KR20060115977A - Therapeutic agent for treating sleep disorder of prader-willi syndrome children containing adiponectin as active ingredient - Google Patents

Abstract

A therapeutic agent for treating sleep disorder of Prader-Willi syndrome in children containing adiponectin as an active ingredient is provided to alleviate sleep disordered breathing(SDB) and induce rapid eye movement(REM). The therapeutic agent for treating sleep disorder of Prader-Willi syndrome in children contains adiponectin as an active ingredient, wherein the dosage of the therapeutic agent in children is 0.1-10 mg/kg per day; and the therapeutic agent further contains pharmaceutically acceptable carriers and auxiliary.

Description

Therapeutic agent for treating sleep disorder of Prader-Willi syndrome children containing adiponectin as active ingredient

FIG. 1A shows plasma adiponectin levels in Prader-Willi syndrome (PWS) children and controls. "-" Represents an average value.

FIG. 1B shows plasma lecithin levels in children with Prader-Willi syndrome (PWS) and control group. "-" Represents an average value.

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1C shows plasma retinol binding protein-4 (RBP4) levels in Prader-Willi syndrome (PWS) children and controls. "-" Represents an average value.

Figure 2 shows the correlation between rapid eye movement (REM) sleep rate and body fat rate in children with Prader-Willi syndrome (PWS).

Figure 3 shows the correlation between REM sleep rate and plasma adiponectin level in children with Prader-Willi syndrome (PWS).

The present invention relates to adiponectin that prolongs rapid eye movement sleep (REM sleep) during sleep in children with Prader-Willi syndrome. More particularly, the present invention relates to the use of adiponectin to treat sleep respiratory disorder in children with Prader-Willi syndrome, which usually leads to sleep respiratory disorder and to induce rapid eye movement sleep (REM sleep).

Prader-Willi syndrome (PWS) is a genetic disorder that predominantly decreases tension in infancy and becomes more pronounced in children and adolescents with developmental delays, obesity and behavioral problems. Because of the development of obesity, craniofacial dysplasia and muscle tone, PWS patients are at risk of sleep disordered breathing (SDB), such as obstructive sleep apnea (OSA), central apnea, or hypoventilation. On the other hand, since growth hormone worsens sleep respiratory disorders, careful examination is required before administration of growth hormone to children with PWS symptoms.

In addition to sleep respiratory disorders in PWS patients, other sleep disorders have been reported, such as excessive sleep, early eye movement (REM) sleep circadian rhythms, and sleep seizure characteristics during REM sleep. In addition, PWS patients have an early abnormal ventilation response to hypoxia and hypercarbonate, which is exacerbated by obesity.

Adipose tissue secretes several factors, including adipokine, supplements, and components of the coagulation chain reaction. The levels of these factors are higher in obese patients. Moreover, plasma adipokin levels are closely related to insulin resistance and obesity. Lecithin is one of the polypeptide hormones produced by adipocytes and has been shown to play an important role in the development of hepatic insulin resistance in rodents.

RBP4 (Retinol Binding Protein-4) is a type of adipokin that limits glucose absorbed by skeletal muscle and increases blood glucose levels by increasing glucose produced by the liver. Moreover, circulating RBP4 levels were substantially higher in humans as well as in several mouse models of obesity and insulin resistance symptoms. Moreover, insulin-sensitive agents have been shown to reduce elevated RBP4 levels in both mouse adipose tissue and serum.

Meanwhile, adiponectin, a kind of adipokin-based hormone, is one of endocrine factors. Adiponectin was first isolated from human adipose tissue by Maeda et al. In 1996 and is a secreted protein consisting of 244 amino acid residues of approximately 30 KDa, which is specifically expressed in animal adipose tissue (K.Maeda et al., Biochem.Biophys.Res.Commun., 221, 286-289 (1996)). To date, several groups of studies have revealed the action of adiponectin. Specifically, anti-obesity action, anti-diabetic action, anti-arteriosclerosis action and inhibitory action of reactive oxygen production are known (AH Berg et al., Nat. Med., 7, 947-953 (2001); T. Yamauchi et al., Nat. Med., 8, 1288-1295 (2002); Iichiro Shimomura et al., Medical Science Digest, 28 (12), 17-20 (2002); Toshimasa Yamauchi, Takashi Kadowaki, Experimental Medicine, 20 (12) (August), 1762-1767 (2002); H. Motoshima et al., Biochemical and Biophysical Research Communications, 315 (2004), 264-271).

This document describes the action of adiponectin secreted from adipose tissue, which inhibits the adhesion of monocytes to vascular endothelial cells, inhibits lipid accumulation and foaming of macrophages, and also inhibits proliferation and migration of smooth muscle cells. It is described that the anti-arteriosclerosis effect is achieved by.

In addition, adiponectin is secreted from adipocytes after meals to promote fatty acid oxidation by activating AMP kinase (AMPK) and burning excess fat or sugar by meals to lower blood sugar levels, but fat accumulates in muscles and liver by obesity. It is known that the function of adipocytes that secrete adiponectin is weakened, the absorption of sugar in the blood is reduced, and as a result, it leads to diabetes. In other words, hyperlipidemia such as obesity and hyperlipidemia and diabetes are closely related to the action of adiponectin.

On the other hand, sleep profiles change with age and change more with REM sleep than with slow-wave sleep. In general, the amount of REM sleep time decreases rapidly during the first year after birth as the REM / non-REM cycle develops. To date, sleep disorders associated with PWS are not well understood and are simply considered "hypothalamic defects".

The present invention measured the relationship between the plasma adipokin level and the polysomnography parameters in children with PWS, the plasma levels of adiponectin, lecithin and RBP4 were measured and the relationship with the polysomnography parameters was calculated accordingly. The relationship between sleep and adipokin was also analyzed according to age, obesity index and insulin resistance.

The present invention determined that REM sleep rate of PWS children is closely related to hormone factors such as plasma adiponectin level as well as age, thereby completing the present invention.

The technical problem to be achieved in the present invention is to develop a therapeutic agent that treats sleep respiratory disorder in children with Prader-Willi syndrome that usually causes sleep respiratory disorder and induces rapid eye movement sleep (REM sleep).

The present invention provides a therapeutic agent for children with Prader-Willi syndrome, which alleviates sleep respiratory disorders containing adiponectin as an active ingredient and induces rapid eye movement sleep (REM sleep).

In addition, the daily dose of the Prader-Willy syndrome pediatric sleep disorder therapeutic agent is to provide a therapeutic agent, characterized in that the body weight of 0.1 to 10 mg / kg.

In addition, Prader-Willy syndrome pediatric sleep disorder treatment of the present invention is characterized in that it further comprises adiponectin and a pharmaceutically acceptable carrier and adjuvant.

Hereinafter, the present invention will be described in more detail.

In the present invention, plasma adiponectin selected for the treatment of sleep respiratory disorder in children with PWS was measured to have a large pharmacological effect on the sleep stage period, especially REM sleep rate, even after adjusting the age, insulin resistance and obesity of the administered patient. This means that PWS children with elevated adiponectin levels in plasma show longer REM sleep during sleep periods than PWS children with reduced adiponectin levels in plasma. Therefore, the present invention is the first to identify the correlation between plasma adiponectin levels and REM sleep rate (or vice versa non-REM sleep rate) and through this attempt to develop a therapeutic agent for sleep respiratory disorder.

To date, plasma adiponectin levels have been reported to be negatively associated with insulin resistance, resulting in high incidence in sleep apnea syndrome subjects with low adiponectin levels. However, in the present invention, it is the first time that REM sleep rate, not the apnea-low apnea index, correlates with plasma adiponectin level even after adjusting age, body fat rate and HOMA-IR (evaluation of homeostasis model for insulin resistance).

This correlation between REM sleep and plasma adiponectin was also significant after HOMA-IR was replaced by WBISI (Whole Body Insulin Sensitivity Index). Moreover, this correlation was significant even after controlling for factors such as age, obesity (body fat percentage), insulin resistance (HOMA-IR) and apnea-low breathing.

It is not clear how the adiponectin of the present invention affects the sleep stage, but it is believed that the sleep stage is affected by cytokines in the brain and that plasma adiponectin levels regulate the production of cytokines.

Recently, a mechanism for the effect of adiponectin on cytokine expression of brain endothelial cells has been elucidated. In animal experiments, adiponectin administered parenterally to mice did not cross the blood brain barrier, but treatment with adiponectin reduced the release of active interleukin-6 from brain endothelial cells. This cerebrovascular effect of adiponectin contrasts with that of other pro-inflammatory cytokines.

Interleukin 1 beta, on the other hand, is known to increase non-REM sleep through increased c-Fos expression in the medial pre-nucleus of the hypothalamus. Moreover, interleukin-6 is known as a vehicle of drowsiness, and its circadian pattern reflects a homeostatic trend for sleep. Mice lacking interleukin-6 spend more time in REM sleep.

Thus, the most reliable explanation for the adiponectin effect on increased REM sleep rate is that increased adiponectin lowers the expression of pro-inflammatory cytokines in brain endothelial cells in PWS children. Thus, adiponectin reduces levels of cytokines such as interleukin 6, which increases REM sleep.

PWS pediatric sleep disorder treatment dose of adiponectin of the present invention may be reduced depending on the symptoms, but can be converted into a daily dose of 0.1 to 10 mg / kg pediatric body weight and about 3% of adiponectin in the blood It seems to increase. This can be seen through the animal experiments confirmed that the 1.0 mg / g body weight / dose to increase the adiponectin blood concentration of the mouse by about 60%.

It should be noted that young PWS children, a group of the present invention, are somewhat specific for insulin resistance. As shown in Table 1, subjects of the present invention had similar body fat percentage but were insulin sensitive. Moreover, high insulin sensitivity has been reported in PWS children. Since there is a high correlation between adiponectin and insulin sensitivity, higher adiponectin levels in PWS children than in controls are closely related to the distribution of fat, and PWS children have been reported to preferentially accumulate fat in subcutaneous tissue.

In contrast to previous studies, there was no correlation between apnea-low apnea index and plasma adiponectin levels in the present invention. PWS subjects in the present invention are somewhat younger (mean age 9.5 years) and there may be a negative correlation between age and adiponectin levels in the PWS children of the present invention, thus showing a higher insulin resistance and a higher apnea and lower apnea experience Is believed to show different results than previous studies in many PWS patients.

In conclusion, plasma adiponectin levels act as an independent function of REM sleep stage duration in children with PWS despite age, obesity or insulin resistance.

The present invention will be described in more detail with reference to the following examples. However, these examples do not limit the scope of the present invention.

Subject

The experiment was conducted in two parts. The first part consists of approximately 100 PWS children in the hospital and 26 PWS children who were invited and agreed to participate in the trial [mean age 9.5 years, quartile range (7.0-11.0 years), BMI: mean 24.1 kg / m2 ( 19.7-27.9 kg / m²) and 13 healthy obese children [mean age 10.0 years, quartile range (7.0-12.0 years), BMI: average 23.3 kg / m² quartile range (21.9-29.6 kg / m²)] A comparative study of plasma adipokin levels was included. The second part included another study of 26 PWS children enrolled in the first part of the study, and a sleep polymorphism test was performed overnight. All subjects of all diabetic symptoms (ie> 126 mg / dl fasting plasma per 2 hour oral glucose tolerance test> 200 mg / dl) were excluded. No growth hormones or other drugs were used for the participants in this study.

All subjects were pre-pubertal. Table 1 lists the clinical characteristics of the patients. Controls were recruited from several elementary and middle schools in southern Seoul. The purpose of the study was explained to the teacher, and a written experimental protocol was sent to all parents, including information on the age, gender, and distribution of BMI in the PWS children who participated in the study. Consent notices were received from participants or parents or guardians of appropriate controls as well as parents or guardians of PWS children.

Statistical analysis

HOMA-IR (evaluation of homeostasis model for insulin resistance) and WBISI (systemic insulin sensitivity index) were calculated as insulin sensitivity index.

All figures are indicated in the table as mean and quartile ranges. T-test with Bonferroni calibration was used if the sample was normally distributed. If the sample was not normally distributed, the Mann-Whitney test with Bonferroni calibration was used to compare hormone levels between the PWS and the control group.

Correlations were measured using Spearman correlation analysis because the samples were not normally distributed. A P value of <0.05 was considered significant. Moreover, plasma adipokin levels were further analyzed to determine whether they were independently related to sleep parameters even after being adjusted for obesity and insulin resistance by Spearman partial correlation analysis. All statistical analyzes were performed using SAS version 8.2 (SAS Corporation, Cary, NC).

Example 1 Design of Experiment for Measuring Insulin Resistance and Adipokin

All subjects were housed in the pediatric ward of Samsung Hospital to guarantee night fasting. After 10-12 hours of night fasting, children were subjected to oral glucose tolerance test (OGTT; 1.75 g / kg, up to 75 g) to measure insulin sensitivity index, including WBISI (Whole Body Insulin Sensitivity Index). Blood samples were taken at 0, 30, 60, 90 and 120 minutes to measure blood glucose and insulin levels. Samples were collected on ice and immediately centrifuged at 4 ° C. and stored at −70 ° C. until needed for analysis. Fasting plasma samples (0 min OGTT samples) were measured for adiponectin, lecithin and RBP4 levels.

Body fat percentage was assessed by dual energy x-ray absorptiometry (DXA) using Expert-XL (Lunar Corp. Madison, Wis.).

Table 1 shows the clinical characteristics of the subject of the present invention. PWS and obese control children had similar age and sex ratios. However, although BMI, BMI SDS and body fat percentage were similar between the two groups, as demonstrated by HOMA-IR and WBISI [HOMA-IR; PWS vs. Control, 3.04 (2.26-5.37): 5.51 (5.23-7.71), P = 0.017, WBISI; PWS vs. Control, 3.51 (1.90-5.26): 2.08 (1.16-2.57), P = 0.032] PWS children were more insulin-sensitive than controls.

Example 2 Hormone Analysis

Plasma levels were measured using a YSI 2300 dual analyzer (Yellow Springs Instrument Co., Yellow Springs, OH), and serum insulin was measured at 1 μU / ml detection limit and intra- and inter-assay coefficients of <10% deviation. Measurements were made using commercially available immunoradiometric kits (BioSource Europe SA).

Plasma adiponectin levels were measured using intra- and inter-assay coefficients of <10% and <15% deviations, and lower and upper detection limits of 0.15 and 10 ng / ml, respectively, using conventional ELISA kits (Phoenix Pharmaceuticals, Belmont, CA). Was measured.

Plasma lecithin levels were determined using conventional ELISA kits (Phoenix Pharmaceuticals, Belmont, CA) with intra- and inter-assay coefficients of <10% and <15% deviations, and lower and upper detection limits of 0.16 and 1 ng / ml, respectively. It was measured using.

Plasma retinol binding protein-4 (RBP-4) levels were measured with intra- and inter-assay coefficients of <5% and <14% deviations, respectively, and lower and upper detection limits of 0.2.03 and 20.7 ng / ml, respectively. It was measured using the kit (Phoenix Pharmaceuticals, Belmont, CA).

Comparison of Plasma Adiponin Levels (Adiponectin, Lecistin and RBP4) in PWS Children and Normal Obesity Controls

Plasma adiponectin levels were significantly higher in PWS children than in controls (P = 0.0006 by Mann-Whitney test) (FIG. 1A). However, lecithin and RBP4 levels were not high in PWS children (P = 0.789 and P = 0.222, respectively, by the Mann-Whitney test) (FIG. 1b, c).

Correlation between adipokin levels and obesity index (body fat percentage and BMI SDS) and between adipokin levels and insulin sensitivity index (HOMA-IR, QUICK and WBISI)

In PWS children, a positive correlation was observed between plasma lecithin and body fat percentage (r = 0.645, P = 0.0004) and between plasma lecithin and BMI SDS (r = 0.637, P = 0.0005). In contrast, plasma RBP4 was associated with body fat percentage (r = 0.632, P = 0.02) and BMI SDS (r = -0.731, P = 0.002) in the control group. However, there was no association between plasma adiponectin levels and obesity index in PWS children or controls. No correlation was observed between adipokin and insulin sensitivity index (P> 0.05 by Spearman correlation analysis).

Example 3 night sleep polymorphism test

The day before the night sleep polyp, the patient was asked not to drink caffeine. Nocturnal sleep studies were recorded on the Alice-3 system (Healthdyne, USA) and the Compumedics S-series (Compumedics, Australia). Standard sleep polygraphs include four-channel electroencephalograms (EEG, C3 / A2; C4 / A1; O1 / A2; O2 / A1) and four-channel safety latitudes (EOG), submandibular, intercostal and anterior tibial muscles ( EMG) and electrocardiogram to surface electrodes. Thermistors (for nasal airflow monitors), nasal air pressure monitors, oxygen meters (for oxygen saturation), piezoelectric bands (for chest and abdominal wall exercises) and body position sensors were also attached to the patient. Patients were recorded on videotape using an infrared video camera and subsequently observed by a polysomnologist. The patient fell asleep between 10:00 and 11:00 PM, and sleep timing was interpreted according to Rechtshaffen and Kales criteria. REM sleep rate is defined as (REM sleep time / total sleep time) x 100%.

Sleep Parameters vs Obesity Index vs Insulin Sensitivity Index

Table 2 shows the mean and quartile ranges of the sleep parameter indices. Sleep polysomnography was performed only in PWS children, not controls. There was a negative correlation between REM sleep rate and body fat rate (r = -0.539, P = 0.005) (FIG. 2). Moreover, REM sleep rate was negatively correlated with the apnea-low breathing index (r = -0.426, P = 0.029). However, body fat percentage was not correlated with apnea-low breathing index (r = 0.364, P = 0.067). Other sleep parameters were not correlated with insulin sensitivity index.

Sleep Parameters vs. Adipokin

Adiponectin levels were correlated with total sleep time (r = 0.399, P = 0.022), REM sleep latency (r = -0.489, P = 0.006) and REM sleep rate (r = 0.675, P = 0.0001) (FIG. 3). However, there was no significant correlation between lecithin and sleep parameters or RBP4 and sleep parameters.

Age vs. Sleep Parameters and Age vs. Adipokin

The patient's age was negatively correlated with total sleep time (r = -0.584, P = 0.0017) and REM sleep rate (r = -0.561, P = 0.003) while positively correlated with arousal index (r = 0.418, P = 0.033). There was a moderate inverse correlation between age and adiponectin levels (r = −0.412, P = 0.036).

Correlation between Adipokin and Sleep Parameters after Modulation of Age, Obesity, and Insulin Resistance

Since body fat percentage and age were correlated with sleep parameters and adiponectin levels were related to body fat percentage and age, plasma adiponectin levels were measured by Spearman partial correlation analysis to determine whether they were independently related to sleep parameters even after controlling age, obesity, and insulin resistance. Further investigation was done to.

After controlling for age, obesity and insulin resistance index, there was no correlation between total sleep time and adiponectin levels (r = 0.202, P = 0.35). However, there was a correlation between adiponectin levels and REM sleep rate (age, body fat rate, and correlation between adiponectin and REM sleep rate after HOMA-IR control: r = 0.578, P = 0.0039). Moreover, the correlation was significant even when HOMA-IR was replaced with WBISI (correlation between age, body fat rate and adiponectin and REM sleep rate after WBISI control: r = 0.584, P = 0.0034).

Since the REM sleep rate was negatively correlated with the apnea-low breath index, we tested the correlation between REM sleep rates, including adiponectin and apnea-low breath index. In addition to age, HOMA-IR and body fat percentage, the correlation was also significant when the apnea-low apnea index was adjusted (r = 0.570, P = 0.005).

The effect of the present invention is to provide a therapeutic agent that treats sleep respiratory disorder in children with Prader-Willi syndrome, which usually causes sleep respiratory disorder, and induces rapid eye movement sleep (REM sleep).

Claims (3)

Prader-Willy Syndrome Children's Sleep Disorders Relieve Sleep Respiratory Disorder with Adiponectin as an Active Ingredient and Induce Rapid Eye Movement Sleep (REM Sleep) The method of claim 1, wherein the daily dose of Prader-Willi syndrome pediatric sleep disorder therapeutic agent is 0.1 to 10 mg / kg pediatric weight disorder therapeutic agent for pediatric sleep disorders The method of claim 1, wherein the Prader-Willy syndrome pediatric sleep disorder treatment agent further comprises adiponectin and a pharmaceutically acceptable carrier and adjuvant

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