FORM 2

2 6 JUN 2002

THE PATENTS ACT, 1970
As Amended by The Patent (Amendment) Act, 1999.
COMPLETE SPECIFICATIO
SEE SECTION 10
Process for preparation of anti-inflammatory phytoconstituent derivatives"
APPLICANT
Ajanta Pharma Limited an Indian Company, Registered under Companies Act, 1956 and having its Registered Office at:
Ajanta House, 98 Govt. Industrial Area, Charkop, Kandivli (W), Mumbai - 400 067, Maharashtra, India
INVENTORS
The inventors under Sec.28 are :
i) BIYANI Milind Kesharlal ;
ii) BANAVALIKER Manisha Manohar;
iii) SUTHAR Ashish Chandrakant;
All working as Research Scientists at,
Research Centre, Ajanta Pharma Limited, Ajanta House,
98 Govt. Industrial Area, Charkop, Kandivli (W),
Mumbai - 400 067, Maharashtra, India.
Nationality : All being Indian nationals.
The following specification particularly describes the nature of the invention and the manner in which it is to be performed:

Field of Invention
The present invention relates to a process for preparation of anti-inflammatory
at
phytoconstituent derivatives more particularly it relates to a process for achieving ameliorated amounts of acetates of keto-beta-boswellic acid and incorporation of the same in oral dosage forms.
Background! of Invention
The gum resin of Boswellia serrata is known in Indian Ayurvedic system of medicine as Salai guggal and has been used for thousands of years for the treatment of various inflammatory diseases, joint problems, respiratory diseases, skin problems, and liver disorders. It has become a standard treatment for arthritic disorders in Ayurvedic medicines.
Recent clinical trial studies have confirmed its anti-inflammatory effects.
Pharmacological studies have established that an alcoholic extract of the gum
displayed marked anti-inflammatory activity in mice and rats. Further studies have
shown that the ethanolic extracts of gum resin of Boswellia serrata significantly
inhibited the in-vitro synthesis of leukotriene B4 in rat peritoneal polymorphonuclear
leukocytes, this indicates its usefulness as anti-inflammatory agent. The chemical
constituents of Boswellia serrata gum resin are reported to be pentacyclic and
tetracyclic triterpene acids. Later studies have shown that that the pentacyclic
tritearpenes commonly called as Boswellic acids are the major component of the gum
resin and are the source of in-vitro and in-vivo activity of the extract. The various
boswellic acids reported are : beta-boswellic acid, alpha-boswellic acid, acetyl-beta-
boswellic acid, acetyl-alpha-boswellic acid, 11-keto-beta-boswellic acid and acetyl-
11-keto-beta-boswellic acid.
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Boswellic acids are found to inhibit elevated human leukocyte elastase (HLE) or plasmin activity and useful in the clinical conditions characterized by the elevated activity of elastase and/or plasmin. HLE is a serine protease, which initiates injury to the tissue, which in turn triggers the inflammation. HLE also stimulates rrfucus secretion and thus may play a role in cystic fibrosis, chronic bronchitis and acute respiratory distress syndrome. Thus suggesting role of boswellic acid in clinical conditions like inflammatory diseases, arthritis, asthma, and other related chronic conditions.
Boswellic acids are now contemplated to be natural NSAIDs (non-steroidal
anti-inflammatory drugs) since they are specific inhibitor of 5-lipoxygenase; an
enzyme responsible for the synthesis of leukotrienes that maintain inflammation much
like the conventional non-steroidal anti-inflammatory drugs (NSAIDs) used for
inflammatory conditions. Also, boswellic acids do not impair the peroxidation of
arachidionic acid by iron ascoarbate. Moreover the toxicity profile of Boswellic acid
has been found to be low with most studies reporting no side effects. At present
treatment of various inflammations include the first line drugs such as non-steroidal
anti-inflammatory drugs (NSAIDs), secondary treatment includes corticosteroids.
These drugs have limited advantages and their effects mainly of short-term duration
and there are many disadvantages in the use of these drugs over extended periods of
time since, these drugs have severe side effects. As opposed to NSAIDs, long-term
use of boswellic acids does not lead to irritation or ulceration of the stomach. On
contrary, it is useful for treating gastric ulceration. Which is evident from the
controlled, double blind studies that boswellia extracts are very helpful for ulcerative
colitis. Therefore Boswellic acids are regarded as non-steroidal anti-inflammatory
compound (drag) NSAID with no side effects.
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Since boswellic acids are specific non-redox inhibitors of leukotrienes, it can have many possible uses apart from treatment of inflammation, in many other conditions in which leukotrienes play role such as asthma, rheumatoid arthritis; cystic fibrosis, astrocytoma (a certain kind of brain tumor), adult respiratory distress syndrome, Croahn's disease, liver cirrhosis, multiple sclerosis, ischaemic heart disease, allergic conjunctivitis, systemic lupus erythaematosus, gout, psoriasis, urticaria, ulcerative colitis, acute pancreatitis, heavy smoking and the lung damage it causes, traumatic injury sever burns etc.
The more recent studies have shown that among the boswellic acids, acetyl 11-keto-beta-boswellic acid (AKBA), is the strongest inhibitor of the formation of leukotrienes. According to one study the ability of the different boswellic acid to inhibit the formation of leukotrienes - LTB4 was determined. The concentration needed to block the activity of LTB4 by 50% was measured and it was observed that for AKBA the concentration needed was 1.5 micromoles. For other boswellic acids, it was 4-7 micromoles. AKBA decreased the activity of human leukocyte elastase (HLE) in vitro with an IC.sub.50 value of about 3 micromole. Acetyl-beta-boswellic acid and beta boswellic acid, both lacking the 11-keto function, inhibit 5-lipoxygenase activity incompletely. At concentration of about 15 micromoles their inhibitory activities reach a maximum (60% inhibition) and can not be increased further. These findings revealed that a hydrophilic group at C4 in combination with an 11-keto-function is essential for 5-lipoxygenase inhibition. Further it was also found that presence of non-inhibitory beta-boswellic acid and acetyl-beta-boswellic acid prevented the AKBA - induced inhibition of 5-lipoxygenase. These findings singled out AKBA as being the most powerful boswellic acids.
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Despite the above mentioned facts that acetates of boswellic acids that is AKBA is the most active derivative among the boswellic acids, the commercially available extracts of Boswellia serrata contains chiefly other boswellic acids h>more quantity with relatively less or negligible amounts of their acetates particularly AKBA. Therefore there is a need for the derivatives of boswellic acids with ameliorated amounts of acetates and less of other inhibitory boswellic acids to get the maximum activity. Accordingly the present invention provides the process for extraction of phytoconstituent derivatives from gum resin of Boswellia serrata with ameliorated amounts of acetates of keto-beta-boswellic acid.
Prior art
US Patent 5064823, granted to Lee, Yue-Wei, et al in 1991, discloses a method for preparation of Pentacyclic triterpenoid compound such as alpha-boswellic acid acetate, beta-boswellic acid acetate, which have an inhibitory effect on topoisomerase -1, and topoisomerase - II.
The prior art method adopts fractionation and chromatographic method to get alpha-boswellic acid acetate, beta-boswellic acids acetate, which is very time consuming and skillful technique, mainly for isolation purpose and not practical to adopt at an industrial level for large scale preparation. Unlike prior art method the present invention does not involve the use of chromatographic technique for separation, rather it is suitable for large-scale preparations of phytoconstituent derivative with ameliorated amounts of acetates of keto-beta-boswellic acid.
Japanese patent JP4288095A2, 1992, discloses a complemental activity -inhibiting agent containing a specific beta-boswellic acid derivative as an active
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ingredaient, eifective for treating autoimmune diseases such as systemic erythematoides and articular rheumatism.
As per the prior art column chromatographic and high performance liquid chromatographic techniques are used to obtain alpha- and beta- boswellic acid acetates. Whereas the present invention does not involve for preparation of keto-beta-boswellic acid acetates by chromatographic or high performance liquid chromatographic techniques, which are not suitable to be employed for industrial preparation, but provides a simple process which is also an appropriate at industrial level.
Indian Patent no. 180516, dated 13.5.94, describes a process for the isolation of a fraction containing a mixture of boswellic acid comprising mainly 6 triterpenoic acids namely beta-boswellic acid (3-alpha-hydroxy-urs-12-ene-24-oic acid) derivatives. US patent 5629351 granted in May 1997 which is equivalent to Indian Patent no. 182615 dated 13.5.94, describes a process for the isolation of a new boswellic acid from gum resin of plant Boswellia serrata.
In the prior arts processes are mainly separation and isolation techniques and fraction contain acetyl-11-keto-beta-boswellic acid in much lesser quantity that is 3 -13 % and other boswellic acids in major quantities that is beta-boswellic acid in the range of 35 - 55% and acetyl-beta-boswellic acid in the range of 25-45%. Also the processes of the prior art and present invention are totally different. Moreover in the prior art acetylation of the fraction of the boswellic acid is carried out at the final stage only for the isolation purpose. Unlike to prior art the present invention involves the acetylation of either crude initially or of an intermediate extract for getting the ameliorated amount of acetates of keto-beta-boswellic acid.
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International publication W00066111, published in November 2000, discloses compositions of boswellic acids derived from Boswellia serrata gum resin, for treating lymphoproliferative and autoimmune conditions by novel industrial process providing standardized compositions.
Prior art embodies entirely different process for extraction of boswellic acids which contains acetyl-11-keto-beta-oswellic acid not less than 4%, 11-keto-beta-boswellic acid not less than 5%, acetyl-beta boswellic acid not less than 10% and beta-boswellic acid not less than 14% by weight. Thus it is for extraction of mixture of boswellic acids referred to as total organic acids and with AKBA in the least quantity.
German patent publication DE 10035591, Jan'2002 relates to process for isolation and characterization of tetra and pentacyclic triterpene acids from biological material containing it by subjecting it to precipitation with one or more calcium and/or magnesium salt to isolate the triterpen acid precipitates.
The prior art process is directed towards isolation of mixture of the tetra and pentacyclic triterpen acids for e.g. acetyl-11-keto-beta-boswellic acid, acetyl beta-boswellic acid, acetyl alpha-boswellic acid, acetyl-11-hydroxy-boswellic acid, keto-boswellic acid, beta-boswellic acid, alpha-boswellic acid, 11-hydroxy-beta-boswellic acid, poly-hydroxy boswellic acid, and further boswellic acid derivatives. Thus the final extract would have other boswellic acids also which could interfere with AKBA activity.
International publication W00215916, published in February 2002 relates to dihydroboswellic acids and the physiologically tolerable salts and derivatives thereof, the physiologically tolerable salts of said derivatives and hydrogenated extracts of
boswellia. The invention also relates to the a method for the production of the same
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and use of dihydroboswellic acids, physiologically tolerable salts, and derivatives thereof, the physiologically tolerable salts of said derivatives or hydrogenated extracts of boswellia for prophylactic and/or therapeutic treatment of undesired physical or mental condition in human or animals.
Th prior art extracts of boswellia contains primarily dihydroboswellic acids and the physiologically tolerable salts and derivatives thereof, and other boswellic acids and does not contain AKBA in the major quantity. Also the process for preparation of extract of boswellia involves catalytic hydrogenation which is a different technique.
The present invention differs from the above prior art as the present invention relates to obtain ameliorated amount of acetates of keto-beta-boswellic acid with less quantity of other boswellic acids by entirely different process than those described by the prior arts for getting boswellic acids mixtures.
Therefore it is apparent from the foregoing that the nature and miens as described in present invention to obtain ameliorated amounts of acetates of keto-beta-boswellic acid in resultant extract with several benefits are altogether different from the prior arts.
Objects of Invention
Therefore it is an object of the present invention to provide process for preparation of phytoconstituent derivatives of Boswellia serrata with ameliorated amount of acetates of keto-beta-boswellic acids.
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It is an object of the present invention to provide process for preparation of phytoconstituent derivatives of Boswellia serrata with ameliorated amount of acetates of keto-beta-boswellic acids and less amount of other boswellic acids for better' activity.
It is yet another object of the present invention to provide process for preparation of phytoconstituent derivatives of Boswellia serrata devoid of other chemical constituents for example resinous constituents not required for particular therapeutic activities, and thereby lessening interference of such impurities for analysis or treatment.
It is still another object of the present invention to provide process for preparation of phytoconstituent derivatives of Boswellia serrata with ameliorated amount of acetates of keto-beta-boswellic acid with increased potency to decrease the dosage or number of dosing for getting the desired therapeutic activity.
It is further object of present invention to provide process for preparation of phytoconstituent derivatives of Boswellia serrata with ameliorated amount of acetates of keto-beta-boswellic acid by a process suitable for industrial scale.
It is also an object of the present invention to provide process for preparation of phytoconstituent derivatives of Boswellia serrata with ameliorated amount of acetates of keto-beta-boswellic acid for treatment of various inflammations without having side effects of conventional NSAIDS or steroidal drugs.
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It is also further object of the present invention to provide a process for preparation of suitable oral dosage forms by incorporation of phytoconstituent derivatives of Boswellia serrata with ameliorated amount of acetates of keto-beta-boswellic acid for treatment of various inflammations.
Summary of the Invention:
The present invention provides process for preparation of anti-inflammatory
phytoconstituent derivatives derived from the gum resin of Boswellia serrata with
ameliorated amounts of acetates of keto-beta-boswellic acid comprising steps of
comprises steps of, fine crushing the crude gum resin of Boswellia; extracting the gum
resin as such or which has been previously acetylated, with organic solvent; separating
the exhausted residue from the liquid extract; concentration of the extract under
reduced pressure to remove organic solvent to obtain resinous mass at temperature 30
- 50° C and at pressure of 25 inch of mercury; fractionation of the above said resinous
mass with non polar solvent or acidified or basified aqueous solution; drying the
concentrated resinous in vacuo; optionally treating the resultant extract with basified
aqueous solution, fractionating with nonpolar solvent, treating with acidified aqueous
solution, subsequently washing several times to make neutral, for further elimination
of non desirable components; optionally subjecting either the crude gum resin or
extract by treating with acetic anhydride under basic condition created by addition of
suitable base like pyridine, sodium acetate, or the like and further processing for
converting hydroxyl group of boswellic acid to acetyl group to get ameliorated
amounts of acetates of keto-beta-boswellic acids; preparing oral dosage forms by
blending anti-inflammatory phytoconstituent derivatives with ameliorated amounts of
acetates of keto-beta-boswellic acid in the range of 100 mg to 400 mg with excipients;
further granukting the blend; tableting the granules or, filling into capsules or, sachets
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useful in various inflammatory conditions in asthma, rheumatoid arthritis, cystic fibrosis, astrocytoma (a certain kind of brain tumor), adult respiratory distress syndrome, Crohn's disease, liver cirrhosis, multiple sclerosis, ischaemic heart disease, allergic conjunctivitis, systemic lupus erythaematosus, gout, psoriasis, urticaria, ulcerative colitis, acute pancreatitis, heavy smoking and the lung damage it causes, traumatic injury sever burns or the like.
Detailed description:
The present invention relates to process for preparation of anti-inflammatory phytoconstituent derivatives. According to the present invention the anti-inflammatory phytoconstituent derivatives are derived from the gum resin of Boswellia serrata with ameliorated amounts of acetates of keto-beta-boswellic acid.
Boswellic acids comprises of various triterpenoic acids like beta-boswellic acid, alpha-boswellic acid, acetyl beta-boswellic acid, acetyl alpha-boswellic acid, acetyl-11-hydroxy-boswellic acid, keto-boswellic acid, 11-hydroxy-beta-boswellic acid, acetyl-11-keto-beta-boswellic acid, and further boswellic acid derivatives. As per the latest clinical research studies AKBA - acetyl 11-keto-beta-boswellic acid is found to be the more potent derivative among the boswellic acids.
However the commercial extracts available contains either less, or negligible amount of AKBA than other boswellic acids. The present invention accordingly provides specifically designed novel process for extraction of phytoconstituent derivatives from gum resin of Boswellia serrata with ameliorated amounts of acetates of keto-beta-boswellic acid and less of other boswellic acids for better activity.
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The present invention process for extraction of phytoconstituent derivatives from gum resin of Boswellia serrata with ameliorated amounts of acetates of keto-beta-boswellic acid and also which can be adopted at an industrial level for large scale preparation, comprises steps of,
a) fine crushing the crude gum resin of Boswellia serrata by commuting or any other known methods;
b) extracting the gum resin as such or which has been previously acetylated, with organic solvent by a soxhlet or suitable method of extraction;
c) separating the exhausted residue from the liquid extract by means of centrifugation or any suitable means of filtration or separation;
d) concentration of the extract under reduced pressure to remove organic solvent to obtain resinous mass at temperature 30 - 50° C and at pressure of 25 inch of mercury;
e) fractionation of the above said resinous mass with non polar solvent or acidified or basified aqueous solution for removing the resinous constituents from the extract and enriching the therapeutically active components by falling drop method or any other suitable method;
f) drying the concentrated resinous extract after dissolving it into chlorinated solvent and evaporating it to dryness in vacuo, at temperature 35-55°C so as to get a powder free flowing in nature;
g) optionally treating the resultant extract with basified aqueous solution, fractionating with nonpolar solvent, treating with acidified aqueous solution, subsequently washing several times to make neutral, for further elimination of non desirable components;
h) optionally subjecting either the crude gum resin or extract by treating with acetic
anhydride under basic condition created by addition of suitable base like pyridine,
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sodium acetate, or the like and further processing for converting hydroxyl group of boswellic acid to acetyl group to get ameliorated amounts of acetates of keto-beta-boswellic acids for the treatment of various inflammations.
In accordance with the present invention process the organic solvents which may be used for extraction are selected from chloroform, dichloromethane, acetone, hexane, or the like and or combination thereof.
In accordance with the present invention process the nonpolar solvents which may be used for fractionation are selected from hexane, petroleum ether, or the like and or combination thereof.
In accordance with the present invention process the bases which may be used for preparing basified solution for fractionation or purification are selected from salts of calcium, sodium, potassium or the like and or combination thereof.
In accordance with the present invention process the acids which may be used for preparing acidified solution for fractionation or purification are selected from mineral acids like hydrochloric acid, sulfuric acid, nitric acid, or organic acids like citric acid, lactic acid, malic acid, ascorbic acid, or the like and or combination thereof.
In accordance with the present invention there has been also provided process for
preparation of oral dosage forms by blending anti-inflammatory phytoconstituent
derivatives with ameliorated amounts of acetates of keto-beta-boswellic acid in the
range of 100 mg to 400 mg with excipients; further granulating the blend; tableting
the granules or, filling into capsules or, sachets useful in various inflammatory
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conditions in asthma, rheumatoid arthritis, cystic fibrosis, astrocytoma (a certain kind of brain tumor), adult respiratory distress syndrome, Crohn's disease, liver cirrhosis, multiple sclerosis, ischaemic heart disease, allergic conjunctivitis, systemic lupus erythaematosus, gout, psoriasis, urticaria, ulcerative colitis, acute pancreatitis, heavy smoking and the lung damage it causes, traumatic injury sever burns or the like.
In accordance with the present invention various oral dosage forms prepared of anti-inflammatory phytoconstituent derivatives with ameliorated amounts of acetates of keto-beta-boswellic acid are selected from of tablets, or capsules or granules or the like.
The following examples illustrate but do not limit the scope of the invention. It is to be understood by those of the ordinary skill in the art that the present discussion is of exemplary embodiments only, and is not intended as limiting the broader aspects •of the present invention, which broader aspects are embodied in the exemplary construction.
Example 1 ;
1 Kg of crude Boswellia serrata gum resin was fine crushed. The crushed
Boswellia serrata gum resin powder was extracted with 2.5 liter of chloroform in
Soxhlet apparatus. The resultant liquid extract was filtered to separate the residue
from the extract. The filtered extract was concentrated in rotary vacuum evaporator at
60°C temperature and under pressure of 25 inch of mercury. The concentrated extract
was added dropwise to 3 liter of cold Hexane under stirring at the speed of 2000 rpm
to get a fine precipitate. Precipitate settled at the bottom was separated from the
supernatant by filtration. Precipitate thus obtained was dried in vacuum at 40°C
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temperature to creamish powdered extract comprising of 60.3% of acetyl-11-keto-beta-boswellic acid - AKBA and 8.2 % of other boswellic acids.
Example 2:
1 Kg of crude Boswellia serrata gum resin was fine crushed. The crushed Boswellia serrata gum resin powder was extracted with 2.5 liter of chloroform in Soxhlet apparatus. The resultant liquid extract was filtered to separate the residue from the extract. The filtered extract was concentrated in rotary vacuum evaporator at 60°C temperature and under pressure of 25 inch of mercury. The concentrated extract was added dropwise to 1 liter of Hexane at the temperature of 30-40° C, under stirring at the speed of 1000 rpm and stirring, after addition of the concentrated extract the stirring was further continued for about 30 minutes to get a fine precipitate. The precipitate settled at the bottom was separated from the supernatant by filtration. The precipitate thus obtained was dried in vacuum at 40°C temperature to creamish powdered extract comprising of 57% of acetyl-11-keto-beta-boswellic acid -AKBA and 5.4% of other boswellic acids.
Example 3:
1 Kg of crude Boswellia serrata gum resin was fine crushed. The crushed
Boswellia serrata gum resin powder was extracted with 2.5 liter of chloroform in
Soxhlet apparatus. The resultant liquid extract was filtered to separate the residue
from the extract. The filtered extract was concentrated in rotary vacuum evaporator at
60°C temperature and under pressure of 25 inch of mercury. The concentrated extract
was added dropwise under stirring at the speed of 2000 rpm to 7 liter of acidified
water with pH adjusted to 3 by addition of dilute hydrochloric acid to get a fine
precipitate. The precipitate thus obtained is collected by discarding the supernatant.
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The precipitate is dried in vacuum at 40°C temperature to creamish powdered extract comprising of 55.8% of acetyl-11-keto-beta-boswellic acid - AKBA and 5.1% of other boswellic acids.
Example 4:
1 Kg of crude Boswellia serrata gum resin was fine crushed. The crushed Boswellia serrata gum resin powder was extracted with 2.5 liter of chloroform in Soxhlet apparatus. The resultant liquid extract was filtered to separate the residue from the extract. The filtered extract was concentrated in rotary vacuum evaporator at 60°C temperature and under pressure of 25 inch of mercury. The concentrated extract was added dropwise to 3 liter of cold Hexane under stirring at the speed of 2000 rpm to get a fine precipitate. Precipitate settled at the bottom was separated from the supernatant by filtration. Precipitate thus obtained was dried in vacuum at 40°C temperature to creamish powdered extract. Acetic anhydride 450 ml and 2 ml of pyridine was added in powder extract, which was then warmed and stirred for 10-30 mins. The reaction mixture was kept overnight. After overnight incubation, precipitates of acetate of boswellic acid were collected and cold water wash was given to remove the traces of acetic anhydride and pyridine. The washed precipitates were then redissolved in chloroform, which was evaporated under vacuum to get —gm creamish powder in dry form comprising of 65% of acetyl-11-keto-beta-boswellic acid - AKBA and 1.4% of other boswellic acids.
Example 5:
1 Kg of crude Boswellia serrata gum resin was fine crushed. The crushed Boswellia serrata gum resin powder was extracted with 2.5 liter of chloroform in Soxhlet apparatus. The resultant liquid extract was filtered to separate the residue

from the extract. The filtered extract was concentrated in rotary vacuum evaporator at 60°C temperature and under pressure of 25 inch of mercury. The concentrated extract was added dropwise to 3 liter of cold Hexane under stirring at the speed of 2000 rpm to get a fine precipitate. Precipitate settled at the bottom was separated from the supernatant by filtration. Acetic anhydride 300 ml and 3 gm of sodium acetate was added in powder extract, which was then warmed and stirred for 10-30 min. The reaction mixture was kept for 1- 6 hr at room temperature. After incubation at room temperature reaction mixture was added in crushed ice with slow addition of Sodium carbonate till pH 8.0 - 8.5. The mixture was stirred immediately till the ice melts. Precipitates of acetate of boswellic acid were collected by centrifugation. Cold water wash was given to precipitates to remove the traces of acetic anhydride. The washed precipitates were then redissolved in methanol which was then evaporated under vacuum to get dried powder comprising of 67% of acetyl-11-keto-beta-boswellic acid - AKBA and 0.4% of other boswellic acids .
Example 6:
1 Kg of crude Boswellia serrata gum resin was fine crushed. The crushed Boswellia serrata gum resin powder was extracted with 2.5 liter of chloroform in Soxhlet apparatus. The resultant liquid extract was filtered to separate the residue from the extract. The filtered extract was concentrated in rotary vacuum evaporator at 60°C temperature and under pressure of 25 inch of mercury. The concentrated extract was added dropwise to 3 liter of cold Hexane under stirring at the speed of 2000 rpm to get a fine precipitate. Precipitate settled at the bottom was separated from the supernatant by filtration. Precipitate thus obtained was dried in vacuum at 40°C temperature to creamish powdered extract Dried extract powder of Boswellia serrata
was suspended in 1.5 liter basified water with pH adjusted to 9.0 - 10 by addition of
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5% calcium hydroxide and was stirred for 4 hours at room temperature to homogenous solution. The resultant solution was fractionated with 1.5 liter of petroleum ether. The aqueous layer was collected and its pH was adjusted to 2.5-3 with concentrated hydrochloric acid to obtain the precipitate. The precipitate was separated by filtration and cold water washes were given to the precipitate for removal of residual acid. The washed precipitate was then dried in vacuum evaporator to get creamish powder comprising of 90% of acetyl-11 -keto-beta-boswellic acid - AKBA and 1.4% of other boswellic acids.
Example 7:
Preparation of tablets of phytoconstituent derivatives with ameliorated amounts of acetates of keto-beta-boswellic acid :
Phytoconstituent derivatives with ameliorated amounts of acetates of keto-beta-boswellic acid from any of the example 1 to 6 - 300mg, colloidal silicone dioxide -234 mg, and sodium starch glycolate 22.8 mg were sifted through 40 mesh BSS sieve, the resultant blend was granulated using cross-linked polyvinyl pyrrolidone - 3 OK -100 mg using sufficient quantity of dichloromethane, granules were air dried the for 30 to 60 minutes, the wet granules were sifted through 8 mesh BSS sieve, sifted granules were dried at 55° C for 40-90 minutes, dried granules were passed through 20 mesh sieve, talc 4 mg, magnesium stearate 4 mg, sodium starch glycolate 16,5 mg, colloidal silicone dioxide 4 mg and microcrystalline cellulose 30 mg were sifted through 60 mesh sieve, the resultant mix was blended with dried granules, the resultant granule blend was compressed into tablets using suitable dies and punch.
Example 8 ;
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Preparation of capsules of phytoconstituent derivatives with ameliorated amounts of acetates of keto-beta-boswellic acid :
Phytoconstituent derivatives with ameliorated amounts of acetates of keto-beta-boswellic acid from any of the example 1 to 6 - 300mg was granulated with isopropyl alcohol, the granules were passed through 8 mesh standard BSS sieve and air dried, dried granules were passed through 18 mesh standard BSS sieve, the resultant granules were blended with talc 3 mg and magnesium stearate 1.5 mg, the resultant granule blend was filled into zero size hard gelatin capsule shell.
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We Claim,
1. A process for preparation of anti-inflammatory phytoconstituent derivatives from gum resin of Boswellia serrata, with ameliorated amounts of acetates of keto-beta-boswellic acid and incorporation of the same in oral dosage forms comprising steps of
a) comminuting the crude gum resin of Boswellia serrata;
b) extracting the gum resin as such or previously acetylated gum resin, with organic non polar solvent;
c) separating the exhausted residue from the liquid extract by means of centrifugation or filtration;
d) concentrating the liquid extract under reduced pressure of 25 inches of mercury and at temperature 30 - 50° C to obtain resinous mass;
e) fractionation of the resinous mass by pouring it dropwise in a non polar solvent under stirring;
f) drying the concentrated resinous extract in vacuo, at temperature 35-55°C so as to get a powder free flowing in nature containing 55-60% of acetates of keto-beta-boswellic acid;
g) optionally treating the resultant extract with basified aqueous solution, fractionating with non polar solvent, treating with acidified aqueous solution, subsequently washing several times to make neutral, for further elimination of non desirable components to get a powder free flowing in nature containing 85-90% of acetates of keto-beta-boswellic acid;
h) optionally subjecting either extract or the crude gum resin to treatment with acetic anhydride under basic condition created by addition of base like pyridine, sodium acetate, or the like and further processing for converting hydroxyl group of

boswellic acid to acetyl group to get ameliorated amounts ranges from 63 - 68% of acetates of keto-beta-boswellic acids;
i) preparing oral dosage forms by blending anti-inflammatory phytoconstituent derivatives with ameliorated amounts of acetates of keto-beta-boswellic acid in the range of 100 mg to 400 mg with pharmaceutically accepted inert safe excipients; granulating the blend; tableting the granules or, filling into capsules or, sachets useful in various inflammatory conditions.
2. Process for preparation of anti-inflammatory phytoconstituent derivatives as claimed in claim 1 wherein the non polar solvents which may be used are selected from hexane, petroleum ether, or the like and or combination thereof.
3. Process for preparation of anti-inflammatory phytoconstituent derivatives as claimed in claim 1 wherein bases which may be used for preparing basified solution are selected from salts of calcium, sodium, potassium or the like and or combination thereof.
4. Process for preparation of anti-inflammatory phytoconstituent derivatives as claimed in claim 1 wherein acids which may be used for preparing acidified solution are selected from mineral acids like hydrochloric acid, sulfuric acid, nitric acid, or organic acids like citric acid, lactic acid, malic acid, ascorbic acid, or the like and or combination thereof.
5. Process for preparation of anti-inflammatory phytoconstituent derivatives as claimed in claim 1 wherein various oral dosage forms prepared of anti-inflammatory
SO
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phytoconstituent derivatives with ameliorated amounts of acetates of keto-beta-boswellic acid are selected from of tablets, or capsules or granules or the like.
6. Process for preparation of anti-inflammatory phytoconstituent derivatives from gum resin of Boswellia serrata, with ameliorated amounts of acetates of keto-beta-boswellic acid and oral dosage forms thereof substantially as herein described with reference to each of the examples.

Dated this 10th day of October month 2003

For Ajanta Pharma Limited


The Controller of Patents,
The Patent Office, Mumbai-400 013.

Dr. JATHAR Shripad R„
Deputy General Manager - FD Ajanta Pharma Limited Ajanta House, 98, Govt. Industrial Area, Charkop, Kandivli (W), Mumbai - 400 067

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