U.S. patent application number 10/704457 was filed with the patent office on 2004-12-09 for fagopyrum cymosum (trev.) meisn composition, method to prepare and analyze the same and uses thereof.
Invention is credited to Chen, Bo, Guo, Qiyu, Han, Pei, Zhu, Hongwu.
Application Number | 20040247701 10/704457 |
Document ID | / |
Family ID | 25744369 |
Filed Date | 2004-12-09 |
United States Patent
Application |
20040247701 |
Kind Code |
A1 |
Han, Pei ; et al. |
December 9, 2004 |
Fagopyrum cymosum (Trev.) meisn composition, method to prepare and
analyze the same and uses thereof
Abstract
This invention provides different compositions extracted from
Fagopyrum cymosum (Trev.) Meisn. Said compositions comprise active
components for therapeutic applications. This invention also
provides a method of preparation of the compositions and a method
of identification and determination of individual components of
said compositions. Finally, this invention provides various uses of
the compositions.
Inventors: |
Han, Pei; (Beijing, CN)
; Guo, Qiyu; (Beijing, CN) ; Chen, Bo;
(Beijing, CN) ; Zhu, Hongwu; (Beijing,
CN) |
Correspondence
Address: |
Albert Wai-Kit Chan
Law Offices of Albert Wai-Kit Chan, LLC
World Plaze, Suite 604, 141-07 20th Avenue
Whitestone
NY
11357
US
|
Family ID: |
25744369 |
Appl. No.: |
10/704457 |
Filed: |
August 9, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10704457 |
Aug 9, 2004 |
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09785953 |
Feb 16, 2001 |
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6726938 |
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09785953 |
Feb 16, 2001 |
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09161251 |
Sep 26, 1998 |
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6451353 |
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Current U.S.
Class: |
424/725 |
Current CPC
Class: |
A61P 35/00 20180101;
A61K 31/70 20130101; G01N 33/50 20130101; G01N 2333/415 20130101;
A61K 31/70 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/725 |
International
Class: |
A61K 035/78 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 29, 1997 |
CN |
97 1 16956.X |
Claims
What is claimed is:
1. A method for identifying by chromatography a composition of
Fagopyrum cymosum (Trev.) Meisn capable of treating cancer
comprising the steps of: a) preparing the assay comprising the
steps of: I. ultrasonically dissolving 0.5 g of Fagopyrum cymosum
(Trev.) Meisn composition with 30 ml. of distilled water for 30
minutes to form a solution; II. removing the solution to the
separator funnel; extracting the solution from the water with
chloroform (30 ml. each time for 3 times) in order to degrease;
III. continuously extracting the water layer from said solution
with acetic ether (30 ml. each time for 5 times) to form a
fraction; IV. collecting and blending the acetic ether fraction to
form a blend; V. drying the blend with anhydrous calcium chloride;
VI. filtering the blend to form a filtrate; VII. drying the
filtrate; VIII. mixing said filtrate with water in constant volume
of 10 ml. to form a suspension; IX. injecting 1 ml. of the
suspension into C.sub.18ods cartridge and eluting it with 5 ml. of
water, 5 ml. of 50% methanol and 5 ml. of methanol respectively; X.
making the eluent of 50% methanol the constant volume of 5 ml.; and
XI. filtering the eluent with 0.45 .mu. filtration membrane; b).
using (-) epicatechin as the standard; and c). performing HPLC
assay under following conditions: I. Gradient mobile phase:
62 Time(minutes) Water(PH = 3.00) Acetonitrile 0 90 10 40 74 26 50
20 80
II. column temperature: room temperature; flow rate: 0.8 ml/min;
III. injection volume: 10 .mu.l; IV. wavelength: 282 nm; and V.
calculating according to the following formula: (-)
epicatechin=As.times.Cst/Ast.time- s.Ws.times.100% wherein As=peak
area of sample, Cst=concentration of standard, Ast=peak area of
standard, Ws=weight of sample.
2. A method for identifying a composition of Fagopyrum cymosum
(Trev.) Meisn capable of treating cancer comprising the steps of:
a) preparing the assay comprising the steps of: I. dissolving a
suitable amount of Fagopyrum cymosum (Trev.) Meisn composition in
distilled water to form a solution; II. extracting said solution
from the water with chloroform to degrease; III. continuously
extracting the water layer from said degreased solution with acetic
ether; IV. collecting said acetic ether fractions to form a blend;
V. drying said blend with anhydrous calcium chloride; VI. filtering
said dried blend to form a filtrate; VII. drying said filtrate;
VIII. mixing said filtrate with water in constant volume of 10 ml.
to form a suspension; IX. injecting 1 ml. of said suspension into a
C.sub.18ods cartridge and eluting it with 5 ml. of water, 5 ml. of
50% methanol and 5 ml. of methanol respectively; X. making the
eluent of 50% methanol the constant volume of 5 ml.; and XI.
filtering the eluent with a 0.45 .mu. filtration membrane; b).
using (-) epicatechin as the standard; c). performing HPLC assay
under the following conditions: I. Gradient mobile phase:
63 Time(minutes) Water(PH = 3.00) Acetonitrile 0 90 10 40 74 26 50
20 80
II. column temperature: room temperature; flow rate: 0.8 ml/min;
III. injection volume: 10 .mu.l; IV. wavelength: 282 nm; and d).
calculating according to the following formula: (-)
epicatechin=As.times.Cst/Ast.time- s.Ws.times.100% wherein As=peak
area of sample, Cst=concentration of standard, Ast=peak area of
standard, Ws=weight of sample.
3. A composition comprising the product that when subjected to
method of claim 1 produces 6 peaks as shown in FIG. 1.
4. A composition comprising the product that when subjected to the
method of claim 1 produces a fingerprint as tabulated herein:
64 Relative Area Peak Retention Retention Apearance Area Ratio #
Time Time Probability Area Ratio Range 6 14.57 0.76 80-100 705156
0.69 0.07-3.45 8 16.81 0.88 80-100 1833159 1.80 0.18-9.0 9 19.07
1.0 100 1018514 1.00 10 21.45 1.12 80-100 208454 0.20 0.02-3.0 14
29.70 1.56 80-100 51306 0.05 0.01-2.0 15 30.91 1.62 80-100 145424
0.14 0.03-3.0
5. A composition as in claim 3, wherein: the occurrence frequency
of the 6 peaks are 80-100%; the retention time ratios of the 6
peaks in comparison with (-) epicatechin are 0.76, 0.88, 1.0, 1.12,
1.56, and 1.62 respectively; the area ratios of the 6 peaks in
comparison with (-epicatechin are 0.69, 1.80, 1.00, 0.20, 0.05 and
0.14 respectively; and the ranges of the area ratios of the 6 peaks
in comparison with (-) epicatechin are 0.07-3.45, 0.18-9.0, 1.0,
0.02-3.0, 0.01-2.0 and 0.03-3.0 respectively.
6. A composition comprising: a). 30-70% tannin content; and b).
0.2-1.0% (-) epicatechin.
7. The composition of claim 6 extracted from Fagopyrum cymosum
(Trev.) Meisn rhizome.
8. The composition of claim 7 wherein the Fagopyrum cymosum (Trev.)
Meisn rhizome is obtained from cultivated plants.
9. A method for obtaining a Fagopyrum cymosum (Trev.) Meisn
composition comprising steps of: a). obtaining, washing and cutting
rhizome of fresh Fagopyrum cymosum (Trev.) Meisn; b). drying the
rhizome; c). crushing the rhizome into particles; d). putting the
particles through a process of hot reflux in a solution selected
from water, methanol, ethanol, acetone, water-methanol mixture,
water-ethanol mixture, water-acetone mixture and ethyl acetate
several times to produce extracts; e). mixing the extracts; f).
separating the extract from precipitate and solid materials; g).
concentrating the separated extract; h). diluting the extract with
water; i). passing the suspension through pretreated macroporous
resin column; j). eluting the chromatographic column; k).
concentrating the eluant at reduced pressure; and l). spray-drying
the extract into amorphous red-brown powder, thereby producing an
extract containing the active constituents of Fagopyrum cymosum
(Trev.) Meisn.
10. A method for obtaining a Fagopyrum cymosum (Trev.) Meisn
composition as in claim 9 further comprising the steps of: I. in
step (b) drying the rhizome to less than 10% moisture; II. In step
(c) crushing the dried rhizome into particles smaller than 5 mm. in
diameter while less than 10% of the particles are smaller than 0.1
mm. in diameter; III. in step (d) putting the particle through a
process of hot reflux in the solution of 10-90% ethanol with a
ratio of 1:10 (w/v) twice (1-3 hours and 1-2 hours respectively) to
produce extracts at the temperature of 50-70.degree. C.; IV. in
step (g) concentrating the separated extract at reduced pressure
under the temperature of 70.degree. C. at a density of 1.3 and a
temperature of 50.degree. C.; V. in step (h) diluting the extract
with 5-10 times the amount of water; VI. in step (i) passing the
suspension through pretreated macroporous resin column (model D101)
and other suitable polystyrene resin, saturated with water
pre-column; VII. in step (j) eluting the chromatographic column
with distilled water and 80-90% ethanol repeatedly until the
effluent does not react positively with ferric chloride reagent;
and VIII. in step (k) concentrating the eluant at reduced is
pressure under 50-70.degree. C. to the density of 1.10-1.13.
11. The method of claim 10, wherein the particle of Fagopyrum
cymosum (Trev.) Meisn rhizome is put through a process of hot
reflux in the solvent of 10-90% ethanol twice (for 1-3 hours and
1-2 hours respectively with the volume of solvent being 3-8 times
and 2-7 times the medicinal material).
12. The method of claim 10, wherein the macroporous resins are
packed in columns.
13. The method of claim 10, wherein the chromatographic material is
selected from porous polymer, silicon gel, aluminum oxide,
polyamide, activated charcoal, cellulose or sephadex in addtion to
macroporous resin (model D101).
14. The method of claim 10, wherein the chromatographic column
eluate is concentrated at reduced pressure under 60.degree. C. to a
relative density of 1.10-1.13.
15. The composition comprising the product produced by the method
of claims 9, 10, 11, 12, 13 or 14.
16. A method for determination of total tannin content in a
Fagopyrum cymosum (Trev.) Meisn composition comprising the steps
of: a). preparing the assay comprising the steps of: I. dissolving
a suitable amount of Fagopyrum cymosum (Trev.) Meisn composition
with 20% ethanol to form a solution; II. separating the solution
from the residue by filtration; III. discarding the first filtrate;
IV. drawing 75 ml. of subsequent filtrate; V. evaporating the
filtrate to the absence of ethanol on a water bath; VI. cooling the
filtrate; VII. transferring the filtrate into a 250 ml. volumetric
flask; and VIII. diluting the filtrate; b). determining the total
water soluble portion by evaporating 25 ml. of the assay to dryness
and drying the precipitate at 105.degree. C. for 3 hours (T.sub.1);
c). determining the water soluble portion not bound with crude
powder comprising the steps of: I. adding 6 g. of crude powder of
Fagopyrum cymosum (Trev.) Meisn composition to 100 ml. of the assay
and shaking for 15 minutes; II. separating the solution from the
residue by filtration; and evaporating 25 ml. of the filtrate to
dryness; and III. drying the precipitate at 105.degree. C. for 3
hours (T.sub.2); and d). determining the water soluble portion of
crude powder comprising the steps of: I. dissolving 6 g. of crude
powder of Fagopyrum cymosum (Trev.) Meisn composition with 100 ml.
of water; II. shaking the solution for 15 minutes; separating the
solution from residue by filtration; III. evaporating 25 ml. of the
filtrate to dryness; and IV. drying the precipitate at 105.degree.
C. for 3 hours (T.sub.0); and The total tannin content in
percentage is calculated according to the following formula: Total
tannin content %=(T.sub.1-T.sub.2+T.sub.0).times.10/W.times.100%
wherein W=quantity of sample (dried substance).
17. A method for determination of the amount of (-) epicatechin in
a Fagopyrum cymosum (Trev.) Meisn composition comprising the steps
of: a). performing adaptability test of the system under following
conditions: I. using octadecyl silicomethane-linked silica gel as
the packing; II. using water (pH 3.0)/acetonitrile for mobile phase
gradient elution; the proportions being 0 minutes: 90/10.fwdarw.40
minutes: 74/26.fwdarw.50 minutes: 20/80.fwdarw.110 minutes: stop;
and making detection at the wavelength 282 nm; and III. the
theoretical number of plate calculated with regard to (-)
epicatechin should be no less than 20,000; b). preparing the
standard comprising steps of: I. dissolving 10 mg. (-) epicatechin
with 20 ml. of mobile phase solution by ultrasonic treatment in a
25 ml. volumetric flask (power not lower than 150 W, frequency not
lower than 25 Hz) for 10 minutes; II. removing the solution after
complete dissolution; cooling the solution to room temperature;
III. diluting the solution with mobile phase solution to the mark;
and IV. shaking the solution to homogeneity, 40 .mu.g/ml (-)
epicatechin is obtained; c). preparing the assay comprising the
steps of: I. dissolving 0.5 g. of the crude powder of Fagopyrum
cymosum (Trev.) Meisn composition with 30 ml. of distilled water by
ultrasonic treatment for 30 minutes; II. transferring the solution
to a separating funnel; extracting and defatting the solution by
chloroform for 3 times; III. extracting the water layer with 30 ml.
of ethyl acetate for 5 times; IV. pooling the ethyl acetate
portions; V. dehydrating the pooled ethyl acetate portions with
anhydrous calcium chloride; VI. separating the residue by
filtration; VII. volatilizing the filtrate to dryness; VIII. making
the precipitate into suspension with water with a constant volume
of 10.0 ml.; IX. passing the assay from step (I) through C.sub.18
ODS cartridge and performing solid phase extraction; X. eluting the
assay with 5 ml. of water, 50% methanol and methanol respectively,
the 50% methanol portion having a constant volume of 5 ml.; and XI.
passing the 50% methanol portion through a filter membrane as the
assay; and d). performing the assay comprising the steps of: I.
pipetting the standard and the assay, 10 .mu.l. each; II.
transferring the pipetted standard and assay into a liquid
chromatograph; and III. performing the calculation and obtaining
the results.
18. The method of claim 16, wherein (-) epicatechin is used as the
standard to determine the amount of (-) epicatechin in a Fagopyrum
cymosum (Trev.) Meisn composition.
19. A pharmaceutical composition comprising an effective amount of
the composition of claims 3, 5, 6, 7 or 8 and a pharmaceutically
acceptable carrier.
20. The formulation of claim 19, wherein the formulation is a pill,
capsule, granule, tablet, suspension, injection, syrup, or
tincture.
21. A method for treating lung cancer in a subject comprising
administering to the subject an effective amount of the
pharmaceutical composition of claim 19.
22. A method for alleviating clinical symptoms in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim
19.
23. A method for alleviating the symptom of cough in a subject
suffering from common cold, bronchitis, pneumonia, pulmonary
tuberculosis, pulmonary abscess, lung cancer, and upper respiratory
track infection comprising administering to the subject an
effective amount of the pharmaceutical composition of claim 19.
24. A method for alleviating the symptom of expectoration in a
subject suffering from lung cancer comprising administering to the
subject an effective amount of the pharmaceutical composition of
claim 19.
25. A method for alleviating the symptom of chest pain in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim
19.
26. A method for alleviating the symptom of bloody sputum in a
subject suffering from lung cancer comprising administering to the
subject an effective amount of the pharmaceutical composition of
claim 19.
27. A method for alleviating the symptom of fever in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim
19.
28. A method for improving blood picture in a subject suffering
from lung cancer comprising administering to the subject an
effective amount of the pharmaceutical composition of claim 19.
29. A method for increasing hemoglobin count in a subject suffering
from lung cancer comprising administering to the subject an
effective amount of the pharmaceutical composition of claim 19.
30. A method for increasing leucocyte count in a subject suffering
from lung cancer comprising administering to the subject an
effective amount of the pharmaceutical composition of claim 19.
31. A method for increasing platelet count in a subject suffering
from lung cancer comprising administering to the subject an
effective amount of the pharmaceutical composition of claim 19.
32. A method for improving clinical signs in a subject suffering
from lung cancer comprising administering to the subject an
effective amount of the pharmaceutical composition of claim 19.
33. A method for increasing body weight in a subject suffering from
lung cancer comprising administering to the subject an effective
amount of the pharmaceutical composition of claim 19.
34. A method for increasing daily food consumption in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim
19.
35. A method for decreasing nausea occurrence in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim
19.
36. A method for decreasing vomiting occurrence in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim
19.
37. A method for decreasing diarrhea occurrence in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim
19.
38. A method for decreasing debility occurrence in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim
19.
39. A method for decreasing blood sedimentation rate in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim
19.
40. A method for decreasing blood urea nitrogen in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim
19.
41. A method for improving Karnofsky performance scores in a
subject suffering from lung cancer comprising administering to the
subject an effective amount of the pharmaceutical composition of
claim 19.
42. A method for treating lung cancer in a subject comprising
administering to the subject an effective amount of the
pharmaceutical composition of claim 19 combined with
chemotherapy.
43. A method for treating lung cancer in a subject comprising
administering to the subject an effective amount of the
pharmaceutical composition of claim 19 combined with
radiotherapy.
44. A method for increasing Complement C3 level in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim 19
combined with radiotherapy.
45. A method for increasing E Rosette level in a subject suffering
from lung cancer comprising administering to the subject an
effective amount of the pharmaceutical composition of claim 19
combined with radiotherapy.
46. A method for increasing lymphocyte transforming factor level in
a subject suffering from lung cancer comprising administering to
the subject an effective amount of the pharmaceutical composition
of claim 19 combined with radiotherapy.
47. A method for increasing IgA level in a subject suffering from
lung cancer comprising administering to the subject an effective
amount of the pharmaceutical composition of claim 19 combined with
radiotherapy.
48. A method for increasing IgG level in a subject suffering from
lung cancer comprising administering to the subject an effective
amount of the pharmaceutical composition of claim 19 combined with
radiotherapy.
49. A method for increasing IgM level in a subject suffering from
lung cancer comprising administering to the subject an effective
amount of the pharmaceutical composition of claim 19 combined with
radiotherapy.
50. A method for increasing macrophage phagocytic rate in a subject
suffering from lung cancer comprising administering to the subject
an effective amount of the pharmaceutical composition of claim 19
combined with radiotherapy.
51. A method for increasing macrophage phagocytic index in a
subject suffering from lung cancer comprising administering to the
subject an effective amount of the pharmaceutical composition of
claim 19 combined with radiotherapy.
52. A method for treating gastric cancer, cervical cancer, sarcoma
and other neoplasms, relieving inflammation, and alleviating toxic
and adverse effects associated with chemotherapy and radiotherapy
in a subject comprising administering to the subject an effective
amount of the pharmaceutical composition of claim 19.
Description
[0001] Throughout this application, various publications are
referenced and full citations for these publications may be found
in the references at the end of the specifications preceding the
claims. The disclosures of these publications are hereby
incorporated by reference into this application in order to more
fully describe the state of the art as known to the skilled therein
as of the date of the invention described and claimed herein.
BACKGROUND OF THE INVENTION
[0002] Fagopyrum cymosum (Trev.) Meisn is a species of buckwheat,
which belongs to the Genus Fagopyrum, family Polygonaceae. There
are about 14 species of buckwheat, either cultivated or wild,
existing around the world. Buckwheat grows mainly in the temperate
area of Asia, Europe and North America. (1)
[0003] China was the earliest country in the world to cultivate
buckwheat. According to archaeological studies, China began to grow
buckwheat about 2,000 years ago. The genetic origins of buckwheat
are believed to occur in Yunnan and Sichuan Provinces of China. (2)
Buckwheat was brought from China to Japan via Korea Peninsula and
then to Europe via Siberia and southern Russia. Germany was the
first country in Europe to grow buckwheat in 1396. Then the crop
was introduced into Belgium, France, Italy and Britain in the 17th
century. Later buckwheat was brought to North America from the
Netherlands.
[0004] At present, there are three best known species of buckwheat:
common buckwheat (Fagopyrum esculentum), tartary buckwheat
(Fagopyrum tataricum) and cymose buckwheat (Fagopyrum cymosum). The
common buckwheat (Fagopyrum esculentum) is widely cultivated in the
Northern Hemisphere. In China, it grows mainly in the northern and
northwestern provinces. The tartary buckwheat (Fagopyrum tataricum)
is cultivated primarily in the Himalayan area and south and
southwest China. The cymose buckwheat (Fagopyrum cymosum) is
basically a wild species, not available for human consumption as a
crop, although some reports indicate that it is also cultivated in
China on a small scale. The cymose buckwheat (Fagopyrum cymosum)
distributes only in south and southwest China.
[0005] Although buckwheat is cultivated mainly as a minor crop
around the world, it has been found recently that certain species
of buckwheat also possess therapeutic properties. Animal tests and
clinical trials in China have indicated that tartary buckwheat
flour is effective in preventing and treating diabetes,
hypertension, cerebral arteriosclerosis and cardiovascular disease.
It also has the function of invigorating the stomach, facilitating
digestion, enhancing the immune system and alleviating
inflammation. (3)
[0006] The most important therapeutic property discovered in
buckwheat is the anti-cancer effect of certain buckwheat species.
For example, Samel, D. et al. reported in 1996 that they had
examined the effect of a purified extract of the flowering herb of
Fagopyrum esculentum on various protein kinases involved in signal
transduction, finding that Fagopyrum esculentum contains red
fluorescent compounds having photosensitizing properties.
Spectrophotometric analysis of the extract indicated structural
similarity to hypericin. Dose- and light-dependent inhibition of
various protein kinases was observed. The purified Fagopyrum
esculentum extract strongly inhibited two receptor-associated
protein tyrosine kinases (EGF-R and Ins-R) and a Ser/Thr kinase
(PK-C) at an ng/ml concentration range. Selectivity was exhibited
as a decreased sensitivity to cytosolic PTKs and protein kinase
CK-2. The protein kinases are important components of the signal
transduction pathway. Aberration of signal transduction is a
hallmark of several proliferative diseases. The researchers
believed that the results of their experiment indicated that
photosensitizing compounds in Fagopyrum esculentum are potential
antiproliferative agents. (4)
[0007] In China, research on anti-cancer effect of buckwheat
species was concentrated on Fagopyrum cymosum (Trev.) Meisn, which
is also termed as Fagopyrum dibotrys (D. Don) Hara.
[0008] Fagopyrum cymosum (Trev.) Meisn has been used as a herbal
medicine in China since ancient times, but mainly in combination
with other medicinal herbs. Modern clinical studies conducted in
China have shown that preparations of Fagopyrum cymosum (Trev.)
Meisn can be used to treat lung abscess, bacillary dysentery and
pyogenic infections. (5) Most importantly it has been found that
preparations of Fagopyrum cymosum (Trev.) Meisn can be applied
effectively for therapeutic purposes to various kinds of
tumors.
[0009] Liu, W F et al. published an article in 1981 on some
pharmacological properties of Fagopyrum cymosum (Trev.) Meisn, the
root of which had been used for the treatment of pulmonary abscess.
Liu pointed out that major active principle of Fagopyrum cymosum
(Trev.) Meisn is flavanol (5, 7, 3', 4'-tetrahydroxyflan-3-01
dipolymers). Neither the extract of Fagopyrum cymosum (Trev.) Meisn
nor flavanol exhibited significant antibacterial action in vitro.
No antibacterial substance was found in the urine of either mice or
human beings nor was it found in the organs of mice given the
extract p. o. Liu indicated that therapeutic effect of Fagopyrum
cymosum (Trev.) Meisn was shown in mice infected I. p. with
staphylococcus aureus only when the extract of Fagopyrum cymosum
(Trev.) Meisn or flavanol was given via the same route. Phagocytic
action of peritoneal macrophages was enhanced, but the number of
macrophages was not increased when the extract of Fagopyrum cymosum
(Trev.) Meisn or flavanol was given I. p. to mice. Antipyretic
action was demonstrated in rabbits. Flavanol was shown to be
anti-inflammatory in mice and rats. Platelet aggregation in rats
induced by ADP and staphylococcus aureus in vitro was inhibited by
flavanol. The same effect on platelet aggregation induced by ADP
and collagen was found when flavanol was given IV to rats. Plavanol
given IV showed also expectorant effect in mice as shown by the
phenol red method, but the bronchial excretion was not increased.
(6)
[0010] Liu, Y L et al. in 1983 reported their research results on
the chemical constituents of Fagopyrum cymosum (Trev.) Meisn. Liu
indicated that they had isolated three components of Fagopyrum
cymosum (Trev.) Meisn. Component A is the main constituent in
Fagopyrum cymosum (Trev.) Meisn and accounts for its pronounced
therapeutic effect on pulmonary abscess. The octamethylether,
octamethylether diacetate and decanacetate derivatives were
prepared from this component. On the basis of spectroscopic
analyses, degradation products and physico-chemical constants,
component A was identified as the dimer of 5, 7, 3',
4'-tetrahydroxyflavan-3-o1 (C4-C8 linked), named dimeric
procyanidin. Components B and C were identified as hecogenin and
.beta.-sitosterol respectively. (7)
[0011] Liu T C published an article in 1983 analyzing the growth
and accumulation of active constituents during different
development stages of Fagopyrum cymosum rhizome. The author
reported that condensed procyanidin, the active constituent of
Fagopyrum cymosum rhizome was observed and studied, including its
existence in different parts of the plant and at different
development stages, and its development in the root, stem and the
above-the-ground part in different growth stages. The paper showed
that the amount of condensed procyanidin is higher in the root and
stem and is highest from mid-October till the withering season. The
content rapidly increases from mid-June and reaches its highest
level in late October. (8)
[0012] Yao R C et al. reported anti-tumor active constituents of
cymose buckwheat in China in 1989. The researchers reported that
they had extracted the active portion (A) from rhizome of cymose
buckwheat using ethanol extraction, macroporous resin (D101) column
chromatography and acetone extraction process. Chemical analysis
revealed that A is a compound of proanthocyanidin, from which, in
comparison with standard substances, (-) epicatechin, 3-galloyl (-)
epicatechin, procyanidin B-2 and B-4, and 3,3'-digalloyl
procyanidin B-2 had been detected. In the meantime, methylated,
acetylated and trimethyl silica etherified derivatives of A had
been prepared. (9)
[0013] Ma Y P et al. published an article in 1989 in China on
prediction of responsiveness of human lung cancer xenograft to
extracts of Fagopyrum cymosum (Trev.) Meisn by SRC assay. The
researchers reported that they had determined responsiveness of
human lung cancer explant to extracts E and CD1 of Fagopyrum
cymosum (Trev.) Meisn by the method of subrenal capsule (SRC)
assay. A total of 12 lung cancers had been tested in which 60
xenografts were implanted. With a mean growth of >5 OMU (1
OMU=0.02 mm) in the control animal an assay was considered
evaluable. This study provided an evaluable assay rate of 83.3%.
Taking <-10 OMU of tumor xenograft size as the standard of
responsiveness extracts E and CD1 offered response rates of 40% and
20% respectively. Squamous cell carcinoma was more sensitive to E
than other types of lung cancer. These findings were similar to the
clinical effects of CD1 phase-I study. The study also confirmed
that extracts E and CD1 had no toxicity or side effects in mice.
(10)
[0014] Liang X Z et al. described using TLC scanning to determine
procyanidin B-2 in Fagopyrum dibotrys's rhizome in an article
published in China in 1990. The authors reported that procyanidin
B-2 in Fagopyrum cymosum was determined by TLC scanning, taking
high efficient silica gel 60F.sup.254 as thin-layer absorbent,
toluene-methylformaic-methanol-formi- c acid (1:2:0.2:0.1) as
developing agent, Rf=0.21, scanned at 280 nm with CAMAG 76510 TLC
scanner. The sample was prepared by leaching for 62 hours in
methanol. The average recovery was 98%. Standard deviation was
0.0092, coefficient of variation is 1.8% (n=10). (11)
[0015] Liang, M D et al. published an article in 1991 on extra
corporeal anti-cancer function of Fagopyrum cymosum rootin. The
authors pointed out that Fagopyrum cymosum rootin (FCR) is a new
anti-cancer medicine screening from Fagopyrum cymosum roots, with
the method of extra corporeal culture of human cancer cell. The
active constituent is large-moleculared condensed tannin D. At the
concentration of 125 .mu.g/ml, its inhibition ratio is 84.5% for
SGC, 78.9% for Hela and 100% for KB, caused by the impairment of
the cancer cell's membrane, RNA, DNA metabolization and cell
nucleus cleavage. (12)
[0016] In the same year, Ma M F et al. reported the results of
mutagenicitic and teratogenicitic tests of Fagopyrum cymosum
(Trev.) Meisn. The results showed that no positive mutation was
induced in four Ames standard bacterial strains (.+-.S9), using
seven different doses of Fagopyrum cymosum (Trev.) Meisn varying
from 1-5,000 .mu.g/plate. Antimutational effect was exhibited
against reversional mutation of TA98 and TA 100 strains induced by
daunorubicin and methyl methane sulphonate. No increase in
micronucleus frequency in bone marrow polychromatophil erythroblast
of NIH mice, no teratogenicity in Chinese hamster oocyte
chromosomes (.+-.S9), no adverse effect on reproductive capacity of
NIH mice and development of mouse fetuses, nor teratogenicity of
appearance, skeleton and internal organs of mouse fetuses were
observed. (13)
[0017] Gao Z. et al. described in 1993 the effects of Fagopyrum
cymosum root extract on proliferation of four human tumor cells in
vitro in comparison with 5-FU. The researchers found that the
proliferation of four human tumor cells were markedly inhibited by
the extract and the inhibition rates were positively proportional
to concentration. At the concentration of 12.5 micrograms/ml, the
inhibition rates were 98.7% for GLC, 82.1% for KB, 65.4% for SGC
and 53.8% for Hela cells. (14)
[0018] Zhang W J et al. discussed phenolic constituents from
Fagopyrum dibotrys in 1994. The researchers reported that six
phenolic constituents had been separated from the alcohol extracted
powder of the dry rhizome of Fagopyrum cymosum (Trev.) Meisn. By
polarimetry, HNMR, .sup.13C-NMR and FAB-MS, they were identified
as: 3,4-dihydroxybenzoic acid, gallic acid, (-) epicatechin, (-)
epicatechin-3-O-gallate, procyanidin B-2 and procyanidin C-2.
(15)
[0019] In the same year Meng F H et al. published an article on
anticancer effect of cymose buckwheat roots on human tumor cells
cultured in vitro. The researchers reported that anticancer effect
of cymose buckwheat roots on various human cancer cells cultured in
vitro had been studied using direct killing method, colony
inhibition test and .sup.3H-TdR incorporation test. The results
showed that the drug at the concentration of 1 g.multidot.L.sup.-1
had a killing rate of over one logasithmic killing against a number
of human cancer cells, and when the concentration was lowered to
0.125 g.multidot.L.sup.-1, its killing rate could still reach
74.3-92.1%, approximating one logasithmic killing. The extract from
the roots of cymose buckwheat had significant anticancer effect,
showing a colony inhibiting rate of 100% against several cancer
cells when its concentration was 0.1 or 0.05 g.multidot.L.sup.-1,
and that of 75.1-89.2% at the concentration of 0.0125
g.multidot.L.sup.-1. .sup.3H-TdR labeling revealed that the drug
could significantly inhibit nucleic acid metabolism in cancer
cells, the inhibitory effect being close to that of positive
control, fluorouracil, at the same concentration. (16)
[0020] In another article published in the same year, Meng F H et
al. reported anticancer effect of Jin E in vitro. Jin E is a
compound of tannin extracted from cymose buckwheat. In the studies
on anticancer effect of Jin E, direct killing method, colony
inhibition test and .sup.3H-TdR radioautography were applied.
Results of the studies showed that the 50% inhibiting
concentrations of Jin E against GLC, Hela, SGC and KB cells were
67.8, 73.1, 79.9 and 83.0 .mu.g/ml respectively, with a definite
concentration and time-response relationship. Jin E at the
concentrations of 100 and 50 .mu.g/ml could completely inhibit
colony formation of several human cancer cells, and Jin E at the
concentration of 25 .mu.g/ml had a colony inhibiting rate as high
as 95% against four kinds of human cancer cells; the best
anticancer activity was observed in Jin E D and alcohol soluble
portion. Jin E at the high concentration (100 .mu.g/ml) had an
inhibiting rate of 87.9% against intracellular nucleic acid
synthesis, the difference being very significant (P<0.01),
compared with blank control group. It is concluded that Jin E has
significant inhibitory effect on growth of multiple human cancer
cells in vitro. (17)
[0021] Peng Yong et al. reported the research and development of
Fagopyrum dibotrys in a paper published in China in 1996. The paper
reviewed new developments in Fagopyrum dibotrys studies in the
areas of the materia medica, the plant, the chemistry, the
pharmacology, the clinic applications and the preparation of the
medicine. The authors also provided suggestions on further
exploitation and utilization of Fagopyrum dibotrys. (18)
SUMMARY OF THE INVENTION
[0022] It is an object of the present invention to provide a
Fagopyrum cymosum (Trev.) Meisn composition.
[0023] It is also an object of the present invention to provide a
method of preparation of Fagopyrum cymosum (Trev.) Meisn
compositions.
[0024] It is another object of the present invention to provide
methods of identification and determination of individual active
constituents of Fagopyrum cymosum (Trev.) Meisn compositions.
[0025] It is an additional object of the present invention to
provide pharmaceuticals containing active constituents of Fagopyrum
cymosum (Trev.) Meisn and apply the pharmaceuticals to treating
lung cancer, gastric cancer, cervical cancer, sarcoma and other
neoplasms, relieving inflammation, and alleviating toxic and
adverse effects associated with chemotherapy and radiotherapy.
[0026] It is an additional object of the present invention to
provide pharmaceuticals containing active constituents of Fagopyrum
cymosum (Trev.) Meisn and apply the pharmaceuticals to treating
coughing, caused by common cold, bronchitis, pneumonia, pulmonary
tuberculosis, pulmonary abscess, lung cancer, and upper respiratory
track infection.
[0027] The present invention relates generally to compositions
extracted from Fagopyrum cymosum (Trev.) Meisn and particularly to
Fagopyrum cymosum (Trev.) Meisn composition containing condensed
tannins and procyanidins, including epicatechin,
3-O-galloyl-epicatechin, procyanidin B-2, B-4 and
3,3'-digalloyl-procyanidin B-2, etc. The invention is also
concerned with methods of preparation of Fagopyrum cymosum (Trev.)
Meisn compositions and methods of identification and determination
of individual effective components of Fagopyrum cymosum (Trev.)
Meisn compositions. In addition, the invention pertains to
pharmaceuticals containing effective components of Fagopyrum
cymosum (Trev.) Meisn and application of the pharmaceuticals.
Accordingly, the present invention will be described in detail with
respect to such fields of endeavor; however, those skilled in the
art will appreciate that such description of the invention is meant
to be exemplary only and should not be viewed as limitative of the
full scope thereof.
[0028] DETAILED DESCRIPTION OF THE INVENTION
[0029] This invention provides a composition comprising 30-70%
tannin content.
[0030] This invention provides a composition comprising 0.2-1.0%
(-) epicatechin.
[0031] This invention provides a composition comprising 30-70%
tannin content and 0.2-1.0% (-) epicatechin.
[0032] This invention provides a composition that when subjected to
High Pressure Liquid Chromatography (HPLC) the 6 peak fingerprint
shown in FIG. 1 and tabulated in Table 4 is produced. In this
fingerprint the occurrence frequency (OF) of the 6 peaks are
80-100%; the retention time (RT) ratios of the 6 peaks in
comparison with (-) epicatechin are 0.76, 0.88, 1.0, 1.12, 1.56,
and 1.62 respectively; the area ratios of the 6 peaks in comparison
with (-) epicatechin are 0.69, 1.80, 1.00, 0.20, 0.05 and 0.14
respectively; and the ranges of the area ratios of the 6 peaks in
comparison with (-) epicatechin are 0.07-3.45, 0.18-9.0, 1.0,
0.02-3.0, 0.01-2.0 and 0.03-3.0 respectively.
[0033] This invention provides the above compositions having
components extracted from Fagopyrum cymosum (Trev.) Meisn
rhizome.
[0034] This invention provides the above compositions having
components extracted from Fagopyrum cymosum (Trev.) Meisn rhizome
that is obtained from cultivated plants.
[0035] This invention provides a method for obtaining a Fagopyrum
cymosum (Trev.) Meisn composition comprising steps of: (1)
obtaining, washing and cutting rhizome of fresh Fagopyrum cymosum
(Trev.) Meisn; (2) drying the rhizome; (3) crushing the rhizome
into particle; (4) putting the particle through a process of hot
reflux in a solution selected from water, methanol, ethanol,
acetone, water-methanol mixture, water-ethanol mixture,
water-acetone mixture and ethyl acetate; (5) mixing the extracts;
(6) separating the extract from precipitate and solid materials;
(7) concentrating the separated extract; (8) diluting the extract
with water; (9) passing the suspension through pretreated macro
porous resin column; (10) eluting the chromatographic column; (11)
concentrating the eluant at reduced pressure; (12) spray-drying the
extract into amorphous red-brown powder, thereby producing an
extract containing the active constituents of Fagopyrum cymosum
(Trev.) Meisn.
[0036] This invention provides a method of extraction wherein the
particle of Fagopyrum cymosum (Trev.) Meisn rhizome is put through
a process of hot reflux. Comparing with other methods of
extraction, such as maceration and percolation, the method of hot
reflux consumes the smallest amount of time (less than 3 hours) and
solvent, and provides highest yield and purity. By comparison, the
method of maceration takes 7-10 days, consumes large quantity of
solvent, and causes oxidation and polymerization of active
constituents due to standing at the solvent state for a long time,
that in turn decreases the therapeutic efficacy of the extract.
[0037] This invention provides a method of extraction wherein the
solvent used in the process of hot reflux is ethanol.
[0038] This invention provides a method of extraction wherein the
solvent used in the process of hot reflux is 10-90% ethanol.
[0039] Since various kinds of solvent can be used to extract the
active constituents of Fagopyrum cymosum (Trev.) Meisn, researchers
of this invention performed the following experiment to identify
the solvent which produces the highest extraction yield.
Considering the characteristics of the constituents of Fagopyrum
cymosum (Trev.) Meisn and the strong polarity of the polyphenol
mixtures, the researchers extracted 1 kg. particle of the rhizome
of Fagopyrum cymosum (Trev.) Meisn respectively with four different
solvents: ethanol, water, acetone and ethyl acetate. The extracts
were dried by evaporation. Then the yields of the dried substances
were calculated. The researchers found that the extract yield of
ethanol was the highest and the others were ranked in the following
order: water>acetone>ethyl acetate (Table 1).
1TABLE 1 Extraction Yield of Different Solvent Solvent Extraction
Yield (%) Ethanol 8.88 Water 3.65 Acetone 1.65 Ethyl acetate
0.83
[0040] In another experiment, researchers of this invention found
that the total tannin content in Fagopyrum cymosum (Trev.) Meisn
extract by ethanol was significantly higher than that in the
extract by acetone, indicating that purity of the former was higher
than that of the latter. In the experiment, total tannin content in
extracts by ethanol and acetone were determined by using a modified
method for determination of tannin content, described in the
appendix to the Pharmacopoeia of the People's Republic of China,
1995 edition. The results are shown in Table 2.
2TABLE 2 Total Tanning Content in Extracts by Different Methods of
Extraction Total Tannin Batch No. Solvents Content 880206 Ethanol
73.47% 880609 Ethanol 73.16% 870513 Ethanol 71.97% 870103 Acetone
67.16% 880305 Acetone 59.93% 880420 Acetone 58.97%
[0041] In yet another experiment, researchers of this invention
found that different solvents yield active constituents of
Fagopyrum cymosum (Trev.) Meisn not only disparate in amount, but
also different in cancer-inhibiting effects. The experiment showed
that tumor-inhibiting rates of water and ethyl acetate extracts
were lower than 30 percent and those of ethanol and acetone
extracts were higher than 30 percent. The experiment also
demonstrated that ethanol extract had higher anti-tumor efficacy
than acetone extract (Table 3).
3TABLE 3 Tumor-inhibiting Rates of Fagopyrum cymosum (Trev.) Meisn
Extracted with Ethanol and Acetone No. of Mice at Tumor-inhibiting
Beginning/End Rate** (%) Dosage* Acetone Ethanol Acetone Ethanol
(mg/kg) Extract Extract Extract Extract 500 10/10 10/9 35.19 37.87
100 10/10 10/9 33.13 56.44 50 10/10 10/10 23.87 41.58 5-FU#10 10/10
31.37 0.5% CMC 14/14 -- Note: *Mode of Administration .times. Times
= p.o. .times. 10; **P < 0.01
[0042] All above experiments indicate that selection of an
appropriate solvent is the key in the extraction of active
constituents from Fagopyrum cymosum (Trev.) Meisn and that ethanol
is the best solvent for this process. Ethanol extraction yields the
highest amount of active constituents from the rhizome of Fagopyrum
cymosum (Trev.) Meisn. It also demonstrates the best anti-cancer
efficacy. In addition, ethanol is cheap, readily available,
recoverable and preferred from the point of view of safety.
[0043] This invention provides a method of extraction wherein the
particle of Fagopyrum cymosum (Trev.) Meisn rhizome is put through
a process of hot reflux in the solvent of 10-90% ethanol with a
ratio of 1:10 (W/V) twice (1-3 hours and 1-2 hours respectively) to
produce extracts at the temperature of 50-70.degree. C.
[0044] This invention provides a method of chromatography wherein
the macroporous resins are packed in columns.
[0045] This invention provides a method of chromatography wherein
the chromatographic material includes, but is not limited to porous
polymer, silicon gel, aluminum oxide, polyamide, activated
charcoal, cellulose or sephadex.
[0046] This invention provides a method of chromatography wherein
the column is eluted with distilled water and 10-90% ethanol. The
column is first eluted with distilled water in order to remove
water soluble impurities, such as saccharides, pigments, organic
acids and inorganic salts still present in the extracts while the
active constituents are absorbed by macroporous resin. Then the
column is eluted with 10-90% ethanol.
[0047] This invention provides a method of chromatography wherein
color developing agent of phenol mixtures is ferric chloride
reagent.
[0048] This invention provides a method of concentration wherein
the chromatographic column eluant is concentrated at reduced
pressure under 60.degree. C. to a relative density of 1.10-1.13
since the tannin content of Fagopyrum cymosum (Trev.) Meisn is
liable to change when heated.
[0049] This invention provides a method for obtaining a Fagopyrum
cymosum (Trev.) Meisn composition comprising steps of: (1)
obtaining, washing and cutting rhizome of fresh Fagopyrum cymosum
(Trev.) Meisn; (2) drying the rhizome to containing less than 10%
moisture; (3) crushing the rhizome into particle of the size
smaller than 5 mm. in diameter while less than 10% of the particle
of the size smaller than 0.1 mm. in diameter; (4) putting the
particle through a process of hot reflux in the solution of 10-90%
ethanol with a ratio of 1:10 (W/V) twice (1-3 hours and 1-2 hours
respectively) to produce extracts at the temperature of
50-70.degree. C.; (5) mixing the extracts; (6) separating the
extract from precipitate and solid materials; (7) concentrating the
separated extract at reduced pressure under the temperature of
70.degree. C. at a density of 1.3 and a temperature of 50.degree.
C.; (8) diluting the extract with 5-10 times the amount of water;
(9) passing the suspension through pretreated macroporous resin
column (model D101 and other suitable polystyrene resin, saturated
with water pre-column); (10) eluting the chromatographic column
with distilled water and 80-90% ethanol repeatedly until the
effluent does not react positively with ferric chloride reagent;
(11) concentrating the eluant at reduced pressure under
50-70.degree. C. to the density of 1.10-1.13; ethanol is completely
recovered and the concentrate is free of alcohol; (12) spray-drying
the extract into amorphous red-brown powder, thereby producing an
extract containing the active constituents of Fagopyrum cymosum
(Trev.) Meisn. The yield of powder totals 4-10% of the medicinal
material. The total tannin content is 30-70% and the content of (-)
epicatechin is 0.2-1.0%.
[0050] This invention provides a method of fingerprint
chromatography for Fagopyrum cymosum (Trev.) Meisn composition
comprising steps of: (1) preparing the assay comprising steps of:
(a) dissolving 0.5 g. of Fagopyrum cymosum (Trev.) Meisn
composition with 30 ml. of distilled water by ultrasonic treatment
for 20 minutes; (b) removing the solution to the separator funnel;
(c) extracting the solution from the water with chloroform (30 ml.
each time for 3 times) in order to degrease; (d) continuously
extracting the water layer with acetic ether (30 ml. each time for
5 times); (e) collecting and blending the acetic ether fractions;
(f) drying the blend with anhydrous calcium chloride; (g) filter
the blend; (h) drying the filtrate; (I) suspending the residue with
water in constant volume of 10 ml.; (j) injecting 1 ml. of the
suspension into C.sub.18ods cartridge and eluting it with 5 ml. of
water, 5 ml. of 50% methanol and 5 ml. of methanol respectively;
(k) making the eluent of 50% methanol the constant volume of 5 ml.
(l) filtering the eluent with 0.45 .mu. filtration membrane. (2)
using (-) epicatechin as the standard; (3) performing HPLC assay
under following conditions: (a) Gradient mobile phase:
4 Time (minutes) Water (PH = 3.00) Acetonitrile 0 90 10 40 74 26 50
20 80
[0051] (b) column temperature: room temperature; (c) flow rate: 0.8
ml/min; (d) injection volume: 10 .mu.l; (e) wavelength: 282 nm; (f)
calculating according to the following formula: (-)
epicatechin=As.times.Cst/Ast.times.Ws.times.100% wherein As=peak
area of sample, Cst=concentration of standard, Ast=peak area of
standard, Ws=weight of sample.
[0052] This invention provides a method for determination of total
tannin content in a Fagopyrum cymosum (Trev.) Meisn composition
comprising steps of: (1) preparing the assay comprising steps of:
(a) dissolving 3 g. of Fagopyrum cymosum (Trev.) Meisn composition
with 20% ethanol in a 100 ml. volumetric flask; (b) diluting the
solution to the mark; (c) separating the solution from the residue
by filtration; (d) discarding the first filtrate; (e) drawing 75
ml. of subsequent filtrate; (f) evaporating the filtrate to the
absence of ethanol on a water bath; (g) cooling the filtrate; (h)
transferring the filtrate into a 250 ml. volumetric flask; (i)
diluting the filtrate to the mark; (2) determining the total water
soluble portion by evaporating 25 ml. of the assay to dryness and
drying the precipitate at 105.degree. C. for 3 hours (T.sub.1); (3)
determining the water soluble portion not bound with crude powder
comprising steps of: (a) adding 6 g. of crude powder of Fagopyrum
cymosum (Trev.) Meisn composition to 100 ml. of the assay and
shaking for 15 minutes; (b) separating the solution from the
residue by filtration; (c) evaporating 25 ml. of the filtrate to
dryness; (d) drying the precipitate at 105.degree. C. for 3 hours
(T.sub.2); (4) determining the water soluble portion of crude
powder comprising steps of: (a) dissolving 6 g. of crude powder of
Fagopyrum cymosum (Trev.) Meisn composition with 100 ml. of water;
(b) shaking the solution for 15 minutes; (c) separating the
solution from residue by filtration; (d) evaporating 25 ml. of the
filtrate to dryness; (e) drying the precipitate at 105.degree. C.
for 3 hours (T.sub.0). The total tannin content in percentage is
calculated according to the following formula: Total tannin content
%=(T.sub.1-T.sub.2+T.sub.0).times.10/W.times.100% wherein
W=quantity of sample (dried substance).
[0053] This invention provides a method for determination of the
amount of (-) epicatechin in a Fagopyrum cymosum (Trev.) Meisn
composition comprising steps of: (1) performing adaptability test
of the system under following conditions: (a) using octadecyl
silicomethane-linked silica gel as the packing; (b) using water (pH
3.0)/acetonitrile for mobile phase gradient elution; the
proportions being 0 minute: 90/10.fwdarw.40 minutes:
74/26.fwdarw.50 minutes: 20/80.fwdarw.110 minutes: stop; (c) making
detection at the wavelength 282 nm; (d) the theoretical number of
plate calculated with regard to (-) epicatechin should be no less
than 20,000; (2) preparing the standard comprising steps of: (a)
dissolving 10 mg. (-) epicatechin with 20 ml. of mobile phase
solution by ultrasonic treatment in a 25 ml. volumetric flask
(power not lower than 150 W, frequency not lower than 25 Hz) for 10
minutes; (b) removing the solution after complete dissolution; (c)
cooling the solution to room temperature; (d) diluting the solution
with mobile phase solution to the mark; (e) shaking the solution to
homogeneity, 40 .mu.g/ml (-) epicatechin is obtained; (3) preparing
the assay comprising steps of: (a) dissolving 0.5 g. of the crude
powder of Fagopyrum cymosum (Trev.) Meisn composition with 30 ml.
of distilled water by ultrasonic treatment for 30 minutes; (b)
transferring the solution to a separating funnel; (c) extracting
and defatting the solution by chloroform for 3 times; (d)
extracting the water layer with 30 ml. of ethyl acetate for 5
times; (e) pooling the ethyl acetate portions; (f) dehydrating the
pooled ethyl acetate portions with anhydrous calcium chloride; (g)
separating the residue by filtration; (h) volatilizing the filtrate
to dryness; (i) making the precipitate into suspension with water
with a constant volume of 10.0 ml.; (j) passing the assay from step
(I) through C.sub.18 ODS cartridge and performing solid phase
extraction; (k) eluting the assay with 5 ml. of water, 50% methanol
and methanol respectively, the 50% methanol portion having a
constant volume of 5 ml.; (l) passing the 50% methanol portion
through a filter membrane as the assay; (4) performing the assay
comprising steps of: (a) pipetting the standard and the assay, 10
.mu.l. each; (b) transferring the pipetted standard and assay into
a liquid chromatograph; (c) performing the calculation and
obtaining the results.
[0054] This invention provides a method wherein (-) epicatechin is
used as the standard to identify (-) epicatechin in a Fagopyrum
cymosum (Trev.) Meisn composition.
[0055] This invention provides a method wherein (-) epicatechin is
used as the standard to determine the amount of (-) epicatechin in
a Fagopyrum cymosum (Trev.) Meisn composition.
[0056] This invention provides a formulation containing the above
compositions.
[0057] This invention provides the above formulation that can take
the form of pill, capsule, granule, tablet, suspension, injection,
syrup, tincture, or adhesive plaster.
[0058] This invention provides a pharmaceutical composition
prepared according to the above methods, which comprises an
effective amount of the above compositions and a pharmaceutically
acceptable carrier.
[0059] For the purposes of this invention, "pharmaceutically
acceptable carriers" means any of the standard pharmaceutical
carriers. Examples of suitable carriers are well known in the art
and may include, but not limited to, any of the standard
pharmaceutical carriers such as a phosphate buffered saline
solutions, phosphate buffered saline containing Polysorb 80, water,
emulsions such as oil/water emulsion and various wetting agents.
Other carriers may include additives used in tablets, coated
tablets, granules and capsules, etc.
[0060] Typically such carriers contain excipients such as starch,
milk, sugar, certain types of clay, gelatin, stearic acid or salts
thereof, magnesium or calcium stearate, talc, vegetable fats or
oils, gums, glycols or other known excipients. Such carriers may
also include flavor and color additives or other ingredients.
Compositions comprising such carriers are formulated by well known
conventional methods.
[0061] This invention provides a pharmaceutical composition
prepared according to the above methods, which comprises 1-99% of
Fagopyrum cymosum (Trev.) Meisn compositions and 99-1%
pharmaceutically acceptable carrier.
[0062] This invention provides a pharmaceutical composition
prepared according to the above methods, which comprises 5-80% of
Fagopyrum cymosum (Trev.) Meisn compositions and 95-20%
pharmaceutically acceptable carrier.
[0063] This invention provides a pharmaceutical composition
prepared according to the above methods, which comprises 10-75% of
Fagopyrum cymosum (Trev.) Meisn compositions and 90-25%
pharmaceutically acceptable carrier.
[0064] This invention provides a pharmaceutical composition
prepared according to the above methods, which comprises 20-70% of
Fagopyrum cymosum (Trev.) Meisn compositions and 80-30%
pharmaceutically acceptable carrier.
[0065] This invention provides a pharmaceutical composition
prepared according to the above methods, which comprises 65% of
Fagopyrum cymosum (Trev.) Meisn compositions and 35%
pharmaceutically acceptable carrier as the optimum ratio.
[0066] This invention provides a method for treating lung cancer in
a subject by administering to the subject an effective amount of
the above pharmaceutical compositions.
[0067] This invention provides a method for alleviating clinical
symptoms in a subject suffering from lung cancer by administering
to the subject an effective amount of the above pharmaceutical
compositions.
[0068] This invention provides a method for alleviating the symptom
of cough in a subject suffering from lung cancer by administering
to the subject an effective amount of the above pharmaceutical
compositions.
[0069] This invention provides a method for alleviating the symptom
of expectoration in a subject suffering from lung cancer by
administering to the subject an effective amount of the above
pharmaceutical compositions.
[0070] This invention provides a method for alleviating the symptom
of chest pain in a subject suffering from lung cancer by
administering to the subject an effective amount of the above
pharmaceutical compositions.
[0071] This invention provides a method for alleviating the symptom
of bloody sputum in a subject suffering from lung cancer by
administering to the subject an effective amount of the above
pharmaceutical compositions.
[0072] This invention provides a method for alleviating the symptom
of fever in a subject suffering from lung cancer by administering
to the subject an effective amount of the above pharmaceutical
compositions.
[0073] This invention provides a method for improving blood picture
in a subject suffering from lung cancer by administering to the
subject an effective amount of the above pharmaceutical
compositions.
[0074] This invention provides a method for increasing hemoglobin
count in a subject suffering from lung cancer by administering to
the subject an effective amount of the above pharmaceutical
compositions.
[0075] This invention provides a method for increasing leucocyte
count in a subject suffering from lung cancer by administering to
the subject an effective amount of the above pharmaceutical
compositions.
[0076] This invention provides a method for increasing platelet
count in a subject suffering from lung cancer by administering to
the subject an effective amount of the above pharmaceutical
compositions.
[0077] This invention provides a method for improving clinical
signs in a subject suffering from lung cancer by administering to
the subject an effective amount of the above pharmaceutical
compositions.
[0078] This invention provides a method for increasing body weight
in a subject suffering from lung cancer by administering to the
subject an effective amount of the above pharmaceutical
compositions.
[0079] This invention provides a method for increasing daily food
consumption in a subject suffering from lung cancer by
administering to the subject an effective amount of the above
pharmaceutical compositions.
[0080] This invention provides a method for decreasing nausea
occurrence in a subject suffering from lung cancer by administering
to the subject an effective amount of the above pharmaceutical
compositions.
[0081] This invention provides a method for decreasing vomiting
occurrence in a subject suffering from lung cancer by administering
to the subject an effective amount of the above pharmaceutical
compositions.
[0082] This invention provides a method for decreasing diarrhea
occurrence in a subject suffering from lung cancer by administering
to the subject an effective amount of the above pharmaceutical
compositions.
[0083] This invention provides a method for decreasing debility
occurrence in a subject suffering from lung cancer by administering
to the subject an effective amount of the above pharmaceutical
compositions.
[0084] This invention provides a method for decreasing blood
sedimentation rate in a subject suffering from lung cancer by
administering to the subject an effective amount of the above
pharmaceutical compositions.
[0085] This invention provides a method for decreasing blood urea
nitrogen in a subject suffering from lung cancer by administering
to the subject an effective amount of the above pharmaceutical
compositions.
[0086] This invention provides a method for improving Karnofsky
performance scores in a subject suffering from lung cancer by
administering to the subject an effective amount of the above
pharmaceutical compositions.
[0087] This invention provides a method for treating lung cancer in
a subject by administering to the subject an effective amount of
the above pharmaceutical compositions combined with
chemotherapy.
[0088] This invention provides a method for treating lung cancer in
a subject by administering to the subject an effective amount of
the above pharmaceutical compositions combined with
radiotherapy.
[0089] This invention provides a method for increasing Complement
C3 level in a subject suffering from lung cancer by administering
to the subject an effective amount of the above pharmaceutical
compositions and combined with radiotherapy.
[0090] This invention provides a method for increasing E Rosette
level in a subject suffering from lung cancer by administering to
the subject an effective amount of the above pharmaceutical
compositions combined with radiotherapy.
[0091] This invention provides a method for increasing lymphocyte
transforming factor level in a subject suffering from lung cancer
by administering to the subject an effective amount of the above
pharmaceutical compositions combined with radiotherapy.
[0092] This invention provides a method for increasing IgA level in
a subject suffering from lung cancer by administering to the
subject an effective amount of the above pharmaceutical
compositions combined with radiotherapy.
[0093] This invention provides a method for increasing IgG level in
a subject suffering from lung cancer by administering to the
subject an effective amount of the above pharmaceutical
compositions combined with radiotherapy.
[0094] This invention provides a method for increasing IgM level in
a subject suffering from lung cancer by administering to the
subject an effective amount of the above pharmaceutical
compositions combined with radiotherapy.
[0095] This invention provides a method for increasing macrophage
phagocytic rate in a subject suffering from lung cancer by
administering to the subject an effective amount of the above
pharmaceutical compositions combined with radiotherapy.
[0096] This invention provides a method for increasing macrophage
phagocytic index in a subject suffering from lung cancer by
administering to the subject an effective amount of the above
pharmaceutical compositions combined with radiotherapy.
[0097] This invention provides a method for treating gastric
cancer, cervical cancer, sarcoma and other neoplasms, relieving
inflammation, and alleviating toxic and adverse effects associated
with chemotherapy and radiotherapy in a subject by administering to
the subject an effective amount of the above pharmaceutical
compositions.
[0098] This invention provides a method for treating coughing,
caused by common cold, bronchitis, pneumonia, pulmonary
tuberculosis, pulmonary abscess, lung cancer, and upper respiratory
track infection.
[0099] The present invention is further explained by way of the
following examples which are to be construed as merely illustrative
and not limitative of the remainder of the disclosure in any way
whatsoever.
EXPERIMENTAL DETAILS
EXAMPLE 1
Preparation of Fagopyrum cymosum (Trev.) Meisn Pharmaceutical
[0100] 1. Preparation of Raw Materials
[0101] 370 kg. of Fagopyrum cymosum (Trev.) Meisn rhizome is
obtained, washed, dried and crushed into 333 kg. of particle of the
size less than 5 mm. in diameter, 20 kg. of that is of the size
less than 0.1 mm in diameter.
[0102] 2. Extraction
[0103] The above mentioned 333 kg. of particle is put into a
stainless steel extractor. Then 1,000 kg. of 70% ethanol is added.
Steam is introduced into the jacket of the extractor for heating to
the constant temperature of 70.degree. C. Stirring from the top is
performed, the solid and liquid phases are fully mixed and the
extraction is performed for 1.5 hours. The extract is drained out
and filtered to remove fine particle impurities of the size larger
than 0.05 mm. in diameter. The extract is kept in a stainless steel
container.
[0104] Then 750 kg. of 70% ethanol is added, heated to the constant
temperature of 70.degree. C. and stirred. The extraction process is
performed for 1 hour. The extract is drained out. The solid
material is compressed, filtered, pooled and kept in a container.
1,600 kg. of extract is obtained.
[0105] 3. Evaporation
[0106] The extract is continuously introduced into an evaporator,
in which a vacuum degree of 500-600 mm Hg is maintained. The
solvent is recovered. 170 kg. of condensed extract is obtained.
[0107] 4. Separation through Macroporous Resin
[0108] The condensed extract is put into a suspension apparatus.
900 kg. of water is added. The suspension is maintained for 2
hours, filtered to remove fine particles of the size greater than
0.05 mm. in diameter, passed through AB-8 macroporous resin C for
absorption and repeatedly washed with water. It is eluted with 700
kg. of 80% ethanol. 650 kg. of eluant is obtained. The solvent is
recovered by vacuum evaporation. 130 g. of condensed eluant is
obtained.
[0109] 5. Drying
[0110] The condensed eluant is spray-dried. The temperature of
influent air is controlled at 130-140.degree. C., and that of
effluent air at 65-70.degree. C. The spray disc revolves at a speed
of 8,000-10,000 rev/min. The spray keeps a speed of 300 kg/h. 17
kg. of red-brown amorphous powder is obtained. The powder comprises
63% of total tannin and 0.4% of (-) epicatechin.
[0111] 6. Preparation of Fagopyrum cymosum (Trev.) Meisn
Pharmaceutical
[0112] 17 kg. of starch is fully mixed with the above mentioned red
brown amorphous powder, then compressed into granules, with which
#1 capsules are filled.
EXAMPLE 2
Preparation of Fagopyrum cymosum (Trev.) Meisn Pharmaceutical
[0113] 1. Preparation of Raw Materials
[0114] 280 kg. of Fagopyrum cymosum (Trev.) Meisn rhizome is
obtained, washed, dried and crushed into 250 kg. of particle of the
size less than 5 mm. in diameter, 18 kg. of that is of the size
less than 0.1 mm in diameter.
[0115] 2. Extraction
[0116] The above mentioned 250 kg. of particle is put into a 2
m.sup.3 stainless steel extractor. Then 1000 kg. of 80% ethanol is
added. Steam is introduced into the jacket of the extractor for
heating to the constant temperature of 70.degree. C. Stirring from
the top is performed, the solid and liquid phases are fully mixed
and the extraction is performed for 2 hours. The extract is drained
out and filtered to remove fine particle impurities of the size
larger than 0.05 mm. in diameter. The extract is kept in a
stainless steel container.
[0117] Then 750 kg. of 80% ethanol is added, heated to the constant
temperature of 70.degree. C. and stirred. The extraction process is
performed for 1.5 hour. The extract is drained out. The solid
material is compressed, filtered, pooled and kept in a container.
1,550 kg. of extract is obtained.
[0118] 3. Evaporation
[0119] The extract is continuously introduced into an evaporator,
in which a vacuum degree of 500-600 mm Hg is maintained. The
solvent is recovered. 170 kg. of condensed extract is obtained.
[0120] 4. Separation through Macroporous Resin
[0121] The condensed extract is put into a suspension apparatus.
900 kg. of water is added. The suspension is maintained for 2
hours, filtered to remove fine particles of the size greater than
0.05 mm. in diameter, passed through AB-8 macroporous resin C for
absorption and repeatedly washed with water. It is eluted with 700
kg. of 80% ethanol. 670 kg. of eluant is obtained. The solvent is
recovered by vacuum evaporation. 130 kg. of condensed eluant is
obtained.
[0122] 5. Drying
[0123] The condensed eluant is spray-dried. The temperature of
influent air is controlled at 130.degree. C., and that of effluent
air at 65.degree. C. The spray disc revolves at a speed of 8,000
rev/min. The spray keeps a speed of 300 kg/h. 13.5 kg. of red-brown
amorphous powder is obtained. The powder comprises 58.6% of total
tannin and 0.98% of (-) epicatechin.
[0124] 6. Preparation of Fagopyrum cymosum (Trev.) Meisn
Pharmaceutical
[0125] 13.5 kg. of starch is fully mixed with the above mentioned
red brown amorphous powder, then compressed into granules, with
which #1 capsules are filled.
EXAMPLE 3
Method of Fingerprint Chromatography for Fagopyrum cymosum (Trev.)
Meian Composition
[0126] This invention provides a method of fingerprint
chromatography for Fagopyrum cymosum (Trev.) Meisn composition
comprising steps of: (1) preparing the assay comprising steps of:
(a) ultrasonically dissolving 0.5 g. of Fagopyrum cymosum (Trev.)
Meisn composition with 30 ml. of distilled water for 20 minutes;
(b) removing the solution to the separator funnel; (c) extracting
the solution from the water with chloroform (30 ml. each time for 3
times) in order to degrease; (d) continuously extracting the water
layer with acetic ether (30 ml. each time for 5 times); (e)
collecting and blending the acetic ether fractions; (f) drying the
blend with anhydrous calcium chloride; (g) filtering the blend; (h)
drying the filtrate; (i) suspending the residue with water in
constant volume of 10 ml.; (j) injecting 1 ml. of the suspension
into C.sub.18ods cartridge and eluting it with 5 ml. of water, 5
ml. of 50% methanol and 5 ml. of methanol respectively; (k) making
the eluent of 50% methanol the constant volume of 5 ml. (2)
filtering the eluent with 0.45 .mu. filtration membrane. (3) using
(-) epicatechin as the standard; (4) performing HPLC assay under
following conditions: (a) Gradient mobile phase:
5 Time (minutes) Water (PH = 3.00) Acetonitrile 0 90 10 40 74 26 50
20 80
[0127] (b) column temperature: room temperature; (c) flow rate: 0.8
ml/min; (d) injection volume: 10 .mu.l; (e) wavelength: 282 nm; (f)
calculating according to the following formula: (-)
epicatechin=As.times.Cst/Ast.times.Ws.times.100% wherein As=peak
area of sample, Cst=concentration of standard, Ast=peak area of
standard, Ws=weight of sample.
[0128] Fingerprint chromatography data and area ratio range are
shown in Table 4.
6TABLE 4 Fingerprint Chromatography Data and Area Ratio Range
Appear- Reten- Relative ance Area Peak tion Retention Proba- Area
Ratio # Time Time bility Area Ratio Range 6 14.57 0.76 80-100
705156 0.69 0.07-3.45 8 16.81 0.88 80-100 1833159 1.80 0.18-9.0 9
19.07 1.00 100 1018514 1.00 10 21.45 1.12 80-100 208454 0.20
0.02-3.0 14 29.70 1.56 80-100 51306 0.05 0.01-2.0 15 30.91 1.62
80-100 145424 0.14 0.03-3.0
[0129] In an embodiment of the above method, (-) epicatechin is
used as the standard to identify (-) epicatechin in a Fagopyrum
cymosum (Trev.) Meisn composition.
EXAMPLE 4
Method for Determination of Total Tannin Content in a Fagopyrum
cymosum (Trev.) Meisn Composition
[0130] This invention provides a method for determination of total
tannin content in a Fagopyrum cymosum (Trev.) Meisn composition
comprising steps of: (l) preparing the assay comprising steps of:
(a) dissolving 3 g. of Fagopyrum cymosum (Trev.) Meisn composition
with 20% ethanol in a 100 ml. volumetric flask; (b) diluting the
solution to the mark; (c) separating the solution from the residue
by filtration; (d) discarding the first filtrate; (e) drawing 75
ml. of subsequent filtrate; (f) evaporating the filtrate to the
absence of ethanol on a water bath; (g) cooling the filtrate; (h)
transferring the filtrate into a 250 ml. volumetric flask; (i)
diluting the filtrate to the mark; (2) determining the total water
soluble portion by evaporating 25 ml. of the assay to dryness and
drying the precipitate at 105.degree. C. for 3 hours (T.sub.1); (3)
determining the water soluble portion not bound with crude powder
comprising steps of: (a) adding 6 g. of crude powder of Fagopyrum
cymosum (Trev.) Meisn composition to 100 ml. of the assay and
shaking for 15 minutes; (b) separating the solution from the
residue by filtration; (c) evaporating 25 ml. of the filtrate to
dryness; (d) drying the precipitate at 105.degree. C. for 3 hours
(T.sub.2); (4) determining the water soluble portion of crude
powder comprising steps of: (a) dissolving 6 g. of crude powder of
Fagopyrum cymosum (Trev.) Meisn composition with 100 ml. of water;
(b) shaking the solution for 15 minutes; (c) separating the
solution from residue by filtration; (d) evaporating 25 ml. of the
filtrate to dryness; (e) drying the precipitate at 105.degree. C.
for 3 hours (T.sub.0). The total tannin content in percentage is
calculated according to the following formula: Total tannin content
%=(T.sub.1-T.sub.2+T.sub.0).times.10/W.times.100% wherein
W=quantity of sample (dried substance).
EXAMPLE 5
Method for Determination of the Amount of (-) Epicatechin in a
Fagopyrum cymosum (Trev.) Meisn Composition
[0131] This invention provides a method for determination of the
amount of (-) epicatechin in a Fagopyrum cymosum (Trev.) Meisn
composition comprising steps of: (1) performing adaptability test
of the system under following conditions: (a) using octadecyl
silicomethane-linked silica gel as the packing; (b) using water (pH
3.0)/acetonitrile for mobile phase gradient elution; (c) the
proportions being 0 minutes: 90/10.fwdarw.40 minutes:
74/26.fwdarw.50 minutes: 20/80.fwdarw.110 minutes: stop; (d) making
detection at the wavelength 282 nm; (e) the theoretical number of
plate calculated with regard to (-) epicatechin should be no less
than 20,000; (2) preparing the standard comprising steps of: (a)
dissolving 10 mg. (-) epicatechin with 20 ml. of mobile phase
solution by ultrasonic treatment in a 25 ml. volumetric flask
(power not lower than 150 W, frequency not lower than 25 Hz) for 10
minutes; (b) removing the solution after complete dissolution; (c)
cooling the solution to room temperature; (d) diluting the solution
with mobile phase solution to the mark; (e) shaking the solution to
homogeneity, 40 .mu.g/ml (-) epicatechin is obtained; (3) preparing
the assay comprising steps of: (a) dissolving 0.5 g. of the crude
powder of Fagopyrum cymosum (Trev.) Meisn composition with 30 ml.
of distilled water by ultrasonic treatment for 30 minutes; (b)
transferring the solution to a separating funnel; (c) extracting
and defatting the solution by chloroform for 3 times; (d)
extracting the water layer with 30 ml. of ethyl acetate for 5
times; (e) pooling the ethyl acetate portions; (f) dehydrating the
pooled ethyl acetate portions with anhydrous calcium chloride; (g)
separating the residue by filtration; (h) volatilizing the filtrate
to dryness; (i) making the precipitate into suspension with water
with a constant volume of 10.0 ml.; (j) passing the assay from step
(i) through C.sub.18 ODS cartridge and performing solid phase
extraction; (k) eluting the assay with 5 ml. of water, 50% methanol
and methanol respectively, the 50% methanol portion having a
constant volume of 5 ml.; (l) passing the 50% methanol portion
through a filter membrane as the assay; (4) performing the assay
comprising steps of: (a) pipetting the standard and the assay, 10
.mu.l. each; (b) transferring the pipetted standard and assay into
a liquid chromatograph; performing the calculation and obtaining
the results.
[0132] In an embodiment of the above method, (-) epicatechin is
used as the standard to determine the amount of (-) epicatechin in
a Fagopyrum cymosum (Trev.) Meisn composition.
The First Series of Experiments: Inhibiting Effect of the Fagopyrum
cymosum (Trev.) Meisn Composition of This Invention on Mouse
Sarcoma 180
Experimental Animals
[0133] Healthy Kunming and ICR mice with a body weight of 18-22 g.
each were obtained from the breeding center of Sichuan Institute of
Antibiotic Industry, Chengdu, Sichuan Province, P. R. China. Both
male and female mice were used in the experiment.
Tumor Line
[0134] Sarcoma 180 was obtained from the Division of Pharmacology,
Sichuan Institute of Antibiotic Industry, Chengdu, Sichuan
Province, P. R. China.
Investigational Drug
[0135] The Fagopyrum cymosum (Trev.) Meisn composition of this
invention was prepared into 0.5% CMC suspensions in concentrations
needed. 5-Fluorouracil (5-FU) was purchased on the market and
diluted to the concentration needed with saline.
Experimental Method
[0136] The experiment was carried out in vivo according to
"Procedures of in vivo Screening of Antineoplastic Drugs"
established in P. R. China in 1978.
Experimental Results
[0137] The inhibiting effect of the Fagopyrum cymosum (Trev.) Meisn
composition of this invention on mouse sarcoma 180 is shown in
Tables 5, 6, 7 and 8.
7TABLE 5 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on ICR Mouse Sarcoma 180 No. of
Change in Average Mice at Mean Value Tumor Tumor Dosage beginning/
of Body Weight Inhibiting (mg/kg/d) end Weight (g) (g) ({overscore
(x)} .+-. SD) Rate (%) P Value 200 10/10 +1.54 0.56 .+-. 0.21 44.55
<0.001 100 10/10 +1.86 0.44 .+-. 0.22 56.44 <0.001 50 10/10
+2.01 0.59 .+-. 0.17 41.58 <0.001 0.5% CMC 15/15 +1.52 1.01 .+-.
0.25 Mode of administration .times. times = p.o. .times. 12
[0138]
8TABLE 6 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on ICR Mouse Sarcoma 180 No. of
Change in Average Mice at Mean Value Tumor Tumor Dosage beginning/
of Body Weight Inhibiting (mg/kg/d) end Weight (g) (g) ({overscore
(x)} .+-. SD) Rate (%) P Value 10 10/9 -0.90 0.60 .+-. 0.26 40.59
<0.01 5 10/10 +1.04 0.66 .+-. 0.84 34.65 <0.05 5-Fu 5 10/10
+2.01 0.49 .+-. 0.25 51.49 <0.001 Mode of administration .times.
times = i.p. .times. 12
[0139]
9TABLE 7 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on Kunming Mouse Sarcoma 180 No. of
Change in Average Mice at Mean Value Tumor Tumor Dosage beginning/
of Body Weight Inhibiting (mg/kg/d) end Weight (g) (g) ({overscore
(x)} .+-. SD) Rate (%) P Value 500 10/9 +1.98 1.96 + 0.29 37.78
<0.001 100 10/9 +1.77 1.71 + 1.07 45.71 <0.01 20 10/8 +3.65
2.11 + 1.15 33.02 <0.05 0.5% CMC 21/20 +4.67 3.15 + 0.73 Mode of
administration .times. times = p.o. .times. 10
[0140]
10TABLE 8 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on Kunming Mouse Sarcoma 180 No. of
Change in Average Dosage Mice at Mean Value Tumor Tumor (mg/
beginning/ of Body Weight Inhibiting kg/d) end Weight (g) (g)
({overscore (x)} .+-. SD) Rate (%) P Value 40 10/8 +0.45 1.63 +
0.60 48.25 <0.001 20 10/9 +2.23 2.14 + 0.55 32.06 <0.001 10
10/8 +3.05 2.15 + 0.67 31.75 <0.01 0.5% CMC 21/20 +4.67 3.15 +
0.73 Mode of administration .times. times = i.p. .times. 12
[0141] Tables 5, 6, 7 and 8 show that the Fagopyrum cymosum (Trev.)
Meisn composition of this invention by either oral or
intraperitoneal administration is very effective in inhibiting the
growth of mouse sarcoma 180.
The Second Series of Experiment: Inhibiting Effect of the Fagopyrum
cymosum (Trev.) Meisn Composition of This Invention on Lewis Lung
Carcinoma
Experimental Animals
[0142] Healthy C57BL/6N mice with a body weight of 18-22 g. each
were obtained from the breeding center of Sichuan Institute of
Antibiotic Industry, Chengdu, Sichuan Province, P. R. China. Both
male and female mice were used in the experiment.
Tumor Line
[0143] Lewis lung carcinoma was obtained from the Division of
Pharmacology, Sichuan Institute of Antibiotic Industry, Chengdu,
Sichuan Province, P. R. China.
Investigational Drug
[0144] The Fagopyrum cymosum (Trev.) Meisn composition of this
invention was prepared into 0.5% CMC suspensions in concentrations
needed. 5-Fluorouracil (5-FU) was purchased on the market and
diluted to the concentration needed with saline.
Experimental Method
[0145] The experiment was carried out in vivo according to
"Procedures of in vivo Screening of Antineoplastic Drugs"
established in P. R. China in 1978.
Experimental Results
[0146] The inhibiting effect of the Fagopyrum cymosum (Trev.) Meisn
composition of this invention on Lewis lung carcinoma is shown in
Tables 9, 10, 11 and 12.
11TABLE 9 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on Lewis Lung Carcinoma No. of Change
in Average Mice at Mean Value Tumor Tumor Dosage beginning/ of Body
Weight Inhibiting (mg/kg/d) end Weight (g) (g) ({overscore (x)}
.+-. SD) Rate (%) P Value 200* 10/10 +0.7 0.69 .+-. 0.37 55.48
<0.05 100* 10/10 +1.3 0.91 .+-. 0.72 41.29 <0.05 50* 10/10
+1.1 0.80 .+-. 0.51 48.39 <0.05 5-Fu 5** 9/9 +0.4 0.74 .+-. 0.49
52.26 <0.05 0.5% CMC* 14/13 +1.0 1.55 .+-. 1.07 *Mode of
administration .times. times = p.o. .times. 10 **Mode of
administration .times. times = i.p. .times. 10
[0147]
12TABLE 10 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on Lewis Lung Carcinoma No. of Change
in Average Mice at Mean Value Tumor Tumor Dosage beginning/ of Body
Weight Inhibiting (mg/kg/d) end Weight (g) (g) ({overscore (x)}
.+-. SD) Rate (%) P Value 100 10/10 -1.94 0.77 .+-. 0.24 41.22
<0.01 50 10/10 -1.74 0.86 .+-. 0.28 34.35 <0.05 0.5% CMC
10/10 -1.31 1.31 .+-. 0.46 Mode of administration .times. times =
p.o. .times. 10
[0148]
13TABLE 11 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on Lewis Lung Carcinoma No. of Change
in Average Mice at Mean Value Tumor Tumor Dosage beginning/ of Body
Weight Inhibiting (mg/kg/d) end Weight (g) (g) ({overscore (x)}
.+-. SD) Rate (%) P Value 500 10/10 -1.77 0.58 .+-. 0.38 25.64
<0.05 100 10/10 -0.52 0.41 .+-. 0.35 47.44 <0.05 20 10/9
-0.15 0.49 .+-. 0.30 37.18 <0.05 0.5% CMC 15/14 -0.85 0.78 .+-.
0.42 Mode of administration .times. times = p.o. .times. 13
[0149]
14TABLE 12 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on Lewis Lung Carcinoma No. of Change
in Average Mice at Mean Value Tumor Tumor Dosage beginning/ of Body
Weight Inhibiting (mg/kg/d) end Weight (g) (g) ({overscore (x)}
.+-. SD) Rate (%) P Value 40 10/7 -1.35 1.33 .+-. 0.43 20.44
<0.05 20 10/9 -1.05 1.09 .+-. 0.43 34.79 <0.05 10 10/9 -0.75
1.57 .+-. 0.47 6.17 <0.05 0.5% CMC 14/14 -0.67 1.67 .+-. 0.83
Mode of administration .times. times = i.p. .times. 10
[0150] Tables 9, 10, 11 and 12 show that the Fagopyrum cymosum
(Trev.) Meisn composition of this invention is very effective in
inhibiting the growth of Lewis lung carcinoma.
The Third Series of Experiment: Inhibiting Effect of the Fagopyrum
cymosum (Trev.) Meisn Composition of This Invention on Mouse
Cervical Carcinoma U14
Experimental Animals
[0151] Healthy Kunming and ICR mice with a body weight of 18-22 g.
each were obtained from the breeding center of Sichuan Institute of
Antibiotic Industry, Chengdu, Sichuan Province, P. R. China. Both
male and female mice were used in the experiment.
Tumor Line
[0152] Uterine carcinoma U14 was obtained from the Division of
Pharmacology, Sichuan Institute of Antibiotic Industry, Chengdu,
Sichuan Province, P. R. China.
Investigational Drug
[0153] The Fagopyrum cymosum (Trev.) Meisn composition of this
invention was prepared into 0.5% CMC suspensions in concentrations
needed. 5-Fluorouracil (5-FU) was purchased on the market and
diluted to the concentration needed with saline.
Experimental Method
[0154] The experiment was carried out in vivo according to
"Procedures of in vivo Screening of Antineoplastic Drugs"
established in P. R. China in 1978.
Experimental Results
[0155] The inhibiting effect of the Fagopyrum cymosum (Trev.) Meisn
composition of this invention on mouse cervical carcinoma U14 is
shown in Tables 13, 14, 15, 16 and 17.
15TABLE 13 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on ICR Mouse Cervical Carcinoma U14
No. of Change in Average Mice at Mean Value Tumor Tumor Dosage
beginning/ of Body Weight Inhibiting (mg/kg/d) end Weight (g) (g)
({overscore (x)} .+-. SD) Rate (%) P Value 200* 10/10 +2.52 1.28
.+-. 0.38 35.03 <0.001 100* 10/10 +2.16 1.14 .+-. 0.40 42.13
<0.001 50* 10/10 +2.38 1.02 .+-. 0.29 48.22 <0.001 5-Fu 5**
10/10 +3.93 1.27 .+-. 0.37 35.53 <0.001 0.5% CMC* 15/15 +1.63
1.97 .+-. 0.45 *Mode of administration .times. times = p.o. .times.
10 **Mode of administration .times. times = i.p. .times. 9
[0156]
16TABLE 14 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on ICR Mouse Cervical Carcinoma U14
No. of Change in Average Mice at Mean Value Tumor Tumor Dosage
beginning/ of Body Weight Inhibiting (mg/kg/d) end Weight (g) (g)
({overscore (x)} .+-. SD) Rate (%) P Value 100 10/10 -0.15 1.15
.+-. 0.21 26.28 <0.01 50 10/9 -0.18 1.01 .+-. 0.27 35.26
<0.001 25 10/10 -0.04 1.09 .+-. 0.21 30.13 <0.001 0.5% CMC
15/15 -0.63 1.56 .+-. 0.40 Mode of administration .times. times =
p.o. .times. 10
[0157]
17TABLE 15 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on Kunming Mouse Cervical Carcinoma
U14 No. of Change in Average Mice at Mean Value Tumor Tumor Dosage
beginning/ of Body Weight Inhibiting (mg/kg/d) end Weight (g) (g)
({overscore (x)} .+-. SD) Rate (%) P Value 500 11/11 +4.26 3.74
.+-. 0.96 32.85 <0.001 100 11/11 +4.56 3.99 .+-. 1.16 28.37
<0.01 0.5% CMC 14/14 +4.86 5.57 .+-. 1.31 Mode of administration
.times. times = p.o. .times. 9
[0158]
18TABLE 16 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on Kunming Mouse Cervical Carcinoma
U14 No. of Change in Average Mice at Mean Value Tumor Tumor Dosage
beginning/ of Body Weight Inhibiting (mg/kg/d) end Weight (g) (g)
({overscore (x)} .+-. SD) Rate (%) P Value 40 10/8 +1.81 3.39 .+-.
0.65 37.34 <0.001 20 10/9 +5.55 3.37 .+-. 0.66 29.05 <0.01 10
10/9 +3.90 4.08 .+-. 1.14 25.14 <0.05 0.5% CMC 10/10 +5.25 5.45
.+-. 1.33 Mode of administration .times. times = i.p. .times.
10
[0159]
19TABLE 17 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on ICR Mouse Cervical Carcinoma U14
No. of Change in Average Mice at Mean Value Tumor Tumor Dosage
beginning/ of Body Weight Inhibiting (mg/kg/d) end Weight (g) (g)
({overscore (x)} .+-. SD) Rate (%) P Value 10 10/8 +0.57 1.56 .+-.
0.37 33.62 <0.001 5 10/9 +0.84 1.69 .+-. 0.40 28.09 <0.01
5-Fu 5 10/10 +1.25 1.55 .+-. 0.23 34.04 <0.001 0.5% CMC 10/13
+0.03 2.35 .+-. 0.40 Mode of administration .times. times = i.p.
.times. 10
[0160] Table 13, 14, 15, 16 and 17 show that the Fagopyrum cymosum
(Trev.) Meisn composition of this invention is very effective in
inhibiting the growth of mouse cervical carcinoma U14.
The Fourth Series of Experiment: Inhibiting Effect of the Fagopyrum
cymosum (Trev.) Meisn Composition of This Invention on Colony
Formation of Human Gastric Carcinoma Cell Line SGC-7901
Tumor Line
[0161] Human gastric cancer cell line SGC-7901 was obtained from
Shanghai Institute of Materia Medica, Academia Sinica, Shanghai, P.
R. China.
Cell Culture Vials
[0162] The round culture vials with a diameter of 40 mm. used in
this experiment were procured from Jiangyin Glassworks, Jiangsu
Province, P. R. China.
Investigational Drug
[0163] The Fagopyrum cymosum (Trev.) Meisn composition of this
invention was weighed by aseptic technique and put into a sterile
agate mortar. A few drops of dimethyl sulfoxide (DMSO) were added
as complex solubilizer. It was finely ground and diluted with
TC-199 culture medium to the concentrations needed.
Experimental Method
[0164] Cells were cultured in vitro according to the normal cell
clone method. Eight experimental groups were established, with 3
vials for each group. The cells were exposed to the Fagopyrum
cymosum (Trev.) Meisn composition of this invention for four time
periods of 0.5, 4 and 8 hours, and 21 days. For the three sets of
cells that were exposed to the drug for 0.5, 4 and 8 hours, the
drug-containing culture medium was discarded. The cells were then
washed with Hank's solution and added with 3 ml. fresh culture
medium. The incubation of the fourth set of cells extended to day
21. The drug-containing culture medium was discarded. The cells
were fixed with methyl alcohol and stained with Wright's and Giemsa
stain. The number of colonies (.gtoreq.50 cells were taken as a
colony) was counted. The colony forming efficiency (CFE) was
calculated according to the equation "CFE=number of colony
formation/total number of inoculated cells." Inhibition of colony
formation (ICF) was calculated according to the equation "ICF
%=(CFE of control group-CFE of exposed group)/CFE of control
group.times.100%."
Experimental Results
[0165] The inhibiting effect of the Fagopyrum cymosum (Trev.) Meisn
composition of this invention on colony formation of Gastric
Carcinoma Cell Line SGC-7901 is shown in Tables 18 and 19.
20TABLE 18 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on Colony Formation of SGC-7901 Cells
(Colony Formation Efficiency) Concentration (.mu.g/ml) 0.5 Hours 4
Hours 8 Hours 21 Days 0 8.5 .times. 10.sup.2 10 .times. 10.sup.2
7.7 .times. 10.sup.2 10 .times. 10.sup.2 0.48 8.2 .times. 10.sup.2
9.7 .times. 10.sup.2 5.9 .times. 10.sup.2 7.4 .times. 10.sup.2 1.92
7.6 .times. 10.sup.2 8.7 .times. 10.sup.2 5.0 .times. 10.sup.2 6.6
.times. 10.sup.2 7.8 6.9 .times. 10.sup.2 8.4 .times. 10.sup.2 4.4
.times. 10.sup.2 5.1 .times. 10.sup.2 31.3 4.9 .times. 10.sup.2 5.4
.times. 10.sup.2 2.5 .times. 10.sup.2 2.8 .times. 10.sup.3 125 3.9
.times. 10.sup.2 4.5 .times. 10.sup.3 1 .times. 10.sup.3 6.7
.times. 10.sup.5 500 2.6 .times. 10.sup.2 2.3 .times. 10.sup.4 4.4
.times. 10.sup.4 2000 9.7 .times. 10.sup.5
[0166]
21TABLE 19 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention on Colony Formation of SGC-7901 Cells
(Inhibition of Colony Formation) Concentration (.mu.g/ml) 0.5 Hours
4 Hours 8 Hours 21 Days 0 0.48 4 3 23 24 1.92 10 13 35 34 7.8 18 16
43 49 31.3 42 46 68 72 125 54 55 99 99.9 500 61 98 99.5 100 2000
99.9 100 100 100
[0167] Table 18 and 19 show that the Fagopyrum cymosum (Trev.)
Meisn composition of this invention is very effective in inhibiting
colony formation of human gastric carcinoma cell line SGC-7901.
The Fifth Series of Experiment: Alleviating Effect of the Fagopyrum
cymosum (Trev.) Meisn Composition of This Invention on
Xylene-induced Mouse Ear Inflammation
Experimental Animals
[0168] 49 healthy male adult Kunming mice were obtained from the
breeding center of Sichuan Institute of Antibiotic Industry,
Chengdu, Sichuan Province, P. R. China.
Investigational Drug
[0169] The Fagopyrum cymosum (Trev.) Meisn composition of this
invention was prepared into 0.5% CMC suspensions in concentrations
needed.
Experimental Method
[0170] Inflammation was induced in both ears of each mouse by local
application of 0.05 ml. of xylene. The mice were then randomized
into 3 groups. Half an hour later, one group received local
application of 0.5% CMC suspension in right ear as the control. Two
other groups were treated by the Fagopyrum cymosum (Trev.) Meisn
composition of this invention in right ear in concentrations of 10
and 50 mg/ml respectively.
[0171] Mice were killed four hours after administration of the
composition. Both left and right ears were excised. Holes 9 mm. in
diameter were punched at the same part of both ears. The sections
were weighed by a precision balance. The difference in weight
between the two ears was used as indication of the intensity of
inflammation.
Experimental Results
[0172] The alleviating effect of the Fagopyrum cymosum (Trev.)
Meisn composition of this invention on xylene-induced mouse ear
inflammation is shown in Table 20.
22TABLE 20 Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention in Alleviating Xylene-induced Mouse
Ear Inflammation Average Weight of No. of Inflamed Parts
Concentration Mice (mg{overscore (x)} .+-. SD) P Value 0.5% CMC 17
6.94 .+-. 3.56 10 mg/ml 16 4.81 .+-. 3.90 <0.05 50 mg/ml 16 3.00
.+-. 2.31 <0.001
[0173] Table 20 shows that the Fagopyrum cymosum (Trev.) Meisn
composition of this invention is very effective in alleviating
xylene-induced mouse ear inflammation.
The Sixth Series of Experiment: Radioprotective Effect of the
Fagopyrum cymosum (Trev.) Meisn Composition of This Invention on
Mice Against .sup.60Co Gamma-ray Whole Body Irradiation
Experimental Animals
[0174] 8-10 weeks old inbred balb/c and Kunming mice with a body
weight of 22.+-.2 g. each were obtained from the breeding center of
Chengdu Institute of Biological Products, the Ministry of Health,
Chengdu, Sichuan Province, P. R. China. Both male and female mice
were used in the experiment.
Investigational Drug
[0175] The Fagopyrum cymosum (Trev.) Meisn composition of this
invention of the batch number of 880208 was used in this
experiment. The composition was finely ground in agate mortar,
dissolved by dimethyl sulfoxide (DMSO) and diluted with saline to a
concentration of 5-10 mg/ml (containing 2.5% DMSO).
Experimental Method
[0176] Mice were divided into 12 groups. The Fagopyrum cymosum
(Trev.) Meisn composition of this invention was administered per os
to mice at a dose of 0.2 ml. once a day. 2.5% DMSO in saline was
administered to the control group. The drug schedule is shown in
Tables 21, 22 and 23.
.sup.60Co Radiation Source
[0177] Cobalt-60 therapy unit.
Irradiation Condition
[0178] Mice were fixed in special irradiation boxes, 5 mice per
box. Then they were irradiated for 14 minutes at a distance of 75
cm. from the .sup.60Co source to the center of the animals in an
irradiation field of 20.times.20 cm. at a dose rate of 0.64-0.59
Gy/min. The whole body total irradiation dosage was 8 Gy.
Observation Parameters
[0179] After 8 GY of whole body irradiation, the number of dead
mice in all groups was counted daily. The number of surviving mice
in all groups was counted 30 days after irradiation. 30 day
survival rates were calculated. The average surviving time of dead
animals in all groups was also calculated.
Experimental Results
[0180] The radioprotective effect of the Fagopyrum cymosum (Trev.)
Meisn composition of this invention on mice against .sup.60Co gamma
ray irradiation is shown by table 21, 22 and 23.
23TABLE 21 Radioprotective Effect of the Fagopyrum cymosum (Trev.)
Meisn Composition of This Invention on mice against Gamma Ray
Irradiation (Balb/c Female Mice, Mode of Administration: p.o.)
Average Dosage 30 Day Increased Surviving No of (mg/ Survival
Survival Time of Protection Mice animal) Schedule Rate (%) Rate (%)
Dead Mice Efficacy P Value 20 0 Once/day; 45 / 17.9 1.00 / 5, 4, 3,
2, (9/20) 20 0.5 1 and 0 65 20.0 10.0 0.98 >0.05 days before
(13/20) 19 1.0 irradiation 89.5 44.5 12.0 1.21 >0.01 (17/19) 21
2.0 66.7 21.7 15.1 1.16 >0.05 (14/21)
[0181]
24TABLE 22 Radioprotective Effect of the Fagopyrum cymosum (Trev.)
Meisn Composition of This Invention on Mice against Gamma Ray
Irradiation (Kunming Male Mice, Mode of Administration: p.o.)
Average Dosage 30 Day Increased Surviving No of (mg/ Survival
Survival Time of Protection Mice Animal) Schedule Rate (%) Rate (%)
Dead Mice Efficacy P Value 20 0 Saline once/day; 5 / 10.4 1.00 / 6,
5, 4, 3, 2 (1/20) and 1 days before irradiation 10 1 Once a day; 7,
20 15 12.6 1.31 >0.05 6, 5, 4, 3, 2 (2/10) and 1 days before
irradiation 10 1 Once a day; 3, 50 45 10.5 1.65 <0.005 2 and 1
days (5/10) before irradiation 20 1 Once a day; 1 10 5 12.6 1.03
>0.05 day before (2/20) irradiation 20 1 Once 5 0 12.1 0.99
>0.05 immediately (1/20) before irradiation
[0182]
25TABLE 23 Radioprotective Effect of the Fagopyrum cymosum (Trev.)
Meisn Composition of This Invention on Mice against Gamma Ray
Irradiation (Kunming Female Mice, Mode of Administration: po)
Average Dosage 30 Day Increased Surviving No of (mg/ Survival
Survival Time of Protection Mice animal) Schedule Rate (%) Rate (%)
Dead Mice Efficacy P Value 20 0 Saline once/ / 13.5 13.5 1.00 /
day; 5, 4, 3, 2 and 1 days before irradiation 20 1 Once a day; 45
45 13 1.49 <0.005 5, 4, 3, (9/20) 2 and 1 days before
irradiation 19 1 Once a day; 45 42 13 1.53 <0.005 3, 2 and 1
(9/20) days before irradiation
[0183] Table 21, 22 and 23 show that the Fagopyrum cymosum (Trev.)
Meisn composition of this invention is very effective in protecting
mouse against .sup.60Co gamma ray irradiation. The tables also show
that gastric instillation of the Fagopyrum cymosum (Trev.) Meisn
composition of this invention at different doses (0.5-2.0 mg/mouse)
once a day 5, 4, 3, 2, 1 and 0 days before radiation exposure can
protect mice against gamma ray irradiation to varying extent. Of
all dose groups, the 1 mg. dose group shows the best effect with an
increased survival rate of 44.5%. With the same dose of 1 mg,
administration once a day, 3, 2 and 1 days before radiation
exposure shows the best radio protective effect.
The Seventh Series of Experiment: Inhibiting Effect of the
Fagopyrum cymosum (Trev.) Meisn Composition of This Invention in
Combination with 5-Fluorouracil and .sup.60Co Gamma Rays on Mouse
Sarcoma 180
Experimental Animals
[0184] 60 Kunming mice were obtained from the breeding center of
Chengdu Institute of Biological Products, the Ministry of Health,
Chengdu, Sichuan Province, P. R. China.
Tumor Line
[0185] Sarcoma 180 was obtained from the Division of Pharmacology,
Sichuan Institute of Antibiotic Industry, Chengdu, Sichuan
Province, P. R. China. The tumor line had been passed on several
times in Kunming mice.
Investigational Drug
[0186] The Fagopyrum cymosum (Trev.) Meisn composition of this
invention was used in the experiment.
5-Fluorouracil
[0187] The drug was prepared by Nantong Pharmaceuticals, Nantong,
Jiangsu Province, P. R. China.
.sup.60Co Radiation Source
[0188] Cobalt-60 therapy unit.
Experimental Method
[0189] Inoculation of tumor line and evaluation of therapeutic
efficacy were performed according to "Procedures of in vivo
Screening of Antineoplastic Drugs" established in P. R. China in
1978.
[0190] Sixty mice were divided into 6 groups, 10 mice for each
group. Group 1 was the control group wherein the mice were treated
only by saline. Groups 2 to 6 were treatment groups using different
therapies. Group 2 was treated with the Fagopyrum cymosum (Trev.)
Meisn composition of this invention. Group 3 was treated with
5-Fluorouracil. Group 4 was treated with .sup.60Co irradiation.
Group 5 was treated with the Fagopyrum cymosum (Trev.) Meisn
composition of this invention combined with 5-Fluorouracil, while
Group 6 was treated with the Fagopyrum cymosum (Trev.) Meisn
composition of this invention combined with .sup.60Co irradiation.
Result of each group was then calculated and compared.
Experimental Results
[0191] The inhibiting effect of the Fagopyrum cymosum (Trev.) Meisn
composition of this invention in combination with 5-fluorouracil
and .sup.60Co gamma ray irradiation on Mouse Sarcoma 180 is shown
by Table 24.
26TABLE 24 Inhibiting Effect of the Fagopyrum cymosum (Trev.) Meisn
Composition of This Invention in Combination with 5-Fluorouracil
and .sup.60Co Gamma Ray Irradiation on Mouse Sarcoma 180 No. of
Mode of Mice at Tumor Tumor Formu- Dosage Administration beginning/
Weight Inhibiting Drug lation (mg/kg) and Schedule end (g) Rate (%)
Saline 2.5% p.o. .times. 10 10/10 1.36 DMSO Fagopyrum Powder 100
p.o. .times. 10 10/10 0.88 35.2 Composition 5-FU Injection 5 .sup.
i.p. .times. 10.sup. 10/10 0.71 47.8 .gamma.-Irradiation 300 rad 3
days after 10/10 0.57 50.7 inoculation Fagopyrum Powder + 100 mg
p.o. .times. 10 10/9 0.62 54.4 Composition + Injection + .sup. i.p.
.times. 10.sup. 5-FU 5 mg Fagopyrum Powder 100 mg p.o. .times. 10
10/9 0.44 67.6 Composition + + 300 rad .times. 1
.gamma.-Irradiation 300 rad
[0192] Table 24 shows that the Fagopyrum cymosum (Trev.) Meisn
composition of this invention when applied in combination with
5-Fluorouracil and .sup.60Co gamma ray irradiation is more
effective than other experimental therapies in inhibiting the
growth of Mouse Sarcoma 180.
The First Series of Clinical Study: Anticancer Effect of the
Pharmaceutical Composition of Fagopyrum cymosum (Trev.) Meisn of
This Invention
Patient Selection
[0193] 136 patients with lung cancer were recruited in the clinical
study. Their age and sex distributions were as follows:
27TABLE 25 Age and Sex Distribution of 136 Lung Cancer Patients
Total Cases Age Sex Cases % Range of Age Mean Value of Age Male 102
75% 34-75 61 Female 34 25% 31-80 60
[0194] The major clinical symptoms of the 136 patients were as
follows:
28TABLE 26 Major Clinical Symptoms of 136 Lung Cancer Patients
Expector- Chest Bloody Total Cough ation Pain Sputum Fever Cases
136 122 118 82 57 37 % 100 89.7 86.8 60.3 41.9 27.2
[0195] The radiographic presentations of the 136 patients were as
follows:
29TABLE 27 Radiographic Presentations of 136 Lung Cancer Patients
Centrally Located Peripheral Total Lesions Lesion Cases 136 118 18
% 100 86.78 13.22
[0196] The 136 patients were staged based on "The Guiding
Principles for Clinical Research of New Drugs (Traditional Chinese
Materia Medica)" established by the Ministry of Health of P. R.
China in 1988. Table 28 shows that the majority of the 136 patients
fell into intermediate and late stages of lung cancer.
30TABLE 28 Clinical Stages of 136 Lung Cancer Patients Total I II
III IV III + IV Cases 136 6 37 61 32 93 % 100 4.41 27.21 44.85
23.53 68.38
[0197] The pathological classifications of the 136 patients were as
follows:
31TABLE 29 Pathological Classifications of 136 Lung Cancer Patients
Small Cell Squamous Cell Adeno- Undiffereniated Large Cell Total
Carcinoma carcinoma Carcinoma Carcinoma Unclassified Cases 136 77
44 12 1 2 % 100% 56.62% 32.35% 8.82% 0.74% 1.47%
[0198] Methods of Diagnosis: All enrolled patients were diagnosed
by detection of lung cancer cells through biopsy or brushing in
fiberopic bronchoscopy, pathological biopsy of lymph nodes, biopsy
by needle aspiration or pathological examination of sputum or
pleural effusion.
Design of the Clinical Study
[0199] All patients were administered orally the pharmaceutical
composition of Fagopyrum cymosum (Trev.) Meisn of this invention in
the formulation of capsule (containing 200 mg. of the active
components), 1.2-1.6 g. each time, 3 times a day. Patients took the
medicine only after meals. The duration of treatment was 2
months.
[0200] No patient received anticancer chemotherapy or radiotherapy
prior to or during the course of the clinical study.
Evaluation of Therapeutic Effectiveness
[0201] Therapeutic effectiveness of the pharmaceutical composition
of Fagopyrum cymosum (Trev.) Meisn of this invention was evaluated
according to the established criteria of four grades: Complete
Remission (CR), Partial Remission (PR), Stableness (S) and
Progression (P).
Therapeutic Results
[0202] The results of clinical study show that the pharmaceutical
composition of Fagopyrum cymosum (Trev.) Meisn of this invention is
effective in treating lung cancer.
[0203] 1. The pharmaceutical composition of Fagopyrum cymosum
(Trev.) Meisn of this invention is effective in treating lung
cancer.
32TABLE 30 Effects of the Pharmaceutical Composition of Fagopyrum
cymosum (Trev.) Meisn of This Invention in Treating Lung Cancer
Patients Total CR PR S P CR + PR Cases 136 1 17 79 39 18 % 100 0.74
12.5 58.08 28.68 13.24
[0204] 2. The pharmaceutical composition of Fagopyrum cymosum
(Trev.) Meisn of this invention is effective in alleviating major
clinical symptoms of lung cancer.
33TABLE 31 Effects of the Pharmaceutical Composition of Fagopyrum
cymosum (Trev.) Meisn of This Invention in Alleviating Major
Clinical Symptoms of Lung Cancer Patients No. of Cases Before After
Change Symptom Severity Treatment Treatment Number % Cough Mild 62
79 +17 +27.4% Moderate 46 18 -28 -60.9% serious 14 7 -7 -50% Total
122 104 -18 -14.8% Expectoration Mild 80 60 -20 -25% Moderate 35 16
-19 -54.3% serious 3 1 -2 -66.7% Total 118 77 -41 -34.7% Chest Pain
Mild 57 46 -11 -19.3% Moderate 19 8 -11 -57.9% serious 6 3 -3 -50%
Total 82 57 -25 -30.5% Bloody Sputum Mild 47 24 -23 -48.9% Moderate
9 4 -5 -55.6% serious 1 0 -1 -100% Total 57 28 -29 -50.9% Fever
Mild 29 13 -16 -55.2% Moderate 8 2 -6 -75% serious 1 2 +1 +100%
Total 38 17 -21 -55.3% P < 0.05 Expectoration: Mild < 50
ml/day, Moderate < 100 ml/day, serious > 100 ml/day
[0205] 3. The pharmaceutical composition of Fagopyrum cymosum
(Trev.) Meisn of this invention is effective in improving blood
picture of lung cancer patients by increasing their hemoglobin,
leucocyte and platelet counts.
34TABLE 32 Effects of the Pharmaceutical Composition of Fagopyrum
cymosum (Trev.) Meisn of This Invention in Improving Blood Picture
of Lung Cancer Patients Blood Total Increased Stable Decreased
Count No. % No. % No. % No. % .chi..sup.2 Hemoglobin 136 100 68 50
25 18.4 43 31.6 6.42 Leucocyte 136 100 14 10.3 120 88.2 2 1.5 8
Platelet 136 100 6 4.4 129 94.9 1 0.7 3.69 Increase or decrease of
hemoglobin count denotes exceeding 0.5% above or below normal
value, while those for leucocyte and platelet counts denote the
values above or below normal range; P < 0.05.
[0206] 4. The pharmaceutical composition of Fagopyrum cymosum
(Trev.) Meisn of this invention is effective in improving clinical
signs of lung cancer patients.
[0207] After treatment by the pharmaceutical composition of
Fagopyrum cymosum (Trev.) Meisn of this invention, the 136 lung
cancer patients generally gained weight and better appetite,
decreased blood sedimentation rate and improved kidney function.
There was no significant change of liver function after the
treatment.
35TABLE 33 Effects of the Pharmaceutical Composition of Fagopyrum
cymosum (Trev.) Meisn of This Invention in Improving Clinical Signs
of Lung Cancer Patients. Clinical Signs Before Treatment After
Treatment Average Weight (kg.) 57.2 57.5 Average Daily Food 588 660
Consumption (g.) Average Blood 24.8 23.5 Sedimentation Rate (mm/h)
Average Blood Urea 4.76 3.97 Nitrogen (mmol/L)
[0208] During the clinical study, the pharmaceutical composition of
Fagopyrum cymosum (Trev.) Meisn of this invention brought out a
combined complete and partial remission rate of 13.24% and a stable
rate of 58.08%. It alleviated to a varying degree various clinical
symptoms of lung cancer, such as cough, expectoration, chest pain,
bloody sputum and fever. The pharmaceutical composition of
Fagopyrum cymosum (Trev.) Meisn of this invention also improved
blood picture of lung cancer patients by increasing their
hemoglobin, leucocyte and platelet counts. In addition, it improved
various clinical signs of lung cancer patients. Of 136 patients,
one achieved complete remission. These results indicate that the
pharmaceutical composition of Fagopyrum cymosum (Trev.) Meisn of
this invention is effective in treating lung cancer.
The Second Series of Clinical Study: Alleviating Effect of the
Pharmaceutical Composition of Fagopyrum cymosum (Trev.) Meisn of
This Invention on the Toxicity and Other Side-effects of Anticancer
Chemotherapeutic Agents
Patient Selection
[0209] 60 patients with lung cancer were recruited in the clinical
study. 35 patients were male and 25 patients were female. They aere
35-76 years old with an average age of 55.5. Their age distribution
is as follows (Table 34):
36TABLE 34 Age Distribution of 60 Lung Cancer Patients Total <40
40-60 61-70 >70 number 60 6 24 22 8 % 100 10 40 36.67 13.33
[0210] All patients were hospitalized with definite diagnosis of
lung cancer confirmed pathologically. Their clinical stages are
shown in Table 35.
37TABLE 35 Clinical Stages of 60 Lung Cancer Patients Total I II
III IV III .+-. IV Cases 60 0 6 28 26 54 % 100 0 10 46.7 43.3
90
[0211] Table 35 shows that patients in stage III and IV accounted
for 90% of the total. The pathological classifications of the
patients are as follows:
38TABLE 36 Pathological Classifications of 60 Lung Cancer Patients
Squamous Cell Adeno- Small Cell Total Carcinoma carcinoma Carcinoma
Cases 60 16 40 4 % 100 26.67 66.67 6.66
[0212] 48 patients (80%) had received radiotherapy or chemotherapy
six or three months before they were enrolled in the clinical
study.
Design of the Clinical Study
[0213] The 60 lung cancer patients were randomly divided into a
treatment group and a control group, 30 patients in each group.
Patients in the treatment group were administered orally of the
pharmaceutical composition of Fagopyrum cymosum (Trev.) Meisn of
this invention at the dose of 1.6 g. (8 capsules), three times a
day. They might take symptomatic drugs at the same time, but no
other anticancer drugs. The control group was treated by
chemotherapy, mainly with cisplatium, VP-16 and CAP program.
Patients with squamous carcinoma were further treated with
daunorubicin and intrapleural therapy mainly by MMC and DDP
programs. Patients with small cell carcinoma were further treated
mainly with CEA program while patients with non-small cell
carcinoma were further treated with CAP program.
[0214] Before the treatment started, patients in both groups took
cardiac, renal, hepatic, bone marrow and immune function
examinations at the same time. Two months after medication, they
were examined again for the same parameters of various
functions.
Evaluation of Therapeutic Effectiveness
[0215] Therapeutic effectiveness of the Pharmaceutical composition
of Fagopyrum cymosum (Trev.) Meisn of this invention was evaluated
according to the established criteria of four grades: Complete
Remission (CR), Partial Remission (PR), Stableness (S) and
Progression (P). Toxic reaction was evaluated according to the
unified criteria for grading acute, subacute and toxic reactions of
anticancer drugs. They are divided into grade 0, I, II, III and
IV.
Therapeutic Results
[0216] 1. The pharmaceutical composition of Fagopyrum cymosum
(Trev.) Meisn of this invention is effective in treating lung
cancer.
[0217] Two months after medication, no complete remission was
observed. The partial remission rate in the control group was
higher than that in the treatment group. The stableness rate in the
treatment group was significantly higher than that in the control
group. The combined rate of PR+S in the treatment group (83.33%)
was higher than that in the control group (63.33%). The number of
patients with progression of tumor in the control group were about
2 times more than that in the treatment group. The data indicate
that the pharmaceutical composition of Fagopyrum cymosum (Trev.)
Meisn of this invention is effective in treating lung cancer.
39TABLE 37 Effects of the Pharmaceutical Composition of Fagropyrum
cymosum (Trev.) Meisn of This Invention in Treating Lung Cancer
Group Total CR(%) PR(%) S(%) P(%) PR + S(%) Treatment 30 0(0)
6(20.00) 19(63.33) 5(16.67) 25(83.33) Control 30 0(0) 8(26.66)
11(36.67) 11(36.67) 19(63.33)
[0218] 2. The pharmaceutical composition of Fagopyrum cymosum
(Trev.) Meisn of this invention achieves its therapeutic effect
with less toxic reaction than chemotherapy.
40TABLE 38 Comparison of Toxic Reactions between the Treatment
Group Receiving the Pharmaceutical Composition of Fagopyrum cymosum
(Trev.) Meisn of This Invention and the Control Group Receiving
Chemotherapy in Terms of Change in Hemoglobin Level Severity of
Reaction Total 0 I II III Cases Before After Before After Before
After Before After Groups and % Treatment Treatment Treatment
Treatment Treatment Treatment Treatment Treatment Treatment 30 22
26 6 4 2 0 0 0 100% 73.3% 86.7% 20% 13.3% 6.7% 0% 0% 0% Control 30
23 17 6 10 1 3 0 0 100% 76.7% 56.7% 20% 33.3% 3.3% 10% 0% 0%
[0219]
41TABLE 39 Comparison of Toxic Reactions between the Treatment
Group Receiving the Pharmaceutical Composition of Fagopyrum cymosum
(Trev.) Meisn of This Invention and the Control Group Receiving
Chemotherapy in Term of Change in Leukocyte Level Severity of
Reaction Total 0 I II III Cases Before After Before After Before
After Before After Groups and % Treatment Treatment Treatment
Treatment Treatment Treatment Treatment Treatment Treatment 30 25
28 4 1 1 1 0 0 100% 83.4% 93.4% 13.3% 3.3% 3.3% 3.3% 0% 0% Control
30 30 20 0 5 0 4 0 1 100% 100% 66.7% 0% 16.7% 0% 13.3% 0% 3.3%
[0220]
42TABLE 40 Comparison of Toxic Reactions between the Treatment
Group Receiving the Pharmaceutical Composition of Fagopyrum cymosum
(Trev.) Meisn of This Invention and the Control Group Receiving
Chemotherapy in Terms of Change in Platelet Level Severity of
Reaction Total 0 I II III Cases Before After Before After Before
After Before After Groups and % Treatment Treatment Treatment
Treatment Treatment Treatment Treatment Treatment Treatment 30 27
29 1 0 1 0 1 1 100% 90% 96.7% 3.4% 0% 3.3% 0% 3.3% 3.3% Control 30
28 23 0 3 1 2 1 2 100% 93.4% 76.6% 0% 10% 3.3% 6.7% 3.3% 6.7%
[0221] Leukocyte and platelet count of patients in both treatment
and control groups is shown by table 41. Leukocyte
count>4000/mm.sup.3 and platelet count>80000/mm.sup.3 are
used as the normal value. Table 41 shows that the pharmaceutical
composition of Fagopyrum cymosum (Trev.) Meisn of this invention
has no inhibiting effect on leukocyte and platelet counts.
43TABLE 41 Comparison of Leukocyte and Platelet Counts of 60 Lung
Cancer Patients after Treatment between the Two Groups Treatment
Group Control Group Parameter (N. = 30) (N. = 30) .chi..sup.2
Leukocyte >4000/mm.sup.3 30 17 16.569 Count <4000/mm.sup.3 0
13 Platelet >80000/mm.sup.3 30 19 13.469 Count
<80000/mm.sup.3 0 11 P < 0.01
[0222] During the clinical study, more patients in the treatment
group gained weight than those in the control group. By contrast,
less patients lost weight in the treatment group than those in the
control group. The difference between the two groups was
statistically significant (P<0.01). The definition for gaining
weight in this study was that the weight of a patient increased by
more than 2 kg. The definition for losing weight was that the
weight of a patient decreased by more than 2 kg. The definition for
stableness was that the weight of a patient increased or decreased
by less than 2 kg.
44TABLE 42 Effect of the Pharmaceutical Composition of Fagopyrum
cymosum (Trev.) Meisn of This Invention on Change of Body Weight of
Lung Cancer Patients Treatment group Control Group Cases % Cases %
.chi..sup.2 Increased 6 20 2 6.7 10.2294 Stable 22 73.3 20 66.7
Decreased 2 6.7 8 26.6 Total 30 100 30 100
[0223] During the clinical study, no patients in the treatment
group had nausea, vomiting and diarrhea, while 9 patients in the
control group had nausea and vomiting (30%) and 3 patients had
diarrhea (10%) occurred in the control group. One patient in the
treatment group developed slight abdominal distention after
treatment with the pharmaceutical composition of Fagopyrum cymosum
(Trev.) Meisn of this invention for one week and the symptom
disappeared in two days.
45TABLE 43 Comparison of Gastrointestinal Reactions between the
Treatment Group Receiving the Pharmaceutical Composition of
Fagopyrum cymosum (Trev.) Meisn of This Invention and the Control
Group Receiving Chemotherapy in Terms of Nausea and Vomiting
Occurrence Severity of Reaction Total 0 I II III Cases Before After
Before After Before After Before After Groups and % Treatment
Treatment Treatment Treatment Treatment Treatment Treatment
Treatment Treatment 30 30 30 0 0 0 0 0 0 100% 100% 100% 0% 0% 0% 0%
0% 0% Control 30 21 21 7 3 2 5 0 1 100% 70% 70% 23.33% 10% 6.67%
16.67% 0% 3.33%
[0224]
46TABLE 44 Comparison of Gastrointestinal Reactions between the
Treatment Group Receiving the Pharmaceutical Composition of
Fagopyrum cymosum (Trev.) Meisn of This Invention and the Control
Group Receiving Chemotherapy in Terms of Diarrhea Occurrence
Severity of Reaction Total 0 I II III Cases Before After Before
After Before After Before After Groups and % Treatment Treatment
Treatment Treatment Treatment Treatment Treatment Treatment
Treatment 30 28 30 1 0 1 0 0 0 100% 93.34% 100% 3.33% 0% 3.33% 0%
0% 0% Control 30 27 27 1 2 2 1 0 0 100% 90% 90% 3.33% 6.67% 6.67%
3.33% 0% 0%
[0225] During the clinical study, patients with debility increased
only 6.7% in the treatment group. By contrast, patients with
debility increased 33.3% in the control group. The difference
between the two groups was statistically significant
(P<0.01).
47TABLE 45 Effect of the Pharmaceutical Composition of Fagopyrum
cymosum (Trev.) Meisn of This Invention on Reducing the Increase of
Debility of Lung Cancer Patients Treatment group Control Group
Cases % Cases % .chi..sup.2 Increased 2 6.7 10 33.3 21.4416 Stable
28 93.3 20 66.7 Total 30 100 30 100
[0226] As for heart and kidney functions, the serum glutamic
pyruvic transaminase (SGPT) level of one patient in the treatment
group increased slightly during the clinical study. This patient
was diagnosed later as having been infected with hepatitis C. No
toxicity effects to heart or kidney functions were found in the
treatment group. In contrast, the SGPT level of 3 patients in the
control group increased during the clinical study while abnormal
renal function with toxicity level I of blood urea nitrogen
occurred in 2 patients of this group. No alopecia or injury to the
nerve system was found in treatment group. The results show that
the pharmaceutical composition of composition of Fagopyrum cymosum
(Trev.) Meisn of this invention is safe for clinical
application.
[0227] 3. The pharmaceutical composition of Fagopyrum cymosum
(Trev.) Meisn of this invention is effective in improving Karnofsky
performance scores of lung cancer patients. Table 46 shows that
Karnofsky performance scores in the treatment group rose
significantly while that in the control group dropped
significantly.
48TABLE 46 Effects of the Pharmaceutical Composition of Fagopyrum
cymosum (Trev.) Meisn of This Invention on Improving Karnofsky
Performance Scores of Lung Cancer Patients Change after Scores
(Mean Value) Treatment (Cases) Before After Score Score Groups
Treatment Treatment Increased Decreased Treatment 60.58 69.44 20 4
Control 67.60 30.00 4 12
The Third Series of Clinical Study: Comparison between Therapeutic
Effect of the Pharmaceutical Composition of Fagopyrum cymosum
(Trev.) Meisn of This Invention Combined with Chemotherapy and the
Effect of Chemotherapy Alone
Patient Selection
[0228] 80 patients with lung cancer were enrolled in the clinical
study. There were 68 male patients and 12 female patients. Their
age distribution is as follows:
49TABLE 47 Age Distribution of 80 Lung Cancer Patients Total 41-50
51-60 61-70 >70 number 80 22 35 18 5 % 100 27.5 43.75 22.5
6.25
[0229] All patients were hospitalized between December, 1990 and
June 1992, definitely diagnosed as having small cell lung carcinoma
by anteroposterior and lateral chest tomography and CT and
cytological examination (sputum or fiberoptic bronchoscopy).
Clinical stages of the patients are shown in Table 48.
50TABLE 48 Clinical Stages of 80 Lung Cancer Patients Total I II
III IV III .+-. IV Cases 80 3 19 35 23 58 % 100 3.75 23.75 43.75
28.75 72.5
[0230] Table 48 shows that patients at stages III and IV accounted
for 72.5% of the total.
Design of the Clinical Study
[0231] Eighty patients were randomly divided into 3 groups: Group A
including 20 patients, was treated by the pharmaceutical
composition of Fagopyrum cymosum (Trev.) Meisn of this invention
alone; Group B including 30 patients was treated by the
pharmaceutical composition of Fagopyrum cymosum (Trev.) Meisn of
this invention combined with chemotherapy as the experiment group;
Group C including 30 patients was treated with chemotherapeutic
agents alone as the control group.
Experimental Results
[0232] 1. Therapeutic Effects
[0233] Effectiveness of the three different regimens was evaluated
according to the established criteria of four grades: Complete
Remission (CR), Partial Remission (PR), Stableness (S) and
Progression (P). Comparison of therapeutic effect of the three
groups is shown in table 49.
51TABLE 49 Comparison of Therapeutic Effects among Three Groups
Group Cases CR PR S P A 20(100%) 0(0%) 3(15%) 12(60%) 5(25%) B
30(100%) 0(0%) 11(36.67%) 14(46.66) 5(16.67%) C 30(100%) 0(0%)
5(16.67%) 12(40%) 13(43.33%)
[0234] Table 49 indicates that the partial remission rate in Group
A treated by the pharmaceutical composition of Fagopyrum cymosum
(Trev.) Meisn of this invention alone (3/20 or 15%) was lower than
that in the control group (5/30 or 16.67%).
[0235] There was no statistically significant difference
(X.sup.2=1.75, P>0.05).
[0236] The partial remission rate and stable rate in Group B (the
experiment group, 11/30 or 36.67% and 14/30 or 46.66% respectively)
were higher than those in Group C (the control group, 5/30 or
16.67% and 12/30 or 40% respectively). The difference was
statistically significant (X.sup.2=5.79, P<0.05).
[0237] The effective rate for Group A was lower than that for Group
C with no statistically significant difference (t=1.56, p>0.05).
The effective rate for Group B was higher than that for Group C
with statistically significant difference (t=1.75, p<0.05) and
also higher than that for Group A with statistically significant
difference (t=1.67, p<0.05).
[0238] The results of this experiment show that therapeutic effect
of the pharmaceutical composition of Fagopyrum cymosum (Trev.)
Meisn of this invention combined with chemotherapeutic agents is
greater than that of Fagopyrum cymosum (Trev.) Meisn composition
alone or chemotherapy alone, suggesting that the pharmaceutical
composition of Fagopyrum cymosum (Trev.) Meisn of this invention
has a synergistic effect with chemotherapy.
[0239] 2. Toxic and Untoward Side Effects
52TABLE 50 Comparison of Toxic and Untoward Effects in Three Groups
Group A Group B Group C (20 Cases) (30 Cases) (30 Cases) Parameters
Cases % Cases % Cases % Bone Marrow Depression 1 5 2 6.67 15 50
Anorexia or Diminution 0 0 1 3.33 26 86.67 of Food Intake Nausea
and Vomiting 0 0 0 0 9 30 Decrease in Immunity 1 5 3 10 8 26.67
Lowering in Functional 5 25 5 16.67 17 56.67 Status
[0240] Table 50 indicates that only 2 patients (2/30) in Group B
(the experiment group) showed reduction in leucocyte and platelet
counts, accounting for only 6.67% of the cases in that group, while
15 patients (15/30) in Group C (the control group) had the same
manifestation, accounting for 50% of the cases in that group; the
difference was statistically significant (t=3.37, p<0.01).
Patients with anorexia and diminution of food intake in Group B
were also much less than those in the control group (1/30 or 3.33%
v. 26/30 or 86.67%). No case with nausea or vomiting appeared in
Group B, while 9 cases developed these symptoms in Group C (0/30 or
0% v. 9/30 or 30%); the difference was also statistically
significant (t=6.48, p=0.01). Cases of decrease of immunity in
group B were less than those in Group C as well. In addition, cases
with lowering in functional status in Group B (5/30 or 16.67%) were
also less than those in the Group C (17/30 or 56.67%). The
difference was statistically significant (t=2.78, p=0.01). From
these experimental results, it is concluded that the pharmaceutical
composition of Fagopyrum cymosum (Trev.) Meisn of this invention
combined with chemotherapy can effectively reduce toxic and
untoward effects of single chemotherapy.
The Fourth Series of Clinical Study: Short Term Therapeutic Effect
of the Pharmaceutical Composition of Fagopyrum cymosum (Trev.)
Meisn of This Invention Combined with Radiotherapy
Patient Selection
[0241] Seventy patients with lung cancer were recruited in the
clinical study. They were randomly divided into 2 groups: 40
patients in the treatment group were treated by the pharmaceutical
composition of Fagopyrum cymosum (Trev.) Meisn of this invention
combined with radiotherapy. 30 patients in the control group were
treated by radiotherapy alone. The age and sex distribution of the
patients are as follows:
53TABLE 51 Age and Sex Distribution of 70 Lung Cancer Patients Mean
Value Mean Value Group Cases Male Age of Age Female Age of Age
Treatment 40 35 28-77 58.2 5 57-68 60.2 Control 30 25 32-68 56.4 5
52-67 60.4
[0242] The major clinical symptoms of the 70 lung cancer patients
are as follows:
54TABLE 52 Major Clinical Symptoms of 70 Lung Cancer Patients
Expec- Chest Bloody Groups Total Cough toration Pain Sputum Fever
Treatment Cases 40 37 25 19 18 12 Group % 100 92.5 62.5 47.5 45 30
Control Cases 30 30 27 12 15 7 Group % 100 100 90 40 50 23.3
[0243] The 70 patients were staged based on the "The Guiding
Principles for Clinical Research of New Drugs (Traditional Chinese
Materia Medica)" established by the Ministry of Health of P. R.
China in 1988. Table 53 shows that the majority of the 70 patients
fell into the intermediate and late stages of lung cancer.
55TABLE 53 Clinical Stages of 70 Lung Cancer Patients Groups Total
I II III IV III .+-. IV Treatment Cases 40 1 7 24 8 32 % 100% 2.5%
17.5% 60% 20% 80% Control Cases 30 0 8 14 8 22 % 100% 0% 26.7%
46.6% 26.7% 73.3%
[0244] The pathological classifications of the 70 lung cancer
patients are as follows:
56TABLE 54 Pathological Classifications of 70 Lung Cancer Patients
Small Cell Squamous Adeno- Undifferentiated Groups Total Carcinoma
carcinoma Carcinoma Treatment Cases 40 25 8 7 Group % 100 62.5 20
17.5 Control Cases 30 17 9 4 Group % 100 56.7 30 13.3
[0245] All 70 patients were definitely diagnosed as having lung
cancer by biopsy or brushing in fiberoptic bronchoscopy, operative
exploration and cytological examination of sputum to find cancer
cells in the tissues.
Methods of Treatment
[0246] Patients in both groups underwent radiotherapy with 150-200
cGy .sup.60Co or 6-8 MV electron accelerator, five times a week, in
a total dose of 4000 cGy over total mediastinum and 6000 cGy for
primary tumors. The dosage varied with a patient's state of
sickness and tolerance.
[0247] Patients in the treatment group was further administered the
pharmaceutical composition of Fagopyrum cymosum (Trev.) Meisn of
this invention, 1.2-1.6 g. each time, 3 times a day for a course of
about two months. The average duration of treatment for patients in
the treatment group was 41 days and the longest 68 days. The
average duration of treatment for patients in the control group was
59 days and the longest 80 days. Patients in the control group did
not take the pharmaceutical composition of Fagopyrum cymosum
(Trev.) Meisn of this invention, but received symptomatic
supportive treatment.
Experimental Results
[0248] 1. Therapeutic Effects
[0249] Therapeutic effectiveness was evaluated according to the
established criteria of four grades: Complete Remission (CR),
Partial Remission (PR), Stableness (S) and Progression (P).
Comparison of therapeutic effect between the treatment and control
groups is shown in Table 55.
57TABLE 55 Comparison of Therapeutic Effects between the Treatment
and Control Groups Total Effective Cases CR PR S P Rate Group Case
% Case % Case % Case % Case % Case % Treat- 40 100 13 32.5 18 45 8
20 1 2.5 31 77.5 ment Control 30 100 1 3.3 14 46.7 12 40 3 10 15 50
.chi..sup.2 11.03 0.02 2.45 0.67 5.75 P <0.01 >0.05 >0.05
>0.05 <0.05
[0250] Table 55 indicates that the complete remission rate in
treatment group (13/40 or 32.5%) was much higher than that in the
control group (1/30 or 3.3%). The effective rate (CR+PR) in the
treatment group (31/40 or 77.5%) was also much higher than that in
the control group (15/30 or 50%).
[0251] At the completion of treatment, major clinical symptoms of
the lung cancer patients in both groups were alleviated to some
extent. By comparison, improvements in the treatment group were
much more prominent. The results are shown in table 56.
58TABLE 56 Alleviation of Clinical Symptoms of 70 Lung Cancer
Patients in the Treatment and Control Groups Treatment N. = 40
Control Group N. = 30 Before After Before After Treatment Treatment
Treatment Treatment Symptoms Case % Case % Case % Case %
.chi..sup.2 P Cough 37 92.5 13 32.5 30 100 27 90 18.4 <0.01
Expectoration 35 87.5 21 52.5 27 90 25 83.3 6.83 <0.01 Chest
Pain 19 47.5 8 20 12 40 10 33.3 4 <0.05 Bloody Sputum 19 47.5 3
7.5 14 46.7 6 20 4.5 <0.05 Fever 12 30 3 7.5 7 23.3 6 20 4.65
<0.05
[0252] At the completion of treatment, reduction of hemoglobin,
leukocyte and platelet values to a varying extent was noted in
patients in both treatment and control groups. Leukocyte count of 5
patients in the control group decreased to less than
40.times.10.sup.9/L. No patient in the treatment group had
leukocyte level decreased to such an extent. The results are shown
in Table 57.
59TABLE 57 Change in Hemoglobin, Leukocyte and Platelet Values of
Lung Cancer Patients in Both Groups Treatment Group Control Group
N. = 40 N. = 30 Before After Before After Parameters Treatment
Treatment Treatment Treatment .chi..sup.2 P Hemoglobin 120.5 121.9
126.8 117.9 1.01 >0.05 (g/L) Leukocytes 8.3 6.5 7.8 5.6 2.587
<0.05 (.times.10.sup.9/L) Platelet 149.7 140.6 155.4 144.1 0.27
>0.05 (.times.10.sup.9/L)
[0253] At the completion of treatment, a majority of patients in
treatment group gained weight and better appetite, and improved
their life quality. However, only a few patients in the control
group exhibited these improvements. The results are shown in Table
58.
60TABLE 58 Increase in Body weight and Food Intake, and Improvement
in Life Quality of Lung Cancer Patients in Both Groups Treatment
Group Control Group N. = 40 N. = 30 Clinical Signs Case % Case %
.chi..sup.2 P Gain in Weight 18/35 51.42 4/30 13.33 7.66 <0.01
Increase of Food 21/32 65.62 1/30 3.33 23.59 <0.01 Intake
Improvement in 23/37 62.16 9/30 30 5.62 <0.05 Life Quality
[0254] At the completion of treatment, all immunological parameters
of patients in the treatment group increased more than those of
patients in the control group, except for IgA. The differences were
statistically significant as shown in Table 59.
61TABLE 59 Increase of Immunological Parameters of Lung Cancer
Patients in Both Groups Treatment Group Control Group N. = 40 N. =
30 Parameters Case % Case % .chi..sup.2 P Complement C3 23/31 74.19
8/30 26.67 11.91 <0.01 E Rosette 22/33 66.67 11/30 36.67 4.51
<0.05 Lymphocyte 11/13 84.62 1/10 10.00 7.54 <0.01
Transforming Factor IgG 20/32 62.50 7/30 23.33 8.13 <0.01 IgA
14/32 43.75 8/30 26.67 1.30 <0.05 IgM 22/32 68.75 12/30 40.00
4.07 <0.05 Macrophage 26/30 86.67 11/28 39.29 12.09 <0.01
Phagocytic Rate (%) Macrophage 18/29 62.07 9/28 32.14 4.83 <0.05
Phagocytic Index
[0255] At the completion of treatment, the value of liver function,
kidney function and carcinoembryonic antigen (CEG) in patients were
also compared between the treatment group and the control group.
The results showed no statistically significant difference
(P>0.05).
[0256] The results of the experiment show that application of
radiotherapy combined with the pharmaceutical composition of
Fagopyrum cymosum (Trev.) Meisn of this invention has a superior
effect than application of radiotherapy alone. They also suggest
that the pharmaceutical composition of Fagopyrum cymosum (Trev.)
Meisn of this invention has a synergistic effect with radiotherapy
and can alleviate the toxic and adverse effects of
radiotherapy.
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* * * * *