THE PATENTS ACT, 1970
PROVISIONAL SPECIFICATION
Section 10
"Lyophillization of Formulation Containing Mannitol”
Serum Institute of India Ltd. a Corporation organized and existing under the laws of India, of 212/2, Off Soli Poonawalla Road, Hadapsar, Pune 411 028 Maharashtra India.
The following specification particularly describes the nature of this invention:

18.00
Provisional Patent Application
Title: Lyophillization of formulation containing mannitol
Objective:
To provide a uniformly lyophilized, stable formulation containing mannitol with zero vial breakage. To provide stable, uniformly lyophilized formulation containing Gemcitabine and Mannitol.
Brief Description:
The formulations containing mannitol are difficult to lyophilize. The critical parameters if not understood and maintained in correct level may result into non-uniform cake, layer separation, ring formation or collapse of the cake. More evidently, the vials break as a result of expansion of mannitol because of crystallization of the cake. The instant invention provides solution to all problems above.
Formulations:
I: 5 ml formulation containing Gemcitabine hydrochloride (200 mg), Mannitol (200 mg), Sodium acetate (12.5 mg) mixed with water (q.s. to make 5 ml). pH was adjusted to 2.5 to 3.5.
II: 25 ml formulation containing Gemcitabine hydrochloride (1000 mg), Mannitol (1000 mg), Sodium acetate (62.5 mg) mixed with water (q.s. to make 25 ml). pH was adjusted to 2.5 to 3.5.
In case of formulation I, the volume can be adjusted to 3.5 to 5 ml.
In case of formulation II, the volume can be adjusted to 15 to 25 ml. Such variation in
volumes can be handled using the process of instant invention.
Vials used: USP type -1,10 ml tubular flint glass vials or 50 ml tubular flint glass vials. Rubber Closures: Fluorocoated, Aluminium seal: Auto fit.
Lyophilization Cycle
Pre-freezed chambers of below -32° C. Preferred temperature -50 to -80° C.
Freezing rate: Rapid, equal to or Above 1° C / min.
Final freezing temperature: below - 25° C. Preferred -25° C to -50° C,
Most preferred -50° C.
The frozen mass was hold for a minimum period of 2 hrs at temperature below -25° C. Preferred -30 to -60° C. Most preferred -50° C.
Primary Drying Specifications:
Temperature below -25° C. Preferred -25° C to -35 ° C, Most preferred -32 ° C. Duration : 15-22 hrs,
Preferred 20 hrs,
Pressure : 50 to 350 millitorr, Preferred 200 mT.

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Stepwise (Optional)
Temperature Between -10 to -5 ° C, Preferred -7 ° C.
Duration : 2-5 hrs, Preferred 3 hrs,
Pressure : 50 to 350 millitorr, Preferred 200 mT.
Secondary Drying: Product is heated at rate of = > 1°C / min.
Stepwise (Optional)
Temperature Between : 5-40 °C, Preferred 18-25 ° C.
Duration: 2-5 hrs, preferred 4 hrs,
Pressure: 10 to 350 millitorr, Preferred 200 mT.
Temperature Between + 40 to 55° C, Preferred 42°C.
Duration: 5-12 hrs, preferred 10 hrs,
Pressure: 0 to 350 millitorr, Preferred 0-100 mT.
High strength formulation
The specifications mentioned above were applied for 1000 mg product batches with a few modifications. In this case the primary drying duration was 15 to 30 hrs. Preferred is 27 hrs.
The cycle time for low strength product was between 30 to 35 hrs. The lyophilization cycle time for high strength product batches was 32 to 42 hrs.
Advantages:
The lyophilization cycle was designed to minimize annealing, crystallization and or expansion of formulation contents. The products were uniformly lyophilized. The lyophilized mass didn't show separation, ring formation or collapse. All the vials of the lyophilized products were intact (not broken). The lyophilization cycle duration was miminum i.e. between 32 to 42 hours. The active pharmaceutical ingredient in the formulation obtained by the instant invention exhibited stability after the process completion.

Dated this 31st day of July 2007