FORM2
THE PATENTS ACT, 1970
(39 of 1970)
&
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10; rule 13)
1. Title of the invention - AN IMPROVED PROCESS FOR THE PREPARATION OF
PENTOSAN POLYSULFATE OR SALTS THEREOF

2. Applicant(s)
(a) NAME:
(b) NATIONALITY :
(c) ADDRESS :

ALEMBIC LIMITED
An Indian Company.
Alembic Campus, Alembic Road, Vadodara - 390 003, Gujarat, India.

3. PREAMBLE TO THE DESCRIPTION
The following specification particularly describes the invention and the manner in which it is to be performed:

Field of Invention:
The present invention relates to an improved process for the preparation of Pentosan polysulfate of formula (I)

or salt thereof, wherein R represents -SO3Y, and Y is at least one member selected from the group consisting of H and a pharmaceutically acceptable cation such as sodium, potassium, and magnesium. It also provides an amorphous form of Pentosan polysulfate sodium and process thereof.
Background of the invention:
Pentosan polysulfate sodium is chemically known as β-D-Xylan, (1-4), 2, 3-bis (hydrogen sulphate) sodium salt having molecular weight range of 4000 to 6000 Dalton. The structural formula of Pentosan polysulfate sodium represents as below:

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The current pharmaceutical product containing Pentosan polysulfate sodium salt is being sold by Ortho McNeil under the tradename Elmiron® in the form of capsules which is used for the treatment of bladder infection, interstitial cystitis and tumors.
Pentosan polysulfate sodium [37319-17-8] is a semi-synthetically produced heparin-like macromolecular carbohydrate derivative which chemically and structurally resembles glycosaminoglycans. It is a white odorless powder, slightly hygroscopic and soluble in water to 50% at pH 6.
Pentosan polysulfate sodium is a mixture of linear polymers of β-l -> 4-linked xylose, usually sulfated at the 2- and 3-positions and occasionally substituted at the 2-position with 4-Omethyl-α-D-glucuronic acid 2, 3-O-suflate.
Pentosan polysulfate is a semi-synthetic compound whose polysaccharide backbone, xylan is extracted from the bark of the beech tree or other plant sources and then treated with sulfating agents such as chlorosulfonic acid or sulfuryl chloride and acid. After sulfation, Pentosan polysulfate is usually treated with sodium hydroxide to yield the sodium salt.
Pentosan polysulfate is disclosed in US patent no. 2689848. The process wherein Xylan is treated with chlorosulfonic acid in the presence of pyridine to obtain sulfuric acid ester salt of xylan followed by oxidative depolymerisation in acidic or neutral aqueous medium to obtain depolymerised product which is dialyzed and followed the fractionation process to obtain desired product. However, this process does not provide end product with desired molecular weight.
US patent 2689848 disclosed the process for the production of salts of sulfuric acid ester of Xylan, the steps which comprise oxidizing the aqueous solution of the a salt of a sulfuric acid ester of highly polymeric xylan in an aqueous solution of H2O2 and H2SO4 to depolymerize such highly polymeric xylan ester partially, dialyzing the depolymerized product and fractionating an aqueous solution of the
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dialyzate with an organic water miscible solvent to obtain a fraction having a Zη value between 0.0030 and 0.015 and sulfur content of 13.5 to 17%.
Moreover, the process as disclosed above for the preparation of Pentosan polysulfate is tedious, costly and provides low yield of final product. Moreover, the use of cellophane tubes for dialysis makes process inapplicable at industrial scale up.
Therefore, it is necessary to develop a process for preparation of Pentosan polysulfate sodium which provides good yield as well as operationally simple and easily applicable at an industrial scale.
Surprisingly, the present inventors have found that carrying out the purification of crude Pentosan polysulfate of formula (I) or its salt by dissolving it in water, followed by filtration through NF membrane system and crystallization of resulting mixture from alcohol provides high yield with good purity of Pentosan polysulfate of formula (I) or its salt of molecular weight of 3000 to 10000 Dalton.
Objects of the invention:
It is therefore an object of the present invention is to provide an improved process for the preparation of Pentosan polysulfate of formula (I) or salt thereof.
Another object of the present invention is to provide the process for the preparation of Pentosan polysulfate of formula (I) or its salt which is operationally simple, easy to handle and applicable at an industrial scale.
A furthermore object of the present invention is to provide an amorphous form of Pentosan polysulfate of formula (I) or its salt thereof.
Another object of the present invention is to provide an amorphous form of Pentosan polysulfate sodium.
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Yet another object of the present invention is to provide an improved process for preparation of Pentosan polysulfate of formula (I) or salt thereof, comprising steps of:
i) treating xylan with chlorosulfonic acid in the presence of pyridine
followed by addition of alkali in the presence of alcoholic solvent to obtain salt of sulfuric acid ester of xylan
ii) oxidizing the salt of sulfuric acid ester of xylan with the mixture of H2SO4 and 30% H2O2 to obtain depolymerized crude Pentosan polysulfate of formula (I) or its salt
iii) purifying depolymerized crude Pentosan polysulfate of formula (I) or its salt by dissolving it in water, followed by filtration through NF membrane system
iv) crystallizing the product obtained in the step (iii) from alcohol to obtain pure Pentosan polysulfate of formula (I) or its salt
A further object of the present invention is to provide an improved process for purification of Pentosan polysulfate of formula (I) or salt thereof, comprising steps of:
i) dissolving crude Pentosan polysulfate of formula (I) or its salt in water ii) filtering the reaction mixture obtain in the step (i) with NF membrane
system
iii) crystallizing the resulting product obtained in the step (ii) from alcohol to obtain pure Pentosan polysulfate of formula (I) or its salt
Yet another object of the present invention is to provide an improved process for preparation of Pentosan polysulfate sodium, comprising steps of:
i) treating xylan with chlorosulfonic acid in the presence of pyridine
followed by addition of sodium hydroxide in the presence of alcoholic solvent to obtain sodium salt of sulfuric acid ester of xylan
ii) oxidizing the sodium salt of sulfuric acid ester of xylan with the
mixture of H2SO4 and 30% H2O2 to obtain depolymerized crude Pentosan polysulfate sodium
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iii) purifying depolymerized crude Pentosan polysulfate sodium, by dissolving it in water, followed by filtration through NF membrane system
iv) lyophilizing the product obtained in the step (iii) to obtain pure Pentosan polysulfate sodium
A further object of the present invention is to provide an improved process for purification of Pentosan polysulfate sodium, comprising steps of:
i) dissolving crude Pentosan polysulfate sodium in water
ii) filtering the reaction mixture obtain in the step (i) with NF membrane
system
iii) lyophilizing the resulting product obtained in the step (ii) to obtain pure Pentosan polysulfate sodium
Summary of the invention:
According to one aspect, present invention provides an improved process for the preparation of Pentosan polysulfate of formula (I),

or salt thereof, wherein R represents -SO3Y, and Y is at least one member selected from the group consisting of H and a pharmaceutically acceptable cation such as sodium, potassium, and magnesium said process
comprising steps of:
i) treating xylan with chlorosulfonic acid in the presence of pyridine
followed by addition of alkali in the presence of alcoholic solvent to obtain salt of sulfuric acid ester of xylan;
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ii) oxidizing the salt of sulfuric acid ester of xylan with the mixture of H2SO4 and 30% H2O2 to obtain depolymerized crude Pentosan polysulfate of formula (I) or its salt;
iii) purifying depolymerized crude Pentosan polysulfate of formula (I) or its salt by dissolving it in water, followed by filtration through
NF membrane system;
iv) crystallizing the product obtained in the step (iii) from alcohol to obtain pure Pentosan polysulfate of formula (I) or its salt.
Further aspect of the present invention is to provide an improved process for purification of Pentosan polysulfate of formula (I) or salt thereof, comprising steps of:
i) dissolving crude Pentosan polysulfate of formula (I) or its salt in
water;
ii) filtering the reaction mixture obtain in the step (i) with NF
membrane system;
iii) crystallizing the resulting product obtained in the step (ii) from alcohol to obtain pure Pentosan polysulfate of formula (I) or its salt.
Further aspect of the present invention is to provide an improved process for preparation of Pentosan polysulfate sodium, comprising steps of:
i) treating xylan with chlorosulfonic acid in the presence of pyridine
followed by addition of sodium hydroxide in the presence of
alcoholic solvent to obtain sodium salt of sulfuric acid ester of
xylan;
ii) oxidizing the sodium salt of sulfuric acid ester of xylan with the
mixture of H2SO4 and 30% H2O2 to obtain depolymerized crude
Pentosan polysulfate sodium;
iii) purifying depolymerized crude Pentosan polysulfate sodium by
dissolving it in water, followed by filtration through NF membrane
system;
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iv) lyophilizing the product obtained in the step (iii) to obtain pure Pentosan polysulfate sodium.
A further object of the present invention is to provide an improved process for purification of Pentosan polysulfate sodium, comprising steps of:
i) dissolving crude Pentosan polysulfate sodium in water
ii) filtering the reaction mixture obtain in the step (i) with NF
membrane system
iii) lyophilizing the resulting product obtained in the step (ii) to obtain pure Pentosan polysulfate sodium
Brief description of the drawings:
Figure-1 represents the powder X-ray diffraction (XRD) pattern of an amorphous form of Pentosan polysulfate sodium.
Detailed description of the Invention
The process of the present invention for preparation of Pentosan polysulfate of formula (I) or salt thereof, is operationally simple, easy to handle and applicable at an industrial scale.
The present invention provides an improved process for preparation of Pentosan polysulfate of formula (I) or salt thereof, comprising steps of:
i) treating xylan with chlorosulfonic acid in the presence of pyridine
followed by addition of alkali in the presence of alcoholic solvent
to obtain salt of sulfuric acid ester of xylan
ii) oxidizing the salt of sulfuric acid ester of xylan with the mixture of
H2SO4 and 30% H2O2 to obtain depolymerized crude Pentosan
polysulfate of formula (I) or its salt
iii) purifying depolymerized crude Pentosan polysulfate of formula (I)
or its salt by dissolving it in water, followed by filtration through
NF membrane system
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iv) crystallizing the product obtained in the step (iii) from alcohol to obtain pure Pentosan polysulfate of formula (I) or its salt
The suitable alkali in step (i) is selected from alkali metal salt of carbonate, bicarbonate, hydroxide or mixtures thereof. The most preferable is sodium hydroxide. The suitable alcoholic solvent in step (i) is selected from methanol, ethanol, propanol, isopropanaol, butanol or mixtures thereof. The most preferable is methanol. The suitable alcohol in step (iv) is selected from methanol, ethanol, propanol, isopropanaol, butanol or mixtures thereof. The most preferable is methanol. Crystallization provides an amorphous form of Pentosan polysulfate of formula (I) or its salt.
The present invention also provides an improved process for preparation of Pentosan polysulfate sodium, comprising steps of:
i) treating xylan with chlorosulfonic acid in the presence of
pyridine followed by addition of sodium hydroxide in the
presence of alcoholic solvent to obtain sodium salt of sulfuric
acid ester of xylan
ii) oxidizing the sodium salt of sulfuric acid ester of xylan with the
mixture of H2SO4 and 30% H2O2 to obtain depolymerized
crude Pentosan polysulfate sodium
iii) purifying depolymerized crude Pentosan polysulfate sodium by
dissolving it in water, followed by filtration through NF
membrane system
iv) lyophilizing the product obtained in the step (iii) to obtain pure
Pentosan polysulfate sodium
The suitable alcoholic solvent in step (i) is selected from methanol, ethanol, propanol, isopropanaol, butanol or mixtures thereof. The most preferable is methanol. Lyophilization provides an amorphous form of Pentosan polysulfate of formula (I) or its salt.
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According to present invention, pyridine is added in chlorosulfonic acid slowly at below 40-50°C. The resulting salt was heated to 60-65°C before addition of xylan and raised the temperature to 75-80°C. After completion of reaction, reaction mixture is cooled to 25-30°C and added methanol to isolate the pyridinium salt. The isolated pyridinium salt is dissolved in water and decolorized with chlorine dioxide, then it is quenched with alkaline methanol. It is then adjusted pH 6.5-7.0 with glacial acetic acid, filtered and dried the material at 50-55°C under reduced pressure to get the sodium salt of sulfuric acid ester of xylan.
Sodium salt of sulfuric acid ester of xylan is dissolved in water and heated to 90-100°C, then added preheated mixture of 5N H2SO4 and 30% hydrogen peroxide. The reaction mixture is maintained at 90-100°C. After completion of reaction, the reaction mixture is cooled to 25-30°C and adjusted pH 7.5-7.3 with 5N sodium hydroxide solution. Then this reaction mixture is precipitated with methanol, filtered and dried to get the depolymerized product i.e. crude Pentosan polysulfate sodium.
The depolymerized product is dissolved in water and subjected to NF Membrane system of cut off molecular weight of 3000 Dalton and monitored the rejection of lower molecular weight material with gel permeation chromatography. After the completion of reaction, the concentrate was subjected to water recovery at 40-50°C. Finally residue was crystallized with methanol to get the pure amorphous form of Pentosan polysulfate sodium having value 0.006 and Molecular weight of 6543 Dalton. X-ray powder diffraction pattern is matching with Figure-1.
Moreover, after the completion of filtration, the concentrate is transferred in a single neck round bottom flask and fitted to Lyophilizer (BT4KZL-105) and starts the operation of lyophilyzation. It takes about 24 hr to complete the lyophilyzation
and provides an amorphous form of Pentosan polysulfate having Zη value -0.007
and Mol weight of 5406 Dalton. X-ray powder diffraction pattern is matching with Figure-1.
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In the present invention, the purification step as discussed hereinabove is carried out by using "PERMA" Pilot Membrane System manufactured by PERMIONIC Membranes Pvt. Ltd., 3/29, 4/19, B.I.D.C, Gorwa Estate, Vadodara-390016, Gujarat, India.
In accordance with the aspects of this invention, purification of crude Pentosan polysulfate of formula (I) or its salt is carried out in following manner: The crude Pentosan polysulfate of formula (I) or its salt is dissolved in water. Feeding the resultant solution and allow it to pass through micron filter membrane system and concentrate flow meter, thereby filtering out the Pentosan polysulfate of formula (I) or its salt with desired molecular weight in the range of 3000 to 10000 Dalton.
For the purpose of this specification, the meaning of the term "NF membrane system" is nano-filtration accompanied by membrane which is capable to pass the undesired product having molecular weight of at least 3000 Dalton.
Advantages of the present invention over prior art:
i) The present invention provides process which is economical,
operationally and industrially applicable,
ii) The present invention provides cheaper as well as less time consuming
process for the preparation of Pentosan polysulfate sodium,
iii) The process is simple and easy to handle and does not require special
handling care or critical conditions,
iv) The process provides the desired Pentosan polysulfate sodium with
molecular weight of 3000 to 10000 Dalton specifically.
The process of the present invention is described by the following examples, which are illustrative only and should not be construed so as to limit the scope of the invention in any manner.
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Example-1
To a three neck 31t round bottom flask the pyridine (800ml) and chlorosulfonic
acid (200ml) was added slowly at below 40-50° C. The resulting salt was heated to 60-65°C then added xylan (l00g) and raised the temperature to 75-80° C. After completion of reaction, reaction mixture was cooled to 25-30 C and methanol was added (2500ml) to isolate the pyridine salt of xylan. The isolated pyridine salt of xylan was dissolved in water 500ml and decolorized with chlorine dioxide (200ml), then it was added a mixture of methanol (2000ml) and 33% Sodium hydroxide (200ml). It was then adjusted to pH to neutral with acetic acid, filtered and dried the material at 50-55° C under reduced pressure to get 240 gm sodium salt of sulfuric acid ester of xylan.
To a three neck 31t round bottom flask charged sodium salt of sulfuric acid ester of xylan (500g) in water (1500ml) and heated to 90-100°C, then added the preheated mixture of 5N sulfuric acid (25ml) and 30% hydrogen peroxide (250ml). Monitored the reaction with HPLC, after completion of reaction till average molecular weight came about 4000-8000 Dalton, cooled the reaction mixture and solution was adjusted pH to neutral with dilute sodium hydroxide solution. Then the reaction mixture was precipitated with methanol (121t), filtered and dried at 50-55 C to get crude Pentosan polysulfate sodium having yield of 450 gm and Zη value -0.006.
The 450 gm of the crude Pentosan polysulfate sodium was dissolved in water (201t) and subjected to the NF membrane of cut off molecular weight of 3000 Dalton, filtration. The NF Membrane system was monitored the rejection of lower molecular weight material with HPLC. After the completion of filtration, the concentrate was subjected to water recovery at 40-50°C under reduced pressure up to 340ml. Finally residue was crystallized with methanol (2.71t) to get the pure Pentosan polysulfate sodium. Filtered the material and dried it at 45-50°C under vacuum. The yield of pure Pentosan polysulfate sodium was 170gm having Zη
value 0.006 and Molecular weight of 6543 Dalton.
X-ray powder diffraction pattern is matching with Figure-1.
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Example-2
To a three neck 3 It round bottom flask the pyridine (800ml) and chlorosulfonic acid (200ml) was added slowly at below 40-50° C. The resulting salt was heated to 60-65°C then added xylan (l00g) and raised the temperature to 75-80° C. After completion of reaction, the reaction mass was cooled 25-30°C and added methanol (2500ml) to isolate the pyridine salt of xylan. The isolated pyridine salt of xylan was dissolved in water 500ml and decolorized with chlorine dioxide (200ml), then it was added a mixture of methanol (2000ml) and 33% Sodium hydroxide (200ml). It was then adjusted to pH to neutral with acetic acid, filtered and dried the material at 50-55° C under reduced pressure to get 240 gm of sodium salt of sulfuric acid ester of xylan.
To a three neck 1 It round bottom flask, charged sodium salt of sulfuric acid ester of xylan (200g) in water (600ml). The reaction was heated to 90-100°C, then
added preheated mixture of 5N sulfuric acid (10ml) and 30% hydrogen peroxide (100ml). Monitored the reaction with HPLC after completion of reaction, till average molecule weight came about 4000-8000 Dalton cooled the reaction mixture and the solution was adjusted pH to neutral with diluted sodium hydroxide solution. Then the reaction mixture was subjected to NF membrane system.
The above reaction mixture was diluted with water (201t) and subjected to the NF membrane of cut off molecular weight of 3000 Dalton, filtration. The NF Membrane system was monitored the rejection of lower molecular weight material with HPLC. After the completion of filtration, the concentrate was transferred in a single neck round bottom flask and fitted to Lyophilizer (BT4KZL-105) and started the operation of lyophilyzation. It took about 24 hr to complete the lyophilyzation. The yield of amorphous form of Pentosan polysulfate was 60gm and Mol wt.-5406 and Zn value -0.007. X-ray powder diffraction pattern is matching with Figure-1.
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We claim:
1. A process for preparation of Pentosan polysulfate of formula (I)

or salts thereof, wherein R represents -SO3Y, and Y is at least one member
selected from the group consisting of H and a pharmaceutically acceptable
cation such as sodium, potassium, and magnesium,
comprising steps of:
i) treating xylan with chlorosulfonic acid in the presence of pyridine
followed by addition of alkali in the presence of alcoholic solvent
to obtain salt of sulfuric acid ester of xylan
ii) oxidizing the salt of sulfuric acid ester of xylan with the mixture of
H2SO4 and 30% H2O2 to obtain depolymerized crude Pentosan
polysulfate of formula (I) or its salt
iii) purifying depolymerized crude Pentosan polysulfate of formula (I)
or its sah by filtration through NF meifrbrane system-
iv) crystallizing the product obtained in the step (iii) from alcohol to
obtain pure Pentosan polysulfate of formula (I) or its salt
2. A process as claimed in claim 1, wherein said alkali in step (i) is selected from alkali metal salt of carbonate, bicarbonate, hydroxide or mixtures thereof.
3. A process as claimed in claim 1, wherein said alcoholic solvent in step (i) is selected from methanol, ethanol, propanol, isopropanaol, butanol or mixtures thereof.
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4. A process as claimed in claim 1, wherein said purification in step (iii) is carried out by dissolving crude Pentosan polysulfate of formula (I) or its salt in water and followed by filtration through NF membrane system.
5. A process as claimed in claim 1, wherein said alcohol in step (iv) is selected from methanol, ethanol, propanol, isopropanaol, butanol or mixtures thereof.
6. A process for preparation of an amorphous form of Pentosan polysulfate Sodium, comprising steps of:
i) treating xylan with chlorosulfonic acid in the presence of pyridine
followed by addition of sodium hydroxide in the presence of methanol to obtain sodium salt of sulfuric acid ester of xylan
ii) oxidizing the sodium salt of sulfuric acid ester of xylan with the mixture of H2SO4 and 30% H2O2 to obtain depolymerized crude Pentosan polysulfate sodium
iii) purifying depolymerized crude Pentosan polysulfate sodium by filtration through NF membrane system
iv) crystallizing the product obtained in the step (iii) from methanol to obtain an amorphous form of Pentosan polysulfate sodium
7. A process as claimed in claim 6, wherein said purification in step (iii) is carried out by dissolving crude Pentosan polysulfate sodium in water and followed by filtration through NF membrane system.
8. An amorphous form of Pentosan polysulfate sodium with molecular weight of range 3000 to 10000 Dalton as shown in figure-1.
9. A process for the purification of Pentosan polysulfate of formula (I) or salts thereof, comprising steps of:
i) dissolving crude Pentosan polysulfate of formula (I) or its salt in
water
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ii) filtering the reaction mixture obtain in the step (i) with NF
membrane system
iii) crystallizing the resulting product obtained in the step (ii) from
alcohol to obtain pure Pentosan polysulfate of formula (I) or its salt
10. A process as claimed in claim 9, wherein said alcohol in step (iii) is selected from methanol, ethanol, propanol, isopropanaol, butanol or mixtures thereof.
11. A process for the purification of Pentosan polysulfate sodium, comprising steps of:
i) dissolving crude Pentosan polysulfate sodium in water
ii) filtering the reaction mixture obtain in the step (i) with NF
membrane system
iii) crystallizing the resulting product obtained in the step (ii) from
alcohol to obtain pure Pentosan polysulfate sodium
12. A process as claimed in claim 11, wherein said alcohol in step (iii) is selected from methanol, ethanol, propanol, isopropanaol, butanol or mixtures thereof.
13. A process for the preparation of Pentosan polysulfate sodium comprising a step of purifying depolymerized crude Pentosan polysulfate sodium by filtration through NF membrane system.
14. A process as claimed in claim 13, wherein said purification is carried out by dissolving crude Pentosan polysulfate sodium in water and followed by filtration through NF membrane system.
15. A process for preparation of amorphous form of Pentosan polysulfate sodium, comprising steps of:
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i) treating xylan with chlorosulfonic acid in the presence of pyridine
followed by addition of sodium hydroxide in the presence of
alcoholic solvent to obtain sodium salt of sulfuric acid ester of
xylan
ii) oxidizing the sodium salt of sulfuric acid ester of xylan with the
mixture of H2SO4 and 30% H2O2 to obtain depolymerized crude
Pentosan polysulfate sodium
iii) purifying depolymerized crude Pentosan polysulfate sodium by
filtration through NF membrane system
iv) lyophilizing the product obtained in the step (iii) to obtain
amorphous form of Pentosan polysulfate sodium
16. A process as claimed in claim 15, wherein said alcoholic solvent in step (i) is selected from methanol, ethanol, propanol, isopropanaol, butanol or mixtures thereof.
17. A process as claimed in claim 15, wherein said purification in step (iii) is carried out by dissolving crude Pentosan polysulfate sodium in water and followed by filtration through NF membrane system.
18. A process for the purification of Pentosan polysulfate sodium, comprising steps of:
i) dissolving crude Pentosan polysulfate sodium in water ii) filtering the reaction mixture obtain in the step (i) with NF membrane system
iii) lyophilizing the resulting product obtained in the step (ii) to obtain pure Pentosan polysulfate sodium
Dated this 6th day of March 2007
Ashwini Sandu
Of S. Majumdar & Co. Applicant's Agent
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ABSTRACT
A process for the preparation of Pentosan polysulfate sodium comprising a step of purifying depolymerized crude Pentosan polysulfate sodium by filtration through NF membrane system.