shown in Figure 6, the composition according to one embodiment was shown to reduce the amount of IBA1 expression in the brain tissue of the mouse fetus.

These results mean that by administering the composition according to one embodiment, brain damage of the fetus, which is a side effect of intrauterine inflammation, can be prevented.

Experimental Example 3. Confirmation of the effect of preventing cardiovascular system damage of the pharmaceutical composition

In order to confirm whether the composition has an effect of preventing cardiovascular damage caused by premature birth in premature infants, the following experiment was conducted.

Pregnant rats were divided into 4 groups: control group, LPS group, melatonin alone administration group, and melatonin and LPS administration group together. LPS, one of the components of the cell wall of Gram-negative bacteria, has been used in mouse models to study complications associated with preterm birth. On day E17, control rats were intraperitoneally injected with PBS, and 30 minutes later, intrauterine PBS was injected. In the LPS group, pregnant mice were intraperitoneally injected with LPS, and 30 minutes later, intrauterine LPS was injected. In the melatonin group, mice were intraperitoneally injected with 10 mg/kg of melatonin (Sigma-Aldrich), followed by intrauterine PBS injection 30 minutes later. In addition, in the group administered with melatonin and LPS, melatonin was injected intraperitoneally, and LPS was injected intrauterinely 30 minutes later.

As shown in Figures 7 and 8, the composition according to one embodiment was found to normalize the Doppler waveform and the Tei index of the fetal heart.

These results mean that by administering the composition according to one embodiment, it is possible to prevent damage to the cardiovascular system of the fetus, which is a side effect of intrauterine inflammation.

Claims (13)

A pharmaceutical composition containing melatonin for reducing, improving, preventing or treating side effects of premature birth,
The side effect is selected from the group consisting of lung damage of premature infants, brain damage of premature infants, and cardiovascular damage of premature infants,
The composition is administered to a subject who is pregnant,
The pharmaceutical composition, wherein the subject has intrauterine inflammatory symptoms during pregnancy. delete delete delete The method of claim 1, wherein the side effect is neonatal respiratory distress syndrome (RDS), bronchopulmonary dysplasia (BPD), intracranial hemorrhage (ICH), congestive heart failure (CHF) Or a pharmaceutical composition that is uteroplacental insufficiency. The pharmaceutical composition according to claim 1, wherein the premature infant is less than 28 weeks of gestation. delete delete The pharmaceutical composition according to claim 1, wherein the composition has an effect of alleviating inflammation and oxidative stress in the uterus. A health functional food composition containing melatonin for reducing, improving or preventing side effects caused by premature birth of premature babies,
The side effect is selected from the group consisting of lung damage of premature infants, brain damage of premature infants, and cardiovascular damage of premature infants,
The composition is administered to a subject who is pregnant,
The subject is a health functional food composition containing melatonin that has intrauterine inflammatory symptoms during pregnancy. delete A method for reducing, improving, preventing or treating side effects of premature birth in premature infants, comprising administering a pharmaceutical composition containing the melatonin of claim 1 to a subject,
The side effect is selected from the group consisting of lung damage of premature infants, brain damage of premature infants, and cardiovascular damage of premature infants,
The subject is an animal other than human,
A method for reducing, improving, preventing or treating side effects of premature birth in which the subject has symptoms of intrauterine inflammation during pregnancy. delete

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