FORM 2
THE PATENTS ACT 1970
(39 of 1970)
AND
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10 and rulel3)
1. TITLE OF THE INVENTION:
"STABLE ETOPOSIDE PREPARATION WITH GOOD ORAL BIOAVAILABILITY"
2.APPLICANT(S):
(a) NAME: NEON LABORATORIES LIMITED
(b) NATIONALITY: Indian Company incorporated under the Companies
Act, 1956
(c) ADDRESS: 140, Damji Shamji Industrial Complex, Mahakali Caves Road,
Andheri (East), Mumbai - 400093, Maharashtra, India.
3.PREAMBLE TO THE DESCRIPTION:
The following specification particularly describes the invention and the manner in which it is to be performed

Technical field:
This invention relates to solid- oral formulation of etoposide, i.e. 4'-demethylepipodophillotoxin-9-(4,6-O-ethylidene- β -D-glucopyranoside ), comprising free flowing granules of Etoposide in hard gelatin capsules with good oral bioavailability.
Background and prior art:
Etoposide is a drug effective for treating lung cancer, malignant lymphoma, and testicular
tumor.
Upon administration, etoposide forms a ternary complex with DNA and the
topoisomerase II enzyme, preventing re-ligatibn of the DNA strands. This causes errors in
DNA synthesis and promotes apoptosis of the cancer cell. Normally, etoposide is
administered intravenously or orally in. capsule form and the reported side effects are low
blood pressure, hair loss, constipation and diarrhea.
US4734284 discloses a pharmaceutical composition comprising etoposide and a water-soluble cellulose ether derivative or polyvinylpyrrolidone. Since, etoposide is water insoluble, the preparation has been designed to show a reduced tendency to separate out crystals of etoposide, and with improved absorbability in a living body.
According to the above patent, the content of etoposide in the preparation is only upto 8%. Since, etopside is practically insoluble in water; it is very difficult to increase the content of etoposide in the formulation due to the possibility of separating out of the crystals. Consequently, it would be difficult to prepare a composition with increased content of etoposide due to excessively large capsule size.
US 4701327 discloses an etoposide soft capsule comprising a soft gelatin shell of about pH 4.0 to about pH 5.5 having enclosed therein, an etoposide solution containing an organic acid having 1 to 3 carboxyl groups and 1-6 carbon atoms; a solvent of said etoposide solution being a polyhydric alcohol which is liquid at room temperatures. However, soft gelatin capsules are sensitive and require critical control in comparison

with hard gelatin capsules.
US5609882 discloses an etoposide solution composition containing (1) etoposide and (2) polyvinylpyrrolidone and a water-soluble cellulose ether derivative to reduce the tendency to separate out its crystals when added into water even at a concentration of as high as 10%, specifically upto 25% and also with slightly improved stability over a period of time. Other existing marketed formulations contain liquid or self -emulsifying liquid formulation filled in soft gelatin capsules.
An article titled "Studies on Enhancement of Dissolution Rate of Etoposide", discloses Solid dispersions of etoposide with various surfactants prepared by using kneading technique and studied the bioavailability. The surfactants such as cetrimide, sodium lauryl sulphate, Tween 80 and Cremophor RH40 were used in different proportions and confirmed that etoposide gave faster dissolution rates when Cremophor RH40 was used. However, cremophor RH 40 was used at 20-40% to achieve USP dissolution rate. Use of surfactant in such a high amount is not desirable for ingestion.
In view of the above it is evident that there exists a need in the art to provide an etoposide formulation, where, larger amount of drug may be incorporated into the formulation with greater stability and improved oral bioavailability, which has been achieved in the instant invention and sought protection for the same.
Therefore, the object of the invention is to eliminate one or more of the above drawbacks and to provide solid oral formulation of Etoposide which contains dry granules of Etoposide with excipients with good oral bioavailability that can be filled into hard gelatine capsules.
Summary of the invention:
In accordance with the above, the present invention provides etoposide preparation as free flowing dry granules with good oral bioavailability. The granules of the invention can be directly filled into hard gelatin capsule or may be compressed into tablet.

In another aspect, the invention provides etoposide preparation as free flowing granules that can be directly compressed into a tablet.
In yet another aspect, the invention provides etoposide preparation as free flowing granules that can be directly filled into a sachet.
In a further aspect, the invention provides process for preparation of free flowing granules of etoposide that can be either filled into hard gelatin or compressed into a tablet. Accordingly, the invention provides process for preparation of free flowing granules of etoposide which comprises:
(a) Sifting and dry mixing of Diluents, Etoposide and intragranular stage of Disintegrant;
(b) Homogeneous shear mixing with mixture of surfactants and Solubilizer followed by granulation;
(c) Drying of wet granules and sizing followed by addition of lubricant, extra granular stage of disintegrant and surfactants to obtain free flowing granules and
d) Filling directly into hard gelatin capsule or a sachet or compressing into a tablet.
Further, the process of the instant invention is robust and cost effective and can easily be adopted on industrial scale.
Detailed description of the invention:
The present invention provides etoposide preparation as free flowing dry granules with good oral bioavailability, comprising etoposide in an amount of 22%; diluents in an amount of 50 to 70% and surfactant as binders in an amount of 5 to 15% together with other pharmaceutical excipients.
The other pharmaceutical excipients are selected from glidants/lubricants, disintegrants and solubilizers. The free flowing granules of Etoposide can be directly filled into hard gelatin capsule or a sachet or can be compressed into a tablet.

Diluents used for the purpose of the invention including sugar derivatives such as lactose, sucrose, glucose, mannitol, sorbitol; starch derivatives such as cornstarch, potato starch, alpha -starch, and dextrin, pregel starch; cellulose derivatives such as microcrystalline cellulose, carbonates such as calcium carbonate; and sulfate derivatives such as calcium sulfate.
Lubricants/glidants used for the purpose of the invention for instance may be selected from metallic salts of stearic acid such as colloidal silicon dioxide, stearic acid, calcium stearate, magnesium stearate; talc; waxes such as beeswax, spermaceti; boric acid; laurylsulphates such as sodium lauryl sulfate, magnesium lauryl sulfate; silicates such as anhydrous silicic acid, silicic acid hydrates; and starch derivatives described above can be listed.
Binders used for the purpose of the invention are a mixture of one or more surfactants
selected from Tween-80,and Sodium Lauryl Sulphate (SLS).
Surfactants used for the purpose of invention are the mixture of one or more surfactants
selected from Tween-80, Polyoxyl-40 Hydroxygenated castor oil (Cremophore-RH-40)
and sodium Lauryl sulphate (SLS).
Polyoxyl-40 Hydroxygenated castor oil (Cremophore-RH-40) can also be used as
solubilizer for the purpose of the instant invention.
Disintegrants used for the purpose of the invention may be selected from cellulose derivatives such as Crospovidone, low-substituted hydroxypropylcellulose and internally bridged-sodium carboxymethylcellulose; chemically modified starch/cellulose derivatives such as carboxymethylstarch, sodium carboxymethylstarch, bridged polyvinylpyrrolidone; and starch derivatives as described above.
In another embodiment, the invention provides etoposide formulation as free flowing granules that can be directly compressed into a tablet. The tablet of the invention may optionally be coated with tablet coatings those are well known in the art for instance film coating.

In a preferred embodiment, the instant invention in the form of free flowing granules comprises:
a) Etoposide present in an amount of 22%;
b) Lactose anhydrous present in an amount of 5% to 20%.
c) Microcrystalline cellulose PH101 present in an amount of 15 % to 35 % preferably 25 %;
d) ' Cross povidone present in an amount of 2% to 5% preferably 4%;
e) Pregelatinised starch present in an amount of 10% to 25 % preferably 20 %;
f) Tween-80 present in an amount of 3% to 6 % preferably 4%;
g) Polyoxyl-40 Hydroxygenated castor oil (Cremophore-RH-40) present in an amount of 3 to 6% preferably 4 %;
h) Sodium lauryl sulphate present in an amount of 1% to 5 % preferably 3%; and i) Colloidal silicon dioxide present in an amount of 1 % to 5% preferably 3 %.
In a further embodiment the invention provides process for preparation of free flowing granules of etoposide which comprises:
a) Sifting and dry mixing of Lactose anhydrous, Microcrystalline Cellulose (Avicel
PH-101), Crospovidone, pregelatinized Starch & Etoposide;
b) Preparation of homogeneous mixture of Polyoxyl-40 Hydroxygenated castor oil
(Cremophore-RH-40), Tween-80 and Sodium Lauryl Sulphate;
c) Shear mixing of mixture of surfactant with dry mix blend of Etoposide, followed by Granulating with Isopropyl Alcohol and Methylene Dichloride solvent to ensure uniform mixing;
d) Drying and sizing the above blend in to granules;
e) Lubricating the free flowing dried granules with Crospovidone, Colloidal Silicon Dioxide and Sodium Lauryl Sulphate; and
f) Finally filling the lubricated granules in to hard gelatin capsule or alternately compressed into a tablet.
The free flowing granules thus obtained can be filled into the capsule or sachet. Alternately, the same may be compressed into a tablet. The tablet may be optionally coated with the coatings well known in the art. The process of the instant invention is advantageous for industrial scale as it is easy,

robust and cost effective.
The following examples, which include preferred embodiments, will serve to illustrate the
practice of this invention, it being understood that the particulars shown are by way of
example and for purpose of illustrative discussion of preferred embodiments of the
invention.
Examples
Examples of Etoposide capsules 50mg

Sr. No Ingredients Rational Example 1 Example 2 Example 3 Example 4
STEP-I DRY MIXING
1. Etoposide USP Active 50 50.00 50 50
2. Lactose anhydrous Diluents 42 32 29 24
3. Microcry stall ine cellulose(Avicel PH 101) Diluent 60.00 60.00 60.00 60.00
4. Crospovidone Disintegrants 10.00 10.00 10.00 10.00
5. Pregelatinised starch Diluents 45.00 45.00 45.00 45.00
STEP-II BINDING
6. Tween-80 Surfactant 10.00 10.00 10.00 10.00
7. Polyoxyl 40 hydroxygenated castor oil(cremophore RH-40) Solubiliser 10.00 10.00 10.00 10.00
8. Sodium Lauryl Sulphate Surfactant 3.00 3.00
9. Isopropyl alcohol Solvent/Bind ing vehicle Q.S. Q.S. Q.S. Q.S.
10. Methylene dichloride Solvent Q.S. Q.S. Q.S. Q.S.
STEP-III DRY MIXING (LUBRICATION)
11. Crospovidone Disintegrant 5.00 5.00 5.00 5.00

12. Colloidal silicon dioxide Lubricant 8.00 8.00 8.00 8.00
13 Sodium Lauryl Sulphate Surfactant 5.00
Manufacturing process:
Stepl Dry mixing and Granulation
Sifted the following material through 40# sieve and mixed uniformly
• Lactose anhydrous,
• Microcrystalline Cellulose (Avicel PH-101),
• Cross Povidone and
• Pregelatinised starch Step2:
Mixed batch quantity of Etoposide with above mixed material of step-1 uniformly
Step 3:
Mixed homogenously the given quantity of Tween-80, cremophorRH40 and Sodium Lauryl Sulphate (SLS) to bind the dry mixed material of step 2
Step 4:
Added the given quantity of Methylene Dichloride (MDC) and Isopropyl Alcohol (IPA) to the above binded material followed by shear mixing to form uniform dispersion of material.
Step 5:
Air dried the material of step 4 to evaporate the solvent and then dried at 50°C fori hour to obtain semi dried material. The semidried material passed through 20# sieve and after fully drying, the material was passed through 30# sieve
Step 6: Dry mixing and Lubrication
Sifted cross povidone, Colloidal Silicon Dioxide (Aerosil-200) and Sodium Lauryl
Sulphate through 60# and mixed with dried granules uniformly.

Step 7: Filled the lubricated granules in size'2' empty hard gelatin capsule.
Alternately, the lubricated free flowing granules either filled into a sachet or compressed into a tablet using suitable tablet punches to obtain the tablets with desired shape and size.
Example 5:
Stability data of etoposide capsule-50mg in HDPE bottle with child resistant screw
packs.

Sr. No. Parameters STD Limit Withdrawal Period

Initial 1M 2M 3M 6M

RT 30°C RT 30°C RT 30°C RT 30°C
1. Description Size '2' Hard gelatin filled capsules, containing almost white to off white granules same same same same same same same same *
2. Identification (ByUV) To comply. Compli
es Compli es Compli
es Compli
es Compli es Compli es Compli es Compli es Compli es
3. Average Weight 295 gm± 5% 296 gm 296 gm 297 gm 292 gm 289 gm 288 gm 293 gm
4. Average Inner content 230 mg± 5%.' 233 mg 239, mg 228 mg 227 mg 236 mg 232 mg 229 mg
5 Disintegration Time N.M.T. 30
minutes 2.31 min 2.19
min 2.25 min 2.35 min 2.37 min 2.33 min. 2.29 min. 4.65 min. 3.32 min.
6. Length 17.0 ±0.2 mm 17.26 mm 17.19
mm 17.18 mm 17.21 mm 17.23 mm 17.25 mm 17.28 mm
7. Related
Compound
(ByHPLC)
I) Picro
Etoposide
II) Total
Impurities NMT2.0% NMT3.0% NIL
0.087% NIL 0.095 NIL 0.098 ' NIL 0.157% NIL 0.17% NIL
0.16% NIL 0.17% NIL
0.18 NIL
0.18
8. Dissolution (ByHPLC) NLT 85% in30 min 98.39% 98.81% 98.90% 99.56% 105.62 % 92.49% 96.58% 97.24% 87.23 %
9. Assay (By HPLC) (OA 2%) NLT 90%
to NMT110% 104.65 % 107.16 % 103.46 104.73 % 107.26
% 104.69
% 98.11%
s 102.
99% 103.44 %

*Average Inner content==Empty capsule weight is around 65mg.
Filled content is 230mg
so total average weight is 295mg.
Conclusion:
The Etoposide capsule developed with this novel method showed successful compliance at RT and intermediate condition (30°C/65%RH) throughout 6 months. Dissolution of this poor water insoluble API proved to be bioavailable for oral route as per the remarkable invitro dissolution results obtained. The shelf life of the product is therefore proved to be 2 years.

We Claim,
1. A stable etoposide preparation with good oral bioavailability comprising free flowing granules of etoposide containing etoposide in an amount of 22%, diluents in an amount of 50 to 70% and one or more surfactant as binders in an amount of 5 to 15% together with other pharmaceutical excipjents.
2. The preparation according to claim 1, wherein said preparation is enclosed in hard gelatin capsule.
3. The preparation according to claim 1, wherein said preparation is compressed into a tablet.
4. The preparation according to claim 1, wherein said preparation is filled into a sachet.
5. The preparation according to claim 1, wherein the diluents are selected from lactose, sucrose, glucose, mannitol, sorbitol; starch derivatives such as cornstarch, potato starch, alpha -starch, and dextrin, pregel starch; cellulose derivatives such as microcrystalline cellulose , carbonates such as calcium carbonate; and sulfate derivatives such as calcium sulfate.
6. The preparation according to claim 1, wherein the Surfactants are selected from one or more of surfactants selected from Tween-80, Polyoxyl-40 Hydroxygenated castor oil (Cremophore-RH-40) and Sodium Lauryl Sulphate (SLS).
7. The preparation according to claim 1, wherein the other pharmaceutical excipients are selected from lubricants/glidants, solubilizers and disintegrants.
8. The preparation according to claim 1, wherein the Disintegrants are selected from Crosspovidone, Croscarmellose sodium and sodium starch glycolate.
9. The preparation according to claim 1, wherein the glidant is Colloidal silicon

dioxide.
10. The preparation according to claim 1, wherein the solubilizer is Polyoxyl 40 hydroxygenated castor oil(cremophore RH-40).
11. The preparation according to claim 1, wherein the free flowing granules of etoposide comprising:

(a) Etoposide present in an amount of 22%
(b) Lactose anhydrous present in an amount of 5% to 20%.
(c) Microcrystalline cellulose PH101 present in an amount of 15 % to 35, more
preferably 25%
(d) Cross povidone present in an amount of 2 % to 5, more preferably 4%
(e) Pregelatinised starch present in an amount of ] 0% to 25 %, more preferably 20%
(f) Tween-80 present in an amount of 3% to 6 %, more preferably 4%.
(g) Polyoxyl-40 Hydroxygenated castor oil (Cremophore-RH-40) present in an
amount f 3 to 6%; more preferably 4%.
(h) Sodium lauryl sulphate present in an amount of 1% to 5 %, and, more preferably
3%
(i) Colloidal silicon dioxide present in an amount of 1 % to 5%., more preferably 3%
12. A process for preparation of etoposide preparation in the form of free flowing
granules comprises:
(a) Sifting and dry mixing of Diluents, Etoposide and intragranular stage of
Disintegrant;
(b) Homogeneous shear mixing with mixture of surfactants and Solubilizer followed by granulation;
(c) Drying of wet granules and sizing followed by addition of lubricant, extra granular stage of disintegrant and surfactants to obtain free flowing granules and
(d) Filling directly into hard gelatin capsule or a sachet or compressing into a tablet.
13. The process according to claim 12, wherein the diluents are selected from Lactose
Anhydrous, Microcrystalline cellulose (Avicel PH 101), and Pregelatinised starch.

14. The process according to claim 12, wherein the surfactants are selected from Tween-80, Polyoxyl-40 Hydroxygenated castor oil (Cremophore-RH-40) and Sodium Lauryl Sulphate (SLS).
15. The process according to claim 12, wherein the solubilizer is Polyoxyl 40 hydroxygenated castor oiI(cremophore RH-40).
16. The process according to claim 12, wherein the intragranular and extra granular disintegrant used is crosspovidone.
17. The process according to claim 12, wherein the glidant is Colloidal silicon dioxide.