Composition for preventing and treating of neuropathic pain containing <i>Nypa fruticans </i>Wurmb extract

Abstract

Provided are a composition for preventing and treating neuropathic pain, comprising a Nypa fruticans Wurmb extract, and a method of treating or preventing a neuropathic pain, comprising administering a composition comprising a Nypa fruticans Wurmb extract to a subject in need thereof.

Claims

1. A method of treating a human suffering from a spinal cord injury consisting essentially of administering a therapeutically effective amount of an extract of Nypa fruticans Wurmb and an extract selected from the group consisting of Cephalotaxus koreana Nakai extract, Sceptridium tematum (Thunb.) Lyon extract, Saururus chinensis (Lora.) Bail, extract and mixtures thereof to the human in need thereof to effectively treat the spinal cord injury in the human.

2. The method according to claim 1, wherein the human suffering from the spinal cord injury also suffers from nerve damage, neuropathy, central neuropathic pain or peripheral neuropathic pain.

3. The method according to claim 1, wherein the method inhibits transient receptor potential vanilloid 1 (TRPV1) to exhibit antinociceptive and nerve regeneration effects in the human suffering from the spinal cord injury.

4. The method according to claim 1, wherein the Nypa fruticans Wurmb is extracted with a solvent selected from the group consisting of water, C.sub.1 to C.sub.6 lower alcohols, and mixtures thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 shows a neuropathic pain model test result of a composition according to one embodiment of the present disclosure.

DETAILED DESCRIPTION

(2) In the following detailed description, reference is made to the accompanying drawing, which forms a part hereof. The illustrative embodiments described in the detailed description, drawing, and claims are not meant to be limiting. Other embodiments may be utilized, and other changes may be made, without departing from the spirit or scope of the subject matter presented here.

(3) Hereinafter, embodiments of the present disclosure are described in detail so that those skilled in the art can easily carry out the present disclosure. However, the present disclosure may be embodied in many different forms and is not limited to the embodiments described herein.

Preparation Example: Preparation of Extract

1. Preparation of Nypa fruticans Wurmb Extract

(4) 100 g of dry pulverized Nypa fruticans Wurmb extract was mixed with 1 L of 80% ethanol at 20 C., and the extraction was carried out at room temperature for 4 hours. After the extraction, a filter cake was used to remove the powder component, and only the liquid component was filtered through the flask. The filtered liquid component was boiled in a water bath at 65 C. to produce pure Nypa fruticans Wurmb extract (N1).

2. Preparation of Other Natural Extracts

(5) 1 kg of the original sample of the dry pulverized Cephalotaxus koreana Nakai extract in a powder form was added to 30% ethanol, and the reflux extraction was performed at 80 C. for 1 to 3 hours. The extraction was repeated twice at the same times and at the same period, and the obtained extract was dried under reduced pressure and then vacuum dried at 60 C. to prepare Cephalotaxus koreana Nakai extract (N2).

(6) Botrychium ternatum Swartz. extract (N3), Saururus chinensis (Lour.) Baill. extract (N4), Pachysandra terminalis extract (X1) and Siraitia grosvenorii extract (X2) were prepared using the same method as the preparation method of Cephalotaxus koreana Nakai extract (N2).

3. Preparation of Complex Extract

(7) The Nypa fruticans Wurmb extract (N1), Cephalotaxus koreana Nakai extract (N2), Botrychium ternatum Swartz. extract (N3), Saururus chinensis (Lour.) Baill. extract (N4), Pachysandra terminalis extract (X1) and Siraitia grosvenorii extract (X2) were mixed in the weight range as shown in the following Table 1 to prepare a composition for preventing and treating neuropathic pain.

(8) TABLE-US-00001 TABLE 1 G1 G2 G3 G4 G5 G6 G7 G8 G9 N1 100 100 100 100 100 100 100 100 100 N2 30 40 60 80 30 40 60 80 N3 30 40 60 80 30 40 60 80 N4 30 40 60 80 30 40 60 80 X1 10 20 30 40 X2 30 40 50 60 (Unit: parts by weight)

Experimental Example 1: Neuropathic Pain Relief Effect of Nypa fruticans Wurmb Extract

(9) The formalin test a representative experimental model for evaluating the inflammatory pain behavior response through injured tissue and inflammatory reaction caused by subcutaneously injecting formalin into the facial surface of mouse. Capsaicin, an activator of TRPV1, can be additionally injected to evaluate neuropathic pain behavior response.

(10) The specific experimental method is described below. The experimental animals were adapted for at least 10 minutes in a laboratory plastic container before evaluation of their pain response. 25 l of 5% formalin was injected using an insulin syringe (0.258 mm), and they were observed to regain consciousness within a few seconds after the injection of formalin. First, to examine the effect of TRPV1 channel activity on the behavior response of formalin-injected experimental animals, the pain behavior responses caused by capsaicin alone at a concentration of 0.1 g/10 L, 1 g/10 L, 10 g/10 L injected through an inserted polyethylene tube were observed in the same manner as after the formalin injection. Next, the pain behavior responses were observed for 45 minutes after injection of capsaicin at a concentration of 0.1 g/10 L and 1 g/10 L on the same injection site one hour before the formalin injection. The solvent (vehicle) of capsaicin was a mixture of 10% ethanol and physiological saline.

(11) It was known that before the formalin injection, 0.1 g/10 L and 1 g/10 L capsaicin was pretreated to show a significant increase in prominent pain behavior response, indicating that the TRPV1 channel activated by capsaicin injected into the face of the white mouse affected the change in the degree of formalin-induced neuropathic pain. Then, in order to examine relief effect of Nypa fruticans Wurmb extract on neuropathic pain, 500 mg/kg Nypa fruticans Wurmb extract (N1) was treated 10 minutes before the injection of capsaicin to observe effects on the change of neuropathic pain behavior response (See FIG. 1).

(12) As a result, it was found that the treatment of Nypa fruticans Wurmb extract (N1) significantly alleviated the increased pain behavior response. It was known that the TRPV1 channel increased neuropathic pain sensation by capsaicin, and Nypa fruticans Wurmb extract (N1) significantly reduced this hyperpathia.

Experimental Example 2: Comparison of Results of Pain Tests for Complex Extract

1. Capsaicin Pain Model Test for Complex Extract

(13) It was confirmed that the N1 had effects of preventing and treating neuropathic pain through experiment of neuropathic pain model for the N1.

(14) Thereafter, the same experiments were conducted on G2 to G5, and the results are described as index with G1 for the purpose of making a relative comparison. The degree of effect of G1 is set to index 5, and the degree of effect of G2 to G5 is expressed as an index of 1 to 10. The higher the number, the better the pain control effect.

(15) TABLE-US-00002 TABLE 2 G1 G2 G3 G4 G5 neuropathic pain model 5 3 4 5 7 (Unit: index)

(16) Table 2 above shows the relative comparison results of the pain control effects of G2 to G5 compared to G1. Compared with G1, G3 and G4 showed equivalent pain control effects, whereas G5 showed relatively good pain control effects.

Experimental Example 3: Sciatic Nerve Regeneration Effect Due to TRPV1 Expression Reduced by Nypa fruticans Wurmb Extract

1. Induction of Crushing Injury on the Sciatic Nerve

(17) 15 mg/100 g ketalar (ketarnin hydrochloride) of Yuhan was intraperitoneally injected to general anesthesia. Then, the hairs from both femurs were removed. Then, the operating field was sterilized with 10% betadine solution. Surgery was performed in the conventional manner under the aseptic operation. After the sciatic nerve was exposed, and the crushing injury was caused for 30 seconds on spot which was 1 cm above rams between the tibial nerve and the total peroneal nerve using hemostatic forceps with 1 mm diameter at the tip. For the analysis of the results, in order to easily recognize the crush area at the time of nerve cutting, the muscles and skin were sutured after ligation with 7-Osilk loosely to the injured area of the epineurium.

2. Evaluation of the Effects of Nypa fruticans Wurmb Extract on Nerve Regeneration

(18) Experimental animals were divided into experimental group (n=15) and control group (n=15). Animals of the experimental group were administered with 500 mg/kg Nypa fruticans Wurmb extract (N1) and divided into 1 week-administered group, 2 week-administered group and 3 week-administered group in which five animals were assigned to each group. Animals of the control group were administered with physiological saline and divided into 1 week-administered group, 2 week-administered group and 3 week-administered group in which five animals were assigned to each group. After the end of each experiment by time, the animals in which the experiment was completed were anesthetized, and then the sciatic nerve was excised and the histomorphological changes were observed by optical microscope and electron microscope.

(19) As a result, the number of total myelinated nerve fibers in the experimental group treated with Nypa fruticans Wurmb extract (N1) changed to 36.03.4, 154.333.8 and 129.312.7, respectively at 1 week, 2 weeks and 3 weeks after administration, and the number of total myelinated nerve fibers tends to be increased rapidly (p<0.01) between 1 and 2 weeks but decreased slightly at 3 weeks. The control group administered with physiological saline tends to be gradually increased from 35.39.1, 74.39.6, and 116.015.1 at 1 week, 2 weeks and 3 weeks, respectively. The number of degenerated myelinated nerve fiber of the control group were 38.38.5, 40.715.0 and 36.38.5 at 1 week, 2 weeks and 3 weeks, respectively. Therefore, it was found that the administration of Nypa fruticans Wurmb extract (N1) after crushing injury to the sciatic nerve reduced the degenerated myelinated nerve fibers produced by the crush injury and recovered the myelinated nerve fibers.

(20) TABLE-US-00003 TABLE 3 1 week 2 weeks 3 weeks Experimental group administered 36.0 3.4 154.3 33.8 129.3 12.7 with N1 Control group administered with 35.3 9.1 74.3 9.6 116.0 15.1 physiological saline

Experimental Example 4: Preference Test

1. Sensuality Evaluation Test

(21) G1, G5 Of complex extracts, and G6 to G9 of fermentation complex extracts were diluted to prepare the tea. The tea was tasted by 10 tasting persons, and the taste and aroma were represented by an index of 1 to 10. The average value (0.5 rounded) was shown in the following Table 4. The higher the number of the index, the higher the preference.

(22) TABLE-US-00004 TABLE 4 G1 G5 G6 G7 G8 G9 Taste 5.5 4.5 6 6 6 5 Aroma 4.5 2.5 4 5 6 4 Complex preference 5.5 4 5.5 6 6.5 5 (average) (Unit: index)

(23) Referring to Table 4, the taste of Nypa fruticans Wurmb extract in G1 was evaluated to be good, but the aroma peculiar to the seaweeds reduced the degree of preference. The G5 mixture added the sour aroma to reduce the degree of preference. In addition, the G9 mixture increased bitter and sour aroma to reduce the degree of preference. On the other hand, it was confirmed that the specific aroma and taste of Nypa fruticans Wurmb extract was neutralized by the complex extracts G7 and G8 to enhance the degree of preference.

(24) Therefore, the complex extracts G7 and G8 may provide the functional food having effects for relieving neuropathic pain having aroma and taste with higher preference.

(25) The preferred embodiments of the present disclosure have been described in detail hereinabove, but the scope of the present disclosure is not limited to the above embodiments. Various modifications and improvements of the present disclosure using the basic concept of the present disclosure by the skilled person in the art, which is defined by the following appended claims, also falls within the scope of the present disclosure.

(26) From the foregoing, it will be appreciated that various embodiments of the present disclosure have been described herein for purposes of illustration, and that various modifications may be made without departing from the scope and spirit of the present disclosure. Accordingly, the various embodiments disclosed herein are not intended to be limiting, with the true scope and spirit being indicated by the following claims.