Industrial production sennosides by plant tissue culture and total synthesis

1. Industrial Production of Sennosides
Pharmacognosy
Rajasekhar Reddy Alavala
M. Pharm., (PhD)
Assistant Professor,
K L COLLEGE OF PHARMACY, Vaddeswaram, Guntur, A.P-522502

2. ◦ Industrial production, estimation and utilization of the following
phytoconstituents:
Forskolin Sennoside
Artemisinin Diosgenin
Digoxin Atropine
Podophyllotoxin Caffeine
Taxol Vincristine and Vinblastine

3. ◦ Industrial production, estimation and utilization of the following
phytoconstituents:
Forskolin Sennoside
Artemisinin Diosgenin
Digoxin Atropine
Podophyllotoxin Caffeine
Taxol Vincristine and Vinblastine

4. Sennosides
◦ Senna and its preparation are used as purgative in habitual constipation.
◦ The anthraquinone glycosides of senna are absorbed first in intestinal
tract after which the aglycone part is separated and excreted in colon.
◦ These excreted anthraquinoness irritate and stimulate the colon thereby
movements are increased due to local action.
◦ The increase in peristalsis also causes reduction in the water absorption.
This results in soft and bulky faeces.
◦ It is believed that griping effect caused by senna is due to its resin or
emodin content. The drug given by parenteral way is secreted in colon
and thereby causes the therapeutic action

5. Sennosides:
◦ The rhizome and root of Rheum
spp. (Polygonaceae), is an
important drug in Chinese
medicine as well as in western
medicine since ancient times.
◦ The main purgative principle of
rhubarb was proved to be
sennosides, identical with those
isolated from Senna leaves.

6. Sennosides
Plant material:
◦ The seeds of a stable hybrid of Rheum palmatum L. and
Rh. Coreanum Nakai were used for this experiment.
◦ The seeds, treated at 4°C for 3 weeks, were placed on
absorbent cotton soaked with water for 24 h, and
sterilized by soaking in sodium hypochlorite solution for
10 mm, then washed with sterilized, distilled water.

7. Sennosides
Medium:
◦ For germination, the following medium was used: sucrose
(20 g/l) and agar (9 g/l) were dissolved in water by heating
on a boiling water bath, and the solution was divided into 20
ml each in the 100 ml conical flasks which were sterilized in
an autoclave at 120°C, under the atmospheric pressure of 1.2
kg/cm2 for 15 mm.
◦ For callus formation and cultivation, the Murashige-Skoog
(MS) medium added with sucrose, glucose or maltose (20
g/l) and some plant hormones, one or two of those among
2,4-dichlorophenoxyacetic acid (2,4-D), a-naphthaleneacetic
acid (NAA), kinetin (K), B-indoleacetic acid (IAA), and N-
(2-chloro-4-pyridyl)-N'-phenylurea (4-PU-30), were used in
suspension culture.
◦ For stationary culture, agar was added to the above medium
solution.

8. Sennosides
Selection of callus cells:
◦ The callus was suspended in sterilized, distilled water in a test
tube, and homogenized under stirring with a small glass stick. The
separated cell pieces were inoculated on an MS medium plate
containing IAA (1ppm) and 4PU-30 (1 ppm), and incubated in the
dark at 21 °C for 1 month.
◦ The colonies were separated and periodically transferred to the
medium at above every 1 month to get a callus of 2-3 cm diameter
after cultivation for 3 months.
◦ Then the content of sennoside in the callus was determined as
indicated below to select a productive strain.

9. Sennosides
Determination of products:
◦ Analyzed by means of high performance liquid
chromatography (HPLC)
Non-glycosidic hydroxyanthraquinones
◦ Column:- Senshu Pak silica gel column
◦ Solvent: n-hexane/AcOH (4:1) as the solvents
Hydroxyanthraquinone glycosides
◦ Aqua-sil SS-112 column using CHCl3/MeOH/H2O
(40:16:3) as the solvent
Sennosides
◦ Senshu Pak SN-112 N column using THF/H2O/AcOH
(8:2:1) as the solvent