FORM 2
THE PATENT ACT, 1970
(39 OF 1970)
AND
THE PATENTS RULES, 2003
PROVISIONAL/COMPLETE SPECIFICATION
(See section 10; rule 13)
1 TITLE OF THE INVENTION: Process of Manufacturing Rapid Reconstituting
Lyophilized Doxorubicin.
2 APPLICANT (S)
(a) Name: Mrs. Anju Sunil Kaudanya
(b) Nationality: Indian
(c) Address: 802/A wing, 8th floor President Park, Plot No. 77 & 77A, Sector 29, Opp. Rajiv Gandhi Udyan, Vashi, Navi Mumbai
APPLICANT (S)
(d) Name: Mr.Baban K. Jogale
(e) Nationality: Indian
(f) Address: Plot No. 19, Ashirwad Bungalow No. 2, Gulmohar Colony, Behind
Weight Bridge, in front of Reliance Petrol Pump, Pipe line road, Nasik-422 007, India
3 PREAMBLE TO THE DESCRIPTION
PROVISIONAL COMPLETE
The following specification desefibes the The following specification
invention particularly describes the invention
and the manner in which it is to be
performed.:
4 DESCRIPTION (Description shall start from next page)
See the attachment
5 CLAIMS (not applicable for provisional specification. Claims should start with
the preamble - "I/We claim" on separate page)
See the attachment
6 DATE AND SIGNATURE (to be given at the end of last page of specification)
See the attachment
7 ABSTRACT OF THE INVENTION (to be given along with complete specification
on separate page)
See the attachment

FIELD OF INVENTION:
The invention relates to a process of manufacturing doxorubicin drug composition, which is then lyophilized. The unique technique of manufacturing and lyophilization process enables the drug to get rapidly reconstituted as well as stay stable for long period of time.
BACKGROUND OF INVENTION:
Lyophilization or freeze-drying has been widely used since decades in pharmaceuticals, food and chemical industries. Lyophilization is particularly desirable in situations where a pharmaceutical or other material is required to be dried or dehydrated but the subject is heat sensitive, Lyophilization is a technique wherein a compound or formulation is dissolved, suspended or emulsified, the resultant solution, suspension or emulsion is freezed and then vacuum is applied thereto to sublimate/ evaporate the solvent and other liquids in the frozen mass used to dissolve, suspend or emulsify the material. A large number of patents have been filed for lyophilization or freeze-drying of various formulations/ compounds.
Doxorubicin is an anticancer drug. It is an anthracycline antibiotic. Doxorubicin is commonly used in the treatment of a wide range of cancers, including hematological malignancies, many types of carcinoma, and soft tissue sarcomas. Doxorubicin is a well-known therapeutically active anticancer drug. Doxorubicin is commonly used to treat some leukemias, Hodgkin's lymphoma, as well as cancers of the bladder, breast, stomach, lung, ovaries, thyroid, soft tissue sarcoma, multiple myeloma, and others. Doxorubicin chemically is 10-(4-amino-5-hydroxy-

6-methyl-oxan-2-yl)oxy-6,8,ll-trihydroxy-8-(2-hydroxyacetyl)-l-methoxy-9,10-dihydro-7H-tetracene-5,12-dione.
The mechanism of action of doxorubicin is complex. It interacts with DNA by intercalation and inhibits macromolecular biosynthesis. The drug intercalates between the bases of DNA and blocks DNA synthesis and transcription. It also inhibits the progression of the enzyme topoisomerase II, which unwinds DNA for transcription. Doxorubicin stabilizes the topoisomerase II complex after it has broken the DNA chain for replication, preventing the DNA double helix from being resealed and thereby stopping the process of replication, and causing cell death. The second mechanism of action is to damage the free radicals in ribose in DNA strands found in malignant or susceptible bacterial cells.
Doxorubicin is a hydroxyl form of daunomycin in structure for the reduction in cardiotoxicity. The most common form of the drug is a crystalline hydrochloric salt and is administered intravenously. Usually the salt form of Doxorubicin i.e. Doxorubicin hydrochloride is prefer of better stability.
It is well known that the anthracycline gylcosides like doxorubicin has lower stability parameters. Doxorubicin undergoes thermal degradation. The potency is lost due to pH of the solvent. For these reasons, doxorubicin is preferred in salt forms. Lyophilization is a best method for reducing the thermal degradation of doxorubicin as well as increasing its shelf life. Lyophilized doxorubicin has elevated stability and gets rapidly reconstituted.

STATEMENT OF THE INVENTION
The present invention relates to a process of manufacturing doxorubicin drug composition, which is then lyophilized. Unique lyophilization cycle is developed for doxorubicin so as the drug can be stable for a longer period of time and gets rapidly reconstituted. The composition of doxorubicin, cryogenic substances, pH buffers and solubilizing agents is produced in lyophilized form.
DESCRIPTION:
The invention relates to a process of manufacturing doxorubicin drug formulation for lyophilization.
In one aspect of the invention, the invention comprises of a lyophilization process for doxorubicin. The lyophilization process for doxorubicin comprises of three essential steps: a] precooling, b] freezing and c] primary drying and secondary drying.
In one embodiment, the lyophilization process comprises of forming a lyophilized composition.
In another embodiment, the lyophilized composition is comprised of cryogenic substances i. e. cryoprotectants such as lactose, dextrose, mannitol, etc.
In another embodiment, pH buffers or pH adjusters may be used for providing stable conditions for lyophilization and drug storage.

In still another embodiment of the invention, optimum temperature and optimum pressure are used in the process to obtain the lyophilized formulation of desired characteristics.
In the second aspect of the invention includes a lyophilized composition or formulation of doxorubicin.
In one embodiment of the invention, the lyophilization composition comprises of cryogenic substance i. e. cryoprotectants such as lactose, dextrose, mannitol, etc.
In another embodiment, pH buffer or adjusters may be used for providing stable conditions for lyophilization and drug storage.
In still other embodiment of the invention, to enhance the solubility of doxorubicin hydrochloride, solubilizing agents are added. Solubilizing agents like propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc. may be used for enhancing the solubility of doxorubicin hydrochloride.
DETAILED DESCRIPTION:
The invention comprises of a process of manufacturing rapidly reconstituting lyophilized doxorubicin. Freeze-drying or lyophilization is a process that removes water from a substance. This dehydration process is performed under vaccum while the substance is in frozen state. Lyophilization technique is used to freeze-dry products such as analytical chemistry moieties and therapeutic biomolecules (eg antibodies, vaccines, drugs and heat sensitive proteins). Lyophilizing such products,

particularly liquid formulations, vastly increases their shelf-life and stability. . Unique lyophilization cycle is developed for doxorubicin. Doxorubicin lyophilized by this process gets reconstituted within 5 to 10 seconds. The lyophilization process comprises of three essential steps: a] precooling, b] freezing and c] primary drying and secondary drying. The process parameters like temperature and pressure play important role for a successful cycle of a lyophilization process.
The present invention describes the lyophilization cycle developed by taking into consideration the critical parameters like temperature and pressure to produce a cake with more number of pores. More pores result in more void space, thereby increasing affinity of the lyophilized formulation towards solvent. The subject of the invention, the lyophilization process described herein, produces very small particles of the drug thereby causing/ resulting in particle size reduction. As particles reduce in size, availability of surface area increases. This in turn increases the wetability of the drug in the solvent. It produces a complete dry cake. Dryness of the cake increases solvent attraction and avoids lump formation that may interfere with drug dissolution. Thus, the process produces a cake having rapid dissolution. Lactose monohydrate used as cryoprotectant enhances the dissolution parameters of the lyophilized cake.
The present invention is directed to a lyophilization cycle developed for doxorubicin. The lyophilization process comprises of three essential steps: a] precooling, b] freezing and c] drying. Drying, further, comprises of primary drying and secondary drying.

In one embodiment, the lyophilization process comprises of forming a lyophilized composition.
In another embodiment, the lyophilized composition is comprised of cryogenic substances i. e. cryoprotectants such as lactose, dextrose, mannitol, etc. Formulation comprising lactose produced better lyophilized cake formation. In preferred embodiment, lactose monohydrate was used as a bulking agent as well as a cryogen.
In another embodiment, pH buffers or pH adjusters may be used for providing stable conditions for lyophilization and drug storage. pH buffers such as sodium hydroxide, sodium carbonate, hydrochloric acid etc. may be used individually or in combination. The formulation in the present invention, preferably, comprises of sodium hydroxide as pH buffer.
In still other embodiment of the invention, to enhance the solubility of doxorubicin hydrochloride, solubilizing agents are added. Solubilizing agents like propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc. may be used for enhancing the solubility of doxorubicin hydrochloride. Precisely, methyl paraben is used for enhancing the solubility of the lyophilized doxorubicin HCL.
In still another embodiment of the invention, optimum temperature and optimum pressure are used in the process to obtain the lyophilized formulation of desired characteristics. This invention designs the lyophilization cycle in such a fashion that the product obtained gets rapidly reconstituted within 5-10 secs as well as the product remains stable for quiet a long time.

In the second aspect of the invention includes a lyophilized composition or formulation of doxorubicin.
In one embodiment of the invention, the lyophilization composition comprises of cryogenic substance i. e. cryoprotectants such as lactose, dextrose, mannitol, etc. Formulation comprising lactose produced better lyophilized cake formation. In preferred embodiment, lactose monohydrate was used as a bulking agent as well as a cryogen.
In another embodiment, pH buffer or adjusters may be used for providing stable conditions for lyophilization and drug storage. pH buffers such as sodium hydroxide, sodium carbonate, hydrochloric acid etc. may be used individually or in combination. The formulation in the present invention, preferably, comprises of sodium hydroxide as pH buffer.
In still other embodiment of the invention, to enhance the solubility of doxorubicin hydrochloride, solubilizing agents are added. Solubilizing agents like propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc. may be used for enhancing the solubility of doxorubicin hydrochloride. Precisely, methyl paraben is used for enhancing the solubility of the lyophilized doxorubicin HCL.
Example 1
The lyophilization process comprises of preparing a formulation comprising of doxorubicin hydrochloride, cryoprotectant, pH buffer, solubilizing agent and other excipients. The next step is precooling the formulation for 30 mins to 4 hrs; precooled formulation is then freezed for 8 - 11 hrs depending upon the weight of the sample formulation to be freezed. Further, the freezed formulation is dried in two steps as primary

drying and secondary drying. Primary drying is performed for 22-32 hrs and secondary drying is performed for 18-24 hrs depending upon the weight of the sample formulation to be dried.
Example 2
The lyophilization process is comprised of forming a lyophilized composition. The lyophilized composition comprises of cryoprotectant lactose and pH adjuster or pH buffer sodium hydroxide. Solubilizing agents such as propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc may be used to increase the solubility of doxorubicin hydrochloride.
Example 3
The lyophilization process is comprised of forming a lyophilized composition. The lyophilized composition comprises of cryoprotectant lactose and pH adjuster or pH buffer sodium carbonate. Solubilizing agents such as propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc may be used to increase the solubility of doxorubicin hydrochloride.
Example 4
The lyophilization process is comprised of forming a lyophilized composition. The lyophilized composition comprises of cryoprotectant lactose and pH adjuster or pH buffer hydrochloric acid. Solubilizing agents such as propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc may be used to increase the solubility of doxorubicin hydrochloride.

Example 5
The lyophilization process is comprised of forming a lyophilized composition. The lyophilized composition comprises of cryoprotectant dextrose and pH adjuster or pH buffer sodium hydroxide. Solubilizing agents such as propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc may be used to increase the solubility of doxorubicin hydrochloride.
Example 6
The lyophilization process is comprised of forming a lyophilized composition. The lyophilized composition comprises of cryoprotectant lactose and pH adjuster or pH buffer sodium carbonate. Solubilizing agents such as propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc may be used to increase the solubility of doxorubicin hydrochloride.
Example 7
The lyophilization process is comprised of forming a lyophilized composition. The lyophilized composition comprises of cryoprotectant lactose and pH adjuster or pH buffer hydrochloric acid. Solubilizing agents such as propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc may be used to increase the solubility of doxorubicin hydrochloride.

Example 8
The lyophilization process is comprised of forming a lyophilized composition. The lyophilized composition comprises of cryoprotectant mannitol and pH adjuster or pH buffer sodium hydroxide. Solubilizing agents such as propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc may be used to increase the solubility of doxorubicin hydrochloride. Example 9
The lyophilization process is comprised of forming a lyophilized composition. The lyophilized composition comprises of cryoprotectant lactose and pH adjuster or pH buffer sodium carbonate. Solubilizing agents such as propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc may be used to increase the solubility of doxorubicin hydrochloride.
Example 10
The iyophilization process is comprised of forming a lyophilized composition. The lyophilized composition comprises of cryoprotectant lactose and pH adjuster or pH buffer hydrochloric acid. Solubilizing agents such as propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc may be used to increase the solubility of doxorubicin hydrochloride.

Example 11
The lyophilization process is comprised of forming a lyophilized composition. The lyophilized composition comprises of cryoprotectant lactose monohydrate and pH adjuster or pH buffer sodium hydroxide. Solubilizing agent methyl paraben may be used to increase the solubility of doxorubicin hydrochloride.
SUMMARY:
Freeze-drying or lyophilization is a process that removes water from a substance. This dehydration process is performed under vacuum while the substance is in a frozen state. Lyophilization technology is used to freeze-dry products such as analytical chemistry moieties, and therapeutic molecules (e.g., antibodies, vaccines, drugs, and heat-sensitive proteins). Lyophilizing such products, particularly liquid formulations, vastly increases their shelf-life and stability.
In the present invention, for lyophilization process of doxorubicin, additives like bulking agents, and pH adjusters are added to the composition for stabilizing the product. Bulking agents like lactose monohydrate. pH adjusters like sodium hydroxide is used. Lactose monohydrate also enhanced the dissolution parameters of the lyophilized cake.
The cycle of lyophilization plays a critical role in formation of optimum product. Specific lyophilization temperature and pressure causes formation of product with desired characteristics. In this invention, the lyophilization cycle is designed in such fashion that the product so

formed gets rapidly reconstituted a within 5-10 seconds as well as the product remains stable for quiet a long time.
The lyophilized doxorubicin has much stability and gets rapidly reconstituted. This invention describes a unique lyophilization cycle of doxorubicin, which enables the dry lyophilized cake to reconstitute within few seconds and also increases the stability of the drug.

WE CLAIM
1] The process of manufacturing rapidly reconstituting lyophilized doxorubicin essentially comprising of:
a] preparing a formulation comprising of doxorubicin hydrochloride,
cryoprotectant, pH buffer, solubilizing agent and other excipients
b] precooling the formulation of step a]
c] freezing the precooled formulation of step b] and
d] drying the freezed formulation of step 3] in two steps as i] primary
drying and ii] secondary drying.
2] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 1, wherein the formulation is precooled for 30 min to 4 hrs.
3] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 2, wherein the formulation is precooled for 1 hr 30 min to 2 hrs 15 min.
4] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 1, wherein the precooled formulation is freezed for 8 hrs to 1l hrs.
5] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 4, wherein the precooled formulation is freezed for 9 hrs to 9 hrs 40 min.

6] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 1, wherein the primary drying is performed for 22 hrs to 32 hrs.
7] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 6, wherein the primary drying is performed for 22 hrs 50 mins to 30 hrs 40 mins.
8] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 1, wherein the secondary drying is performed for 18 hrs to 24 hrs.
9] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 8, wherein the secondary drying is performed for 19 hrs 40 min to 21 hr 40 min.
10] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 1, wherein said cryogenic substance or cryoprotectant is selected from lactose, dextrose, maltose, etc.
11] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 10 wherein said cryogenic substance or cryoprotectant is lactose monohydrate.
12] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 1 wherein said pH buffer is selected from sodium hydroxide, sodium carbonate, hydrochloric acid, etc.

13) The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 12 wherein said pH buffer is sodium hydroxide.
14] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 1 wherein said solubilizing agent is selected from propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc.
15] The process of manufacturing rapidly reconstituting lyophilized doxorubicin as claimed in claim 14 wherein said solubilizing agent is methyl paraben.
16] The process of manufacturing rapidly reconstituted lyophilized doxorubicin essentially comprising of a] addition of lactose monohydrate as cryogenic substance or cryoprotectant, b] addition of sodium hydroxide as pH buffer and c] addition of methyl paraben as solubilizing agent.
17] The rapidly reconstituting lyophilized formulation of doxorubicin, comprising of a cryogenic substance or cryoprotectant, pH buffer and a solubilizing agent.
18] The rapidly reconstituting lyophilized formulation of doxorubicin as claimed in claim 17, wherein, said cryogenic substance or cryoprotectant is selected from lactose, dextrose, maltose, etc.
19] The rapidly reconstituting lyophilized formulation of doxorubicin as claimed in claim 18, wherein, said cryogenic substance or cryoprotectant is lactose monohydrate.

20] The rapidly reconstituting lyophilized formulation of doxorubicin as claimed in claim 17, wherein, said pH buffer is selected from sodium hydroxide, sodium carbonate, hydrochloric acid, etc.
21] The rapidly reconstituting lyophilized formulation of doxorubicin as claimed in claim 20, wherein, said pH buffer is sodium hydroxide.
22] The rapidly reconstituting lyophilized formulation of doxorubicin as claimed in claim 17, wherein, said solubilizing agent is selected from propylene glycol, ethylene glycol, methyl paraben, propyl paraben, etc.
23] The rapidly reconstituting lyophilized formulation of doxorubicin as claimed in claim 22, wherein, said solubilizing agent is methyl paraben.
24] The rapidly reconstituting lyophilized formulation of doxorubicin comprising of lactose monohydrate as cryogenic substance or cryoprotectant, sodium hydroxide as pH buffer and methyl paraben as solubilizing agent.