Title of Invention

A PROCESS OF PREPARING PEGYLATED LIPOSOMAL DRUG DELIVERY SYSTEM FOR DOXORUBICIN HYDROCHLORIDE

Abstract

The delivery by liposome technology gives us the advantage to deliver toxic drugs to the target cells without causing damage to the normal cells. Also, many drugs with low aqueous solubility and/or combination of both water-soluble and lipid soluble drugs can be formulated in the liposomes, increasing the bioavailability of the drug, reducing dosage and also reducing systemic toxicity. Doxorubicin is a toxic antineoplastics agent used in the treatment of Metastatic ovarian cancer refractory to Paclitaxel and Platinum chemotherapy regimen and AIDS related Kaposi syndrome. It is available both in the normal and stealth liposomal injection form. Liposomal doxorubicin delivers the drug 5 to 11 times more to the sarcoma lesions, and also requires frequent dose modification, reduced toxicity to normal cells, reduced number to dosage administration and better patient compliance. The Pegylatation was carried out by Poly Ethylene Glycol (PEG). Pegylated Doxorubicin Liposomal Injection has a good chemical stability which is much better then the innovator's product in terms of physical and chemical stability as per the in-house specifications.

Full Text

FORM 2 THE PATENTS ACT, 1970 (39 of 1970) & THE PATENTS RULES, 2003 COMPLETE SPECIFICATION [See section 10 and rule 13] Title: PREPARATION OF PEGYLATED LIPOSOMAL DRUG DELIVERY SYSTEM FOR DOXORUBICIN HYDROCHLORIDE Applicant: Claris Lifesciences Limited, Claris Corporate Headquarters. Nr.Parimal Crossing, Ellisbridge, Ahmedabad-380 006, Gujarat, India. THE FOLLOWING SPECIFICATION PARTICULARLY DESCRIBES THE INVENTION AND THE MANNER IN WHICH IT IS TO BE PERFORMED. Description FIELD OF INVENTION [0001] Preparation of Pegylated Liposomal Drug Delivery System for Doxorubicin Hydrochloride Drug. BACKGROUND OF THE INVENTION [0002] A. Pegylation Pegylation is the covalent coupling of non-toxic, hydrophilic polyethylene glycol (PEG) to active or inactive pharmaceutical ingredients. It is a well-accepted method for the delivery of biopharmaceuticals especially peptide and protein drugs. Simple modification with polyethylene glycol (PEG) is not only capable of improving the pharmacological properties of a drug but has also to be considered with regard to its life cycle extension. Polyethylene glycol, or PEG, is a polymer that is nontoxic, nonimmunogenic, highly water soluble, and readily cleared from the body. PEG has many different applications. Pharmaceutical grade PEGs are approved for use in the united states by the FDA and widely used as biopharmaceutical carriers, given their high degree of biocompatibility. [0003] B. Liposomal Doxorubicin hydrochloride Intravenous Therapy [0004] Liposomal drug delivery systems have been proposed for a variety of drugs, particularly those that are administered by parenteral routes. Liposome has potential to prove drug release mainly by two mechanism and that are by Endocytosis and deport formation. Doxorubicin hydrochloride is mainly used as antineoplastic drug. The fermentation products of streptomyces peucetius caesius know as doxorubicin hydrochloride drug. [0005]C. Potential Side Effects- of liposomal Doxorubicin hydrochloride Intravenous Therapy. The potential side effect of doxorubicin hydrochloride drug is the cardiactoxicity. The other side effects are malaise, nausea, vomiting, myelosuppression and sever alopecia liposomal doxorubicin hydrochloride, decreases all the side effects due to increase in targeting the drug delivery at the site of action. [0006] Liposomes are completely close lipid bilayer membranes containing an entrapped aqueous volume of drug. The doxorubicin hydrochloride is entrapped in liquid form in liposome. As liposomes are site specific so it will decrease the dose of doxorubicin and by that way it will decrease the side effect of doxorubicin hydrochloride. Liposome in the body can worked mainly by two mechanisms and that is "endosytosis" and "deport" formation. Due to it's drug releasing patent it will give sustain release of doxorubicin hydrochloride in the body. [0007] A composition of liposomal doxorubicin drug delivery complex comprising a narrower side effect and may yield a safer and more efficacious therapy. There exists a need for a liposomal doxorubicin hydrochloride drug delivery complex preparative method that results in a product with low cost, narrower side effect and distributed at a targeted sites as compared to existing compositions. [0008] SUMMARY OF THE INVENTION [0009] A very common object is to make a pegylated liposome doxorubicin hydrochloride complex, which decreases the side effect of doxorubicin hydrochloride and increases its efficiency by attaching on the targeted cell on the site of action. [0010] Definitions [0011] The term "Homogenization" as employed herein refers to a processes that is use to reduce the particle size of any type of solution. [0012] The term "Hydration" as employed herein refers to a chemical reaction in which a hydroxyl group (OH") and a hydrogen cation (an acidic proton) are added to the two carbon atoms bonded together in the carbon-carbon double bond which makes up an alkene functional group. The reaction usually runs in a strong acidic, aqueous solution. [0013] The term "Liposome" as employed herein refers to a membrane composed of a phospholipid and cholesterol bilayer (see on the right). Liposomes can be composed of naturally-derived phospholipids with mixed lipid chains (like egg phosphatidylethanolamine), or of pure surfactant components like DSPE (1,2-disteroyl-sn-glycero-3-phosphoethanolamine). Liposomes, usually but not by definition, contain a core of aqueous solution; lipid spheres that contain no aqueous material are called micelles, however, reverse micelles can be made to encompass an aqueous environment. [0014] The term "Membrane Extrusion" as employed herein refers to a membrane composed of a method of reducing the size of liposomes is to pass them through a membrane filter of defined pore size. The defined pore size can be accheved at much lower pressures than required for the french pressure cell. There are mainly two ways one is "Tortuous Path" and other is "Nucleation Track" to do membrane extrusion. The process which is used in this patent is "Nucleation Track" method. [0015] The term "Spray drying" as used herein refers to Spray drying is a commonly used method of drying a liquid feed through a hot gas. Typically, this hot gas is air but sensitive materials such as pharmaceuticals, and solvents like ethanol require oxygen-free drying and Nitrogen gas is used instead. The liquid feed varies depending on the material being dried and is not limited to food or pharmaceutical products. This process of drying is a one step rapid process and eliminates additional processing. [0016] DETAILED DESCRIPTION OF THE INVENTION [0017] The present invention provides a process for the preparation of pegylated liposomal doxorubicin hydrochloride complex. [0018] A. Cholesterol Cholesterol is a soft, waxy substance found in all parts of the body. This includes the nervous system, skin, muscle, liver, intestines, and heart. It is made by the body and also obtained from animal products in the diet. [0019] A. Histidine Histidine is one of the 20 most common natural amino acids present in proteins. In the nutritional sense, in humans, histidine is considered an essential amino acid, but mostly only in children. Histidine is found in fruits such as bananas and grapes, meat and poultry, and milk and milk products. It is also found in root vegetables and all green vegetables. though in lesser quantities. [0020] A process of preparing pegylated liposomal doxorubicin Hydrochloride injection and use their of can be describe as following steps: [0021] The first step is preparation of organic mixture by mixing chloroform and methanol in a ratio of 0.5 to 5. [0022] The other solvents are Ethanol.acetone and isopropyl alcohol. [0023] The 30% to 80% of chloroform and methanol is mixed in 500 to 1000 ml volume of bottle. [0024] The bottle with solution is sonicated for 10 minutes by using ultrasonic bath. [0025] The second step is the preparation of lipid solution by taking weighed quantity of Hydrogenated phosphatidylcholine (HSPC). N-(Carbonyl-methoxypolyehtylene glycol 2000)-l,2-disteroyl-sn-glycero-2-phosphoethanolamine sodium salt (MPEG 2000-DSPE) and cholesterol, which is dissolve in organic mixture and after that it is passed through 0.2 micron filter for the filtration process. [026] The concentration of lipid solution is ranged between 10 and 30 percentages. [0027]The solution is taken into bottle and then closed with rubber stopper or aluminum foil. [0028]The bottle is stored at 10 to 35 °Ctemperature. [0029]The selected solution is taken for spray dry. [0030] The third step is spray drying [003 l]The spray drying is carried out at selected inlet and outlet temperature, air pressure. airflow and pump flow. [0032]The inlet temperature is 60 °C. [0033]The inlet temperature is ranged from 50 °C to 90 °C. [0034]The outlet temperature is 36 °C. [0035]The range of outlet temperature is 30°C to 60 °C. [0036]The selected compressed air pressure is ranging from 0.6 to 0.7 bars. [0037]The selected compressed air pressure is ranging from 0.2 to 1.0 bars. [0038] As per the [0037], the airflow is ranging from 6 to 8 meter 2/hour. [0039] As per the [0038], the airflow is ranging from 3 to 10 meter 2/hour. [0040] As per the [0031], the pump flow is 0.2 ml/minute. [0041] As per the [0040], the pump flow is 0.1 to 0.6 ml/minute. [0042] As per the [0031], allowing the assembly along with the spray-dried powder to cool down considerably in an air-conditioned environment. [0043] As per the [0042], there will be an extreme loss of dried powder due to its sticky nature, which may interfere during removal and collection. [0044] As per the [0043], the spray-dried powder is collected in dry glass bottle and check the moisture content of this spray dried powder. [0045] The forth step is preparation of ammonium sulphate solution. [0046] As per the [0045], Ammonium sulphate is taken in flask in a quantity of 1 to 10 mg/ml and it is dissolved with water for injection and the pH is adjusted by using selected solution. [0047] As per the [0046], the selected solution is Hydrochloric acid (HC1) and Sodium Hydroxide (NaOH) to adjust the pH at 5.40. [0048] As per the [0047], the pH is ranged from 4.0 to 6.5. [0049] As per the [0048], after adjusting pH the volume is adjusted with Water for injection again the pH is adjusted as per the standard instruction mention in 4.2 and 4.3 and after that the solution is passed through 0.2 micron filter. [0050] As per the [0049], the concentration of ammonium sulphate solution is ranged from 0.01 Mto0.4M. [0051 ] The fifth step is hydration of spray-dried lipid. [0052] As per the [0030], weighing of spray dried lipid powder accurately and mixing it by using of 0.15M-ammonium sulphate solution for hydration is done at selected 60 0C. [0053] As per the [0052], the temperature is ranged from 35 °C to 70 °C. [0054] As per the [0052], the solution is taken for sonication for 1 hour with frequent shaking. [0055] As per the [0054], the time is ranging from 30 minute to 120 minute. [0056] The sixth step is Homogenization [0057] As per the [0056]. the clean homogenizer is heated in water bath at 80 °C. [0058] As per the [0057], the temperature is maintained between 50 °C & 90 °C. [0059] As per the [0057], first homogenize with 2% sodium hydroxide. [0060] As per the [0059], the concentration is ranging between 1 & 6 %. [0061] As per the [0062], wash homogenizer with water for injection till the required pH is obtained in the washed solution. [0062] As per the [0061], after water for injection washing; it is washed with ammonium sulphate solution until the pH is below 7. [0063] As per the [0061], Homogenized hydrated solution of liposome at 6000 PSI to 20000 PSI for 4 passes. [0064] As per the [0063], the number of passes of homogenization is ranging from 2 to 8. [0065] As per the [0063], after homogenization the liposomal solution is checked for particle size, which is ranged from 80 nm to 150 nm; and the pH is ranged from 4.0 to 6.5. [0066] The seventh step is preparation of 0.9% sodium chloride [0067] As per the [0066], take sodium chloride in volumetric flask and dissolved it with water for injection to get 1 liter final volume and then it is passed through 0.2-micron filter. [0068] As per the [0067], the concentration of sodium chloride is ranging from 0.5 to 1.5 %. [0069] The 8th step is preparation of Sephadex slurry [0070] As per the [0069], sephadex G 50 is added in 0.9% sodium chloride solution and the solution is put at room temperature for 5 hours. [0071] As per the [0070], the time is ranged from 2 to 10 hours. [0072] As per the [0071], the solution is sterile at 121 °C for 20 minutes and stores it between the 2 to 8 °C temperatures. [0073] As per the [0072], the slurry is used for the sephadex column preparation and before using for the column preparation the slurry temperature is at room temperature. [0074] The 8th step is preparation of 0.2M Histidine buffer solution. [0075] As per the [0074], L-Histidine is taken in dried flask and dissolved with water for injection and the pH is adjusted at 6.50 with 5 M sodium hydroxide solution and the final volume is make up with water for injection and then passed it from 0.2 micron filter. [0076] As per the [0075], the pH is ranged from 5.0 to 7.50 and the concentration of preparation is ranging from 0.05 to 0.5 M. [0077] The 10th step is removal of external ammonium sulphate. [0078] As per the [0077]. the sephadex slurry is taken in glass syringe and by that way column is prepared. [0079] As per the [0074] & [0077], the dephadex column is first saturated with 0.2 M Histidine buffer solution and after that is saturated with lipilipid solution and keep a side for 3 minutes. [0080] As per the [0079], the time shall be ranging from 1 to 15 minutes. [0081] As per the [0079], the histidine solution is again added in the sephadex column and then the liposome solution is collected. [0082] As per the [0081], the liposome solution is started to collect when the slightly hazy solution is came out. [0083] As per the [0081] & [0082], repeat both steps to get more liposome solution till the hazy solution came out. [0084] As per the [0085], after completion of collection of liposome solution, the sephadex column is regenerated with 0.9% sodium chloride solution. [0085] As per the [0084]. all the steps are followed thrice for the removal of external ammonium sulphate. [0086] As per the [0085], the solution is checked for the ammonium sulphate content, for pH and the final volume is measured. [0087] The 11th step is for the preparation of sucrose solution. [0088] As per the [0087], take sucrose and dissolved it in water for injection and filter the solution from 0.2 micron sterile filter and pass through autoclaved. [0089] As per the [0088], the sucrose solution is added in liposome solution and then measures the total volume of liposome solution. [0090] As per the [0089], the solution is passed through 1.2 and 0.45 sterile filter by membrane extrusion. [0091] The 12th step is for the preparation of doxorubicin hydrochloride solution. [0092] As per the [0091], take Doxorubicin Hydrochloride and mixed with liposome solution in 0.5:10 ratio in dry flask and dissolved with water for injection and filter it from sterile 0.2 micron filter. [0093] The 13th step is the drug loading. [0094] As per the [0093], the liposome solution is taken in flask and keeps it at 52 °C in water bath. [0095] As per the [0094], the temperature is ranging between 35 °C & 60 °C. [0096] As per the [0095], when the temperature is reached add the drug in the flask and slightly shake it and after that check the pH of the solution and the keep the solution for 4 hours at 52 °C. [0097] As per the [0096], the time is ranged from 30 minutes to 24 hours and the temperature is ranged from 35 °C to 60 °C. [0098] As per the [0097], the solution is cool at room temperature and starts the filtration from 0.2-micron filter by membrane extrusion. [0099] As per the [0098], take final drug-loaded solution in 5- liter flask, and start the filtration from 0.2-micron sterile filter. Example 1: Prepare the organic mixture of chloroform and methanol (ethanol, acetone and isopropyl alcohol (ratio 0.5 to 5), take 175 ml chloroform and 175ml methanol HPLC grade in 500 to 1000 ml bottle. Dry glass bottle and sonicate them 10 minutes in ultrasonicate bath. First weight lipids as per batch size (on anhydrous basis) HSPC, mPEG-DSPE 2000 and cholesterol dissolve it in organic mixture and filter it from sterile 0.2 micron filter (lipid solution cone. For spray drying: 10 to 30 %). Close the bottle with rubber stopper or aluminium foil and store it below 10 to 35 °C, after preparation lipid solution it. (Dry lipid ratio 0.5 to 10). Take lipid solution in round flask and attached it rotary evaporator, keep the water batch temperature 60 °C to 70 °C, evaporate the organic solvent thin film formed in round bottom flask. After thin film formation allow the round bottom along with the lipid film to cool down considerably in an air-conditioned enviroment. Take of ammonium sulphate in flask, dissolve it with Water For Injection (WFI) and adjust pH 5.40 with 1M HC1 or 1M NaOH, make volume with WFI, check the pH of the solution and filter the solution from sterile 0.2 micron filter. Add 0.15 M ammonium sulphate solution in round flask for hydration is done at 60 °C with sonication for 1 hour during sonication frequently shaking slightly after then start homogenization. before homogenization measure the volume of liposome solution. Keep Homogenizer at 80°c in water bath and clean the homogenizer, first wash with 2% sodium hydroxide and then repeated washings of WFI till neutral pH of washing is obtained. After then pass ammonium sulphate solution from homogenizer and check pH the solution (if pH of solution above 7 than again it should be cleaned). Start homogenization of hydrated liposome solution. Homogenize it for 4 passes of 6000 to 20000psi. After homogenization measure the volume of liposome solution. After homogenization check particle size. pH and volume of liposome solution for following parameters. Take sodium chloride in volumetric flask and dissolve it with WFI and make up the volume 1 liter with WFI and filter it from sterile 0.2-micron filter. Take Sephadex G 50 and mix it with 0.9% sodium chloride solution, keep it at room temperature for 5 hours after then sterile it 121°c for 20 min and store it 2 to 8°c. when it used for sephadex column preparation, slurry temperature should be room temperature. Take L-Histidine in dry flask, dissolve it with WFI, adjust pH 6.50 with 5 M sodium hydroxide solution and make up the volume 3 lit with WFI, check the pH and filter it from sterile 0.2 micron filter. Take sephadex slurry in glass syringe and make a slurry column. Prepare the sephadex column, sephadex column first saturate with 10 ml of 0.2 M Histidine buffer solution after then add 22 ml of lipid solution in column and keep it 3 min. in sephadex column, after then add 20 ml of histidine buffer in middle of the sephadex column, collect the liposome solution comes out from sephadex column, liposome collection start when slightly solution is comes out from column, it is collected till hazy solution comes out, when start the clear solution stop the collection of the solution (Approx 22 ml Histidine buffer is require for 10 ml lipid solution). After then regenerate the sepahdex column with 0.9 % sodium chloride solution, for one column 50 ml of 0.9 % sodium chloride solution is require for regeneration. Three times used this column for external ammonium sulphate removal after then prepare new sephadex column. Check the ammonium sulphate content form liposome solution and pH. Measure the volume of total liposome solution. Take sucrose and dissolve it in WFI and filter it from 0.2 micron sterile filter and add this solution in liposome solution and measure the total liposome solution and filter it from 1.2 and 0.45 sterile filter. Take Doxorubicin (as per liposome concentration in liposome solution, lipid drug ration keep 10 : 0.5 ) in dry flask and dissolve it with WFI and filter it from sterile 0.2 micron filter. First take the liposome solution in flask and keep it at 52° c in water bath, when reach the temperature 52°c add the drug solution in 5 lit flask, slightly shake it, after then check the pH of the solution and keep the solution 4 hours (range 30 min. to 24 hours) at 52°c. batch, when reach the solution at 52°c after then cool it at room temperature. Take drug-loading solution in flask, and start the filtration from 0,2-micron sterile filter, fill the solution in 10 ml USP type I glass vial under nitrogen gas condition. Exaraple2: A process of preparing Pegylated Liposomal Drug Delivery System for'Doxorubicin Hydrochloride Drug which can be describe as follows: 1. The first step is preparation of organic mixture by mixing chloroform and methanol in a ratio of 0.5 to 5. 1.1. According to step 1, the other solvents are Ethanol, acetone and isopropyl alcohol. 1.2. According to step 1.1, the 30% to 80% of chloroform and methanol is mixed in 500 to 1000 ml volume of bottle, 1.3. According to 1.3, the bottle with solution is sonicated for 10 minutes by using ultrasonic bath. 2. The Second step is the preparation of lipid solution by taking weighed quantity of Hydrogenated phosphatidylcholine (HSPC); N-(Carbonyl- methoxypolyehtylene glycol 2000)-l,2-disteroyl-sn-glycero-2- phosphoethanolamine sodium salt (MPEG2000-DSPE) and cholesterol which is dissolve in organic mixture andafterthatitispassedthrough0.2 micron filter for the filtration process. 2.1. According to step 2, the concentration of lipid solution is ranged between 10 and 30 percentages. 2.2. According to 2.1, the solution is taken into bottle and then closed with rubber stopper or aluminum foil. 2.3. According to2.2, the bottle is stored at 10 to 35 °C temperature. 2.4. According to 2.3, the selected solution is taken for spray dry. 3. The third step is spray drying 3.1 As per the 3, the spray drying is carried out at selected inlet and outlet temperature, air pressure, airflow and pump flow. 3.2 As per the 3.1. the inlet temperature is 60°C. 3.3 As per the 3.2. the inlet temperature is ranged from 50°C to 90°C. 3.4 As per the 3. the outlet temperature is 36°C. 3.5 As per the 3.4. the range of outlet temperature is 30°C to 60°C. 3.6 As per the 3, the selected compressed air pressure is ranging from 0.6 to 0.7 bars. 3.7 As per the 3.6, the selected compressed air pressure is ranging from 0.2 to 1.0 bars. 3.8 As per the 3.7, the airflow is ranging from 6 to 8 meterVhour. 3.9 As per the 3.8, the airflow is ranging from 3 to 10 meterVhour. 3.10 As per the 3, the pump flow is 0.2 ml/minute. 3.11 Asperthe 3.10, the pump flow is 0.1 to 0.6 ml/minute. 3.12 As per the step 3, allowing the assembly along with the spray-dried powder to cool down considerably in an air-conditioned environment. 3.13 As per the 3.12, there will be an extreme loss of dried powder due to its sticky nature, which may interfere during removal and collection. 3.14 As per the 3.13, the spray-dried powder is collected in dry glass bottle and check the moisture content of this spray dried powder. 4. The forth step is preparation of ammonium sulphate solution. 4.1 As per the 4, Ammonium sulphate is taken in flask in a quantity of 1 to 10 mg/ml and it is dissolved with water for injection and the pH is adjusted by using selected solution. 4.2 As per the 4.1, the selected solution is Hydrochloric acid (HC1) and Sodium Hydroxide (NaOH) to adjust the pH at 5.40. 4.3 As per the 4.2, the pH is ranged from 4.0 to 6.5. 4.4 As per the 4.3, after adjusting pH the volume is adjusted with Water for injection again the pH is adjusted as per the standard instruction mention in claim 4.2 and 4.3 and after that the solution is passed through 0.2 micron filter. 4.5 As per the 4.4; the concentration of ammonium sulphate solution is ranged from 0.01 M to 0.4M. 5. The fifth step is hydration of spray-dried lipid. 5.1 As per the 3, weighing of spray dried lipid powder accurately and mixing it by using of 0.15M-ammonium sulphate solution for hydration is done at selected 60 0C. 5.2 As per the 5.1, the temperature is ranged from 35 °C to 70 °C. 5.3 As per the 5.1. the solution is taken for sonication for 1 hour with frequent shaking. 5.4 As per the 5.3, the time is ranging from 30 minute to 120 minute. 6. The sixth step is Homogenization 6.1 As per the 6, the clean homogenizer is heated in water bath at 80 °C. 6.2 As per the 6.1, the temperature is maintained between 50 °C & 90 °C. 6.3 As per the 6.1, first homogenize with 2% sodium hydroxide. 6.4 As per the 6.3, the concentration is ranging between 1 & 6 %. 6.5 As per the 6.3, wash homogenizer with water for injection till the required pH is obtained in the washed solution. 6.6 As per the 6.5, after water for injection washing, it is washed with ammonium sulphate solution until the pH is below 7. 6.7 As per the 6.5, Homogenized hydrated solution of liposome at 6000 PSI to 20000 PSI for 4 passes. 6.8 As per the 6.7, the number of passes of homogenization is ranging from 2 to 8. 6.9 As per the 6.7, after, homogenization the liposomal solution is checked for particle size, which is ranged from 80 nm to 150 nm, and the pH is ranged from 4.0 to 6.5. 7. The seventh step is preparation of 0.9% sodium chloride 7.1 As per the 7, take sodium chloride in volumetric flask and dissolved it with water for injection to get 1 liter final volume and then it is passed through 0.2-micron , filter. 7.2 As per the 7, the concentration of sodium chloride is ranging from 0.5 to 1.5 %. 8. The 8th step is preparation of Sephadex slurry 8.1 As per the 8, sephadex G 50 is added in 0.9% sodium chloride solution and the solution is put at room temperature for 5 hours. 8.2 As per the 8.1, the time is ranged from 2 to 10 hours. 8.3 As per the 8.2, the solution is sterile at 121 °C for 20 minutes and stores it between the 2 to 8 °C temperatures. 8.4 As per the 8.3. the slurry is used for the sephadex column preparation and before using for the column preparation the slurry temperature is at room temperature, 9. The 8th step is preparation of 0.2M Histidine buffer solution. 9.1 As per the 9, L-Histidine is taken in dried flask and dissolved with water for injection and the pH is adjusted at 6.50 with 5 M sodium hydroxide solution and the final volume is make up with water for injection and then passed it from 0.2 micron filter. 9.2 As per the 9.1, the pH is ranged from 5.0 to 7-50 and the concentration of preparation is ranging from 0.05 to 0.5 M. 10. The tenth step is removal of external ammonium sulphate. 10.1 As per the 10. the sephadex slurry is taken in glass syringe and by that way column is prepared. 10.2 As per the 9 & 10.1. the sephadex column is first saturated with 0.2 M Histidine buffer solution and after that it is saturated with lipid solution and keep a side for 3 minutes. 10.3 As per the 10.2, the time shall be ranging from 1 to 15 minutes. 10.4 As per the 10.2; the histidine solution is again added in the sephadex column and then the liposome solution is collected. 10.5 As per the 10.4, the liposome solution is started to collect when the slightly hazy solution is came out. 10.6 As per the 10.4 & 10.5, repeat both steps to get more liposome solution till the hazy solution came out. 10.7 As per the 10.6, after completion of collection of liposome solution, the sephadex column is regenerated with 0.9% sodium chloride solution. 10.8 As per the 10.7, all the steps are followed thrice for the removal of external ammonium sulphate. 10.9 As per the 10.8, the solution is checked for the ammonium sulphate content, for pH and the final volume is measured. 11. The eleventh step is for the preparation of sucrose solution. 11.1 As per the 11, take sucrose and dissolved it in water for injection and filter the solution from 0.2 micron sterile filter and pass through autoclaved. 11.2 As per the 11.1, the sucrose solution is added in liposome solution and then measures the total volume of liposome solution. 11.3 As per the 11.2, the solution is passed through 1.2 and 0.45 sterile filter by membrane extrusion. 12. The twelfth step is for the preparation of doxorubicin hydrochloride solution. 12.1 As per the 12, take Doxorubicin Hydrochloride and mixed with liposome solution in 0.5:10 ratio in dry flask and dissolved with water for injection and filter it from sterile 0.2 micron filter. 13. The thirteenth step is the drug loading. 13.1 As per the 13, the liposome solution is taken in flask and is to be kept at 52 °C in water bath. 13.2 As per the 13.1, the temperature is ranging between 35 °C & 60 °C. 13.3 As per the 13.2. when the temperature is reached add the drug in the flask and slightly shake it and after that check the pH of the solution and to keep the solution for 4 hours at 52 °C. As per the 13.3, the time is ranged from 30 minutes to 24 hours and the temperature is ranged from 35 °C to 60 °C. 13.4 As per the 13.4, the solution is cool at room temperature and starts the filtration from 0.2-micron filter by membrane extrusion. 14 As per the 13.5. take final drug-loaded solution in 5-liter flask, and start the filtration from 0.2-micron sterile filter. We claim: 1. A process of preparing Pegylated Liposomal Drug Delivery System for Doxorubicin Hydrochloride Drug comprising the following steps: i) Preparing the organic mixture of chloroform & methanol, ii)preparation of lipid solution by taking weighed quantity of Hydrogenated phosphatidylcholine (HSPC), N-(Carbonyl- methoxypolyehtylene glycol 2000)-l,2-disteroyl-sn-gIycero-2-phosphoethanolamine sodium salt (MPEG 2000-DSPE) and cholesterol, which is dissolve in organic mixture . iii) spray drying the selected lipid solution , iv) preparing the ammonium sulphate solution separately, v)carrying out the hydration of spray-dried lipid wherein spray dried lipid powder mix ammonium sulphate solution , vi) carrying out the homogenization of the solution , vii) preparing the sodium hydrochloride solution , viii) preparing Sephadex slurry, ix) preparing Histidine buffer solution , x) removing the external ammonium sulphate , xi) preparing sucrose solution , xii) preparing doxorubicin hydrochloride solution wherein Doxorubicin Hydrochloride is mixed with liposome solution . xiii),finally , drug loading the solution. 2. Process as claimed in claim 1 wherein organic mixture of chloroform & methanol is prepared by mixing chloroform and methanol in a ratio of 0.5 to 5. 3. Process as claimed in claim 1 wherein preparation lipid solution comprises y taking weighed quantity of Hydrogenated phosphatidylcholine (HSPC), N-(Carbonyl- methoxypolyehtylene glycol 2000)-l, 2-disteroyl-sn-glycero-2-phosphoethanolamine sodium salt (MPEG2000-DSPE) and cholesterol, which is dissolve in organic mixture and passingthrough0.2 micron filter for the filtration process concentrating lipid solution between 10 and 30 percentages, stored at 10 to 35 °C temperature .selected solution is taken for spray dry. 4. Process as claimed in claim 1 wherein step of Spray drying comprises selecting parameters like inlet temperature,outlet temperature, compressed air pressure, air flow and pump flow etc. , (a) the selected inlet temperature is ranged from 50 °C to 90 T; (b) the selected outlet temperature is ranged from 30 °C to 60 T; (c) the selected compressed air pressure is is ranged from 0.1 to0.7; 5. Process as claimed in claim 1 wherein preparation of the ammonium sulphate solution comprises taking ammonium sulphate in a quality of 1 to 10 mg/ml and it is dissolved with water for injection adjusting pH 4-6.5 by Hydrochloric acid (HC1) and Sodium Hydroxide (NaOH) xoncentrating ammonium sulphate solution from 0.01 M to 0.4 M. 6. Process as claimed in claim 1 wherein hydration of spray dried mixture are carried by mixing spray dried lipid powder with 0.15M-ammonium sulphate solution at selected at 35 °C to 70 °C. the solution is taken for sonication for 30 minute to 120 minute with frequent shaking. 7. Process as claimed in claim 1 wherein step of preparation of sodium hydrochloride solution comprises sodium chloride dissolved in water and then it is passed through 0.2-micron filter, the concentration of sodium chloride is ranging from 0.5 to 3.5%. 8. Process as claimed in claim 1 wherein preparation Sephadex slurry comprises sephadex G 50 is added in 0.9% sodium chloride solution and the solution is put at room temperature for 2 to 10 hours , solution is sterile at 121 °c for 20 minutes and stores it between the 2 to 8 °C temperatures. 9. Process as claimed in claim 1 wherein preparation of Histidine buffer solution comprises dissolving L-Histidine with water and the pH is adjusted at 5.0 to 7.50 with 5 M sodium hydroxide solution. 10. Process as claimed in claim 1 wherein mixing of Doxorubicin Hydrochloride with liposome solution comprises taking sucrose and dissolved it in water and filter the solution from 0.2 micron sterile filter and pass through autoclaved, the sucrose solution is added in liposome solution and then measures the total volume of liposome solution.

Documents:

1892-MUM-2007-ABSTRACT(21-1-2011).pdf

1892-MUM-2007-ABSTRACT(21-9-2011).pdf

1892-MUM-2007-ABSTRACT(8-9-2010).pdf

1892-MUM-2007-ABSTRACT(GRANTED)-(18-10-2011).pdf

1892-mum-2007-abstract.doc

1892-mum-2007-abstract.pdf

1892-mum-2007-assignment.pdf

1892-MUM-2007-CANCELLED PAGES(12-9-2011).pdf

1892-MUM-2007-CLAIMS(AMENDED)-(21-1-2011).pdf

1892-MUM-2007-CLAIMS(AMENDED)-(21-9-2011).pdf

1892-MUM-2007-CLAIMS(AMENDED)-(8-9-2010).pdf

1892-MUM-2007-CLAIMS(GRANTED)-(18-10-2011).pdf

1892-mum-2007-claims.doc

1892-mum-2007-claims.pdf

1892-mum-2007-correspondence(13-3-2008).pdf

1892-MUM-2007-CORRESPONDENCE(21-9-2011).pdf

1892-MUM-2007-CORRESPONDENCE(8-9-2010).pdf

1892-MUM-2007-CORRESPONDENCE(IPO)-(18-10-2011).pdf

1892-mum-2007-correspondence(ipo)-(20-1-2010).pdf

1892-mum-2007-correspondence-received.pdf

1892-mum-2007-description (complete).pdf

1892-MUM-2007-DESCRIPTION(GRANTED)-(18-10-2011).pdf

1892-MUM-2007-FORM 18(13-3-2008).pdf

1892-MUM-2007-FORM 2(GRANTED)-(18-10-2011).pdf

1892-MUM-2007-FORM 2(TITLE PAGE)-(21-1-2011).pdf

1892-MUM-2007-FORM 2(TITLE PAGE)-(21-9-2011).pdf

1892-MUM-2007-FORM 2(TITLE PAGE)-(27-9-2007).pdf

1892-MUM-2007-FORM 2(TITLE PAGE)-(8-9-2010).pdf

1892-MUM-2007-FORM 2(TITLE PAGE)-(GRANTED)-(18-10-2011).pdf

1892-MUM-2007-FORM 9(19-3-2008).pdf

1892-mum-2007-form-1.pdf

1892-mum-2007-form-2.doc

1892-mum-2007-form-2.pdf

1892-mum-2007-form-3.pdf

1892-mum-2007-form-5.pdf

1892-MUM-2007-REPLY TO EXAMINATION REPORT(21-1-2011).pdf

1892-MUM-2007-REPLY TO EXAMINATION REPORT(8-9-2010).pdf

1892-MUM-2007-REPLY TO HEARING(21-9-2011).pdf

1892-MUM-2007-SPECIFICATION(AMENDED)-(21-1-2011).pdf

1892-MUM-2007-SPECIFICATION(AMENDED)-(21-9-2011).pdf

1892-MUM-2007-SPECIFICATION(AMENDED)-(8-9-2010).pdf