CA2317113A1 - Pharmaceutical composition having antitumor activity and process for the preparation thereof - Google Patents
Pharmaceutical composition having antitumor activity and process for the preparation thereof Download PDFInfo
- Publication number
- CA2317113A1 CA2317113A1 CA002317113A CA2317113A CA2317113A1 CA 2317113 A1 CA2317113 A1 CA 2317113A1 CA 002317113 A CA002317113 A CA 002317113A CA 2317113 A CA2317113 A CA 2317113A CA 2317113 A1 CA2317113 A1 CA 2317113A1
- Authority
- CA
- Canada
- Prior art keywords
- powdered
- radix
- mixture
- pharmaceutical composition
- extract
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Abstract
A pharmaceutical composition having antitumor activity comprises a lyophilized powder derived from an extract of Pulsatillae Radix or Ulmaceae Cortex, or a mixture thereof, together with a pharmaceutically acceptable carrier therefor, the extract having been obtained by extracting Pulsatillae Radix or powdered Ulmaceae Cortex, or a mixture thereof, in a solvent at a temperature below 60°C. The pharmaceutical composition according to the invention can stored for several years, without losing its pharmaceutical activity.
Description
-1_ Pharmaceutical composition having antitumor activity and process for the preparation thereof The present invention relates to a pharmaceutical composition having andtumor activity and containing active herb ingredients, as well as to a process for the preparation thereof.
Applicant has already disclosed in Korean Patent No. 72982 a pharmaceutical composition having antitumor activity containing active herb ingredients, as well as a process for the preparation thereof.
The pharmaceutical composition described in the above patent contains as main active ingredients Pulsatillae Radix (Pulsatilla koreana Nakai, P. cerna, P.
danurica, P. ratensis, Chinese Pulsatillae, Mongolian Pulsatillae) and/or Clematis Chinensis Osbeclo (also called Chinese clematis), and optionally Ulmaceae Cortex, An-neniacae Semen, Ginseng Radix and Glycyrrhizae Radix.
Pulsatillae species are grown wild all over the world, and Pulsatillae Radix has been used as an antiphlogistic agent, astringent and hemostatic agent for the treatment of dysentery in Korea. It is known that Pulsatillae Radix contains anemonin, protoanemonin and saponin. Protoanemonin is the precursor of anemonin, and both may be dissolved in water, alcohol, chloroform, methylene chloride and the like.
Clematis Radix contains anemonin, anemonol and saponin. It has been used as an agent for gout, diuretic agent and agent for difficult menstruum.
Ulmaceae Cortex contains mucin and tannin, and has been used as a lenitive and adhesive.
Ginseng Radix has been known as a marvellous medicine in the Far East. It has been used as a tonic, agent for acute gastritis and agent for various bleeding diseases. Recently, it has been reported that Ginseng Radix has antitumor activity and contains Ginseng alkaloids, Ginseng saponins, essence oil, etc.
Applicant has already disclosed in Korean Patent No. 72982 a pharmaceutical composition having antitumor activity containing active herb ingredients, as well as a process for the preparation thereof.
The pharmaceutical composition described in the above patent contains as main active ingredients Pulsatillae Radix (Pulsatilla koreana Nakai, P. cerna, P.
danurica, P. ratensis, Chinese Pulsatillae, Mongolian Pulsatillae) and/or Clematis Chinensis Osbeclo (also called Chinese clematis), and optionally Ulmaceae Cortex, An-neniacae Semen, Ginseng Radix and Glycyrrhizae Radix.
Pulsatillae species are grown wild all over the world, and Pulsatillae Radix has been used as an antiphlogistic agent, astringent and hemostatic agent for the treatment of dysentery in Korea. It is known that Pulsatillae Radix contains anemonin, protoanemonin and saponin. Protoanemonin is the precursor of anemonin, and both may be dissolved in water, alcohol, chloroform, methylene chloride and the like.
Clematis Radix contains anemonin, anemonol and saponin. It has been used as an agent for gout, diuretic agent and agent for difficult menstruum.
Ulmaceae Cortex contains mucin and tannin, and has been used as a lenitive and adhesive.
Ginseng Radix has been known as a marvellous medicine in the Far East. It has been used as a tonic, agent for acute gastritis and agent for various bleeding diseases. Recently, it has been reported that Ginseng Radix has antitumor activity and contains Ginseng alkaloids, Ginseng saponins, essence oil, etc.
Glycyrrhizae Radix contains glycyrrhizin, liquiritin, licoricidin and liquiritoside and has been used as a cough remedy, expectorant, diaphoretic and agent for gastritis.
The pharmaceutical composition described in Korean Patent No. 72982 has excellent antitumor activity and contains an extract or powder of Pulsatillae Radix and/or Clematis Radix as main ingredients, and optionally an extract of Ulmaceae Cortex, Armeniacae Semen, Ginseng Radix and Glycyirhizae Radix. as described in this patent, such a composition may be prepared by drying and finely powdering each herb ingredients, extracting the herb ingredients in a solvent such as water, a lower alkanol, chloroform or methylene chloride, which can dissolve the active ingredients of the herbs at the temperature of 0°C to the boiling point of the solvent used for a period of 30 minutes to 24 hours, and then vaporizing the solvent to obtain the desired extract; which can be dissolved in water, alcohol or a mixture thereof. When extracting the active ingredients, each herb may be extracted independently or two or more herbs may be extracted together. Then, the extract is powdered and formulated into a pharmaceutical preparation by using pharmaceutically acceptable carriers such as lactose, various starches, sucrose, mannitol, sorbitol and inorganic salts such as calcium phosphate, calcium sulfate, aluminium silicate and calcium carbonate ; binders such as sucrose, glucose, starch, gelatin, carboxymethylcellulose, methylcellulose, gum arabic, gum tragacanth, ethylcellulose, sodium alginate, hydroxypropylmethylceilulose, polyvinylpyrrolidone and soluble cellulose; disintegrators such as starch, carboxymethylcellulose, methylcellulose and crystalline cellulose; lubricants such as magnesium stearate and calcium stearate; wetting agents such as glycerine, propylene glycol and sorbitol; preservatives such as sodium benzoate, methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, benzalkonium chloride, chlorobutanol and sodium dehydroacetate; dissolving agents such as soluble alcohols and derivatives thereof, and various surfactants; antioxidants such as sodium sulfite, sodium pyrosulfate, sodium metasulfate, sodium bisulfate, rongalite and ascorbic acid; isotonic agents such as sodium chloride and dextrose;
indolent agents such as benzylalcohol and chlorobutanol; and ointment bases such as vaseline, fluid paraffin, various vegetable oils, waxes and lanoline.
Pursuant to continued intensive study to improve the above antitumor composition, Applicant has found that such a composition is very unstable since it easily loses its pharmaceutical activity in 3 to 6 months.
It is therefore an object of the present invention to provide an improved pharmaceutical composition having antitumor activity, which is stable and maintains its pharmaceutical activity even when it is stored for several years.
Applicant has found quite unexpectedly that by extracting the herb ingredients described in Korean Patent No. 72982 in a solvent at a temperature below 60°C, promptly lyophilizing the extract and formulating the lyophilized powder into a pharmaceutical composition, the resulting composition can be stored for a long period of time without losing its pharmaceutical activity.
According to the present invention, there is thus provided a pharmaceutical composition having antitumor activity and comprising a lyophilized powder derived from an extract of Pulsatillae Radix or Ulmaceae Cortex, or a mixture thereof, together with at least one pharmaceutically acceptable earner therefor, the extract having been obtained by extracting powdered Pulsatillae Radix or powdered Ulrnaceae Cortex, or a mixture thereof, in a solvent at a temperature below 60°C.
According to a preferred embodiment of the invention, the lyophilized powder is derived from an extract of Pulsatillae Radix or Ulmaceae Cortex, or a mixture thereof, and Ginseng Radix or Glycyrrhizae Radix, or a mixture thereof, the extract having been obtained by extracting a mixture containing more than weight % of powdered Pulsatillae Radix or powdered Ulmaceae Cortex, or a mixture thereof, and less than 70 weight % of powdered Ginseng Radix or powdered Glycyrrhizae Radix, or a mixture thereof, in a solvent at a temperature below 60°C.
The present invention also provides, in another aspect thereof, a process for preparing a pharmaceutical composition as defined above. The process according S to the invention comprises the steps of:
a) extracting powdered Pulsatillae Radix or powdered Ulmaceae Cortex, or a mixture thereof, in a solvent at a temperature below 60°C
to obtain an extract solution;
b) filtering and lyophilizing the extract solution to obtain a lyophilized powder; and c) admixing the lyophilized powder with at least one pharmaceutically acceptable carrier; or b') admixing the extract solution obtained in step (a) with at least one pharmaceutically acceptable carrier; and c') filtering and lyophilizing the resulting mixture to obtain a lyophilized powder.
Examples of suitable solvents which may be used for the extraction of the herb ingredients include water, lower alkanol, acetone, ethyl acetate hexane and mixtures thereof.
The herb ingredients are extracted in a solvent at the temperature of below 60°C, and immediately lyophilized. The lyophilized powder may be filled into a vial and distilled water may be added thereinto so as to render the composition suitable for injection, or the lyophilized powder may be formulated into a capsule, tablet or ointment.
In the case where the composition of the invention is to be administered by injection, before lyophilization, the extract solution is preferably admixed with conventional adjuvants including preservatives such as methylparaben, ethylparaben and propylparaben, an isotonic agent such as sodium chloride and an indolent agent such as benzylalcohol.
In the case where the composition is to be used in the form of capsule, tablet, ointment or the like, the extract solution is first lyophilized and then the S lyophilized powder is admixed with conventional adjuvants such as those described in Korean patent No. 72,982.
About 100mg to Sg of the composition according to the invention, on the basis of the lyophilized powder, may be administered in a day, once a week to 3 times for a day. The dose may be varied depending on the sex, age, condition of disease, etc. of the patient.
The following non-limiting examples illustrate the invention.
Comparative examxy 1 6.26g of powdered Pulsadllae Radix were admixed with 90m1 of purified water. The mixture was warmed to 60°C and stirred for 60 minutes, and then centrifuged at 3,500 tpm for 30 minutes. SOmI of the centrifuged solution were filtered in a sterilized room at below 60°C. The resulting solution was rendered isotonic by adding NaCI under aseptic conditions, then sterile-filtered once again and divided into portions of 2.5 ml each, which were introduced into ampoules of 3m1 under aseptic conditions. The ampoules were sealed to provide injection ampoules.
Comrarative example 2 4g of powdered Pulsatillae Radix, 2g of powdered Ulmaceae cortex, 2g of powdered Ginseng Radix and 1 g of powdered Glycyrrhizae Radix were admixed with 90m1 of purified water and the mixture was stirred for 60 minutes at about 80°C while adding purified water in an amount corresponding to the amount of water which distilled off. The resulting solution was cooled to room temperature, centrifuged at 3,500 rpm for 30 minutes to obtain 46m1 of extracted solution.
NaCI
was added to the extract to obtain an isotonic solution. The isotonic solution was filtered in a sterilized room, sterile-filtered and divided into portions of 2m1 each, which were introduced into ampoules of 3m1. The ampoules were sealed and stored in a refrigerator.
Comparative example 3 62.6g of powdered Pulsatillae Radix, 31.3g of powdered Ginseng Radix and log of powdered Glycyrrhizae Radix were admixed with 900m1 of purified water and the mixture was stirred for 60 minutes at about 60°C while adding purified water in an amount corresponding to the amount of water which distilled off.
The resulting solution (40m1) was filtered and concentrated to give 26.4g of concentrated extract.
Example 1 6g of powdered Pulsatillae Radix was added to 100m1 of distilled water and extracted for 60 minutes at a temperature below 60°C under stirring.
The extract solution was then centrifuged at 5000 rpm, and 900mg of an isotonic agent consisting of NaCI and 160mg of a preservative consisting of methyl paraoxybenzoate were added thereto. The resulting mixture was sterile-filtered in a sterilized room, divided into 20 vials of Sml each and promptly lyophilized at a temperature below -40 C to obtain an injectable powder. The vials were thereafter sealed.
Example 2 6g of powdered Pulsatillae Radix, 4g of powdered Ulmaceae cortex and 0.9g of powdered Glycyrrhizae Radix were added to 100m1 of distilled water and extracted for 60 minutes at a temperature below 60°C under stirring.
The extract solution was then centrifuged at 5000 rpm, and 900mg of NaCI and 160mg of methyl paraoxybenzoate were added thereto. The resulting mixture was sterile-filtered in a sterilized room, divided into 20 vials of Sml each and promptly lyophilized at a temperature below -40°C , to obtain an injectable powder. The vials were thereafter sealed.
_7_ Example 3 6g of powdered Pulsatillae Radix, 3g of powdered Ginseng Radix and 0.9g of powdered Glycyrrhizae Radix were added to 100m1 of distilled water and extracted for 60 minutes at a temperature below 60°C under stirring, while adding distilled water in an amount corresponding to the amount of water which distilled off. The extract solution was then centrifuged at 5000 rpm, 900mg of NaCI and 160mg of a preservative consisting of propyl paraoxybenzoate were added thereto.
The resulting mixture was sterile-filtered in a sterilized room, divided into 20 vials of 5ml each and promptly lyophilized at a temperature below -40°C, to obtain an injectable powder. The vials were thereafter sealed.
Example 4 60g of powdered Pulsatillae Radix, 40g of powdered Ulmaceae Cortex and 9g of powdered Glycyrrhizae Radix were added to 1000m1 of distilled water and extracted for 60 minutes at a temperature below 60°C under stirring, while adding distilled water in an amount corresponding to the amount of water which distilled off. The extract solution was then centrifuged at 5000 rpm and promptly lyophilized at a temperature below -40°C to give 38.15g of lyophilized powder.
Example 5 60g of powdered Pulsatillae Radix, 60g of powdered Ulmaceae Cortex and 9g of powdered Glycyrrhizae Radix were added to 1000m1 of 50%(v/v) ethanol and extracted for 60 minutes at a temperature of 50 - 60°C under stirring, while adding ethanol in an amount corresponding to that which distilled off. The extract solution was then centrifuged at 5000 rpm and promptly lyophilized at a temperature below -40°C to give 45.15g of lyophilized powder.
Example 6 60g of powdered Pulsatillae Radix, 30g of powdered Ginseng Radix and 9g of powdered Glycyrrhizae Radix were added to 1004m1 of solvent consisting of a 50%(v/v) aqueous solution of . acetone and extracted for 60 minutes at a temperature of 50 - 60°C under stirring, while adding solvent in an amount corresponding to the amount of solvent which distilled off. The extract solution was centrifuged at 5000 rpm and promptly lyophilized at a temperature below -40°C to give 34.65g of lyophilized powder.
Example 7 6g of powdered Pulsatillae Radix, 4g of powdered Ulmaceae Cortex and 0.9g of powdered Glycyrrhizae Radix were added to 1 OOM 1 of 70%(v/v) ethanol and extracted for 60 minutes at a temperature below 60°C under stirring, while adding ethanol in an amount corresponding to the amount of ethanol which distilled off. The extract solution was then centrifuged at 5000 rpm, and 900mg of NaCI and 160mg of methyl paraoxybenzoate were added thereto. The resulting mixture was sterile-filtered in a sterilized room, divided into 20 vials of 5m1 each and promptly lyophilized at a temperature below -40°C to obtain an injectable powder. The vials were thereafter sealed.
Example 8 lOg of powdered Ulmaceae Cortex was added to 100m1 of 50%(v/v) ethanol and extracted for 60 minutes at a temperature below 60°C under stirring.
The extract solution was centrifuged at 5000 rpm, and 900mg of NaCI and 160mg of methyl paraoxybenzoate were added thereto. The resulting mixture was sterile-filtered in a sterilized room, divided into 20 vials of 5m1 each and promptly lyophilized at a temperature below -40°C, to obtain an inj ectable powder. The vials were thereafter sealed.
Example 9 60g of powdered Pulsatillae Radix, 60g of powdered Ulmaceae Cortex and 9g of powdered Glycyrrhizae Radix were added to 1000m 1 of hexane and extracted for 90 minutes at a temperature below 60°C under stirnng, while adding hexane in an amount corresponding to the amount of hexane which distilled off.
The extract solution was centrifuged at 5000 rpm and the resulting solution was promptly lyophilized at a temperature below -40°C to obtain a lyophilized powder.
Example 10 Lyophilized powder obtained in Example 4 1 SOmg Crystalline cellulose SOmg Lactose SOmg Magnesium stearate 3mg The above ingredients were cast into tablets and sealed with aluminium foil.
Example 11 Lyophilized powder obtained in Example 6 1 SOmg Lactose 30mg Corn starch 30mg Talc Smg Magnesium stearate 3mg The above ingredients were filled into a hard capsule of gelatin and sealed with aluminium foil.
Example 12 Lyophilized powder obtained in Example 5 1000mg Conventional ointment base q,s, The above ingredients were formulated into lOg of ointment, and filled and sealed into an aluminium tube.
Experiment l: Acute toxicity The lyophilized powder obtained in Example 1 was administered to 8 rats of 234-276g, whereby the LDSO was 800mg/kg.
Experiment 2: Antitumor effect O.lml of suspension of Sarcoma 180 cells(1x106 cells) was injected (s.c.) into 30 rats of about 25g to develop tumors. After 6 days, O.15m1 of an injectable solution prepared by dissolving the injectable powder of Example 1 in Sml of distilled water for injection was injected (s.c.) to 10 rats once a day and O.15m1 of the injectable solution obtained in Comparative Example 1 was injected (s.c.) to another 10 rats once a day. In addition, O.lSml of a physiological saline solution was injected to the other 10 rats for 10 days, as a control group.
9 rats of the group which was treated with the injectable solution of Example 1 and 9 rats of the group which was treated with the injectable solution of Comparative Example 1 were cured after 15 days. Only 1 rat of each of these groups died on the 16th day, while the 10 rats of the control group all died between the 10th day and the 15th day.
Experiment 3: Antitumor effect O.lml of suspension of Sarcoma 180 cells(1x106 cells) was injected (s.c.) into 30 rats each weighing about 25g to develop tumors. After 6 days, O.15m1 of an injectable solution prepared by dissolving the injectable powder obtained in Example 1 in Sml of distilled water was injected (s.c.) to 10 rats (group 1) once a day, and O.15m1 of the injectable solution obtained in Comparative Example 1 preserved for 6 months in a refrigerator was injected (s.c.) to another 10 rats (group 2) once a day, and O.15m1 of physiological saline solution was injected to the other 10 rats for 10 days as a control group (group 3).
9 rats of group 1 were cured by the injectable solution after 15 days, and 1 rat of group 1 died on the 17th day. 3 rats of group 2 were cured by the injectable solution after 15 days, 1 rat died on the 12th day, 3 rats died on the 15th day, and 3 rats died on the 17th day. 10 rats of group 3 all died between the 10th day and the 15th day.
Experiment 4: Antitumor effect O.lml of suspension of Sarcoma 180 cells(1x106 cells) was injected (s.c.) into 48 rats each weighing about 25g to develop tumors. Beginning on the 9th day(terminal stage of cancer), O.15m1 of a sample injection prepared by dissolving the injectable powder obtained in Example 3 in Sml of distilled water was injected (s.c.) to 7 rats once a day (group 1), O.15m1 of the same sample injection was injected (s.c.) to another 7 rats twice a day (group 2), O.lSml of a sample injection obtained in Comparative Example 1 was injected (s.c.) to another 7 rats once a day (group 3), O.15m1 of a sample injection obtained in Comparative Example 1 was S injected (s.c.) to another 7 rats twice a day (group 4), O.15m1 of a sample injection obtained in Comparative Example 1 which was preserved for 3 months in a refrigerator was injected (s.c.) to another 7 rats once a day (group 5), O.15m1 of a sample injection obtained in Comparative Example 1 which was preserved in a refrigerator for 6 months was injected (s.c.) to another 7 rats twice a day (group 6), and the other 6 rats were used as a control group.
In each of groups 1 and 3, 1 rat died on the 15th day from carcinogenesis, 1 rat died on the 17th day, 1 rat died on the 18th day, and 1 rat died on the 20th day, while 3 rats were cured by the injection after 20 days (on the 29th day from carcinogenesis).
1 S In each of groups 2 and 4, 1 rat died on the 16th day from carcinogenesis, rat died on the 18th day, 1 rat died on the 19th day, and 1 rat died on the 21 st day, while 3 rats were cured by the injection after 19 days (on the 28th day from carcinogenesis).
In group 5, 1 rat died on the 15th day of carcinogenesis, 1 rat died on the 17th day, 1 rat died on the 18th day, 1 rat died on the 19th day, and 1 rat died on the 21st day, while 2 rats were cured by the injection after 21 days.
In group 6, 1 rat died on the 15th day of carcinogenesis, 1 rat died on the 16th day, 1 rat died on the 18th day, 1 rat died on the 19th day, and 2 rats died on the 20th day, while 1 rat was cured by the injection after 20 days.
In group 7, all rats died on the 15th day of carcinogenesis.
Experiment 5: Clinical Test on a volunteer by administration of an injectable solution prepared by dissolving the injectable powder obtained in Example 1 in distilled water.
Subj ect Name: KIM, Myung-Won ( 36 years old at the treatment, male ) Kind of disease: Progressed thyroid cancer Diagnosis: General Hospital affiliated to the Youngnam University in Taegu on May 7, 1992.
Period of medication: from May 16, 1992 to October 20, 1992 Medication: 12m1 of the injectable solution were injected (i.v.) once a day for 4 days, while lOml of the injectable solution were injected into the protruded tumor twice a day for 8 days (totally 15 times), and then no injection was performed for 25 days. After that, the medication was repeated by i.v. injection and direct injection into the protruded tumor as the same method above, whereby the tumor disappeared completely. After 5 years, the General Hospital affiliated to the Youngnam University and the General Hospital affiliated to the Chungnam University determined that the patient was completely cured.
Experiment 6: Clinical Test on a volunteer by administration of an injectable solution prepared by dissolving the injectable powder obtained in Example 2 in distilled water.
Subject:
Name: KIM, Chul-Ki (SO years old at the treatment, male) Kind of disease: progressed lung cancer. Weight loss, tussis, bloody phlegm, dyspnea due to residual cancer after operation.
Diagnosis: Christian Hospital in Wonju affiliated to Yonsei University on May 18, 1989.
Period of medication: from September 3, 1989 to March 5, 1990 Medication: lOml of the injectable solution were injected (i.v.) once a day for 4 days, and then no treatment was performed for 3 weeks. The same medication was repeated for 7 months, whereby the tumor and related symptoms disappeared completely. After 6 years, the General Hospital affiliated to the Yonsei University and the General Hospital affiliated to the Chungnam University determined that the patient was completely cured.
Experiment 7: Clinical Test on a volunteer by administration of ane injectable solution prepared by dissolving the injectable powder obtained in Example 2 in distilled water.
Subject:
Name: SUH, Sang-Bong (35 years old at the treatment, male) Kind of disease: Progressed rectal cancer. Weight loss and severe pain due to residual cancer after partial removal of tumor by operation.
Diagnosis: Goshin Medical Center in Pusan on July 19, 1991.
Period of medication: from November 19, 1991 to May 12, 1992 Medication: 12m1 of the injectable solution were injected (i.v.) once a day for 4 days, and then no treatment was performed for 3 weeks. The same medication was repeated for 7 months, whereby the residual tumor disappeared completely.
After 6 years, the Goshin Medical Center in Pusam and the General Hospital affiliated to the Chungnam University determined that the patient was completely cured.
Experiment 8: Clinical Test on a volunteer by administration of an injectable solution prepared by dissolving the injectable powder obtained in Example 3 in distilled water.
Subj ect:
Name: PARK, Ju-Sang (45 years old at the treatment, male) Kind of disease: progressed stomach cancer. Feeling heavy on the stomach, dyspepsia and weight loss.
Diagnosis: Goshin Medical Center in Pusan on December 29, 1989 Period of medication: from March 1, 1990 to September 29, 1990 S
Medication: 12m1 of the injectable solution were injected (i.v.) once a day for 4 days, and simultaneously 173mg of the sample were taken internally four times a day. The i.v. injection was performed in such a manner that the injection was i.v.
injected for 4 days and then no injection was made for 4 weeks, while the internal administration was continued. As a result, the symptoms disappeared completely and the patient was ascertained to be normal by biopsy. After 6 years, the Goshin Medical Center determined that the patient was completely cured.
Experiment 9: Clinical Test on a volunteer by administration of an injectable solution prepared by dissolving the injectable powder obtained in Example 5 in distilled water.
Subj ect:
Name: LEE, Bok-Do (65 years old at the treatment, male) Kind of disease: progressed liver cancer. Abdominal dropsy, dyspepsia and weight loss due to residual cancer after partial removal of tumor by operation.
Diagnosis: Goshin Medical Center in Pusan on April 8, 1994.
Period of medication : from May 31, 1994 to December 30, 1994 Medication: 9m1 of the injectable solution were injected (i.v.) once a day for days, and simultaneously 4m1 of the sample were injected directly into the residual tumor in the liver once a day for 3 days, and then no treatment was performed for 3 weeks. The same medication was performed three times, and then from the 4th cycle of medication, only i.v. injection was performed, whereby the residual tumor disappeared completely. After 3 years, the Goshin Medical Center and the General I~iospital affiliated to Chungnam University determined that the patient was completely cured.
Stability test 1 Samples of the lyophilized powder obtained in Examples 1 to 9 were preserved for 2 years and then dissolved in distilled water for injection. The resulting solution had a transparent light brown color and no precipitate settled.
Stability test 2 Samples of injectable solution obtained in Comparative Example 2 were stored for 1 month, 2 months and 3 months, respectively. A precipitate settled after I month of storage and after 3 months the solutions had turbid color, whereby the samples could not be used for injection.
As it is apparent from the above experiments, the antitumor composition to Korean Patent No. 72,982 is sensitive to moisture, and even if stored in a refrigerator, it is easily deteriorated and its efficacy is severely reduced so that it cannot be preserved for a long period of time and thereafter be used as a medicine.
In contrast, the pharmaceutical composition according to the present invention exhibits no change in quality and efficacy after long preservation, so that it may be used safely.
The pharmaceutical composition described in Korean Patent No. 72982 has excellent antitumor activity and contains an extract or powder of Pulsatillae Radix and/or Clematis Radix as main ingredients, and optionally an extract of Ulmaceae Cortex, Armeniacae Semen, Ginseng Radix and Glycyirhizae Radix. as described in this patent, such a composition may be prepared by drying and finely powdering each herb ingredients, extracting the herb ingredients in a solvent such as water, a lower alkanol, chloroform or methylene chloride, which can dissolve the active ingredients of the herbs at the temperature of 0°C to the boiling point of the solvent used for a period of 30 minutes to 24 hours, and then vaporizing the solvent to obtain the desired extract; which can be dissolved in water, alcohol or a mixture thereof. When extracting the active ingredients, each herb may be extracted independently or two or more herbs may be extracted together. Then, the extract is powdered and formulated into a pharmaceutical preparation by using pharmaceutically acceptable carriers such as lactose, various starches, sucrose, mannitol, sorbitol and inorganic salts such as calcium phosphate, calcium sulfate, aluminium silicate and calcium carbonate ; binders such as sucrose, glucose, starch, gelatin, carboxymethylcellulose, methylcellulose, gum arabic, gum tragacanth, ethylcellulose, sodium alginate, hydroxypropylmethylceilulose, polyvinylpyrrolidone and soluble cellulose; disintegrators such as starch, carboxymethylcellulose, methylcellulose and crystalline cellulose; lubricants such as magnesium stearate and calcium stearate; wetting agents such as glycerine, propylene glycol and sorbitol; preservatives such as sodium benzoate, methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, benzalkonium chloride, chlorobutanol and sodium dehydroacetate; dissolving agents such as soluble alcohols and derivatives thereof, and various surfactants; antioxidants such as sodium sulfite, sodium pyrosulfate, sodium metasulfate, sodium bisulfate, rongalite and ascorbic acid; isotonic agents such as sodium chloride and dextrose;
indolent agents such as benzylalcohol and chlorobutanol; and ointment bases such as vaseline, fluid paraffin, various vegetable oils, waxes and lanoline.
Pursuant to continued intensive study to improve the above antitumor composition, Applicant has found that such a composition is very unstable since it easily loses its pharmaceutical activity in 3 to 6 months.
It is therefore an object of the present invention to provide an improved pharmaceutical composition having antitumor activity, which is stable and maintains its pharmaceutical activity even when it is stored for several years.
Applicant has found quite unexpectedly that by extracting the herb ingredients described in Korean Patent No. 72982 in a solvent at a temperature below 60°C, promptly lyophilizing the extract and formulating the lyophilized powder into a pharmaceutical composition, the resulting composition can be stored for a long period of time without losing its pharmaceutical activity.
According to the present invention, there is thus provided a pharmaceutical composition having antitumor activity and comprising a lyophilized powder derived from an extract of Pulsatillae Radix or Ulmaceae Cortex, or a mixture thereof, together with at least one pharmaceutically acceptable earner therefor, the extract having been obtained by extracting powdered Pulsatillae Radix or powdered Ulrnaceae Cortex, or a mixture thereof, in a solvent at a temperature below 60°C.
According to a preferred embodiment of the invention, the lyophilized powder is derived from an extract of Pulsatillae Radix or Ulmaceae Cortex, or a mixture thereof, and Ginseng Radix or Glycyrrhizae Radix, or a mixture thereof, the extract having been obtained by extracting a mixture containing more than weight % of powdered Pulsatillae Radix or powdered Ulmaceae Cortex, or a mixture thereof, and less than 70 weight % of powdered Ginseng Radix or powdered Glycyrrhizae Radix, or a mixture thereof, in a solvent at a temperature below 60°C.
The present invention also provides, in another aspect thereof, a process for preparing a pharmaceutical composition as defined above. The process according S to the invention comprises the steps of:
a) extracting powdered Pulsatillae Radix or powdered Ulmaceae Cortex, or a mixture thereof, in a solvent at a temperature below 60°C
to obtain an extract solution;
b) filtering and lyophilizing the extract solution to obtain a lyophilized powder; and c) admixing the lyophilized powder with at least one pharmaceutically acceptable carrier; or b') admixing the extract solution obtained in step (a) with at least one pharmaceutically acceptable carrier; and c') filtering and lyophilizing the resulting mixture to obtain a lyophilized powder.
Examples of suitable solvents which may be used for the extraction of the herb ingredients include water, lower alkanol, acetone, ethyl acetate hexane and mixtures thereof.
The herb ingredients are extracted in a solvent at the temperature of below 60°C, and immediately lyophilized. The lyophilized powder may be filled into a vial and distilled water may be added thereinto so as to render the composition suitable for injection, or the lyophilized powder may be formulated into a capsule, tablet or ointment.
In the case where the composition of the invention is to be administered by injection, before lyophilization, the extract solution is preferably admixed with conventional adjuvants including preservatives such as methylparaben, ethylparaben and propylparaben, an isotonic agent such as sodium chloride and an indolent agent such as benzylalcohol.
In the case where the composition is to be used in the form of capsule, tablet, ointment or the like, the extract solution is first lyophilized and then the S lyophilized powder is admixed with conventional adjuvants such as those described in Korean patent No. 72,982.
About 100mg to Sg of the composition according to the invention, on the basis of the lyophilized powder, may be administered in a day, once a week to 3 times for a day. The dose may be varied depending on the sex, age, condition of disease, etc. of the patient.
The following non-limiting examples illustrate the invention.
Comparative examxy 1 6.26g of powdered Pulsadllae Radix were admixed with 90m1 of purified water. The mixture was warmed to 60°C and stirred for 60 minutes, and then centrifuged at 3,500 tpm for 30 minutes. SOmI of the centrifuged solution were filtered in a sterilized room at below 60°C. The resulting solution was rendered isotonic by adding NaCI under aseptic conditions, then sterile-filtered once again and divided into portions of 2.5 ml each, which were introduced into ampoules of 3m1 under aseptic conditions. The ampoules were sealed to provide injection ampoules.
Comrarative example 2 4g of powdered Pulsatillae Radix, 2g of powdered Ulmaceae cortex, 2g of powdered Ginseng Radix and 1 g of powdered Glycyrrhizae Radix were admixed with 90m1 of purified water and the mixture was stirred for 60 minutes at about 80°C while adding purified water in an amount corresponding to the amount of water which distilled off. The resulting solution was cooled to room temperature, centrifuged at 3,500 rpm for 30 minutes to obtain 46m1 of extracted solution.
NaCI
was added to the extract to obtain an isotonic solution. The isotonic solution was filtered in a sterilized room, sterile-filtered and divided into portions of 2m1 each, which were introduced into ampoules of 3m1. The ampoules were sealed and stored in a refrigerator.
Comparative example 3 62.6g of powdered Pulsatillae Radix, 31.3g of powdered Ginseng Radix and log of powdered Glycyrrhizae Radix were admixed with 900m1 of purified water and the mixture was stirred for 60 minutes at about 60°C while adding purified water in an amount corresponding to the amount of water which distilled off.
The resulting solution (40m1) was filtered and concentrated to give 26.4g of concentrated extract.
Example 1 6g of powdered Pulsatillae Radix was added to 100m1 of distilled water and extracted for 60 minutes at a temperature below 60°C under stirring.
The extract solution was then centrifuged at 5000 rpm, and 900mg of an isotonic agent consisting of NaCI and 160mg of a preservative consisting of methyl paraoxybenzoate were added thereto. The resulting mixture was sterile-filtered in a sterilized room, divided into 20 vials of Sml each and promptly lyophilized at a temperature below -40 C to obtain an injectable powder. The vials were thereafter sealed.
Example 2 6g of powdered Pulsatillae Radix, 4g of powdered Ulmaceae cortex and 0.9g of powdered Glycyrrhizae Radix were added to 100m1 of distilled water and extracted for 60 minutes at a temperature below 60°C under stirring.
The extract solution was then centrifuged at 5000 rpm, and 900mg of NaCI and 160mg of methyl paraoxybenzoate were added thereto. The resulting mixture was sterile-filtered in a sterilized room, divided into 20 vials of Sml each and promptly lyophilized at a temperature below -40°C , to obtain an injectable powder. The vials were thereafter sealed.
_7_ Example 3 6g of powdered Pulsatillae Radix, 3g of powdered Ginseng Radix and 0.9g of powdered Glycyrrhizae Radix were added to 100m1 of distilled water and extracted for 60 minutes at a temperature below 60°C under stirring, while adding distilled water in an amount corresponding to the amount of water which distilled off. The extract solution was then centrifuged at 5000 rpm, 900mg of NaCI and 160mg of a preservative consisting of propyl paraoxybenzoate were added thereto.
The resulting mixture was sterile-filtered in a sterilized room, divided into 20 vials of 5ml each and promptly lyophilized at a temperature below -40°C, to obtain an injectable powder. The vials were thereafter sealed.
Example 4 60g of powdered Pulsatillae Radix, 40g of powdered Ulmaceae Cortex and 9g of powdered Glycyrrhizae Radix were added to 1000m1 of distilled water and extracted for 60 minutes at a temperature below 60°C under stirring, while adding distilled water in an amount corresponding to the amount of water which distilled off. The extract solution was then centrifuged at 5000 rpm and promptly lyophilized at a temperature below -40°C to give 38.15g of lyophilized powder.
Example 5 60g of powdered Pulsatillae Radix, 60g of powdered Ulmaceae Cortex and 9g of powdered Glycyrrhizae Radix were added to 1000m1 of 50%(v/v) ethanol and extracted for 60 minutes at a temperature of 50 - 60°C under stirring, while adding ethanol in an amount corresponding to that which distilled off. The extract solution was then centrifuged at 5000 rpm and promptly lyophilized at a temperature below -40°C to give 45.15g of lyophilized powder.
Example 6 60g of powdered Pulsatillae Radix, 30g of powdered Ginseng Radix and 9g of powdered Glycyrrhizae Radix were added to 1004m1 of solvent consisting of a 50%(v/v) aqueous solution of . acetone and extracted for 60 minutes at a temperature of 50 - 60°C under stirring, while adding solvent in an amount corresponding to the amount of solvent which distilled off. The extract solution was centrifuged at 5000 rpm and promptly lyophilized at a temperature below -40°C to give 34.65g of lyophilized powder.
Example 7 6g of powdered Pulsatillae Radix, 4g of powdered Ulmaceae Cortex and 0.9g of powdered Glycyrrhizae Radix were added to 1 OOM 1 of 70%(v/v) ethanol and extracted for 60 minutes at a temperature below 60°C under stirring, while adding ethanol in an amount corresponding to the amount of ethanol which distilled off. The extract solution was then centrifuged at 5000 rpm, and 900mg of NaCI and 160mg of methyl paraoxybenzoate were added thereto. The resulting mixture was sterile-filtered in a sterilized room, divided into 20 vials of 5m1 each and promptly lyophilized at a temperature below -40°C to obtain an injectable powder. The vials were thereafter sealed.
Example 8 lOg of powdered Ulmaceae Cortex was added to 100m1 of 50%(v/v) ethanol and extracted for 60 minutes at a temperature below 60°C under stirring.
The extract solution was centrifuged at 5000 rpm, and 900mg of NaCI and 160mg of methyl paraoxybenzoate were added thereto. The resulting mixture was sterile-filtered in a sterilized room, divided into 20 vials of 5m1 each and promptly lyophilized at a temperature below -40°C, to obtain an inj ectable powder. The vials were thereafter sealed.
Example 9 60g of powdered Pulsatillae Radix, 60g of powdered Ulmaceae Cortex and 9g of powdered Glycyrrhizae Radix were added to 1000m 1 of hexane and extracted for 90 minutes at a temperature below 60°C under stirnng, while adding hexane in an amount corresponding to the amount of hexane which distilled off.
The extract solution was centrifuged at 5000 rpm and the resulting solution was promptly lyophilized at a temperature below -40°C to obtain a lyophilized powder.
Example 10 Lyophilized powder obtained in Example 4 1 SOmg Crystalline cellulose SOmg Lactose SOmg Magnesium stearate 3mg The above ingredients were cast into tablets and sealed with aluminium foil.
Example 11 Lyophilized powder obtained in Example 6 1 SOmg Lactose 30mg Corn starch 30mg Talc Smg Magnesium stearate 3mg The above ingredients were filled into a hard capsule of gelatin and sealed with aluminium foil.
Example 12 Lyophilized powder obtained in Example 5 1000mg Conventional ointment base q,s, The above ingredients were formulated into lOg of ointment, and filled and sealed into an aluminium tube.
Experiment l: Acute toxicity The lyophilized powder obtained in Example 1 was administered to 8 rats of 234-276g, whereby the LDSO was 800mg/kg.
Experiment 2: Antitumor effect O.lml of suspension of Sarcoma 180 cells(1x106 cells) was injected (s.c.) into 30 rats of about 25g to develop tumors. After 6 days, O.15m1 of an injectable solution prepared by dissolving the injectable powder of Example 1 in Sml of distilled water for injection was injected (s.c.) to 10 rats once a day and O.15m1 of the injectable solution obtained in Comparative Example 1 was injected (s.c.) to another 10 rats once a day. In addition, O.lSml of a physiological saline solution was injected to the other 10 rats for 10 days, as a control group.
9 rats of the group which was treated with the injectable solution of Example 1 and 9 rats of the group which was treated with the injectable solution of Comparative Example 1 were cured after 15 days. Only 1 rat of each of these groups died on the 16th day, while the 10 rats of the control group all died between the 10th day and the 15th day.
Experiment 3: Antitumor effect O.lml of suspension of Sarcoma 180 cells(1x106 cells) was injected (s.c.) into 30 rats each weighing about 25g to develop tumors. After 6 days, O.15m1 of an injectable solution prepared by dissolving the injectable powder obtained in Example 1 in Sml of distilled water was injected (s.c.) to 10 rats (group 1) once a day, and O.15m1 of the injectable solution obtained in Comparative Example 1 preserved for 6 months in a refrigerator was injected (s.c.) to another 10 rats (group 2) once a day, and O.15m1 of physiological saline solution was injected to the other 10 rats for 10 days as a control group (group 3).
9 rats of group 1 were cured by the injectable solution after 15 days, and 1 rat of group 1 died on the 17th day. 3 rats of group 2 were cured by the injectable solution after 15 days, 1 rat died on the 12th day, 3 rats died on the 15th day, and 3 rats died on the 17th day. 10 rats of group 3 all died between the 10th day and the 15th day.
Experiment 4: Antitumor effect O.lml of suspension of Sarcoma 180 cells(1x106 cells) was injected (s.c.) into 48 rats each weighing about 25g to develop tumors. Beginning on the 9th day(terminal stage of cancer), O.15m1 of a sample injection prepared by dissolving the injectable powder obtained in Example 3 in Sml of distilled water was injected (s.c.) to 7 rats once a day (group 1), O.15m1 of the same sample injection was injected (s.c.) to another 7 rats twice a day (group 2), O.lSml of a sample injection obtained in Comparative Example 1 was injected (s.c.) to another 7 rats once a day (group 3), O.15m1 of a sample injection obtained in Comparative Example 1 was S injected (s.c.) to another 7 rats twice a day (group 4), O.15m1 of a sample injection obtained in Comparative Example 1 which was preserved for 3 months in a refrigerator was injected (s.c.) to another 7 rats once a day (group 5), O.15m1 of a sample injection obtained in Comparative Example 1 which was preserved in a refrigerator for 6 months was injected (s.c.) to another 7 rats twice a day (group 6), and the other 6 rats were used as a control group.
In each of groups 1 and 3, 1 rat died on the 15th day from carcinogenesis, 1 rat died on the 17th day, 1 rat died on the 18th day, and 1 rat died on the 20th day, while 3 rats were cured by the injection after 20 days (on the 29th day from carcinogenesis).
1 S In each of groups 2 and 4, 1 rat died on the 16th day from carcinogenesis, rat died on the 18th day, 1 rat died on the 19th day, and 1 rat died on the 21 st day, while 3 rats were cured by the injection after 19 days (on the 28th day from carcinogenesis).
In group 5, 1 rat died on the 15th day of carcinogenesis, 1 rat died on the 17th day, 1 rat died on the 18th day, 1 rat died on the 19th day, and 1 rat died on the 21st day, while 2 rats were cured by the injection after 21 days.
In group 6, 1 rat died on the 15th day of carcinogenesis, 1 rat died on the 16th day, 1 rat died on the 18th day, 1 rat died on the 19th day, and 2 rats died on the 20th day, while 1 rat was cured by the injection after 20 days.
In group 7, all rats died on the 15th day of carcinogenesis.
Experiment 5: Clinical Test on a volunteer by administration of an injectable solution prepared by dissolving the injectable powder obtained in Example 1 in distilled water.
Subj ect Name: KIM, Myung-Won ( 36 years old at the treatment, male ) Kind of disease: Progressed thyroid cancer Diagnosis: General Hospital affiliated to the Youngnam University in Taegu on May 7, 1992.
Period of medication: from May 16, 1992 to October 20, 1992 Medication: 12m1 of the injectable solution were injected (i.v.) once a day for 4 days, while lOml of the injectable solution were injected into the protruded tumor twice a day for 8 days (totally 15 times), and then no injection was performed for 25 days. After that, the medication was repeated by i.v. injection and direct injection into the protruded tumor as the same method above, whereby the tumor disappeared completely. After 5 years, the General Hospital affiliated to the Youngnam University and the General Hospital affiliated to the Chungnam University determined that the patient was completely cured.
Experiment 6: Clinical Test on a volunteer by administration of an injectable solution prepared by dissolving the injectable powder obtained in Example 2 in distilled water.
Subject:
Name: KIM, Chul-Ki (SO years old at the treatment, male) Kind of disease: progressed lung cancer. Weight loss, tussis, bloody phlegm, dyspnea due to residual cancer after operation.
Diagnosis: Christian Hospital in Wonju affiliated to Yonsei University on May 18, 1989.
Period of medication: from September 3, 1989 to March 5, 1990 Medication: lOml of the injectable solution were injected (i.v.) once a day for 4 days, and then no treatment was performed for 3 weeks. The same medication was repeated for 7 months, whereby the tumor and related symptoms disappeared completely. After 6 years, the General Hospital affiliated to the Yonsei University and the General Hospital affiliated to the Chungnam University determined that the patient was completely cured.
Experiment 7: Clinical Test on a volunteer by administration of ane injectable solution prepared by dissolving the injectable powder obtained in Example 2 in distilled water.
Subject:
Name: SUH, Sang-Bong (35 years old at the treatment, male) Kind of disease: Progressed rectal cancer. Weight loss and severe pain due to residual cancer after partial removal of tumor by operation.
Diagnosis: Goshin Medical Center in Pusan on July 19, 1991.
Period of medication: from November 19, 1991 to May 12, 1992 Medication: 12m1 of the injectable solution were injected (i.v.) once a day for 4 days, and then no treatment was performed for 3 weeks. The same medication was repeated for 7 months, whereby the residual tumor disappeared completely.
After 6 years, the Goshin Medical Center in Pusam and the General Hospital affiliated to the Chungnam University determined that the patient was completely cured.
Experiment 8: Clinical Test on a volunteer by administration of an injectable solution prepared by dissolving the injectable powder obtained in Example 3 in distilled water.
Subj ect:
Name: PARK, Ju-Sang (45 years old at the treatment, male) Kind of disease: progressed stomach cancer. Feeling heavy on the stomach, dyspepsia and weight loss.
Diagnosis: Goshin Medical Center in Pusan on December 29, 1989 Period of medication: from March 1, 1990 to September 29, 1990 S
Medication: 12m1 of the injectable solution were injected (i.v.) once a day for 4 days, and simultaneously 173mg of the sample were taken internally four times a day. The i.v. injection was performed in such a manner that the injection was i.v.
injected for 4 days and then no injection was made for 4 weeks, while the internal administration was continued. As a result, the symptoms disappeared completely and the patient was ascertained to be normal by biopsy. After 6 years, the Goshin Medical Center determined that the patient was completely cured.
Experiment 9: Clinical Test on a volunteer by administration of an injectable solution prepared by dissolving the injectable powder obtained in Example 5 in distilled water.
Subj ect:
Name: LEE, Bok-Do (65 years old at the treatment, male) Kind of disease: progressed liver cancer. Abdominal dropsy, dyspepsia and weight loss due to residual cancer after partial removal of tumor by operation.
Diagnosis: Goshin Medical Center in Pusan on April 8, 1994.
Period of medication : from May 31, 1994 to December 30, 1994 Medication: 9m1 of the injectable solution were injected (i.v.) once a day for days, and simultaneously 4m1 of the sample were injected directly into the residual tumor in the liver once a day for 3 days, and then no treatment was performed for 3 weeks. The same medication was performed three times, and then from the 4th cycle of medication, only i.v. injection was performed, whereby the residual tumor disappeared completely. After 3 years, the Goshin Medical Center and the General I~iospital affiliated to Chungnam University determined that the patient was completely cured.
Stability test 1 Samples of the lyophilized powder obtained in Examples 1 to 9 were preserved for 2 years and then dissolved in distilled water for injection. The resulting solution had a transparent light brown color and no precipitate settled.
Stability test 2 Samples of injectable solution obtained in Comparative Example 2 were stored for 1 month, 2 months and 3 months, respectively. A precipitate settled after I month of storage and after 3 months the solutions had turbid color, whereby the samples could not be used for injection.
As it is apparent from the above experiments, the antitumor composition to Korean Patent No. 72,982 is sensitive to moisture, and even if stored in a refrigerator, it is easily deteriorated and its efficacy is severely reduced so that it cannot be preserved for a long period of time and thereafter be used as a medicine.
In contrast, the pharmaceutical composition according to the present invention exhibits no change in quality and efficacy after long preservation, so that it may be used safely.
Claims (11)
1. A pharmaceutical composition having antitumor activity, comprising a lyophilized powder derived from an extract of Pulsatillac Radix or Ulmaceae Cortex, or a mixture thereof, together with at least one pharmaceutically acceptable carrier therefor, the extract having been obtained by extracting powdered Pulsatillae Radix or powdered Ulmaceae Cortex, or a mixture thereof, in a solvent at a temperature below 60°C.
2. A pharmaceutical composition according to claim 1, wherein the lyophilized powder is derived from an extract of Pulsatillae Radix or Ulmaceae Cortex, or a mixture thereof, and Ginseng Radix or Glycyrrhizae Radix, or a mixture thereof, the extract having been obtained by extracting a mixture containing more than 30 weight % of powdered Pulsatillae Radix or powdered Ulmaceae Cortex or a mixture thereof, and less than 70 weight % of powdered Ginseng Radix or powdered Glycyrrhizae Radix, or a mixture thereof, in a solvent at a temperature below 60°C.
3. A pharmaceutical composition according to claim 1 or 2, wherein the solvent is selected from the group consisting of water, alcohol, acetone, ethyl acetate and mixtures thereof.
4. A pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutically acceptable carrier is selected from the group consisting of diluents, binding agents, disintegrators, preservatives, indolents, isotonic agents and lubricants.
5. A pharmaceutical composition according to any one of claims 1 to 4, wherein the composition is in granule, tablet, capsule, injectable solution or ointment form.
6. A process for preparing a pharmaceutical composition having antitumor activity, comprising the steps of:
a) extracting powdered Pulsatillae Radix or powdered Ulmaceae Cortex, or a mixture thereof, in a solvent at a temperature below 60°C to obtain an extract solution;
b) filtering and lyophilizing the extract solution to obtain a lyophilized powder; and c) admixing the lyophilized powder with at least one pharmaceutically acceptable carrier.
a) extracting powdered Pulsatillae Radix or powdered Ulmaceae Cortex, or a mixture thereof, in a solvent at a temperature below 60°C to obtain an extract solution;
b) filtering and lyophilizing the extract solution to obtain a lyophilized powder; and c) admixing the lyophilized powder with at least one pharmaceutically acceptable carrier.
7. A process for preparing a pharmaceutical composition having antitumor activity, comprising the steps of a) extracting powdered Pulsatillae Radix or Ulmaceae Cortex, or a mixture thereof, in a solvent at a temperature below 60°C to obtain an extract solution;
b) admixing the extract solution with at least one pharmaceutically acceptable carrier; and c) filtering and lyophilizing the resulting mixture to obtain a lyophilized powder.
b) admixing the extract solution with at least one pharmaceutically acceptable carrier; and c) filtering and lyophilizing the resulting mixture to obtain a lyophilized powder.
8. A process according to claim 6 or 7, wherein in step (a) a mixture containing more than 30 weight % of powdered Pulsatillae Radix or powdered Ulmaceae Cortex, or a mixture thereof, and less than 70 weight % of powdered Ginseng Radix or powdered Glycyrrhizae Radix, or a mixture thereof, is extracted in a solvent at a temperature below 60°C.
9. A process according to claim 6, 7 or 8, wherein the solvent is selected from the group consisting of water, alcohol, acetone, ethyl acetate and mixtures thereof.
10. A process according to any one of claims 6 to 9, wherein the pharmaceutically acceptable carrier is selected from the group consisting of diluents, binding agents, disintegrators, preservatives, indolents, isotonic agents and lubricants.
11. A process according to any one of claims 6 to 10, wherein after step (c) the composition is formulated into a granule, tablet, capsule, injectable solution or ointment.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR19980047025 | 1998-11-03 | ||
| KR1998/47025 | 1998-11-03 | ||
| KR1998/48277 | 1998-11-11 | ||
| KR1019980048277A KR100312622B1 (en) | 1998-11-03 | 1998-11-11 | Stabilized anticancer composition mainly composed of herbal medicine and its manufacturing method |
| PCT/KR1999/000659 WO2000025802A1 (en) | 1998-11-03 | 1999-11-03 | Pharmaceutical composition having antitumor activity and process for the preparation thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2317113A1 true CA2317113A1 (en) | 2000-05-11 |
Family
ID=26634281
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002317113A Abandoned CA2317113A1 (en) | 1998-11-03 | 1999-11-03 | Pharmaceutical composition having antitumor activity and process for the preparation thereof |
Country Status (11)
| Country | Link |
|---|---|
| EP (1) | EP1044011A1 (en) |
| JP (1) | JP2002528511A (en) |
| KR (1) | KR100312622B1 (en) |
| CN (1) | CN1179729C (en) |
| AU (1) | AU1080400A (en) |
| BR (1) | BR9907168A (en) |
| CA (1) | CA2317113A1 (en) |
| HU (1) | HUP0101256A3 (en) |
| PL (1) | PL341650A1 (en) |
| RU (1) | RU2201759C2 (en) |
| WO (1) | WO2000025802A1 (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002531513A (en) * | 1998-12-07 | 2002-09-24 | エコスマート テクノロジーズ，インコーポレーテッド | Cancer therapeutic formulations and methods using natural plant essential oils with signaling modulators |
| US7378408B2 (en) * | 2001-11-30 | 2008-05-27 | Pfizer Inc. | Methods of treatment and formulations of cephalosporin |
| KR100568607B1 (en) * | 2002-07-22 | 2006-04-07 | 김송배 | Use of hederagenin 3-O-?-L-rhamnopyranosyl1?2-[?-D-glucopyranosyl1?4]-?-L-arabinopyranoside or extracts from Pulsatillae radix containing the same as therapeutic agents for solid tumors |
| KR100515206B1 (en) * | 2002-09-23 | 2005-09-16 | 김종석 | Cosmetic composition having whitening effect comprising extract of Pulsatilla Radix as main ingredient |
| MD2231C2 (en) * | 2003-03-05 | 2004-02-29 | Ион КАЛМЫК | Composition for treatment of oncologic pathologies |
| KR100628334B1 (en) * | 2004-07-30 | 2006-09-27 | 김송배 | A method of improving anticancer effect of Pulsatillae radix and a composition prepared by the method |
| JP4892833B2 (en) * | 2004-12-17 | 2012-03-07 | 大正製薬株式会社 | Fat absorption inhibitor |
| JP2012246311A (en) * | 2012-09-05 | 2012-12-13 | Sk Chemicals Co Ltd | Pulsatilla radix extract having brain function improving effect |
| RU2759382C2 (en) * | 2017-07-07 | 2021-11-12 | Ци Лю | Drug for injection based on saponin b4 pulsatilla |
| CN107468747A (en) * | 2017-10-10 | 2017-12-15 | 刘建明 | A kind of ginseng composition for being advantageous to anticancer, anti-cancer and preparation method thereof |
| CN111466484A (en) * | 2020-04-15 | 2020-07-31 | 四川中地油科技有限公司 | Black soldier fly soft rot prevention and control compound formula and preparation method thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR940000234B1 (en) * | 1989-09-04 | 1994-01-12 | 김송배 | Novel pharmaceutical composition having an antitumor activity and a process for preparation thereof |
| KR100205045B1 (en) * | 1996-12-05 | 1999-06-15 | 김송배 | Novel triterpene glycosidic compound process thereof |
-
1998
- 1998-11-11 KR KR1019980048277A patent/KR100312622B1/en not_active IP Right Cessation
-
1999
- 1999-11-03 CN CNB998019763A patent/CN1179729C/en not_active Expired - Fee Related
- 1999-11-03 WO PCT/KR1999/000659 patent/WO2000025802A1/en not_active Application Discontinuation
- 1999-11-03 EP EP99954463A patent/EP1044011A1/en not_active Withdrawn
- 1999-11-03 JP JP2000579242A patent/JP2002528511A/en active Pending
- 1999-11-03 PL PL99341650A patent/PL341650A1/en not_active Application Discontinuation
- 1999-11-03 CA CA002317113A patent/CA2317113A1/en not_active Abandoned
- 1999-11-03 BR BR9907168-1A patent/BR9907168A/en not_active IP Right Cessation
- 1999-11-03 HU HU0101256A patent/HUP0101256A3/en unknown
- 1999-11-03 AU AU10804/00A patent/AU1080400A/en not_active Abandoned
- 1999-11-03 RU RU2000120474/14A patent/RU2201759C2/en not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| EP1044011A1 (en) | 2000-10-18 |
| CN1179729C (en) | 2004-12-15 |
| KR100312622B1 (en) | 2002-02-28 |
| JP2002528511A (en) | 2002-09-03 |
| HUP0101256A3 (en) | 2006-02-28 |
| WO2000025802A1 (en) | 2000-05-11 |
| AU1080400A (en) | 2000-05-22 |
| KR20000031989A (en) | 2000-06-05 |
| RU2201759C2 (en) | 2003-04-10 |
| CN1287490A (en) | 2001-03-14 |
| BR9907168A (en) | 2000-10-17 |
| PL341650A1 (en) | 2001-04-23 |
| HUP0101256A2 (en) | 2004-05-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1679058B1 (en) | Pharmaceutical composition for the treatment of cardiovascular and cerebrovascular diseases | |
| KR102193363B1 (en) | New salvianolic acid compound t, preparation method therefor, and use thereof | |
| CN100421680C (en) | Extractive of ginkgo tree leaves, prepn. method and application thereof | |
| CA2317113A1 (en) | Pharmaceutical composition having antitumor activity and process for the preparation thereof | |
| AU2017422167B2 (en) | Preparation of Pulsatilla saponin B4 for injection | |
| KR940000234B1 (en) | Novel pharmaceutical composition having an antitumor activity and a process for preparation thereof | |
| JP2009542634A (en) | Pharmaceutical composition for the prevention and treatment of inflammatory diseases comprising extract | |
| GB2476070A (en) | Subcritical water extraction of Hawthorn Crategus monogyna, Pueraria lobata and Centella asiatica | |
| US4460578A (en) | Pharmaceutical composition for topical use based on a total extract of Hedysarum fructescens willd | |
| CN1943569B (en) | Chinese medicine active component composition and its preparing method and use | |
| TWI527587B (en) | Use of boehmeria species extract in manufacturing drug for treating liver fibrosis | |
| CN101974012B (en) | Novel compound ethyl brevicate with pharmaceutical activity | |
| CN109078072A (en) | It is a kind of for preventing the oral preparation of coronary heart disease | |
| MXPA00006377A (en) | Pharmaceutical composition having antitumor activity and process for the preparation thereof | |
| CN101019878B (en) | Injection medicine composite containing salvianolic acid A and its preparation | |
| JPS625126B2 (en) | ||
| KR101026524B1 (en) | A medicinal herbs composition for protective stomach and intestines | |
| CN112472765B (en) | Preparation method of heat-clearing eight-ingredient extract | |
| CN114736182B (en) | Compound for resisting myocardial ischemia reperfusion injury, dai medicine composition and application thereof | |
| KR100213667B1 (en) | The composition showing antidiabetes | |
| CN108743654B (en) | Traditional Chinese medicine composition for treating ischemic heart disease and preparation method and application thereof | |
| KR930004935B1 (en) | Extracts from acanthopanax elata having antacidic effects | |
| CN101979021B (en) | Compound extractive with antioxidation activity | |
| CN101380347A (en) | Ainsliaea fragrans Champ total flavone extract and extraction method and use thereof | |
| CN1961911A (en) | Lyophilized powder injection containing red sage root and dalbergia wood volatile oil and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Discontinued |