WO2009057975A2

WO2009057975A2 - Pharmaceutical composition for the prevention and treatment of cell proliferative diseases comprising of the feathers of birds as the active ingredient

Info

Publication number: WO2009057975A2
Application number: PCT/KR2008/006446
Authority: WO
Inventors: Sang Moon Lee
Original assignee: Sang Moon Lee
Priority date: 2007-11-02
Filing date: 2008-10-31
Publication date: 2009-05-07
Also published as: KR20090045478A; KR100989249B1; WO2009057975A3

Abstract

The present invention relates to a pharmaceutical composition for preventing and treating cell proliferative diseases comprising feathers of birds as an active ingredient. More particularly, the present invention relates to a pharmaceutical composition for preventing and treating cell proliferative diseases comprising whole feathers of birds, a powder of a calamus or a rachis of bird feathers, as an active ingredient. A composition for preventing and treating cell proliferative diseases of the present invention has the activity of inhibiting and preventing proliferation of tumor cells. The inventive compositions may be used for preventing and treating cell proliferative diseases, and particularly, used for anticancer purpose such as prevention, reduction, and treatment (or control) of cancer.

Description

Invention Title

PHARMACEUTICAL COMPOSITION FOR THE PREVENTION AND TREATMENT OF CELL PROLIFERATIVE DISEASES COMPRISING OF THE FEATHERS OF BIRDS AS THE ACTIVE INGREDIENT

Technical Field

The present invention relates to a pharmaceutical composition for preventing and treating cell proliferative diseases comprising feathers of birds as an active ingredient. More particularly, the present invention relates to a pharmaceutical composition for preventing and treating cell proliferative diseases comprising whole feather, a powder of a calamus or a rachis as an active ingredient.

Background Art

Cancer is a class of diseases resulting from uncontrolled proliferation and non-systemic growth of transformed cells. Mostly, cancers are caused by mutations in oncogenes and tumor suppressor genes which occur due to environmental, genetic or other causes. In the early stages, cancer cells proliferate and invade and destroy nearby tissues. In the later stages, they invade the circulatory system and spread to distant areas, ultimately resulting in the death of the victim.

In order to treat cancers, surgeries, radiotherapies, chemotherapies, and the like are applied, and various substances known to have anticancer effect are used. Although most chemical anticancer agents exhibit the effect of controlling cell growth, they are without selectivity for cancer cells and tend to show toxicity to normal cells. Accordingly, development of new anticancer agents which provide superior selectivity cancer cells, have less toxicity and are capable of overcoming resistance is needed.

Feathers are one of the epidermal variants that form the distinctive outer covering on birds. The major component of feathers is keratin. Embryologically, it corresponds to the hair of mammals or the scale of reptiles. The feathers of birds consists of a calamus (or hollowshaft) which inserts into a follicle in the skin, a rachis which is connected to the calamus and serves as the main shaft, and a plurality of barbs which are branched from the rachis. Small barbs are entangled with each other to form a vane. In general, feathers have brown or black melanin pigments and thus has a color. Typically, it is composed of fibrous hard proteins insoluble in water or aqueous salt solution. The feathers are derived from the bone and is rich in calcium. Connected by calcium chains, the feathers are air-permeable and structurally adequate for flying.

The inventors of the present invention have carried out researches to find substances from natural products that can prevent and inhibit proliferation of cancer cells. As a result, we have found that a composition comprising whole feather of birds, or a calamus and a rachis of birds has superior anticancer effect and completed the present invention.

Disclosure Technical Problem

Accordingly, an object of the present invention is to provide a composition for preventing and treating cell proliferative diseases.

Technical Solution

In order to attain the aforesaid object, in an aspect, the present invention provides a composition for preventing and treating cell proliferative diseases comprising feathers of birds in the form of powder.

In another aspect, the present invention provides a method for preparing a powder of feathers of birds. In another aspect, the present invention provides a method for orally administering the composition of the present invention.

In another aspect, the present invention provides a use of feathers of birds for preparing agent for treating cell proliferative diseases

In another aspect, the present invention provides a method for preventing and treating cell proliferative diseases comprising administering to a subject in need thereof an effective amount of feathers of birds of the present invention.

Hereinafter, the present invention is described in more detail.

The pharmaceutical composition of the present invention is characterized by comprising a powder of feathers of birds as an active ingredient.

The feathers of birds used to prepare the pharmaceutical composition of the present invention are not particularly limited. But, preferably, feathers of chicken, duck, turkey, goose or ostrich may be used. More preferably, feathers of chicken may be used. As described above, feathers of birds comprise a calamus, a rachis and barbs. The feathers of birds may be used as a whole. But, preferably, only the calamus and the rachis of the feathers of birds may be used. In particular, the portion of the feathers of birds which exhibit color includes a lot of brown or black melanin pigments. According to experiments, the compositions including these portions exhibited superior effect. Therefore, it is preferred to use the portion containing a lot of melanin pigments. The separation of the calamus, the rachis and the portions containing a lot of melanin pigments from the feathers of birds can be carried out without particular limitation. Preferably, they may be separated by hydrating the feathers of birds by immersing in water followed by applying physical force, for example, using a knife . The water used for immersing the feathers is not particularly limited. Preferably, common tap water or distilled water may be used, and one having a temperature from 1O⁰C to 5O⁰C is preferable. If the water temperature exceeds 50⁰C, the epidermal tissue may be deformed. And, if the water temperature is below 1O⁰C, hydration may be insufficient. The immersing time may be controlled depending on the condition of the materials so as to attain sufficient hydration. Preferably, the immersing time may be from 5 minutes to 3 hours. If the immersing time is less than 5 minutes, hydration may be insufficient. And, an immersing time exceeding 3 hours is uneconomical and may result in deformation of tissues.

The pharmaceutical composition of the present invention comprises a powder of whole feather of birds or a powder of a calamus or rachis of feathers of birds prepared by the steps of follows. And a powder of whole feather of birds may be prepared by the steps of : (a) immersing feathers of birds in water of 10°C -5O⁰C for 5 minutes to 3 hours; (b) sterilizing the immersed feathers in brine with a salinity from 5.0 to 8.0 for 3 minutes to 90 minutes; (c) washing the feathers with flowing water to remove impurities; (d) immersing and sterilizing the feathers in 0.2-0.5 weight% (w/v) sodium chloride solution for 1 minute to 1 hour; (e) immersing and sterilizing the feathers in 1-4 weight% (v/v) acetic acid solution for 15 minutes to 30 minutes; (f) immersing and sterilizing the feathers in water of 30°C -77⁰C for

1 minute to 3 hours; (g) sterilizing and drying the immersed and sterilized feathers of the above at 15⁰C ~40°C for 1 hour to 96 hours; (h) sterilizing the dried feathers by irradiation in 290-400 nm UV for 5 minutes to

2 hours; (i) cutting the sterilized feathers to 5-15 mm for easier grinding; (j) sterilizing the cut feathers by- irradiation in 290-400 nm UV for 5 minutes to 2 hours; (k) grinding the cut and sterilized feathers; (1) sterilizing the resulting ground powder by irradiation in 290-400 nm UV for 5 minutes to 2 hours; (m) sterilizing the resulting sterilized ground powder by irradiation in 200-290 nm UV for 30 seconds to 20 minutes. And a powder of a calamus or a rachis of feathers of birds prepared by the steps of: (a) immersing feathers of birds in water of 10⁰C ~50°C for 5 minutes to

3 hours; (b) sterilizing the immersed feathers in brine with a salinity from 5.0 to 8.0 for 3 minutes to 90 minutes; (c) washing the feathers with flowing water to remove impurities; (d) separating a calamus or a rachis of feathers of birds from which the impurities were removed; (e) immersing and sterilizing the separated calamus or the rachis in 0.2~0.5 weight% (w/v) sodium chloride solution for 1 minute to 1 hour; (f) immersing and sterilizing the calamus or the rachis in 1-4 weight%

(v/v) acetic acid solution for 15 minutes to 30 minutes; (g) immersing and sterilizing the calamus or the rachis in water of 3O⁰C ~77°C for 1 minute to 3 hours; (h) sterilizing and drying the immersed and sterilized the calamus or the rachis of the above at 15°C ~40°C for 1 hour to 96 hours; (i) sterilizing the dried calamus or rachis by irradiation in 290-400 nm UV for 5 minutes to 2 hours; (j) cutting the sterilized and dried calamus or rachis to 5-15 mm for easier grinding; (k) sterilizing the cut calamus or rachis by irradiation in 290-400 nm UV for 5 minutes to 2 hours; (1) grinding the cut and sterilized calamus or rachis; (m) sterilizing the resulting ground powder by irradiation in 290-400 nm UV for 5 minutes to 2 hours; (n) sterilizing the resulting sterilized ground powder by irradiation in 200-290 nm UV for 30 seconds to 20 minutes.

In the above preparation method, the water used for immersing and hydrating is not particularly limited. Preferably, common tap water or distilled water may be used, and one having a temperature from 10°C to 50⁰C is preferable. If the water temperature exceeds 50⁰C, the epidermal tissue may be deformed. And, if the water temperature is below 1O⁰C, hydration may be insufficient. The immersing time may be controlled depending on the condition of the materials so as to attain sufficient hydration. Preferably, the immersing time may be from 5 minutes to 3 hours. If the immersing time is less than 5 minutes, hydration may be insufficient. And, an immersing time exceeding 3 hours is uneconomical and may result in deformation of tissues.

Meanwhile, Sterilization is performed in order to remove the saprophytes proliferating during the immersion from the feathers. The sterilization method may be any one known in the art, as long as the feathers are not deformed. Preferably, the feathers are immersed in brine with a salinity from 5.0 to 8.0 for 3 minutes to 90 minutes. Then, after removing the impurities, 0.2-0.5 weight% (w/v) of NaCl solution is added, and immersing and sterilization are carried out for 1 minute to 1 hour. For additional sterilization, the said feathers are immersed and sterilized in 1~4 weight% (v/v) acetic acid solution for 15 minutes to 30 minutes. Further, the feathers of birds is immersed and sterilized in water of 3O⁰C -77⁰C for 1 minute to 3 hours. After completion of the immersion and prior to drying, the feathers are washed using tap water or distilled water so as to remove remaining NaCl. The removal of impurities is performed by applying physical force after the feathers are sufficiently hydrated by the immersion. The method for drying the washed feathers of birds is not particularly limited. Preferably, the feathers are dried in a well ventilated place, out of the direct rays of the sun. Preferably, the feathers are dried completely for 1 hour to 96 hours at 15°O40°C, preferably at room temperature of about 25⁰C. Then, UV irradiation is carried out as described below.

The separation of the quill and the rachis is performed as follows. The whole of the dried feathers may be used. But, more preferably, the quill portion with well-developed optical properties and multilayer structure and rich in keratin, and the rachis quill rich in keratin and melanin pigments are screened out from the feathers. From the dried feathers, the portions other than the quill and rachis portions, that is, barbs and vanes are completely removed (taken off) from the rachis by applying physical force. If the separated quill or rachis is in bad condition or, if the lower portion of the quill or the upper portion of the rachis is too thin, only the portions in good condition may be selected fro use. Then, UV irradiation is carried out as described below.

The whole of the separated feathers or the selected calamus or rachis portion is ground into a powder using a grinding machine. If necessary, they may be cut finely (5~15 mm) in advance for easier grinding.

The resultant powder is sterilized using UV. UV with a wavelength of 290-400 nm which is UVA and UVB wavelength range is irradiated using a UV sterilizer without heat emission, at a distance of 10-50 cm from the powder. The irradiation time is not particularly- limited, but 5 minutes to 2 hours is preferred. In addition, finally UV with a wavelength of 200~290 nm which is UVC wavelength range is irradiated for 30 seconds to 20 minutes.

In one examples of the present invention, calamus and rachises of chicken, duck, turkey and goose were used as the feathers of birds in order to prepare the inventive compositions.

In one test example of the present invention, the inventive composition was administered to a nude mouse to which colon cancer cells had been transplanted, in order to confirm the anticancer effect. Then, the effect of inhibiting the cancer cells was measured. As a result, about 50% or more cancer cell growth inhibition effect was confirmed, as compared to the control group to which nothing was administered.

Accordingly, the present invention provides a pharmaceutical composition for preventing and treating cell proliferative diseases which is able to inhibit and prevent the growth of cancer cells.

The pharmaceutical composition according to the present invention may comprise the material prepared by the present invention singly with a pharmaceutically effective amount or may further comprise at least one pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically effective amount" refers to an amount resulting in a better effect as compared to the control group and, preferably, an amount effective in treating or preventing cell proliferative diseases.

The pharmaceutically effective amount of the pharmaceutical composition according to the present invention may be 10~1000 mg/day/kg body weight, preferably 150-600 mg/day/kg body weight. However, the pharmaceutically effective amount may be varied adequately, depending on various factors, including particular disease and severity thereof, age, body weight, physical condition and sex of the patient, administration route, period of treatment, and the like.

The diseases to which the pharmaceutical composition of the present invention can be applied are immunodeficiency-related diseases and cell proliferative diseases. The cell proliferative diseases are neoplastic diseases caused by tumors, and may be, for example, cancer diseases. The cancer diseases include, although not limited thereto, large intestine cancer, spleen cancer, colon cancer, lung cancer, liver cancer, stomach cancer, esophageal cancer, pancreatic cancer, gallbladder cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, cervical cancer, endometrial cancer, choriocarcinoma, ovarian cancer, breast cancer, thyroid cancer, brain cancer, head and neck cancer, malignant melanoma, lymphoma, bone cancer, soft tissue sarcoma, spinal cancer, and the like.

As used herein, the phrase "pharmaceutically acceptable" refers that the carrier is physiologically acceptable, does not inhibit the action of the active ingredient when administered to human, and is non-toxic without allergic reactions such as gastroenteric trouble and dizziness or other adverse reactions. The carrier includes all types of solvent, dispersive medium, oil-in- water or water-in-oil emulsion, aqueous composition, liposome, microbead and microsome.

Meanwhile, the pharmaceutical composition according to the present invention may be formulated along with an adequate carrier depending on the administration route. No particular limitation is imposed on the administration route of the inventive pharmaceutical composition, but it may be administered orally.

In case the pharmaceutical composition of the present invention is administered orally, the pharmaceutical composition of the present invention may be formulated along with an adequate carrier for oral administration into powder, granule, tablet, pill, sugarcoated tablet, capsule, liquid, gel, syrup, suspension, wafer, or the like by a method known in the art. Examples of the adequate carrier may include a saccharide such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, etc., a starch such as corn starch, wheat starch, rice starch, potato starch, etc., a cellulose such as cellulose, methylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, etc., and a filler such as gelatin, polyvinylpyrrolidone, etc. Further, crosslinked polyvinylpyrrolidone, agar, alginic acid, sodium alginate, etc. may be added as disintegrant . In addition, the pharmaceutical composition may further comprise an anticoagulant, a surfactant, a wetting agent, a fragrance, an antiseptic, or the like. Besides, those described in the following literature may be used as the pharmaceutically acceptable carrier: Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995.

Further, the pharmaceutical composition according to the present invention may comprise at least one buffer

(e.g., saline or PBS), carbohydrate (e.g., glucose, mannose, sucrose or dextran) , antioxidant, bacteriostat, chelating agent (e.g., EDTA or glutathione), adjuvant (e.g., aluminum hydroxide), suspension agent, thickener and/or preservative.

And, the pharmaceutical composition of the present invention may be prepared into a formulation which can provide immediate, sustained or delayed release of the active ingredient after being administered to a mammal by a method known in the art.

Further, the pharmaceutical composition of the present invention may be administered in combination with a compound known to have an effect of preventing or treating cancer diseases.

Meanwile, the present invention provides a method for preparing a powder of feathers of birds. As describe above, the preparation method comprises the steps of: (a) immersing feathers of birds in water of 10°C~50°C for 5 minutes to 3 hours; (b) sterilizing the immersed feathers with a salinity from 5.0 to 8.0 for 3 minutes to 90 minutes; (c) washing the feathers well with flowing water to remove impurities; (d) immersing and sterilizing the feathers in 0.2~0.5 weight% (w/v) Sodium

Chloride (NaCl) solution for 1 minute to 1 hour; (e) immersing and sterilizing the feathers in 1~4 weight% (v/v) acetic acid solution for 15 minutes to 30 minutes;

(f) immersing and sterilizing the feathers in water of

30°C~77°C for 1 minute to 3 hours; (g) sterilizing and drying the immersed and sterilized feathers at 15°C -4O⁰C for 1 hour to 96 hours; (h) sterilizing the dried feathers by irradiation in with 290-400 nm UV for 5 minutes to 2 hours; (i) cutting the sterilized feathers to 5~15 mm for easier grinding; (j) sterilizing the cut feathers by irradiation in 290-400 nm UV for 5 minutes to

2 hours; (k) grinding the cut and sterilized feathers;

(1) sterilizing the resulting ground powder by irradiation in 290-400 nm UV for 5 minutes to 2 hours; and (m) sterilizing the resulting sterilized ground powder by irradiation in 200-290 nm UV for 30 seconds to 20 minutes. And a powder of a calamus or a rachis of feathers of birds prepared by the steps of: (a) immersing feathers of birds in water of 1O⁰C -5O⁰C for 5 minutes to 3 hours; (b) sterilizing the immersed feathers in brine with a salinity from 5.0 to 8.0 for 3 minutes to 90 minutes; (c) washing the feathers with flowing water to remove impurities; (d) separating a calamus or a rachis of feathers of birds from which the impurities were removed; (e) immersing and sterilizing the separated calamus or the rachis in 0.2-0.5 weight% (w/v) sodium chloride solution for 1 minute to 1 hour; (f) immersing and sterilizing the calamus or the rachis in 1-4 weight% (v/v) acetic acid solution for 15 minutes to 30 minutes; (g) immersing and sterilizing the calamus or the rachis in water of 30⁰C -77⁰C for 1 minute to 3 hours; (h) sterilizing and drying the immersed and sterilized the calamus or the rachis of the above at 15⁰C -4O⁰C for 1 hour to 96 hours; (i) sterilizing the dried calamus or rachis by irradiation in 290-400 nm UV for 5 minutes to 2 hours; (j) cutting the sterilized and dried calamus or rachis to 5~15 mm for easier grinding; (k) sterilizing the cut calamus or rachis by irradiation in 290-400 nm UV for 5 minutes to 2 hours; (1) grinding the cut and sterilized calamus or rachis; (m) sterilizing the resulting ground powder by irradiation in 290-400 nm UV for 5 minutes to 2 hours; (n) sterilizing the resulting sterilized ground powder by irradiation in 200-290 nm UV for 30 seconds to 20 minutes. The procedures for immersion, washing, drying, sterilization, and UV treatment are the same as described above.

Further, the present invention provides a method for oral administration of the inventive compositions, and the components of the inventive compositions are described above. And the method for oral administration may performed by well known in the art.

Further, the present invention provides a method for sterilization of a powder of feathers of birds of the present invention comprising the steps of: (a) sterilizing with a salinity from 5.0 to 8.0 for 3 minutes to 90 minutes; (b) immersing and sterilizing in 0.2-0.5 weight% (w/v) sodium chloride solution for 1 minute to 1 hour; (c) immersing and sterilizing in 1~4 weight% (v/v) acetic acid solution for 15 minutes to 30 minutes; (d) immersing and sterilizing the feathers of birds in water of 30⁰C ~77°C for 1 minute to 3 hours; (e) sterilizing and drying the immersed and sterilized feathers at 15°C -4O⁰C for 1 hour to 96 hours; (f) sterilizing the dried feathers by irradiation in 290-400 nm UV for 5 minutes to 2 hours; and (g) sterilizing the resulting sterilized ground powder by irradiation in 200~290 nm UV for 30 seconds to 20 minutes. Every step is described above.

Based on the effect of the inventive compositions, the present invention provides a method for preventing and treating cell proliferative diseases comprising administering the feathers of birds to subject in need thereof.

As used herein, the "effective amount" refers to the amount of composition of feathers of birds of the present invention effective in preventing and treating cell proliferative diseases, and the "subject" refers to mammals, particularlly, animals comprising human. The subject may be patient in need of treatment.

Further, the compositions of the feathers of birds may be administered until effect of the above mentioned is derived, and administered orally via various conventional routes.

Further, the present invention provides a use of the feathers of birds for preparing reagent for treating cell proliferative diseases. The cell proliferative diseases, the feathers of birds of the present invention and effect thereof is described above.

Advantageous Effects Accordingly, the present invention provides a composition for preventing and treating cell proliferative diseases which inhibit and prevent proliferation of tumor cells. The inventive compositions may be used for preventing and treating cell proliferative diseases, and particularly, used for anticancer purpose such as prevention, reduction, and treatment (or control) of cancer.

Description of Drawings Fig. 1 is a graph showing the change of the body weight of a nude mouse in which human colon cancer cells (HCT-15) were transplanted, where

Control: negative control group (Gl) Doxorubicin: doxorubicin treated group (positive control, G2) BIC-22: test group (G3) Fig. 2 is a graph showing the change of the tumor volume of a nude mouse in which human colon cancer cells (HCT-15) were transplanted.

Mode for Invention

Hereinafter, the present invention is described in more detail through examples.

However, the following examples are provided for illustrative purposes only, and they are not intended to limit the present invention. Since they may be performed through modified forms, they are also comprised to the present invention.

Example 1: Preparation of inventive composition with the feathers of chicken

Chicken feathers were immersed in water of 50⁰C for 20 minutes, and then in brine with a salinity of 6.5 for 30 minutes. After washing well with flowing water, the feathers were immersed in 0.4% (w/v) Sodium Cloride (NaCl) solution for 10 minutes and impurities were removed. Subsequently, the feathers were immersed and sterilized in 2% (v/v) acetic acid solution for 20 minutes, and then immersed in water of 70⁰C for 2 minutes. And the feathers were dried for 48 hours at room temperature, in a well ventilated place. The dried feathers were irradiated with UV of 320 nm wavelength for 30 minutes, using a UV lamp (Birtcher, U.S.; no heat emission type) at a distance of 30 cm from the feathers, and portions excluding the calamus or the rachis were removed by applying physical force (with hand) . After irradiating with UV of 320 nm wavelength for 30 minutes, using a UV lamp at a distance of 30 cm, they were cut to a size of about 6 mm using a cutter. After cutting, the cut feathers were irradiating with UV of 320 nm wavelength for 30 minutes, followed by grinding into fine powder using a grinding machine. The resulting ground powder was prepared by irradiation in it with UV of 320 nm wavelength for 30 minutes, using a UV lamp at a distance of 30 cm, and irradiating it again with UV of 250 nm wavelength for 1 minute.

Example 2: Preparation of inventive composition with the feathers of duck

A composition of the present invention was prepared in the same manner as in Example 1, except for using feathers of duck in Example 1.

Example 3: Preparation of inventive composition with the feathers of turkey

A composition of the present invention was prepared in the same manner as in Example 1, except for using feathers of turkey in Example 1. Example 4: Preparation of inventive composition with the feathers of goose

A composition of the present invention was prepared in the same manner as in Example 1, except for using feathers of goose in Example 1.

Test Example: Measurement of anticancer effect of inventive anticancer composition

The anticancer effect of the inventive composition was tested at the Chemon Preclinical Research Center (approved by the KGLP) using a nude mouse transplanted with human colon cancer cells.

7-weeks-old specific pathogen free (SPF) athymic BALB/C nude mice (Orient Ltd., Korea) were weighed and grouped to 3 groups, 5 per each group, as follows: negative control group, positive control group and test groups. The mice were identified by the identification label of the cage and using ear punching.

Each group was designed as in Table 1.

Table 1

Gl: negative control group (excipient) G2: positive control group (abdominal administration of positive control substance once in 2 days)

G3 : test substance administration groups (test substance had been administered 225mg/kg from 28^th day)

Colon cancer cells (HCT15) were thawed in a constant-temperature water bath of 37⁰C as soon as possible. After mixing well with 10 mL of RPMI1640 culture medium (Sigma Aldrich, USA) , centrifuge was performed at 1000 rpm for 10 minutes. The resulting cell pellet was mixed well with 5 mL of RPMI1640 culture medium containing 10% fetal bovine serum (FBS), and the cells were cultured in a cell culture flask under the condition of 37⁰C and 5% CO₂. The cultured cancer cells were suspended in physiological saline to a concentration of IxIO⁷ cells/mL and transplanted subcutaneously to the mice, 0.3 mL (3xlO⁶ cells) each.

The composition prepared in Example 1 was suspended at various concentrations in sterilized, distilled water for injection (Choongwae Pharma Corporation, Korea) as excipient, ground using a homogenizer, and orally administered to the mice twice a day, for 5 weeks.

In the negative control group (Gl) , the excipient was orally administered twice a day, for 5 weeks, instead of the inventive anticancer composition. In the positive control group (G2), doxorubicin was abdominally administered at a dose of 2 mg/kg body weight for 5 weeks, once in 2 days, instead of the inventive anticancer composition. The doxorubicin solution was prepared by dissolving in the excipient by a method known in the art.

During the test, the mice were kept under the condition of 23±3°C and relative humidity of 55+15%, and access to water and feed was allowed freely.

Tumor volume was measured on days 9, 13, 16, 20,

23, 27, 30, and 34 after the inoculation of the cancer cells. Length, width and height of the tumor were measured using a vernier caliper, and the tumor volume was calculated by the following equation:

Tumor volume = (Length x Width x Height) ÷ 2 The tumor volume measurement result is given in Table 2 (unit: mm³) .

Table 2

As can be seen from the result of Table 2 and Fig. 2, when the composition according to the present invention is administered, 40-50% of tumor volume was reduced compared with negative control group (Gl) , and it shows excellent antitumor activity compared with the positive control group (G2) which was administered doxorubicin. 0

Abnormal symptoms related with the administration of the inventive composition were not observed, and no significant change in body weight was observed, as shown in Fig 1. Although, the body weight was decreased 5 temporarily on 7^th day, it is regarded not from the administration of the inventive composition.

From the above, it can be confirmed that the composition according to the present invention is 0 effective in inhibiting the proliferation of tumor cells.

Preparation Example: Preparation of capsule comprising inventive composition

100.0 mg of the composition prepared in Example 1,

83.0 mg of cornstarch, 175.0 mg of lactose and 2.0 mg of magnesium stearate were mixed homogeneously, and filled in a gelatin capsule, so that the weight of the contents per one capsule was 360 mg.

Industrial Applicability

As can be seen from the foregoing, a composition for preventing and treating cell proliferative diseases of the present invention has the activity of inhibiting and preventing proliferation of tumor cells. The inventive compositions may be used for preventing and treating cell proliferative diseases, and particularly, used for anticancer purpose such as prevention, reduction, and treatment (or control) of cancer.

Claims

CLAIMS Claim 1

A pharmaceutical composition for preventing and treating cell proliferative diseases comprising feathers of birds as an active ingredient.

Claim 2

The composition of claim 1, wherein the feathers of birds are selected from the group consisting of whole feather of birds, a calamus and a rachis.

Claim 3

The composition of claim 1, wherein the feathers of birds are selected from the group consisting of a powder of whole feather, a powder of a calamus and a powder of a rachis .

Claim 4

The composition of Claim 3, wherein the powder of whole feather is prepared by a method comprising the steps of:

(a) immersing feathers of birds in water of 1O⁰C ~50°C for 5 minutes to 3 hours;

(b) sterilizing the immersed feathers in brine with a salinity from 5.0 to 8.0 for 3 minutes to 90 minutes;

(c) washing the feathers with flowing water to remove impurities; (d) immersing and sterilizing the feathers in 0.2-0.5 weight% (w/v) sodium chloride solution for 1 minute to 1 hour;

(e) immersing and sterilizing the feathers in 1-4 weight% (v/v) acetic acid solution for 15 minutes to 30 minutes;

(f) immersing and sterilizing the feathers in water of 30⁰C -77⁰C for 1 minute to 3 hours;

(g) sterilizing and drying the immersed and sterilized feathers of the above at 15⁰C -4O⁰C for 1 hour to 96 hours;

(h) sterilizing the dried feathers by irradiation in 290-400 nm UV for 5 minutes to 2 hours;

(i) cutting the sterilized feathers to 5-15 mm for easier grinding;

(j) sterilizing the cut feathers by irradiation in 290-400 nm UV for 5 minutes to 2 hours;

(k) grinding the cut and sterilized feathers;

(1) sterilizing the resulting ground powder by irradiation in 290-400 nm UV for 5 minutes to 2 hours; and

(m) sterilizing the resulting sterilized ground powder by irradiation in 200-290 nm UV for 30 seconds to 20 minutes.

Claim 5

The composition of Claim 3, wherein the powder of a calamus or a rachis is prepared by a method comprising the steps of:

(a) immersing feathers of birds in water of 10°C -5O⁰C for 5 minutes to 3 hours; (b) sterilizing the immersed feathers in brine with a salinity from 5.0 to 8.0 for 3 minutes to 90 minutes;

(c) washing the feathers with flowing water to remove impurities;

(d) separating a calamus or a rachis of feathers of birds from which the impurities were removed;

(e) immersing and sterilizing the separated calamus or the rachis in 0.2-0.5 weight% (w/v) sodium chloride solution for 1 minute to 1 hour;

(f) immersing and sterilizing the calamus or the rachis in 1~4 weight% (v/v) acetic acid solution for 15 minutes to 30 minutes;

(g) immersing and sterilizing the calamus or the rachis in water of 3O⁰C -77⁰C for 1 minute to 3 hours;

(h) sterilizing and drying the immersed and sterilized the calamus or the rachis of the above at 15°C ~40°C for 1 hour to 96 hours;

(i) sterilizing the dried calamus or rachis by irradiation in 290-400 nm UV for 5 minutes to 2 hours;

(j) cutting the sterilized and dried calamus or rachis to 5-15 mm for easier grinding;

(k) sterilizing the cut calamus or rachis by irradiation in 290-400 nm UV for 5 minutes to 2 hours; (1) grinding the cut and sterilized calamus or rachis;

(m) sterilizing the resulting ground powder by irradiation in 290~400 nm UV for 5 minutes to 2 hours; and

(n) sterilizing the resulting sterilized ground powder by irradiation in 200-290 nm UV for 30 seconds to 20 minutes.

Claim 6

The composition of Claim 1, wherein the birds are selected from the group consisting of chicken, duck, turkey, goose and ostrich.

Claim 7

The composition of Claim 1, wherein the cell proliferative diseases are neoplastic diseases caused by tumor.

Claim 8

The composition of Claim 7, wherein the neoplastic diseases are cancer.

Claim 9 The composition of Claim 8, wherein the cancer is selected from the group consisting of large intestine cancer, spleen cancer, colon cancer, lung cancer, liver cancer, stomach cancer, esophageal cancer, pancreatic cancer, gallbladder cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, cervical cancer, endometrial cancer, choriocarcinoma, ovarian cancer, breast cancer, thyroid cancer, brain cancer, head and neck cancer, malignant melanoma, lymphoma, bone cancer, soft tissue sarcoma, and spinal cancer.

Claim 10 A method for preparing a powder of whole feathers of birds comprising the steps of:

(a) immersing feathers of birds in water of 10⁰C ~50°C for 5 minutes to 3 hours;

(c) washing the feathers with flowing water to remove impurities;

(d) immersing and sterilizing the feathers in 0.2-0.5 weight% (w/v) sodium chloride (NaCl) solution for 1 minute to 1 hour;

(e) immersing and sterilizing the feathers in 1~4 weight% (v/v) acetic acid solution for 15 minutes to 30 minutes;

(f) immersing and sterilizing the feathers in water of 3O⁰C -77⁰C for 1 minute to 3 hours;

(g) sterilizing and drying the immersed and sterilized feathers at 15°C ~40°C for 1 hour to 96 hours; (h) sterilizing the dried feathers by irradiation in 290-400 ran UV for 5 minutes to 2 hours;

(i) cutting the sterilized feathers to 5~15 mm for easier grinding; (j) sterilizing the cut feathers by irradiation in 290-400 nm UV for 5 minutes to 2 hours;

(k) grinding the cut and sterilized feathers;

Claim 11

A method for preparing a powder of a calamus or a rachis comprising the steps of:

(a) immersing feathers of birds in water of 10⁰C ~50°C for 5 minutes to 3 hours; (b) sterilizing the immersed feathers in brine with a salinity from 5.0 to 8.0 for 3 minutes to 90 minutes;

(c) washing the feathers with flowing water to remove impurities;

(f) immersing and sterilizing the calamus or the rachis in 1-4 weight% (v/v) acetic acid solution for 15 minutes to 30 minutes; (g) immersing and sterilizing the calamus or the rachis in water of 30⁰C ~77°C for 1 minute to 3 hours;

(h) sterilizing and drying the immersed and sterilized the calamus or the rachis of the above at 15⁰C ~40°C for 1 hour to 96 hours; (i) sterilizing the dried calamus or rachis by irradiation in 290~400 nm UV for 5 minutes to 2 hours;

(k) sterilizing the cut calamus or rachis by irradiation in 290-400 nm UV for 5 minutes to 2 hours;

(1) grinding the cut and sterilized calamus or rachis;

(m) sterilizing the resulting ground powder by irradiation in 290-400 nm UV for 5 minutes to 2 hours; and

Claim 12

An administrative method of the composition of claim 1, wherein the method comprises oral administration of the composition of claim 1 to a subject.

Claim 13

The method for sterilizing of the feathers of birds of anyone selected from the groups consisting of claims 4, 5, 10, and 11, wherein the method comprises steps of:

(a) sterilizing with a salinity from 5.0 to 8.0 for 3 minutes to 90 minutes;

(b) immersing and sterilizing in 0.2-0.5 weight% (w/v) sodium chloride solution for 1 minute to 1 hour;

(c) immersing and sterilizing in 1~4 weight% (v/v) acetic acid solution for 15 minutes to 30 minutes;

(d) immersing and sterilizing the feathers of birds in water of 30⁰C ~77°C for 1 minute to 3 hours; (e) sterilizing and drying the immersed and sterilized feathers at 15°C -4O⁰C for 1 hour to 96 hours;

(f) sterilizing the dried feathers by irradiation in 290-400 ran UV for 5 minutes to 2 hours; and

(g) sterilizing the resulting sterilized ground powder by irradiation in 200-290 nm UV for 30 seconds to

20 minutes.

Claim 14

A use of feathers of birds for preparing reagent for treating cell proliferative diseases.

Claim 15 A method for preventing and treating cell proliferative diseases comprising administering to a subject in need thereof an effective amount of feathers of birds.