Title of Invention

A PHARMACEUTICAL COMPOSITION ESSENTIALLY COMPRISING (-)-SECOISOLARICIRESINOL OBTAINED FROM STEREOSPERMUM PERSONATUM USEFUL AS AN ANTIOXIDANT

Abstract This invention relates to a pharmaceutical composition comprising an effective amount of (-)-Secoisolariciresinol together with or associated with an additive useful as an antioxidant; a process for isolating the (-)-Secoisolariciresinol from the pant Stereospermum Personatum and also relates to use of the active fraction (-)-Secoisolariciresinol as an antioxidant or free radical scavenger.
Full Text Field of the Invention
This invention relates to the isolation of compound namely (-)-Secoisolariciresinol from a
new plant source Stereospermum personatum. The compound (-)-Secoisolariciresinol
{2,3 Bis - [4-hydroxy-3- (methoxyphenyl) methyl]-1,4-butanediol} is isolated from new
source in good yield.
It is proved to be useful and better antioxidant molecule than the presently used
medicinally important lipophilic antioxidants Prubucol and a-Tocopherol. It may have
better therapeutic potential in inflammatory disease conditions, atherosclerosis, diabetic
complications, cancer, hepatotoxicity and variety of disease conditions mediated through
or fostered by oxidative stress and / or overt oxidative burden due to increased generation
or under scavenging of free radicals.
Background and Prior Art References
There is a considerable amount of epidemiological evidence indicating association
between diet rich in fruits and vegetables and a decreased risk of cardiovascular disease
and certain forms of cancer. It is generally assumed that the active principles contributing
to these protective effects are nothing but primarily, the antioxidant phytochemicals.
Research in the past decades have accumulated enough evidence to show the beneficial
effect of free-radical scavengers/antioxidants as antimutagenic, anti-inflammatory,
antiamerosclerotic, antidiabetic, antihepatotoxic, antiageing and in a variety of
neurological disorders. The search for new antioxidant principles is becoming therefore,
essential to improve the pharmacological treatment of pathological conditions such as
cataract, rheumatic diseases, atherosclerosis, Alzheimer's disease and other
neurodegenarative conditions.
The pharmacological approaches therefore have focused on the search of potential
resources rich in with antioxidant principles. The medicinal importance of plants bearing
rich proportion of antioxidant principles is therefore becoming hot item.
Stereospermum personatum is a medicinal plant used in traditional Indian System of
Medicine and is widely advocated in the preparations for diuretic, lithontriptic,
expectorant, cardiotonic, aphrodisiac, appetizer, anti-inflammatory, antibacterial.
dyspepsia, diarrhea, renal and vesical calculi, cough, asthma, hyperdipsia, haemorrhoids
and hyperacidity disease conditions ( Indian Medicinal Plants, Vol 5 p 192). It is further
reported to possess antibacterial, antifungal, hypoglycemic activity, and against p388
lymphocytic leukemic cells ( Ind. Jour, of Exp. Biol., 1971,9, 100). Hence, it becomes
pertinent to look for the molecules possessing such important biological properties. In this
connection, the phytochemical investigation of Stereospermum personatum has been taken
up. The applicants have isolated (-)-Secoisolariciresinol in good yields. (-)-
Secoisolariciresinol is known to possess several activities, which are shown in Table 1.
(Table Removed)
Objects of the Invention
The main object of the invention is to provide novel composition containing (-)-
Secoisolariciresinol in combination with or associated with additives and useful as an
antioxidant.
Another object is to provide a new source for obtaining (-)-Secoisolariciresinol in
significant yields.
Yet another object of the invention is to provide a process for producing (-)-
Secoisolariciresinol from a plant namely Stereospermum personatum.
One more object of the invention relates to use of (-)-Secoisolariciresinol as an
antioxidant.
Summary of Invention
In accordance with this invention, it has been found that (-)-Secoisolariciresinol is isolated
from a new plant source, Stereospermum Personatum in significant yield. Also, it has been
found that (-)-Secoisolariciresinol_sho\v antioxidant property.
Detailed description of the invention
Accordingly, the invention provides a novel composition containing (-)-
Secoisolariciresinol and useful as antioxidant. The invention further provides a method for
the isolation of (-)-Secoisolariciresinol from a new source namely Stereospermum
Personatum.
Antioxidant compounds recently have attracted the attention due to their broad spectrum
of activities in disorders of multiple origin viz., coronary heart disease, cancer, diabetes,
rheumatic disorders and inflammatory conditions where free radicals play important role.
Much attention is being directed now to harness and harvest the antioxidant compounds
from natural resources.
The compound (-)-Secoisolariciresinol is used in pure form. Hence, the usage may be
more advantageous than a mixture of compounds having similar properties, which are in
current use. It is also important to note that the process of isolation of (-)-
Secoisolariciresinol is highly economical.
Accordingly, the present invention provides a useful source, Stereospermum Personatum,
comprising antioxidant principle of which (-)-Secoisolariciresinol is isolated as pure and
potent antioxidant molecule. (-)-Secoisolariciresinol has been compared with existing
pharmacologically / therapeutically accepted antioxidant Probucol and alpha-tocopherol.
It is found that (-)-Secoisolariciresinol is better than the above mentioned reference drugs
and hence may be used with pharmaceutically / therapeutically acceptable additives.
In an embodiment of the present invention, (-)-Secoisolariciresinol may be effective in
much less amount than the reference drugs mentioned above.
In an embodiment of the present invention relates to a composition useful as antioxidant,
said composition comprising an effective amount of (-)-Secoisolariciresinol in
combination with or associated with a pharmaceutically acceptable additives.
In another embodiment of the invention, the pharmaceutically acceptable additive is
selected in such a manner that it does not affect or interfere with the efficacy of (-)-
Secoisolariciresinol.
In still another embodiment of the invention, the additive is selected from nutrients such as
carbohydrates, sugar, proteins, fats and pharmaceutically acceptable carrier.
In yet another embodiment of the invention, the amount of (-)-Secoisolariciresinol
administered is in the range between 200 mg to 250mg per dose at least twice a day.
In an embodiment of the present invention, the ratio of (-)-Secoisolariciresinol to the
additive is in the range between 0.1:10 to 3:10, preferably 0.4:10 to 2:10.
In yet another embodiment of the invention, the composition is administered through oral
route in the form of tablets, capsules, syrup or powder.
In one, more embodiment of the invention relates to process for isolation of (-)-
Secoisolariciresinol from the plant Stereospermum personatum said process comprising
the steps of:
a) extracting the dried wood powder of Stereospermum Personatum with
hexane;
b) further extracting the residue from step (a) with chloroform;
c) concentrating the chloroform solution from step (b) under vacuum;
d) absorbing the dark brown extract on a silicagel (60-120mesh) and loaded
on silicagel (60-120 mesh) column (4 cm dia to height of 100 cm);
e) eluting the column with chloroform methanol gradient, and
f) collecting the eluted fraction with 3 % methanol in chloroform and
concentrating the fraction to obtain pure (-^-Secoisolariciresinol.
hi another embodiment of the present invention, the solvents used are selected from
hexane, chloroform and methanol.
hi another embodiment of the present invention, the yield of (-)-Secoisolariciresinol
obtained is about 0.03% of the plant dried material.
One more embodiment relates to use of (-)-Secoisolariciresinol as an antioxidant or freeradical
scavenger animals and human beings.
In yet another embodiment relates to use of (-)-Secoisolariciresinol for the manufacturing
of a composition useful as antioxidant.
hi yet another embodiment of the present invention, (-)-Secoisolariciresinol is
administered orally.
Yet another embodiment of the invention relates to a method for the treatment of free
radical scavenging, comprising the steps of administering an effective amount of (-)-
Secoisolariciresinol to a subject in need thereof, preferably, (-)-Secoisolariciresinol is
administered in the range of 200 mg- 250mg/dose, at least twice a day.
Stereospermum personatum hence is a new source for (-)-Secoisolariciresinol and its
presence in this plant in good yields makes this invention more important. A comparison
of yield (-^-Secoisolariciresinol from different plants is given in Table.2
(Table Removed)
The present invention embodies isolation of (-)-Secoisolariciresinol, as antioxidant
principle from a new plant source and identify its free radical scavenging property
compared with medicinally important antioxidant drug molecules.
Brief description of the accompanying drawing:
Figure 1 shows antioxidant (Anti-free radical, DPPH) activity of (-)-Secoisolariciresinol
from Stereospermum personatum and reference compounds
Figure 2 shows the structural formula of (-)-Secoisolariciresinol
In our copending patent application no. NF368/2001 we have disclosed a process for
preparation of (-)-Secoisolariciresinol from a new plant source Stereospermum
personatum.
The invention is further described in the following examples that are given by the way of
illustration and therefore should not be construed to limit the invention in any manner.
Example-1
Experimental protocol: A process for the isolation of above lignans.
The dried wood powder of Stereospermum personatum (3 Kg) was loaded on a Soxhlet
apparatus. The powder was first extracted with hexane. The residue from the extraction of
hexane was further extracted with chloroform. The chloroform solution was concentrated
under vacuum. The dark brown extract (22 g) was adsorbed on silica gel (60-120 mesh)
and loaded on silica gel (60-120 mesh) column. (4 cm dia to a height of 100 cms).
The column is subjected to elution with chloroform-methanol gradient. The chloroformmethanol
gradient is so selected to obtain specific fraction and thereby the desired
compound. In the present case, the fractions eluted at 3% methanol in chloroform are
collected separately and concentrated.
The above fractions are subjected to further purification using silica gel column
( >200mesh, 2.5 cm dia and 50 cm length) using chloroform-methanol gradient. The
eluent at 3% methanol in chloroform gave pure (-)-Secoisolariciresinol (0.9 g). The
spectrochemical data of (-)-Secoisolariciresinol are given below:
(-)-Secoisolariciresinol
1. Molecular formula: CaoHzeOe
2. 'HNMR : 81.82(2H, m), 2.6(2H, dd), 2.7(2H, dd), 3.55 (2H, dd), 3.80(6H, s),
3.85(2H, d), 5.4 (Ar-OH,brs), 6.55 (2H, s), 6.6(2H,d), 6.8(2H, d).
3. I3CNMR: 6 34.02(C-7), 42.55(C-8), 55.51 ( 2xOCH3) 60.32(C-9), 113.07(C-2),
115.09(C-5), 121.17(C-6), 132.26(C-1), 144.3 l(C-4),147.26(C-3).
4. MS: 362(M+)
5. IR: 3422 cm'1 (OH)
Example 2
In vitro evaluation of free radical scavenging antioxidant potency:
Antioxidant activity of the compounds was tested for its capacity/potency to scavenge
most widely used free radical, l,l-diphenyl-2-picryl hyrazyl radical (DPPH). The wellaccepted
and tested antioxidants namely probucol and a-tocopherol were taken as
reference compounds. Img/ml DMSO concentration of the compounds were prepared and
subsequently, diluted to lower concentrations with DMSO. 200 ul of test compounds were
reconstituted to 1 ml in tris-HCl buffer (pH 7.4). Equal volume of 500 uM of DPPH
radical dissolved in ethanol was reacted with this. After incubation for 45 minutes in dark,
the absorbency at 517 nm was recorded. Percent radical scavenging activity was calculated
accordingly. All the readings were taken in triplicate. Result (Fig. 1 and Table 3) shows
that compound under consideration possess potent antioxidant/free- radical scavenging
property.
Table 3
50% radical scavenging concentratio of compounds
(Table Removed)



WE CLAIM:
1. A pharmaceutical composition comprising an effective amount of (-)-
Secoisolariciresinol together with or associated with an additive useful as an
antioxidant.
2. A composition as claimed in claim 1 wherein, the pharmaceutically acceptable
additive is selected in such a manner that it does not affect or interfere with the
efficacy of (-)-Secoisolariciresinol
3. A composition as claimed in claim 1 wherein, the additive is selected from
nutrients such as carbohydrates, sugar, proteins, fats and pharmaceutically
acceptable carrier.
4. A composition as claimed in claim 1 wherein, the (-)-Secoisolariciresinol is
effective at lower amount.
5. A composition as claimed in claim 1 wherein, the amount of (-)-
Secoisolariciresinol administered is in the range between 200 mg to 250mg per
dose at least twice a day.
6. A composition as claimed in claim 1 wherein, the composition is administered in
the form of tablets, capsules, syrup and powder.
8. Use of (-)-Secoisolariciresinol as an antioxidant or free-radical scavenger for
animals and human beings.
9. Use of (-)-Secoisolariciresinol for the manufacturing of a composition useful as
antioxidant.
10. Use as claimed in claim 8 whereia (-)-Secoisolariciresinol is administered orally.
11. Use as claimed in claim 9 wherein, the composition comprising additives selected
from nutrients such as carbohydrates, sugar, proteins, fats and pharmaceutically
acceptable carrier.
12. A method for the treatment of radical scavenging, comprising the steps of
administration of an effective amount of (-)-Secoisolariciresinol to a subject in
need thereof.
13. A method as claimed in claim 12 wherein, (-)-Secoisolariciresinol is administered
in the range of 200mg-250mg per dose, at least twice a day.
14. A novel pharmaceutical composition useful as antioxidant substantially as herein
described with reference to the examples and drawing accompanying this
specification.
WE CLAIM:
1. A pharmaceutical composition comprising an effective amount of (-)-
Secoisolariciresinol together with or associated with an additive useful as an
antioxidant.
2. A composition as claimed in claim 1 wherein, the pharmaceutically acceptable
additive is selected in such a manner that it does not affect or interfere with the
efficacy of (-)-Secoisolariciresinol
3. A composition as claimed in claim 1 wherein, the additive is selected from
nutrients such as carbohydrates, sugar, proteins, fats and pharmaceutically
acceptable carrier.
4. A composition as claimed in claim 1 wherein, the (-)-Secoisolariciresinol is
effective at lower amount.
5. A composition as claimed in claim 1 wherein, the amount of (-)-
Secoisolariciresinol administered is in the range between 200 mg to 250mg per
dose at least twice a day.
6. A composition as claimed in claim 1 wherein, the composition is administered in
the form of tablets, capsules, syrup and powder.

8. Use of (-)-Secoisolariciresinol as an antioxidant or free-radical scavenger for
animals and human beings.
9. Use of (-)-Secoisolariciresinol for the manufacturing of a composition useful as
antioxidant.
10. Use as claimed in claim 8 wherein, (-)-Secoisolariciresinol is administered orally.
11. Use as claimed in claim 9 wherein, the composition comprising additives selected
from nutrients such as carbohydrates, sugar, proteins, fats and pharmaceutically
acceptable carrier.
12. A method for the treatment of radical scavenging, comprising the steps of
administration of an effective amount of (-)-Secoisolariciresinol to a subject in
need thereof.
13. A method as claimed in claim 12 wherein, (-)-Secoisolariciresinol is administered
in the range of 200mg-250mg per dose, at least twice a day.
14. A novel pharmaceutical composition useful as antioxidant substantially as herein
described with reference to the examples and drawing accompanying this
specification.


Documents:

861-DEL-2001-Abstract.pdf

861-DEL-2001-Claims.pdf

861-DEL-2001-Correspondence-Others.pdf

861-DEL-2001-Correspondence-PO.pdf

861-DEL-2001-Description (Complete).pdf

861-DEL-2001-Drawings.pdf

861-DEL-2001-Form-1.pdf

861-DEL-2001-Form-18.pdf

861-DEL-2001-Form-2.pdf

861-DEL-2001-Form-3.pdf

861-del-2001-petition-137.pdf


Patent Number 211486
Indian Patent Application Number 861/DEL/2001
PG Journal Number 07/2008
Publication Date 15-Feb-2008
Grant Date 01-Nov-2007
Date of Filing 16-Aug-2001
Name of Patentee COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH
Applicant Address RAFI MARG, NEW DELHI-110001, INDIA.
Inventors:
# Inventor's Name Inventor's Address
1 JANASWAMY MADHUSUDANA RAO INDIAN INSTITUTE OF CHEMICAL TECHNOLOGY, UPPAL ROAD, HYDERABAD (A.P)-500007, INDIA.
2 ASHOK KUMAR TIWARI INDIAN INSTITUTE OF CHEMICAL TECHNOLOGY, UPPAL ROAD, HYDERABAD (A.P)-500007, INDIA.
3 UPPARAPALLI SAMPATH KUMAR INDIAN INSTITUTE OF CHEMICAL TECHNOLOGY, UPPAL ROAD, HYDERABAD (A.P)-500007, INDIA.
4 JHILLU SINGH YADAV INDIAN INSTITUTE OF CHEMICAL TECHNOLOGY, UPPAL ROAD, HYDERABAD (A.P)-500007, INDIA.
5 KONDAPURAM VIJAYA RAGHAVAN INDIAN INSTITUTE OF CHEMICAL TECHNOLOGY, UPPAL ROAD, HYDERABAD (A.P)-500007, INDIA.
PCT International Classification Number C07C 37/00
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA