DESC:FIELD OF THE INVENTION:
The present invention relates to a novel high volume and strength parenteral pharmaceutical composition of Liposomal Amphotericin B or a pharmaceutically acceptable salt thereof. The present invention further relates to improved lyophilisation cycle for manufacturing high volume and strength of Liposomal Amphotericin B.

BACKGROUND OF THE INVENTION:
Amphotericin B is a macrocyclic, polyene, antifungal antibiotic produced from a strain of Streptomyces nodosus. It is designated chemically as [1R-(1R*,3S*,5R*,6R*,9R*,11R*,15S*,16R*,17R*,18S*,19E,21E,23E,25E,27E,29E, 31E,33R*,35S*,36R*,37S*)]-33-[(3-Amino-3,6-dideoxy-ß-Dmannopyranosyl)oxy]-1,3,5,6,9,11,17,37-octahydroxy-15,16,18-trimethyl-13-oxo-14,39¬dioxabicyclo [33.3.1] nonatriaconta-19,21,23,25,27,29,31-heptaene-36-carboxylic acid (CAS No. 1397-89-3). It has a molecular formula of C47H73NO17 and a molecular weight of 924.09.

The structure of amphotericin B is shown below:

AMPHOTERICIN B

Amphotericin B binds to ergosterol, the sterol component of the fungi cell membrane resulting in cell death caused by alterations in cell permeability through which monovalent ions (NA+, K+, H+, and Cl-) leak out of the cell. Amphotericin B is also known to cause cell cytotoxicity, since it bind to the cholesterol component of the mammalian cell.

AmBisome®, the liposomal preparation of amphotericin B is indicated for the treatment of
• Empirical therapy for presumed fungal infection in febrile, neutropenic patients.
• Treatment of Cryptococcal Meningitis in HIV-infected patients
• Treatment of patients with Aspergillus species, Candida species and/or Cryptococcus species infections refractory to amphotericin B deoxycholate, or in patients where renal impairment or unacceptable toxicity precludes the use of amphotericin B deoxycholate.
• Treatment of visceral leishmaniasis. In immunocompromised patients with visceral leishmaniasis treated with AmBisome®, relapse rates were high following initial clearance of parasites.

Amphotericin B, the active ingredient of AmBisome®, acts by binding to the sterol component, ergosterol, of the cell membrane of susceptible fungi. It forms transmembrane channels leading to alterations in cell permeability through which monovalent ions (NA+, K+, H+, and Cl-) leak out of the cell resulting in cell death. While amphotericin B has a higher affinity for the ergosterol component of the fungal cell membrane, it can also bind to the cholesterol component of the mammalian cell leading to cytotoxicity. AmBisome®, the liposomal preparation of amphotericin B, has been shown to penetrate the cell wall of both extracellular and intracellular forms of susceptible fungi.

AmBisome® for injection is a sterile, non-pyrogenic lyophilized product for intravenous infusion. Each vial contains 50mg of amphotericin B, USP, intercalated into a liposomal membrane consisting of approximately 0.64mg alpha tocopherol, USP; 52mg cholesterol, NF; 84mg distearoyl phosphatidylglycerol, sodium salt; 213mg hydrogenated soy phosphatidylcholine. In addition, 27mg disodium succinate hexahydrate and 900mg sucrose, NF are used as a buffer. AmBisome® may contain hydrochloric acid and/or sodium hydroxide as pH adjusters. Following reconstitution with Sterile Water for Injection, USP, the resulting pH of the suspension is between 5¬ and 6. All other approved generics currently marketed are also available as 50mg/vial.
Liposomal Amphotericin B dosing and the rate of infusion is individualized based on the age, weight and body size of the patient. Multiple dose vials administration is required to achieve the desirable therapeutic efficacy. Caution is imperative to minimize systemic toxicities or adverse events and antibiotic resistance. Often, dose adjustment results in pharmaceutical wastage. For example, for delivery of maximum dose of Liposomal Amphotericin B for Injection (420mg), 8.4 vials (09 vials) of 50mg Liposomal Amphotericin B is utilized, whereas only 4.2 vials (05 vials) of 100mg will be utilised for the same dose delivery as per the present invention. For dose delivery and administration, the reopening of vial time, duration and administration procedure is reduced as compared to 50mg strength. Further, there are high chances of under or over utilized drug product that needs to be discarded. Furthermore, extra reconstituted product concentrate is required to be stored for upto 24hours at 2°-8° C (36°-46° F) following reconstitution with sterile water for injection, USP.

This ultimately results in higher cost to the patient due to excess, risk of microbiological contamination during aseptic handling, unusable product and disposal cost to the healthcare provider. Hence, there is a need for a commercial formulation that results in fewer vials required per dose that improves patient compliance and reduces costs and importantly, leads to the desired therapeutic effect.

To overcome problems of pharmaceutical waste and mitigating the number of vials needed per dose by addressing differences in patient weight, body size, or age, the inventors of the present invention have arrived at novel high dosage and volume strength parenteral pharmaceutical composition of Amphotericin B or a pharmaceutically acceptable salt thereof. In a low drug formulation, the physical properties, manufacturing and scale-up are dependent on the excipients in the formulation. However, at high drug loading, the contribution of the physical and chemical properties of the active pharmaceutical ingredient to the manufacturability and scale-up of a formulation becomes predominant. Maintaining the necessary chemical and physical properties, such as particle size in order to obtain high dosage strength parenteral pharmaceutical composition requires inventive merit. Largely, pharmacokinetics depends on the composition and particle size of the liposomes. The present invention relates to a composition that is comparable to the existing RLD (AmBisome®).

The inventors of the present invention have encountered the following manufacturing challenges:
1. Liposomal Amphotericin B is a complex injectable product. Hence, slight change in manufacturing process affects the performance of the product.
2. The manufacturing process contains complex procedures like hydration and lyophilisation. For higher strength, hydration process time of product increases.
3. The curing phase time and temperature is required to be improved
4. Increase in volume of Liposomal Amphotericin B per vial compared to 50mg/vial requires increase in primary drying time to in line with CQAs, such as particle size, dissolution, impurity and free drug etc., which requires further improvement in the lyophilisation cycle.
5. The finished product obtained by the lyophilisation technology, such that composition is required to match the RLD (AmBisome®) and quality parameters thereof.

The inventors of the present invention composition have overcome the short comings associated with the prior-art and have arrived at high dosage and volume strength parenteral pharmaceutical composition of Amphotericin B comparable to approved Liposomal Amphotericin B injections (RLD).

The present inventors have prepared parenteral formulations of Amphotericin B Liposome by using pharmaceutically acceptable excipients that is 30 to 50% cost effective to patients. Further, the parenteral pharmaceutical composition of present invention is in the form of an injection, which can be administered to patients having difficulty in oral administration, such as, but not limited to pediatric and geriatric patients. The parenteral formulations of Amphotericin B Liposome is used in the treatment of a wide range of opportunistic and serious fungal pathogens.

OBJECT OF THE INVENTION
The present invention discloses a novel strength and volume of liposomal amphotericin B composition. The composition of the present invention includes lyophilized powder or cake composition for injection and reconstituted injection, while maintaining the concentration of the parenteral drug.

Another object of the present invention to provide an improved process for the manufacture of high dosage strength parenteral pharmaceutical composition of Amphotericin B or a pharmaceutically acceptable salt thereof and procedure for encapsulating amphiphilic drug.

Another of the present invention is to provide composition wherein the average particle size of liposomal amphotericin is having average particle size of not more than 150nm, preferably less than 130nm.

Yet another object of the present invention is to provide a commercially feasible process and improved lyophilization composition for the production of liposomal Amphotericin B without affecting the drug to lipid ratio.

Yet another object of the present invention is to provide a high dosage strength not less than 50mg/vial, preferably 75mg/vial, more preferably 100mg/vial parenteral pharmaceutical composition of Liposomal Amphotericin B in vial having body diameter of 32 to 40mm.

Another object of the present invention is to provide various parenteral pharmaceutical composition dosage for ready to use injection, lyophilized powder composition and reconstituted injections. The parenteral formulations of Amphotericin B Liposome is used in the treatment of a wide range of opportunistic and serious fungal pathogens.

Another object of the present invention is to provide a process for the manufacture of high dosage strength with high fill volume of parenteral pharmaceutical composition of Amphotericin B prepared by spray drying process, hydration, particles size reduction and lyophilisation.

SUMMARY OF THE INVENTION
The present invention discloses a new strength or dose of liposomal amphoteric B not less than 50mg/vial, preferably 75mg/vial, more preferably 100mg/vial. The present invention compositions include ready to use injection, lyophilized powder composition for injection and reconstituted injection.

The pharmaceutical composition according to the present invention is economical and advanced dosage strength over existing dosage strength without resulting in loss of therapeutic efficacy and quality.

The present invention relates to improved methods of liposomes Amphotericin B preparation by commercially feasible process and managing resources.

The present invention provides a composition comprising liposomal amphotericin having average particle size of not more than 150nm, preferably less than 130 nm.

The present invention also discloses the process of preparation of pharmaceutical composition that is time efficient, especially for large-scale production, wherein the pharmaceutical composition is comparable to the approved 50mg/vial dosage form.

The present invention relates high dosage strength not less than 50mg/vial, preferably 100mg/vial, parenteral pharmaceutical composition of Liposomal Amphotericin B in vial having body diameter of 32mm to 40mm.

Another object of the present invention is to provide pharmaceutical composition comprising Amphotericin B and mixture of phospholipid and pharmaceutically acceptable excipients.

Embodiments of the pharmaceutical formulation may include Amphotericin B and one or more phospholipid, cholesterol, anti-oxidant, lyoprotectant and bulking agent, buffering agent, pH adjusting agent, vehicle and additional pharmaceutically acceptable excipients.

In a general aspect, the pharmaceutical formulation as per the present invention is prepared by spray drying process and lyophlisation technology.

The details of one or more embodiments of the invention are set forth in the description below. Other features of the invention will be apparent from the description.

DETAILED DESCRIPTION OF THE INVENTION:
The present invention will now be disclosed by describing certain preferred and optional embodiments, to facilitate various aspects thereof.

The present invention provides a stable parenteral composition comprising therapeutically effective amount of Amphotericin B or a pharmaceutically acceptable salt thereof; and one or more pharmaceutically acceptable excipients.

According to the present invention, the volume of the parenteral composition required to be administered to the patients can be tailored as per prescription.

References to “an”, “one”, or “various” embodiments in this disclosure are not necessarily to the same embodiment, and such references contemplate more than one embodiment. The following detailed description is, therefore, not to be taken in a limiting sense, and the scope is defined only by the appended claims, along with the full scope of legal equivalents to which such claims are entitled.

The term "drug" or “active ingredient” or “active pharmaceutical ingredient (API)” herein refers to Amphotericin B or a pharmaceutically acceptable salt thereof.

The term “composition” or “pharmaceutical composition” or “parenteral composition” or “dosage form” or “pharmaceutical formulation” or “formulation” or “parenteral formulation” as used herein synonymously include the pharmaceutical compositions in a form suitable for parenteral administration such as but not limited to sterile aqueous, hydro-alcoholic, or non-aqueous suspension, aqueous, hydro-alcoholic, or non-aqueous solution, or emulsion, liposomes, microparticles, or the like or any other suitable dosage form meant for parenteral administration. Further, parenteral pharmaceutical composition includes pharmaceutical composition for intramuscular, intravenous, sub-cutaneous, and intrathecal administration. In one embodiment, the pharmaceutical composition may be in the form of a unit dose 10 composition or multi-dose composition

The term "Amphotericin B" as used herein according to the present invention includes, Amphotericin B in the form of free base, a pharmaceutically acceptable salt thereof, amorphous, crystalline, any isomer, derivative, hydrate, solvate or prodrug or a combination thereof.

The term "drug solution" as used herein according to the present invention includes solution obtained by dissolving Amphotericin B or its pharmaceutically acceptable salt thereof in solvent and (or) mixture of solvents.

In accordance with the present invention, an injectable formulation of Amphotericin B comprising Amphotericin B as an active ingredient with pharmaceutically acceptable excipients, preferably herein present invention in the form of intramuscular, subcutaneous, intravenous, and intradermal injection.

In accordance with the present invention an oral solid pharmaceutical formulation of Amphotericin B comprising of Amphotericin B as an active ingredient with pharmaceutically acceptable excipients is provided.

The term "pharmaceutically acceptable excipients" as used herein, refers to excipients those are routinely used in pharmaceutical formulations. The pharmaceutically acceptable excipients may comprise one or more Phospholipid, cholesterol, Anti-Oxidant, Lyoprotectant and bulking agent, Buffering agent, pH adjusting agent, Vehicle and combinations thereof.

The list of excipients are listed below, although it is not limited to the said excipients.

Name of Ingredients Component
Distearoyl Phosphatidylglycerol sodium salt (DSPG-Na) Phospholipid
Hydrogenated Soy Phosphatidylcholine (HSPG) Phospholipid
Alpha Tocopherol Anti-Oxidant
Cholesterol Phospholipid,
Sucrose Lyoprotectant and bulking agent
Disodium Succinate Hexahydrate Buffering agent
Hydrochloric Acid pH adjusting agent
Sodium Hydroxide pH adjusting agent
Water for Injection Vehicle

Suitable phospholipid may include but are not limited to one or more from phosphoglycerides such as Cholesterol, dipalmitoylphosphatidylcholine, distearoyl phosphatidylcholine, diarachidoylphosphatidylcholine, dibehenoylphosphatidylcholine, Distearoyl Phosphatidylglycerol sodium salt (DSPG-Na), diphosphatidyl glycerols, short-chain phosphatidylcholines, hydrogenated phosphatidylcholine, Hydrogenated Soy Phosphatidylcholine, E-100-3 (available from Lipoid KG, Ludwigshafen, Germany), long-chain saturated phosphatidylethanolamines, long-chain saturated phosphatidylserines, long-chain saturated phosphatidylglycerols, long-chain saturated phosphatidylinositols, phosphatidic acid, phosphatidylinositol, and sphingomyelin. The

Suitable antioxidant may include but are not limited to one or more from Ascorbyl palmitate, Ascorbate, Bisulfite sodium, Butylated hydroxy anisole (BHA), Butylated hydroxy toluene (BHT), Cystein/ cysteinate HCl, Dithionite sodium (Na hydrosulfite, Na sulfoxylate), Gentisic acid, Gentisic acid ethanolamine, Glutamate monosodium, Glutathione, Formaldehyde sulfoxylate sodium, Metabisulfite potassium, Metabisulfite sodium, Methionine, Monothioglycerol (Thioglycerol), Propyl gallate, Sulfite sodium, Alpha Tocopherol hydrogen succinate, Thioglycolate sodium, Tocopherol alpha, Sodium thioglycolate and the like.

In one aspect of the present invention, the pH regulator and/or the buffer may be selected from the group consisting of but not limited to acids such as hydrochloric acid, phosphoric acid, sulfuric acid, methanesulfonic acid, acetic acid, lactic acid, maleic acid, citric acid, tartaric acid, ascorbic acid and benzoic acid; salts such as sodium bicarbonate, sodium carbonate, sodium dihydrogenphosphate, potassium dihydrogenphosphate, disodium hydrogenphosphate, dipotassium hydrogenphos phate, trisodium phosphate, disodium citrate and sodium sulfite, Disodium Succinate Hexahydrate; and bases such as sodium hydroxide, trometamol. monoethanolamine, diethanolamine and triethanolamine and the like.

In one aspect of the present invention, the lyoprotectant may be selected from the group consisting of but not limited to sugars such as sucrose and any other sugars.

Solvents play key roles in designing drug delivery systems (DDSs). Specifically, organic solvents are commonly used in the pharmaceutical industry as reaction media. Solvents are uniquely able to dissolve drugs safely and effectively into these medicinal formulations. Further, solvents suitable for preparing binding solution may include one or more of water, organic solvents such as ethanol, methanol, isopropyl alcohol (IPA), acetone, propylene glycol, glycerin, methylene dichloride, trichloromethane and the like.

In one aspect of the present invention, water for injection, any suitable solvent, etc. is used as a vehicle.

In one general embodiment, a pharmaceutical formulation comprising Amphotericin B as active pharmaceutical ingredient and one or more pharmaceutically acceptable excipients and method of preparation thereof.

In one general embodiment, an injectable composition comprising 100mg of Amphotericin B and one or more phospholipid, cholesterol, anti-oxidant, lyoprotectant and bulking agent, buffering agent, pH adjusting agent, vehicle and pharmaceutically acceptable excipients.

In another embodiment, an injectable formulation as per the present invention may be in the form of intramuscular, subcutaneous, intravenous, and intradermal injection.

In another embodiment, the pharmaceutical formulation as per the present invention is prepared as follows:

Spray drying Process
1. Dissolve DSPG-Na (Distearoyl Phosphatidylglycerol sodium salt) in Chloroform: Methanol (1:1) solvent mixture at 60±5oC.
2. Adjust the pH 1.8 to 2.0 by using HCl and remove the precipitation by filtration.
3. Mix Amphotericin B and Alpha Tocopherol in Chloroform: Methanol (1:1) mixture at 60 ±5oC to make a slurry (the API phase).
4. Mix DSPG phase in API phase and stir to obtain a clear solution.
5. The pH of tea colored clear solution is checked for range between pH 3.6 and 4.6). If required, the pH is adjusted using Methanolic sodium hydroxide solution.
6. HSPC (Hydrogenated Soy Phosphatidylcholine) and Cholesterol are dissolve in Chloroform at 65°C to yield clear solution (the phospholipid phase).
7. The above phospholipid phase is mixed into the API phase and stirred to obtain clear solution. The pH is checked and filtered.
8. The filtered bulk solvent is evaporated through spray drying process.

Hydration phase
Take WFI of total batch size. Maintain the temperature at 45 ± 5 °C and allow for nitrogen purging.
1. Sucrose and buffer are mixed and allowed to stir for obtaining clear solution.
2. The Liposomal spray dried powder is added to the above solution and maintained at 45°C to 65 °C and allowed to further stir for 4hrs. The pH of the solution is checked and if required, the pH of bulk solution is adjusted to 5.3 using 2.5 M sodium hydroxide solution.
3. The above suspension is homogenization using high pressure homogenization to achieved desire particle size of liposomes.
4. Make up the volume and the bulk suspension is maintained at temperature of 60 ±5°C.
5. Filter the bulk with 0.2µ filter.
6. Fill the vials as per target fill volume and lyophilize.

According to a further preferred embodiment of the present invention, it is provided that the solution is suitable for parenteral administration, preferably for intravenous and / or subcutaneous administration.

In another embodiment of the present invention, injectable formulations are packaged into containers made of glass or plastic, container systems include ampoules, vials, syringes, cartridges, bottles, and bags as per the requirement.

The present invention has been described by way of example only. It is to be recognized that modifications falling within the scope and spirit of the claims, which would be obvious to a person skilled in the art based upon the disclosure herein, are also considered to be included within the scope of this invention. The scope of the invention is in no manner limited by the disclosed example.

The invention will be further described with respect to the following examples. However, the scope of the invention is not limited thereby. All percentages stated in this specification are by weight, unless otherwise specified. While the present invention has been described in terms of its specific embodiments, certain modifications and equivalents will be apparent to those skilled in the art and are intended to be included within the scope of the present invention. The following examples are provided for illustrative purpose only and these examples are in no way limitative on the present invention.

EXAMPLE 1
Table 1: Amphotericin B Liposome for Injection 100mg/vial
Sr.No Amount (mg) Ingredient
1. 100 Active substance
2. 151 to 185 Phospholipid
3. 383 to 469 Phospholipid
4. 1.15 to 1.41 Anti-Oxidant
5. 94 to114 Phospholipid
6. 1620 to 1980 Lyoprotectant and bulking agent
7. 48.6 to 59.4 Buffering agent
8. q.s pH adjusting agent
9. q.s pH adjusting agent
10. q.s Vehicle

Table 2: Different bulk concentration of Amphotericin B Liposome for Injection 100mg/vial

Sr.
No Name of Ingredients Amphotericin B Liposome for Injection mg/mL
(1) (2) (3)
1. Amphotericin B 8.33 5.55 4.17
2. Distearoyl Phosphatidylglycerol sodium salt (DSPG-Na) 14 9.33 7
3. Hydrogenated Soy Phosphatidylcholine (HSPG) 35.5 23.66 17.75
4. Alpha Tocopherol 0.106 0.071 0.053
5. Cholesterol 8.66 5.77 4.33
6. Sucrose 150 100 75
7. Disodium Succinate Hexahydrate 4.5 3.0 2.25
8. Hydrochloric Acid q.s q.s q.s
9. Sodium Hydroxide q.s q.s q.s
10. Water for Injection q.s to 12.0 ml q.s to 18.0 ml q.s to 24 ml

Manufacturing process
Spray drying Process
1. Dissolve DSPG-Na (Distearoyl Phosphatidylglycerol sodium salt) in Chloroform: Methanol (1:1) solvent mixture at 60±5oC.
2. Adjust the pH 1.8 to 2.0 by using HCl and remove the precipitation by filtration.
3. Mix Amphotericin B and Alpha Tocopherol in Chloroform: Methanol (1:1) mixture at 60 ±5oC to make a slurry (the API phase).
4. Mix DSPG phase in API phase and stir to obtain a clear solution.
5. The pH of tea colored clear solution is checked for range between pH 3.6 and 4.6). If required, the pH is adjusted using Methanolic sodium hydroxide solution.
6. HSPC (Hydrogenated Soy Phosphatidylcholine) and Cholesterol are dissolve in Chloroform at 65°C to yield clear solution (the phospholipid phase).
7. The above phospholipid phase is mixed into the API phase and stirred to obtain clear solution. The pH is checked and filtered.
8. The filtered bulk solvent is evaporated through spray drying process.

Hydration phase
Take WFI of total batch size. Maintain the temperature at 45 ± 5 °C and allow for nitrogen purging.
1. Sucrose and buffer are mixed and allowed to stir for obtaining clear solution.
2. The Liposomal spray dried powder is added to the above solution and maintained at 45°C to 65 °C and allowed to further stir for 4hrs. The pH of the solution is checked and if required pH of bulk solution is adjusted to 5.3 using 2.5 M sodium hydroxide solution.
3. The above suspension is homogenization using high pressure homogenization to achieved desire particle size of liposomes.
4. Make up the volume and the bulk suspension is maintained at temperature of 60 ±5°C.
5. Filter the bulk with 0.2µ filter.
6. Fill the vials as per target fill volume and lyophilize.

Example 2
Comparative Lyophilization cycle of 100mg/vial to 50mg/vial
Lyophilization Process
(100 mg/vial) Lyophilization Process
(50 mg/vial)
Vial size 37mm diameter Vial size 30mm diameter
Stage Vacuum Shelf Temp
(°C) RAMP Time (min) Hold Time
(min) Vacuum Shelf Temp
(°C) RAMP Time (min) Hold Time
(min)
Freezing NA -30 to
-50 70 to 110 410 to 540 NA 5 5 5
-25 120 30
-45 240 300
Primary drying 100 m Torr/133 µ bar -0 to
-40 25 to 200 180 to 3500 90 m Torr/120 µ bar -45 10 10
75 m Torr/100 µ bar 90 m Torr/120 µ bar -30 240 300
75 m Torr/100 µ bar 90 m Torr/120 µ bar -20 240 560
75 m Torr/100 µ bar 90 m Torr/120 µ bar -15 60 560
90 m Torr/120 µ bar -10 120 420
90 m Torr/120 µ bar 0 240 120
90 m Torr/120 µ bar 10 120 120
90 m Torr/120 µ bar 20 60 240
Secondary drying 75 m Torr/100 µ bar -0 to
-35 105 to 140 410 to 540 90 m Torr/120 µ bar 28 120 1140
Total Cycle (Hr.). ~100 to ~150 Total Cycle (Hr.) ~89

The invention described herein comprises in various objects as mentioned above and their description in relation to characteristics, compositions and process adopted. While these aspects are emphasised in the invention, any variations of the invention described above are not to be regarded as departure from the spirit and scope of the invention as described. The above-mentioned examples are provided for illustrative purpose only and these examples are in no way limitative on the present invention. ,CLAIMS:We claim:
1. A lyophilized injectable composition comprising 100mg of Amphotericin B Liposome and one or more pharmaceutically acceptable excipients.

2. The injectable composition as claimed in claim 1 wherein the solution has a concentration of Amphotericin B of 3mg per/ml to 10mg per ml.

3. The injectable composition as claimed in claim 1, wherein the pharmaceutical excipients are selected from phospholipid, anti-oxidant, lyoprotectant and bulking agent, buffering agent, pH adjusting agent, vehicle and mixtures thereof.

4. The injectable composition as claimed in claim 1, wherein the phospholipid is selected from cholesterol, distearoyl phosphatidylglycerol sodium salt, hydrogenated soy phosphatidylcholine or a combination thereof, the anti-oxidant is alpha tocopherol, lyoprotectant and bulking agent is sucrose and the buffering agent is disodium succinate hexahydrate.

5. The injectable composition as claimed in claim 1
a) 100mg of Amphotericin B,
b) 628 to 768mg of phospholipid or a mixture thereof,
c) 1.15 to 1.41mg of anti-oxidant,
d) 1620 to 1980mg of lyoprotectant and bulking agent,
e) 48.6-59.4 mg of buffering agent
f) solvent or a mixture thereof, and optionally other excipients.

6. The injectable composition as claimed in claim 1 comprising liposomal amphotericin having average particle size of not more than 150nm, preferably less than 130nm.

7. The injectable composition as claimed in claim 1, comprising spray drying technique and lyophilisation technology for preparing injectable composition of Amphotericin B Liposome.

8. The injectable composition as claimed in claim 1 prepared by lyophilisation comprising the total time of about 100hours to about 150hours and wherein the method comprise
a. Freezing wherein the RAMP time is about 70mins to about 110mins and the hold time is about 410mins to about 540mins,
b. Primary drying wherein the RAMP time is about 25mins to about 200mins and the hold time is about 180mins to about 3500mins,
c. Secondary drying wherein the RAMP time is about 105mins to about 140mins and the hold time is about 410mins to about 540mins.