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THE PATENTS ACT, 1970 (39 of 1970)
COMPLETE SPECIFICATION (See section 10 and rule 13)
1. TITLE OF THE INVENTION
"Process for production of Tri Choline Citrate and Composition thereof


2. APPLICANTS)
1 .NAME : Bharat Shantilaf Shah
2.NAT10NALITY: Indian
3-ADDRESS: B/20,108 Ashwini, Apna Ghar, Unit # 5,Lokhandwala Complex Andheri (West), Mumbai-400053, India
The following specification particularly describes the invention and the manner in which it is to be performed

24 JUN 2008

FIELD OF THE INVENTION
The present invention is related to novel process for production of Tricholine Citrate by reacting citric acid with choline base at high temperature to obtained more than 65 percent pure of Tricholine Citrate. Such a obtained Tricholine Citrate is used in various composition for used as an effective iron chelator and used to stabilize animal growth hormone preparations also.
The use of Tricholine Citrate and its composition as therapeutic agents has increased considerably and these agents now occupy an important place both in therapy and as a component along with other vitamins in various dietary supplements. However, because of the nature of choline and its salts it has been difficult to prepare choline at high temperature and its composition with the high purities of Tricholine Citrate necessary when these products are intended for human application Various physical properties and state of matter are also requisite when the products are to be incorporated into therapeutic use along with other materials. Efficient and economical means of producing Tricholine Citrate and its composition with pure Tricholine Citrate, therefore, desirable and necessary.
BACKGROUND OF THE INVENTION
Tricholine citrate (hereinafter "TCC") is an ester of citric acid with three choline molecules TCC is a common inexpensive chemical used in the past as an effective iron chelator. In U.S. Pat. No. 2,865,938. TCC has been used to stabilize animal growth hormone preparations and to treat iron deficiency anemia in U.S. Pat. No. 3,395,229. No toxicity or adverse effects have been associated with this molecule since its individual components, citric acid and choline, are naturally occurring metabolites in cells.
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Variety of stabilizers Compositions have been disclosed in the art which avoid the breakdown of the structures of proteins. For example, glycerol has been used to stabilize the activity of various proteins. Gekko, et al, Biochemistry (1981), 20, pp. 4666-76. Examples of proteins which are described in this article include chymotrypsinogen A (from bovine pancreas), ribonuclease A (from bovine pancreas), .beta.-lactoglobin (from milk), bovine serum albumin, insulin (bovine pancreatic), egg-white lysozyme and .alpha.-chymotrypsin.
U.S. Pat. No. 4,179,337 discloses a process for coupling a polypeptide such as enzymes and insulin to polyethylene glycol or polypropylene glycol having a molecular weight of 500 to 20,000 daltons. The polyethylene glycol or polypropylene glycol is described as protecting the polypeptide from loss of activity and the composition can be injected without any immunogenic response.
While the prior art has taught a number of various stabilizers for specific proteins, unfortunately, the fact a particular stabilizer is effective with a particular protein does not necessarily mean that the particular stabilizer is appropriate for the stabilization of a growth promoting hormone. Therefore, there exists a need for a method of stabilization of a growth promoting hormone which decreases the formation of insolubles and preserves the soluble bioactivity of the hormone.
US patent 2774759 discloses process by which an aqueous solution of trimethylamine is reacted with ethylene oxide, the resulting aqueous solution of choline base can be used directly to prepare various cho- line salts by neutralization of the choline base with an appropriate acid. In the described procedure, several steps are involved and the use of costly solvents is required which is an economic disadvantage in viw of low conversion and low purity of product as reaction temperature is below 30 degree Celsius.
US 4,917,685 patent discloses delivery device designed to be implanted in an animal and for the administration of a stabilized growth promoting formulation. This device has a wall formed at least in part by a porous membrane which surrounds and defines a
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reservoir. The reservoir contains a growth promoting hormone and a stabilizer. Examples of such stabilizers include certain polyols, amino acids, polymers of amino acids having a charged side group at physiological pH, and choline salts.
US 5,209,858 patent discloses The stabilization of aqueous or alkanohc solutions of choline and derivative of choline against discoloration on aging by the addition of an unsubstituted hydroxylamine salt to the choline solutions at levels of less than 1 percent.
US 4,816,568 patent discloses that Growth hormones are admixed with various stabilizers to provide for the decreased formation of insolubles and preservation of the soluble bioactivity of the growth hormone in aqueous environments. Examples of such stabilizers include certain polyols, amino acids, polymers of amino acids having a charged side group at physiological PH, and choline salts.
OBJECIVE OF INVENTION
The objective of the present invention to give an economic advantage over the standard procedures since not only is an aqueous procedure more economical than other process, but it is not necessary to go through any steps to isolate the choline base as was necessary heretofore, nor are any costly solvents required since water is the only solvent used. It has been proposed to prepare tricholine citrate by reacting approximately stoichiometric amounts of choline base with citric acid.
Another objective of the present invention is to disclose process wherein the choline base is reacted with citric acid at high temperature means above 50 degree Celsius to achieve high conversion of tricholine citrate. The tricholine citrate thus formed is with law contaminated which is formed as a consequence of the neutralization of the choline base with citric acid.
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Another objective of the present invention is to disclose process wherein the choline base is reacted with citric acid at high temperature means above 50 degree Celsius and product is distilled out in distillation column to give high yield of tricholine citrate . The tricholine citrate is obtained with distillation process to give high-end yield. In a preferred embodiment of the present invention, we have discovered that by the use of single step of reaction between Citric Acid and Choline Base and distillation of formed Tricholine citrate in only one step. Thus reaction between the reactants and distillation is carried out in same vessel or in single step
Another objective of the present invention is to disclose process wherein the choline base is reacted with citric acid at high temperature to give mixture of tricholine citrate which is filter inside or outside of distillation column . If solution of tricholine citrate is filtered outside of Distillation column then it is re fed to distillation column.
Another objective of the present invention is to disclose process wherein the choline base is reacted with citric acid at high temperature to give mixture of tricholine citrate and distillation of filtered tricholine citrate is done distillation column to reduce the heat loss in the reaction and reuse the heat of distillation for carrying out reaction at high temperature above 50 degree Celsius
Another objective of the present invention is to disclose process wherein the choline base is reacted with citric acid at high temperature to give mixture of tricholine citrate and distillation of filtered tricholine citrate is done distillation column to increase the efficiency of the reaction by again recycling unreacted reactants in same reacting zone.
Another objective of the present invention is to disclose process wherein the choline base is reacted with citric acid at high temperature to give mixture of tricholine citrate and distillation of filtered tricholine citrate is done distillation column to obtained at least 65% of tricholine citrate.
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Another preterred embodiment of the present invention is that to provide composition ot Tricholine citrate in high purest form which by the use of single step of reaction between Citric Acid and Choline Base for use in stabilizers Compositions of a growth promoting hormone, therapeutic composition and in Cosmetic Compositions.
SUMMARY OF THE INVENTION
The invention provides a novel process for of production of Tricholine Citrate by reacting citric acid with choline base solution at high temperature and distillation of Tricholine Citrate at the same time in single step.
In the process, citric acid is reacted with choline base solution at above 50 degree Celsius by adjusting pH of solution above 8.5 de-colorizing the solution obtained by de colorizing agent. The obtained product is filtered at temperature below the 40 degree Celsius in filtration unit and then distilled in Distillation Column to obtained high purity of Tricholine Citrate
In this Process, reaction between Citric Acid and Choline base solution and distillation of final product are carried out in same Distillation Column which is maintained at variable temperature.
Thus high purity product obtained is used in Composition comprising from 0.1 to 70 wt. % by weight of the composition of 65 % Tricholine Citrate as in stabilizers Compositions of a growth promoting hormone, therapeutic composition and in Cosmetic Compositions.
BRIEF DESACRIPTION OF DRAWING
In Figure 1, R shows Variable Temperature Distillation Column and A is Choline Base Solution Feedline whereas B is Citric acid feedline . L shows reaction Zone where as C
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and D are Decolorizing Zone and Filtering Zone Respectively. W is waste line for purging waste where as E is Feedline to distillation column in Distillation Zone Q. P is product out let from which pure Tricholine Citrate is taken out.
DETAILED DESCRIPTION
The present invention is based upon the novel process for production of Tricholine Citrate by reacting Citric Acid with Choline base solution at high temperature and distillation of Tricholine Citrate at the same time in single step.
The citric acid is reacted at high temperature above 50 degree Celsius as follows
Citic" acid + tCholine baser above 56 degree Calsius Thonoline citrate -+ water
The Choline base can be prepared by solution of trimethylamne is reacted with ethylene oxide , the resulting solution of choline base solution, The choline base solution thus obatained can be use directally for preparation of Tritholine Citrate. But choline base solution must be purified for such reaction and should be with minimum traces of trimethylamne. If the percentage of impurities of trimethylamine is more then final base solution will give low productivity of the Tricholine Citrite.
The Tricholine Citrate is denoted by formula I as:
CH,CQO" [(CH,)— N!—CH3CH20H0).H3O— C— CQQ-
CH7COo-
In the present invention citric acid and choline base can be used directly in 30% and 60% by w/w respectively during reaction overall pH 0f the solution is basic and this
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are maintain by choline base only in s.s reactor and or in distillation column what ever pH of the solution is maintain above 8.5 pH.
The above process is carried out a above the 50 degree Celsius for better conversion Choline base solution to Tricholine Citrate so that all the choline base solution is coverted to tricholine Citrate. It is found that even if there is impuritied of trimethylamne in Choline base solution is high even then there is best conversion to Tricholine Citrate in Distillation Column which is designed for purification. Thus reaction between Citric acid and Choline base solution is carried out above 50 degree Celsius. But preferred range of working of reaction temperature is in the range of 56 to 90 degree Celsius and more prefired range of working for carrying put reaction is between 61 to 85 degree Celsius.
The reaction between citric acid and choline base can be taken place in distillation column which is maintained at variable temperature. The advantage of the above reaction which takes place in the distillation column is that reaction between Citric acid and choline base as well as distillation of Formed Tricholine Citrate solution is taken place in same column thus distillation heat of the distillation column is used . The distillation temperature which is more than the reaction temperature fall in the same line of Variable Distillation Column.
Thus Tricholine Citrate solution Formed is then decolorized by means of decolorizing agent. In discoloring process, unwanted color of the solution is removed. Decolorisation can be done after distillation of the Tricholine Citrate.
In distillation column decolorisation is done below the reaction zone in distillation column. The decolorization can be done in full manner or in partial manner. This means decolorisation is done of full down flow in the distillation column or in partial down flow in the Distillation Column.
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In procedure solution obtained in previous step is coloured can be decolourised by activated charcoal in form of separate section in the lower portion of the Distillation Column. Thus decolorizing unit may be in form of compact bed form which contains activated charcoal. Thus activated Charcoal is good decolorizing agent.
After discoloring unit, formed Tricholine Citrate solution is filtered through Alteration unit to remove un reacted particle and trac es citric acid and or any other impurities. The filtration unit can be out side of distillation column or in bed form in side the distillation column. When the filtration unit is in bed form in side the column, all down flow in the distillation column can be filtered or only partial down flow or only output of de-coloring unit is filtered. Filtration unit as per conventional procedure to remove carbon cake and other unreacted solid particles.
Distillation of product done to get high purity tricholine citrate which must be without any traces of trimethylamne and citric acid . Thus output of Filtration is loaded in upper section of distillation column. Thus in distillation column, high volatile compound and low volatile compounds are formed. High volatile compound is rich in high purity of Tricholine Citrate which is collected from top of the Distillation Column. Low volatile compound contains unreacted trimethylamne , ethylene oxide, unreacted choline base and citric acid traces which form part of down flow of Distillation Column.
In above Process of production of Tricholine Citrate wherein reaction between Citric Acid and Choline base solution is carried out in Distillation Column which is maintained at variable temperature. Thus there will be four section at large distillation zone, reaction zone , decolouring zone and filtration zone. Thus distillation zone is maintained above 90 degree Celsius and reaction zone is maintained above 50 degree Celsius but below 90 degree Celsius for earring out reaction between choline base solution and citric acid . Thus decoluring unit and filtration unit or zones will be at lower temperature giving the whole unit variable temperature unit.
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In above Process of production of Tricholine Citrate wherein Citric Acid and Choline
base solution are fed at different feed line. Thus Choline base solution feed line must be
above the feedline of Citric acid to dissolve the citric acid fully and to give high
conversion of choline base solution and citric acid. Hence Choline base solution is
always fed above the Citric Acid feed line.
In above Process, decolorisation is done and then filtration is done inside Distillation
column or outside the distillation column and when filtration is done outside the
distillation column, filtered solution is re-fed above the Choline base solution feed line in
Distillation Column.
In process, distillation of the final product at above 90 degree Celsius to obtain pure
Tricholine Citrate can be made.
Process of production of Tricholine Citrate wherein choline base solution is directly fed
to distillation column through Choline base solution feed line without loosing any heat
energy. Thus use of heat of distillation column is made to carry out the reaction. Thus
high purity Tricholine Citrate is obtained from distillation column at the end of the
process with at least 65% of tricholine citrate.
The tricholine citrate is obtained with distillation process to give high-end yield.
In the process, citric acid is reacted with choline base solution at above 50 degree Celsius
by adjusting pH of solution above 8.5 de-colorizing the solution obtained by de
colorizing agent. The obtained product is filtered at temperature below the 40 degree
Celsius in filtration unit and then distilled in Distillation Column to obtained high purity
of Tricholine Citrate
In this Process, reaction between Citric Acid and Choline base solution and distillation of
final product are carried out in same Distillation Column which is maintained at variable
temperature.
Thus high purity product obtained is used in Composition comprising from 0.1 to 70 wt. % by weight of the composition of 65 % Tricholine Citrate as in stabilizers Compositions of a growth promoting hormone, therapeutic composition and in Cosmetic Compositions.
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In general, the amount of 65 % Tricholine Citrate in the present compositions is in the range of from about 0.1% to 70% by weight of the composition. Preferably, in order to lower cost and maximize the effect, the amount of Tricholine Citrate is in the range of from about 0.1% to about 10%, most preferably in the range of from 0.1% to 5%.
Compositions of the invention are intended for topical application to mammalian skin which is already in dry, flaky, wrinkled, aged, photodamaged condition or which suffers from a skin disorder, or, in the alternative, the inventive compositions may be applied prophylactically to normal healthy skin to prevent or reduce the deteriorative changes.
In another Composition is formed comprising from 0.1 to 70 wt % by weight of the composition of 65 % Tricholine Citrate, a cosmetically acceptable vehicle, and an additional cosmetic ingredient selected from the group consisting of an alpha hydroxy acid, a sunscreen, a tanning agent, a skin anti-wrinkling agent, an anti-dandruff agent, an anti-acne agent, a hair growth stimulant and a perfume.
Another use of tricholine citrate as of stabilizers of 0.1 to 70 wt. % by weight of the composition of 65 % in amounts but in preferable way of from about 2.5 to 55 percent by weight of the total weight of the overall formulation prior to introduction into an aqueous environment. Preferably, the choline derivative is present in amounts of from about 15 to about 60 percent by weight of the total weight of the overall formulation
Thus Composition comprising from 0.1 to 70 wt. % by weight of the composition of 65 % Tricholine Citrate, also comprises of growth hormone and a stabilizing amount of a stabilizer.
Thus Examples can be given for carrying out process and composition thereof for end use but there these are not limited to disclosed herein
Example 1
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To 302.5 parts of a 40% aqueous choline base solution is added 284 parts of citric acid,. After stirring in static mixer inline to feed to distillation column in reaction zone , the solution of choline state is given a decolorization treatment with activated charcoal and then filtered the clarified solution is then distilled to slurry . this is poured on to dryer trays and then dred, giving the final Tricholine Citrate is obtained
Example 2
To 600.5 parts of a 40% aqueous choline base solution is added 560 parts of citric acid ,. After stirring in static mixer inline to feed to distillation column in reaction zone , the solution of choline state is given a decolorization treatment with activated charcoal and then filtered the clarified solution is then distilled to slurry . this is poured on to dryer trays and then dred , giving the final Tricholine Citrate is obtained
Example 3
Aqueous solutions of choline base which had been previously decolorized using Darco G-60 decolorizing carbon. The spent carbon was filtered off, leaving a water-white filtrate of choline. To portions of this filtrate were added the color stabilizer component, e.g., a salt of hydroxylamine in the indicated amounts. Sample vials containing the indicated percentages of stabilizer were stored at room temperature as well as in block heaters held at 50degree. and 70.degree. C.
Example 4
To 1500 parts of a 40% aqueous choline base solution is added 1280 parts of citric acid , After stirring in static mixer inline to feed to distillation column in reaction zone , the solution of choline state is given a decolorization treatment with activated charcoal and
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then filtered the clarified solution is then distilled to slurry . this is poured on to dryer trays and then dried , giving the final Tricholine Citrate is obtained
Example 5
Composition comprising from 35 wt. % by weight of the composition of 65 % Tricholine Citrate, a cosmetically acceptable vehicle 3 wt. % by , and an additional cosmetic ingredient selected from the group consisting of an alpha hydroxy acid 13 wt. % by, a sunscreen, a tanning agent, a skin anti-wrinkling agent, an anti-dandruff agent, an anti¬acne agent, a hair growth stimulant and a perfume all make to 100 percent.
Example 6
Composition comprising from 15 wt. % by weight of the composition of 65 % Tricholine Citrate, also comprises of growth hormone 35 wt. % and a Tricholine Citrate is incorporated into phospholipids of keratinocytes, cells were labelled with 1 uCi .sup.3 H choline/ml medium for 48 hours in the presence of different concentrations of the different citrate esters, citric acid or choline. The amount of .sup.3 H choline incorporated into cellular phospholipids was determined and expressed as % of controls. The amount of .sup.3 H choline incorporated into phospholipid fraction in the control was 252 .+-.cpm/.mu.g protein
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I/We Claim
1. Process of production of Tricholine citrate by reacting citric acid with choline base solution at high temperature to form Tricholine Citrate comprising steps of
a) reacting citric acid with choline base solution at above 50 degree Celsius by adjusting pH of solution above 8.5
b) de-colorizing the solution obtained by de colorizing agent
c) filtering the obtained material at temperature below the 40 degree Celsius in filtration unit
d) distillating the solution obtained in Distillation Column designed to obtained high purity of Tricholine Citrate
2. Process of production of Tricholine Citrate wherein reaction between Citric Acid and
Choline base solution is carried out in Distillation Column which is maintained at
variable temperature.
3. Process of production of Tricholine Citrate wherein Citric Acid and Choline base
solution are fed at different feed line.
4. Process of production of Tricholine Citrate wherein Choline base solution is fed above
the Citric Acid feed line.
5. Process of production of Tricholine Citrate wherein filtration is done inside Distillation
column or outside the distillation column
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6. Process of production of Tricholine Citrate wherein when filtration is done outside
the distillation column, filtered solution is re-fed above the Choline base solution feed line in Distillation Column.
7. Process of production of Tricholine Citrate wherein distillating the final product at
above 90 degree Celsius to obtain pure Tricholine Citrate
8. Process of production of Tricholine Citrate wherein choline base solution is directly fed to distillation column through Choline base solution feed line without loosing any heat energy.
9. Composition comprising from 0.1 to 70 wt. % by weight of the composition of 65 %
Tricholine Citrate, a cosmetically acceptable vehicle, and an additional cosmetic ingredient selected from the group consisting of an alpha hydroxy acid, a sunscreen, a tanning agent, a skin anti-wrinkling agent, an anti-dandruff agent, an anti-acne agent, a hair growth stimulant and a perfume.
10. Composition comprising from 0.1 to 70 wt. % by weight of the composition of 65 %
Tricholine Citrate, also comprises of growth hormone and a stabilizing amount of a
stabilizer.
Date: 20th June 2008 (Signature of applicant])
Name of Applicant: Bharat Shantilal Shah
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