DE102009005059B4 - Cough suppressant composition and method of making the same - Google Patents

Abstract

A composition comprising a root bark extract of Morus australis Poir. and a pharmaceutically acceptable carrier or excipient for the treatment or relief of cough, wherein the root bark extract of Morus australis Poir. has been prepared by a method comprising the steps of: extracting root cortices of Morus australis Poir. with water to obtain a crude extract; Filtering the crude extract successively through a filter and a membrane having a pore size of 2 to 10 μm; Loading the resulting filtrate on a reverse phase column; successively eluting the reverse phase column with water and a 35-95% ethanol solution to obtain the root bark extract.

Description

Background of the invention

Field of the invention

The present invention relates to a pharmaceutical composition, and more particularly to a cough suppressant composition comprising an extract of Morus australis Poir. as an active cough suppressant component and the method for producing the same.

Description of the Prior Art

Morus alba L. belongs to the mulberry family. White mulberry root bark is a dry root bark of the Morus alba L. known as SANGBAIPI (translated here according to the Chinese Romanization Pinyin Standard) in traditional Chinese medicine.

In the Chinese Pharmacopoeia of Medicine, a white mulberry root bark is usually spooled, ridged or flattened, twisted, variable in length and width, having a thickness of 1.5 to 4 mm. The outer surface is white or pale yellowish-white, relatively uniform, some with orange yellow or brownish yellow remnants of flaky bark, the inner surface is yellowish white or greyish yellow with fine elongated striations. The texture is light and sticky, strong fibrous, hard to break, but easy to peel off on the side, dusting when peeling off. Smell is weak, the taste is slightly sweet.

In traditional Chinese medicine, the fruit of the white mulberry is used to treat premature graying of the hair to "tonify" the blood and to treat constipation and diabetes. The bark of the white mulberry is used to treat a wheeze, an edema and to promote bladder emptying.

White mulberry root bark is a traditional Chinese medicine for an anti-inflammatory, diuretic and anti-cough application. However, the natural amount of Mors alba L. is not widely used in Taiwan. In order to produce a cough medicine with commercial value, the inventors try the widespread in Taiwan Mors australis Poir. to provide a new cough suppressant composition.

Brief summary of the invention

The invention provides a cough suppressant composition comprising an appropriate amount of the Mors australis poir. Extract and a pharmaceutically acceptable carrier or excipient, wherein the Mors australis Poir. Extract is extracted with water.

The invention also provides a cough suppressant composition for treating cough containing a suitable amount of Mors australis poir. Extract and a pharmaceutically acceptable carrier or excipient, wherein the Mors australis Poir. Extract prepared by the steps: Extracting the roots (or root-bark) of the Mors australis poir. with water to obtain a crude extract, sequentially filtering the crude extract through a filter and a membrane having a pore size of 2 to 10 μm to obtain a filtrate, loading the filtrate thus obtained onto a reverse phase column, eluting the reversed phase column with water and a 35-95% ethanol solution.

The invention also provides a method of making a cough suppressant composition, extracting roots (or root bark) from Morus australis Poir. with water to obtain a crude extract.

The invention further provides a method of alleviating, preventing and / or treating a cough comprising administering the cough suppressant composition and a pharmaceutically acceptable carrier or excipient.

A detailed description of the following embodiments will be given with reference to the accompanying drawings.

Detailed description of the invention

The following description explains the invention.

The invention provides a cough suppressant composition for relieving, preventing and / or treating a cough containing an appropriate amount of a Morus australis poir. Extract as an active component includes. The effective Morus australis Poir. Extract is made by extracting Morus australis poir. made with water. After administration of 2 g / kg of body weight, the cough is reduced by more than 30%, preferably 40% -75%.

The invention further provides a method of making a cough suppressant composition comprising extracting the roots (or root bark) of the Morus australis poir. with water to obtain a crude extract.

Sliced pieces of the Morus australis Poir. Root (or root bark) are self-made. As for the manufacturing steps, they include vertically cutting the roots of Morus australis poir., Peeling the root bark, and then drying the cut roots.

In one embodiment, the crude extract of Morus australis Poir. by a filter and a membrane having 2 to 10 .mu.m, preferably 5 .mu.m pore width are sequentially filtered to obtain a filtrate.

The resulting filtrate is loaded onto a reversing column, with the reverse phase column being eluted sequentially with water and a 35-95% ethanol solution to collect an ethanol eluate.

The pore size of a filter is not limited. In general, the pore size may be about 100 to 500 mesh, preferably about 350 mesh (mesh per inch (25.4 mm)).

The column of the reverse phase column may be packed with a polystyrene resin or an ion exchange resin including Diaion HP20 resin, Amberlite XAD-7HP resin or MCI GEL CHP20P resin. The column may be packed in a ratio of 1 g dry basis of the crude extract (or concentrate) / 5 g to 100 g resin.

The invention further provides a pharmaceutical composition for alleviating, preventing and / or treating cough comprising a suitable amount of the cough suppressant composition of the invention and a pharmaceutically acceptable carrier or excipient.

Examples

Example 1: Evaluation Model

The cough suppressant activity of different extracts of Morus australis Poir. is here evaluated according to the method described by Winter C.A. et al., (J. Pharmacol. Exp. Ther. 112: 99, 1954) with modification.

Duncan Hartley male and female guinea pigs weighing 450 ± 50 g were used. Each guinea pig was placed in a sealed 4 liter chamber equipped with an ultrasonic nebulizer to provide a cough inducing stimulant by aerosol. A microphone has been set to increase the coughing sounds of guinea pigs. The animals were exposed to an aerosolized solution of 10% citric acid for 10 seconds and were selected if 9-15 coughing fits occurred in the following 5 minutes. The next day, solvents, that is, distilled water or extracts, were orally administered to the animals twice a day (10:00 AM and 4:00 PM). The animals were again exposed to aerosolized 10% citric acid 60 minutes after the second dose administration. The inhibitory activity of the extracts on the lemons-induced cough was evaluated as follows: inhibition (%) = [(number of coughing attacks before administration) - (number of coughing fits after administration)] / (number of coughing fits before administration) × 100%.

Sample preparation and cough agent evaluation thereof

Example 2: Water extraction and column separation

One kilogram of dry root bark of Morus australis Poir. was once boiled or heated twice with 10 L of water for one hour under reflux. The crude extracts were then filtered with a 350 mesh screen or the filtrates were collected. The residues were each mixed with ten times the weight water for a second extraction (one hour), and then filtered with a 350 mesh screen to obtain another filtrate. The two filtrates were combined and then filtered through a membrane having a pore size of 5 microns to obtain corresponding water extracts.

2.1 column separation (1)

The above-mentioned water extracts were each carried by a Diaion HP20 column (Diaion, Mitsubishi Chemistry Inc.), wherein the ratio of the dry basis of the extract and the resin was 1/30. After loading the extracts onto the columns, the Diaion HP20 columns were sequentially charged with four times the volume of RO water and three times the volume of a 35-95% ethanol solution to obtain corresponding ethanol eluates (samples 1-1 to 1-6) , The ethanol eluates are concentrated and freeze-dried to remove the ethanol in the eluates and to obtain dry powders. The yield of the above-mentioned eluates is shown in Table 1. The evaluation of the cough medicine of the above-mentioned eluates is shown in Table 2. Table 1 Sample No.. Ratio of dry basis and resin Ethanolkonz. (%) Yield (%) 2-1 1:31 35 4.0 2-2 1:29 50 3.5 2-3 1:29 60 - 2-4 1:29 95 4.2 2-5 1:35 50 4.2 2-6 1:41 50 6.4 Yield: (eluate weight / root bark weight) × 100% Table 2 Sample No.. Amount of administration (dose per body weight) Inhibition rate (%) 1-1 2 g / kg 46 1-2 2 g / kg 61 1-3 2 g / kg 50 1-4 2 g / kg 40 1-5 2 g / kg 43 1-6 2 g / kg 69

2.2 column separation (2)

The above-mentioned water extracts were each separated by a Diaion HP20 column (Diaion, Mitsubishi Chemistry In.), Whereby the dry base ratio of the extract and the resin was 1/20 to 1/30. After loading the extracts onto the columns, the Diaion HP20 columns were sequentially eluted each time by 2 to 4 volumes by volume of RO water and 2 to 3 times by volume of a 50% ethanol solution to obtain ethanol eluates (samples 1-7 to 1-14) , The ethanol eluates were then concentrated and lyophilized to remove the ethanol in the eluates and to obtain dry powders. The yield of the above-mentioned eluates is shown in Table 3. The evaluation of the cough medicine of the above-mentioned eluates is shown in Table 4. Table 3 Sample No.. Pore size of the membrane Ratio of dry basis and resin Vol. Of eluting water Vol. Of eluting ethanol Yield (%) 1-7 5 μm 1:30 4x 3 times 3.7 1-8 5 μm 1:25 2.5x 2- 3.8 1-9 5 μm 1:20 4x 3 times 3.8 1-10 5 μm 1:20 4x 3 times 4.2 1-11 5 μm 1:23 2- 3 times 5.0 1-12 - 1:24 2- 3 times 2.3 1-13 - 1:19 2- 3 times 1.4 1-14 - 1:20 2- 3 times 1.5 Yield: (eluate weight / root bark weight) × 100%. Table 4 Sample No.. Amount of administration (dose per body weight) Inhibition rate (%) 1-7 2 g / kg 48 1-8 2 g / kg 46 1-9 2 g / kg 69 1-10 2 g / kg 57 1-11 2 g / kg 74 1-12 2 g / kg 39 1-13 2 g / kg 48 1-14 Note: Samples 1-13 and 1-14 united by weight 1: 1.

Example 3: 95% ethanol extraction (not according to the invention)

It was one kilogram of dry root bark of Morus australis Poir. once boiled and refluxed twice with 10 L of a 95% ethanol solution for 1 hour to obtain a crude extract. The crude extract was then filtered with a 350 mesh screen collecting the filtrate. The residue was mixed with 10 times the amount of water-water weight for a second extraction (1 hour) and then filtered with a 350 mesh screen to obtain another filtrate. The two filtrates were combined to obtain an ethanol extract (Sample 2-1). The ethanol extract was concentrated and freeze-dried to obtain a dry powder. The yield and evaluation of the cough medicine of the extract are in Table 5 is shown. Table 5 Sample No.. Yield (%) Amount of administration (dose per body weight) Inhibition rate (%) 3-1 9.2 2 g / kg 33 Yield: (eluate weight / root bark weight) × 100%

Example 4: 95% ethanol extraction and column separation (not according to the invention)

It became the dry root bark of Morus australis Poir. used. It became one kilogram of the dry root bark of the Morus australis Poir. once boiled and refluxed twice with 10 L of a 95% ethanol solution for 1 hour to obtain crude extracts. The crude extracts were then filtered with a 350 mesh screen and the appropriate filtrates were collected. The residues were mixed with ten times the weight of water (by weight with ten times the amount of water) for a second extraction procedure (one hour) and then filtered with a 350 mesh screen to obtain other filtrates. The two filtrates were combined to obtain corresponding ethanol extracts.

The ethanol extracts were concentrated 10-fold under reduced pressure and then an equal amount of water was added by weight to obtain appropriate concentrates.

The above-mentioned concentrates were each separated by Diaion HP20 columns (Diaion, Mitsubishi Chemistry, In.), Whereby the ratio of the dry basis of the concentrate and the resin was 1/20. After loading the concentrates on the columns, the Diaion HP20 columns were each eluted with 2 times the volume of a 50% ethanol solution to obtain ethanol eluates (samples 3-1 to 3-3). The ethanol eluates were then concentrated and freeze-dried to remove the ethanol in the eluates and to obtain dry powder. The yield and the evaluation of the cough medicine of the above-mentioned eluates are shown in Table 6. Table 6 Sample No.. Ratio of dry basis and resin Yield (%) Administration amount (dose per body weight) Inhibition rate (%) 3-1 1:20 6.3 0.5 g / kg 55 1 g / kg 61 2 g / kg 63 3-2 1:20 7.3 2 g / kg 48 3-3 1:20 5.6 2 g / kg 48 Yield: (eluate weight / root bark weight) × 100%

Example 5: Ethyl acetate extraction (not according to the invention)

It became the dry root bark of Morus australis Poir. used. It became one kilogram of the root bark of Morus australis Poir. once boiled and refluxed twice with 10 L of an ethyl acetate solution for 1 hour to obtain a crude extract. The crude extract was then filtered with a 350 mesh screen and the filtrate collected. The residue was mixed by weight with ten times the amount of water for a second extraction procedure (one hour) and then filtered with a 350 mesh screen to obtain another filtrate. The two filtrates were combined and then filtered through a membrane having a pore size of 5 μm to obtain an ethyl acetate extract (Sample 4-1). The extract was concentrated under reduced pressure and dried to obtain a dry powder.

The yield and anti-cough assessment of the extract are shown in Table 7. Table 7 Sample No.. Yield (%) Administration amount (dose per body weight) Inhibition rate (%) 4-1 2.1 2 g / kg 48 Yield: (eluate weight / root bark weight) × 100%

Claims (4)

Composition comprising a root bark extract from Morus australis Poir. and a pharmaceutically acceptable carrier or excipient for the treatment or alleviation of cough, wherein the root bark extract of Morus australis Poir. has been prepared by a process comprising the following steps: Extracting root cortices of Morus australis Poir. with water to obtain a crude extract; Filtering the crude extract successively through a filter and a membrane having a pore size of 2 to 10 μm; Loading the resulting filtrate on a reverse phase column; successively eluting the reverse phase column with water and a 35-95% ethanol solution to obtain the root bark extract. A composition according to claim 1, wherein the reverse phase column is packed with a polystyrene resin or an ion exchange resin. A composition according to claim 2, wherein the column is packed in a ratio of 1 g dry basis of the crude extract / 5 g to 100 g resin. A method of making a composition for treating or alleviating coughing, comprising: Extraction of root cortices of Morus australis Poir. with water to obtain a crude extract; Filtering the crude extract in succession through a filter and a membrane having a pore size of 2 to 10 μm; Loading the resulting filtrate on a reverse phase column; Elute the reverse phase column sequentially with water and a 35-95% ethanol solution to obtain the root bark extract.