FORM 2
THE PATENTS ACT, 1970
(39 OF 1970)
&
PATENTS RULES, 2006
COMPLETE SPECIFICATION
(SECTION 10; RULE 13)
Title:
"STABLE PHARMACEUTICAL COMPOSITION OF ACTIVE FRAGMENTS OF BASIC FIBROBLAST GROWTH FACTOR (BFGF) FOR TREATING VITILIGO AND PROCESS OF PREPARATION THEREOF"
Applicants:
ALKEM LABORATORIES LIMITED, A COMPANY INCORPORATED UNDER THE COMPANIES ACT, 1956, HAVING ITS CORPORATE OFFICE AT ALKEM HOUSE, DEVASH1SH, ADJACENT TO MATULYA CENTRE, S.B.MARG, LOWER PAREL, MUMBAI - 400013, MAHARASHTRA, INDIA
THE FOLLOWING SPECIFICATION PARTICULARLY DESCRIBES THE NATURE OF THE INVENTION AND THE MANNER IN WHICH IT IS TO BE PERFORMED:

FIELD OF THE INVENTION
The present invention relates to a stable pharmaceutical composition of active fragments of basic fibroblast growth factor (bFGF) for treating Vitiligo and process of preparation thereof.
BACKGROUND OF THE INVENTION
MelanocytIC cells are responsible for pigmentation of the skin, a greater number of melanocytes present per/cm of skin surface area causes a greater pigmentation. Vitiligo is a disorder caused by a deficiency or absence of functional melanocyte cells in the dermal-epidermal junction. Vitiligo/Leucoderma is an acquired depigmentation or absence of functional melanocyte cells in the dermal-epidermal junction of skin characterized by patchy loss of pigment that becomes progressive with time. This disorder affects about 1% of the world population. Traditional therapies for Vitiligo mainly include photo chemotherapy with topical/oral psoralens followed by exposure to ultra violet A radiation (PUV/A) or topical/oral steroids, PUV-A therapy is perhaps the main stay in the treatment of Vitiligo. However only about 50% of cases get repigmenatation. Moreover in a patient in response to PUV, many Vitiligo patches may repigment only partially and the rest of the patches may remain unresponsive to PUV-A therapy even after long duration of treatment. The repigmenatation in the above therapies is a result of multiplication of melanocytes, the cells, which produce the pigment melanin in the skin. The multiplications of melanocytes in response to the above therapies occur from the margins of the Vitiligo patch or at the pigmented hair follicles and their migration/spread to the Vitiligo patch.
Basic fibroblast growth factor (bFGF) also known as FGF2 so named because it contains a high number of basic amino acid residues (Lysine, Arginine and Histidine) is a potent mitogen for a variety of cell types including melanocytes. Both human and bovine bFGF were isolated and the gene expressing this product were sequenced and cloned. In addition bFGF was found to be expressed in a wide variety of tissue types including placenta, keratinocyles and fibroblasts.

The bFGF or its against peptides were tested on human volunteers in the various phases of clinical trials in India and found to be successful in repigmenting about 80% of volunteers with stable generalized Vitiligo and segmental Vitiligo.
Vitiligo is a pigmentary disorder with patchy loss of skin pigment melanin that deprivation of a mitogen(s) like basic fibroblast growth factor (bFGF) for melanocytes or its decreased level in the skin of Vitiligo patients for use at unknown reason could result in the loss of melanin producing cells melanocytes in skin resulting in Vitiligo. Basic fibroblast growth factor (bFGF) also known as FGF2 is a potent mitogen for variety of cell types including melanocytes. Both human and bovine bFGF have been isolated and the gene expressing tin product have been sequences and cloned. In addition bFGF has been found to be expressed in a wide variety of tissue types including pituitary, brain and adrenal gland corpusluteum, retina, kidney, placenta and keratinocytes fibroblasts.
Patents of interest describing bFGF or against peptides and the formulation for their penetrations through intact skin include US Patent 6143723, AU722626, IN185613, IN186437, IN185703, IN186437, IN217659 and 42/DEL/2004.
US6143723 and INI 85613 disclosed a composition for treatment of pigmentation disorders and tanning applications of a skin surface without the necessity of exposure to sunlight comprising 10-50% of solvent mixed with 10-40% of glycols, 10-40% of a penetration enhancing agent and 0.5%. of selective active fragments of basic fibroblast growth factor (bFGF) mixed therein. Further, according to this invention there is provided a process for the preparation of a composition comprising mixing said solvent with said glycol at ambient temperature and pressure, adding penetration enhancing agent therein.
AU722626 disclosed a composition for treatment of pigmentation disorders and tanning application of a skin surface without the necessity of exposure to sun light comprising of 10-50 % of solvent mixed with 10-40 % of glycols, 10-40 % of a penetration enhancing agent and 0.5' % of selective active fragments of basic fibroblast growth factor (bFGF) mixed therein.
IN185703 disclosed a process for the preparation of a composition for the treating Vitiligo and tanning white skin comprising mixing 10-90% solvent (for example

alcohol or glycols) with 10-90 % sebacate at the ambient temperature and 1 atmospheric pressure and then adding 0.02-0.5% selective active fragments of bFGF thereto and mixing thoroughly therewith to get the above synergistic composition.
INI 86437 disclosed a composition for active agents for application other than tanning of skin surface or pigmentary disorder and this composition capable of administrating an active agent comprising 10-50 % solvent mixed with 10-40 % glycol and 10-40% of penetrating enhancing agent being mixed therewith.
IN217659 disclosed a composition for treating Vitiligo and tanning white skin comprising 10-90% solvent, 10-90% sebacate wherein the said sebacate is sodium or potassium salt of the fatty acid or other cation salt of fatty acid and 0.02-0.5% active fragments of bFGF. In accordance with this invention, the composition has 10-90% weight by weight of solvent, 10-90% weight by weight of sebacate wherein the sebacate is sodium or potassium salt of the fatty acid or other cation salt of fatty acid at the ambient temperature preferably at a temperature around 25°C and 0.02 to 0.5% weight by weight selective active fragment of basic fibroblast growth factor (bFGF) mixed with each other thoroughly to obtain the above mentioned composition. The solvent comprises alcohols or glycols. The alcohols are for example, isopropyl alcohol, propyl alcohol or ethyl alcohol. The glycols are for example propylene glycol or polyethylene glycol.
42/DEL /2004 disclosed a composition for combinatorial synergistic treatment of generalized stable/non-stable and segmental Vitiligo comprising of 0.02 to 5% w/w of at least one peptide selected from a group comprising of bFGF amino acids 106-115 (SEQ ID NO I), bFGF amino acids 106-120 (SEQ ID NO 5), bFGF amino acids 1-24 (SHQ.ID NO 6), SEQ H): 2. SEQ 1D:3, cyclic bFGF amino acids 106-115 (SEQ ID NOl). cyclic bFGF amino acids 106-120 (SEQ ID N05), cyclic bFGF amino acids 1-24 (SEQ ID N06), cyclic SEQ ID:2 and cyclic SEQ 1D:3 in 10-15% w/w of solvent. 10-40% w/w of glycols, and 10-40% w/w of at least one penetration enhancing agent for local application, 10-40 mgs of 8methoxy psoralens and 1-5 mgs of ncopsoralens for oral intake. After 2 hours of oral administration, Vitiligo patches of the patient are exposed to morning sunlight for 5-10 minutes till the patches show erythma /red or to UV-A of 2-8 Joules/Cm .

2022/MUM/2012 disclosed a composition of active fragment of bFGF, decapeptide in aqueous acidulated medium incorporating acetate ions using sodium acetate in solution having pH 4.5 to 5.5, co-penetrating agent in the range of 5-9%w/v, emulsifier in the range of 0.35-5.00%w/v, non-ionic surfactant in the range of 0.0001% to 1% w/v and film forming agent in the range of 5-8%v/v.
OBJECT OF THE INVENTION
It is an object of the present invention to provide a clear, stable pharmaceutical composition in the form of solution for topical administration in the treatment of Vitiligo comprising:
a) Active fragment of bFGF in the range 0.02 to 0.5 % w/w, as an active ingredient;
b) at least one penetration enhancing agent in the range of 10-40 % w/v;
c) at least one non-ionic surfactant in the range of 0.001% to 2 % w/v and acidulated medium in the range of 5 to 20 % v/v, using sodium acetate.in solution having pH 4.00 to 5.00;
Wherein the pH of the final solution for topical administration is about 7.00 and said pharmaceutical composition does not contain a glycol.
It is also an object of the present invention to provide a process to prepare a clear, stable pharmaceutical composition in the form of solution for topical administration in the treatment of Vitiligo, said composition comprising;
a) Active fragment of bFGF in the range 0.02 to 0.5 % w/w, as an active ingredient;
b) at least one penetration enhancing agent in the range of 10-40 % w/v;
c) at least one non-ionic surfactant in the range of 0.001% to 2 % w/v and acidulated medium in the range of 5 to 20 % v/v, using sodium acetate in solution having pH 4.00 to 5.00;
Wherein the pH of the final solution for topical administration is about 7.00 and said pharmaceutical composition does not contain a glycol.
It is also an another object of this invention is to provide a topical pharmaceutical composition which can advantageously be employed to a skin surface for the treatment of Vitiligo, without the necessity of an exposure to sunlight or causing unnecessary skin irritation due to glycols.

It is yet another object of the present invention to provide a clear, stable pharmaceutical composition in the form of solution for topical administration in the treatment of Vitiligo comprising: active fragment of bFGF, which uses a new and different beneficial formulation but is still more stable and non hazy solution for topical administration as compared to the commercially available compositions in the India.
SUMMARY OF THE INVENTION
The present invention relates to a stable pharmaceutical composition of active fragments of basic fibroblast growth factor (bFGF) for treating Vitiligo and process of preparation thereof. More particularly it relates to:
A. A stable pharmaceutical composition in the form of solution for topical administration
in the treatment of Vitiligo comprising:
a) Active fragment of bFGF in the range of 0.02 to 0.5 % w/w, as an active ingredient;
b) at least one penetration enhancing agent in the range of 10-40 % w/v;
c) at least one non-ionic surfactant in the range of 0.001% to 2.0 % w/v and acidulated medium in the range of 5 to 20 % v/v, using sodium acetate in solution having pH 4.00 to 5.00;
Wherein the pH of the final solution for topical administration is about 7.00 and said pharmaceutical composition does not contain a glycol.
B. The pharmaceutical composition as in A above, wherein the Active fragment of bFGF
is selected from a group comprising of bFGF amino acid 106-115 (SEQ ID No 1),
bFGF amino acid 106-120 (SEQ ID No 5), bFGF amino acid 1-24 (SEQ ID No
6),SEQ ID 2, SEQ ID 3, cyclic bFGF amino acid 106-115 (SEQ ID No 1) , cyclic
bFGF amino acid 106-120 (SEQ ID No 5) , cyclic bFGF amino acid 1-24 (SEQ ID
No 6), cyclic (SEQ ID 2) and cyclic (SEQ ID 3).
C. The pharmaceutical composition as in A above, wherein the penetrating enhancing
agent is selected from propyl ethyl myristate , isopropyl ethyl myristate , propyl ethyl
stearate, isopropyl ethyl stearate , propyl ethyl palmitate , isopropyl ethyl palmitate
and mixtures thereof.

D. The pharmaceutical composition as in A above, wherein the non-ionic surfactant is
selected from polyoxylated derivatives such as polysorbate 80, ethoxylated fatty
alcohol and derivatives thereof such as Steareth-2 and Laureth, and mixtures thereof.
E. The pharmaceutical composition as in A above, wherein the aqueous
acidulated medium incorporating acetate ions uses sodium acetate in solution having
pH 4.5 to 5.00.
F. A process to prepare a stable pharmaceutical composition in the form of solution for
topical administration in the treatment of Vitiligo, said composition comprising;
a) Active fragment of bFGF in the range of 0.02 to 0.5 % w/w, as an active ingredient;
b) at least one penetration enhancing agent in the range of 10-40 % w/v;
c) at least one non-ionic surfactant in the range of 0.001% to 2.0 % w/v and acidulated medium in the range of 5 to 20 % v/v, using sodium acetate in solution having pH4.00 to 5.00;
Wherein the pH of the final solution for topical administration is about 7.00 and said pharmaceutical composition does not contain a glycol; Wherein the process comprises:
a) Dissolving non-ionic surfactant in acidulated water,
b) Adding active fragment of bFGF in step a) with continuous stirring to get dissolved and /or adding solvent with continuous stirring to make a clear solution,
c) Adding non-ionic surfactant in the solvent with continuous stirring to get dissolved,
d) Adding step c) to step b) with continuous stirring,
e) Adding penetration enhancing agent into step (d) with continuous stirring,
f) Making up the volume with solvent,
. g) Filtering the solution with the nylon cloth.
G. The pharmaceutical composition as in F above, wherein the solvents) is selected from an alcohol such as isopropyl alcohol, propyl alcohol or ethyl alcohol, an acid such as caprylic acid and mixtures thereof.

H. The pharmaceutical composition as in G above, wherein the solvent(s) is used in the range of 10-90 % of total composition.
DETAILED DESCRIPTION OF THE INVENTION
Before the present process and methods are described, it is to be understood that this invention is not limited to particular compounds, formulas or steps described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.
Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges is also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either both of those included limits are also included in the invention.
Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the publications are cited.
It must be noted that as used herein and in the appended claims, the singular forms "a", "and", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example", reference to "a compound" includes a plurality of such compounds and reference to "the step" includes reference to one or more step and equivalents thereof known to those skilled in the art, and so forth.
The publications discussed herein are provided solely for their availability to the applicant prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of

prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed
The term "stable" as used herein refers to chemical stability of Active fragment of bFGF in topical solution forms wherein there is no change in assay values and dissolution and/or the total impurity remains less than 1%, when the dosage form is kept at room temperature for 3 months.
Preparations for treatment of Vitiligo are known in the art, but which were not satisfactory. Such preparations were taken orally or applied to the surface of the skin of a patient. Another treatment known in the art is the Psoralens and UVA therapy. However, such known methods required a prolonged treatment as long as one too many years. In addition, Psoralen are toxic to liver and carcinogenic. Moreover, such known treatments were more often found to be ineffective.
The composition reported in the prior arts IN185613, IN186437, IN185703, IN186437, IN217659 and 42/DEL/2004 contain three excepients, comprising of solvent mixed with glycols and a penetration enhancing agent and the marketed preparation prepared in accordance with these patents is hazy and unstable for a long time at room temperature. And further the glycol used in this composition may cause skin irritation and contact dermatitis associated with propylene glycol when used for a long time.
2022/MUM/2012 disclosed a composition of active fragment of bFGF with the same three excepients as disclosed above, comprising of solvent, glycols and a penetration enhancing agent with addition of more excepients such as emulsifier, nonionic surfactant and stabilizer and employs a hot melt potential technique for preparation of said composition. Hot melt technology are often related to high energy input mainly related to shear forces and temperature and the compound must be melted or dissolved in molten polymer at high temperatures. Thus, it is less applicable to compounds with higher melting temperatures or those that are thermally labile and a second disadvantage is that the formulation can be affected by process parameters such as temperature, high pressure, which have significant impact on product quality and degradation of Peptide and polymers. This aspect, combined

with the relatively large minimum batch size, results in increased cost and risk during early development.
As the above mentioned, stability problems are inherent in the compositions based on prior art, there still exists a need for an topical pharmaceutical composition in the form of solution, of the active fragment of bFGF which is capable of providing a clear stable formulation, not causing skin irritation and not using compacted hot melt technology along with additional emulsifier, nonionic surfactant with combination of stabilizer as used by the prior art.
It has now been found, surprisingly, that a clear, stable pharmaceutical composition, wherein the active fragment of bFGF cannot be converted into a form which causes the instability, is achieved by using an acid diluted water and non ionic surfactant with simple conventional mixing process of solution.
The active fragment of bFGF used in the ranges of 0.05%w/v to5%w/v can be selected from a group comprising of bFGF amino acid 106-115 (SEQ ID No 1), bFGF amino acid 106-120 (SEQ ID No 5), bFGF amino acid 1-24 (SEQ ID No 6),SEQ ID 2, SEQ ID 3, cyclic bFGF amino acid 106-115 (SEQ ID No 1) , cyclic bFGF amino acid 106-120 (SEQ ID No 5) , cyclic bFGF amino acid 1-24 (SEQ ID No 6), cyclic (SEQ ID 2) and cyclic (SEQ ID 3).
The penetrating enhancing agents(s) used in the range of 10-80 % w/v can be selected from propyl ethyl myristate, isopropyl ethyl myristate, propyl ethyl stearate, isopropyl ethyl stearate/propyl ethyl palmitate, isopropyl ethyl palmitate and mixtures thereof.
The aqueous acidulated medium in range of 5-20 % v/v, incorporating acetate ions using sodium or potassium acetate of the solution having pH 4.0 -5.0 and the pH of the final solution for topical administration is about 7.00, means it may be 7.00 to 7.5.
The solvent(s) used in the range of 10 -90 % of total composition, and is selected from alcohols such as isopropyl alcohol, propyl alcohol or ethyl alcohol, acid such as caprylic acid and mixtures thereof.

The non-ionic surfactant(s) used in the range of 0.001% to 5 % w/v is selected from polyoxylated derivatives such as polysorbate 80, the ethoxylated fatty alcohol and derivatives thereof such as Steareth-2 and Laureth, and mixtures thereof.
Another object of invention is a process to prepare a clear, stable pharmaceutical composition in the form of solution for topical administration in the treatment of Vitiligo, said composition comprising;
a) Active fragment of bFGF in the range of 0.02 to 0.5 % w/w, as an active ingredient; ' b) at least one penetration enhancing agent in the range of 10-40 % w/v;
c) at least one non-ionic surfactant in the range of 0.001% to 2 % w/v and acidulated medium in the range of 5 to 20 % v/v, using sodium acetate in solution having pH 4.00 to 5.00;
Wherein the pH of the final solution for topical administration is about 7.00 and said pharmaceutical composition does not contain a glycol. Wherein the process comprises:
a) Dissolving non-ionic surfactant in acidulated water,
b) Adding active fragment of bFGF in step a) with continuous stirring to get dissolved and /or adding solvent with continuous stirring to make a clear solution,
c) Adding non-ionic surfactant in the solvent with continuous stirring to get dissolved,
d) Adding step c) to step b) with continuous stirring,
e) Adding penetration enhancing agent into step (d) with continuous stirring,
f) Making up the volume with solvent,
g) Filtering the solution with the nylon cloth.
The composition of the present invention may be made by any process known in the art. A simple conventional mixing process of solution or dispersion process can be used. In a preferred example, addition or dispersion of active fragment of bFGF in the solvent, followed by addition of nonionic surfactants), penetrating enhancing agents and finally the volume is made up with the solvent.

The following examples are intended to illustrate the scope of the present invention in all its aspects but not to limit it thereto.
Example 1 % Range in Formulation

Sr.No Ingredients % Range in formulation
1) Active fragment of bFGF 0.02-0.5 %
2) Acidulated water 5-20 %
3) Polysorbate 80 0.5-2.0 %
4) Iso Propyl Alcohol 30-90 %
5) Brij 72 (Polyoxyl 21 stearyl ether) 0.20-2.0 %
6) Iso Propyl Myristate 10-40%
Final pH about 7.00
The polysorbate 80 was dissolved in acidulated water, followed by addition of active fragment of bFGF with continuous stirring. The Polyoxyl 21 stearyl ether was dissolved in isopropyl alcohol with continuous stirring. This solution was added to the prepared solution of active fragment of bFGF, followed by addition of iso propyl myristate with continuous stirring, and finally the volume was made up with isopropyl. The solution was filtered with nylon cloth and checked if the pH was about 7.0.
Example 2

Sr.No Ingredients mg/ml
1) Active fragment of bFGF 6.173 mg
2) Acidulated water 0.40 ml
3) Polysorbate 80 32.50 mg
4) Iso Propyl Alcohol 3.60 ml
5) Brij 72 (Polyoxyl 21 stearyl ether) 17.50 mg
6) Iso Propyl Myristate 1.0 ml
Final pH about 7.00

Manufacturing Procedure:
The polysorbate 80 was dissolved in acidulated water, followed by addition of active fragment of bFGF with continuous stirring. The Polyoxyl 21 stearyl ether was dissolved in isopropyl alcohol with continuous stirring. This solution was added to the prepared solution of active fragment of bFGF, followed by addition of iso propyl myristate with continuous stirring, and finally the volume was made up to 5 ml with isopropyl. The solution was filtered with nylon cloth and checked if the pH was about 7.0.
Example 3 % Range in Formulation

Sr.No Ingredients % Range in formulation
1) Active fragment of bFGF 0.02-0.5 %
2) Acidulated water 5-20 %
3) polysorbate 80 0.5-2.0 %
4). Iso Propyl Alcohol 30-90 %
5) Brij 72(Polyoxyl 21 stearyl ether) 0.20-2.0 %
6) Iso Propyl Myristate 10-40%
7) Caprylic acid 5-10%
8) Final pH about 7.00
Manufacturing Procedure:
The polysorbate 80 was dissolved in acidulated water, followed by addition of active
fragment of bFGF with continuous stirring, followed by addition of caprylic acid.
The Polyoxyl 21 stearyl ether was dissolved in isopropyl alcohol with continuous stirring.
This solution was added to the prepared solution of active fragment of bFGF, followed by
addition of iso propyl myristate with continuous stirring, and finally the volume was made up
to with isopropyl. The solution was filtered with nylon cloth and checked if the pH was about
7.0.

Example 4

Sr.No Ingredients mg/ml
1) Active fragment of bFGF 6.173 mg
2) Acidulated water 0.40 ml
3) polysorbate 80 32.50 mg
4) Iso Propyl Alcohol 3.30 ml
5) Brij 72(Polyoxyl 21 stearyl ether) 17.50 mg
6) Iso Propyl Myristate 1.00 ml
7) Caprylic acid 0.30 ml
8) Final pH about 7.00
Manufacturing Procedure:
The polysorbate 80 was dissolved in acidulated water, followed by addition of active fragment of bFGF with continuous stirring, followed by addition of caprylic acid. The Polyoxyl 21 stearyl ether was dissolved in isopropyl alcohol with continuous stirring. This solution was added to the prepared solution of active fragment of bFGF, followed by addition of iso propyl myristate with continuous stirring, and finally the volume was made up to 5 ml with isopropyl. The solution was filtered with nylon cloth and checked if the pH was about 7.0.
Example 5: Stability Data
The compositions prepared in example 2 and 4 were subjected to stability testing at 25° C/ 65 % Rh for a period of 3M.

S. No. Batch No. Assay value at stability condition

Initial 3 Months at 25°C/65%Rh
1. B2 (based on Example no. 2) 97.74% 97.00%
2. Bl (based on Example no. 4) 101.72% 100.40%
A THUS it can be seen that the compositions of the present invention were stable.

Although the invention has been described in terms of particular embodiments and applications, one of ordinary skill in the art, in light of this teaching, can generate additional embodiments and modifications without departing from the spirit of or exceeding the scope of the claimed invention. It should be emphasized that the above-described embodiments of the present invention, particularly any "preferred" embodiments, are merely possible examples of the invention of implementations, merely set forth for a clear understanding of the principles of the invention. Accordingly, it is to be understood that the drawings-and descriptions herein are preferred by way of example to facilitate comprehension of the invention and should not be construed to limit the scope thereof.

Claims:
1. A stable pharmaceutical composition in the form of solution for topical administration
in the treatment of Vitiligo comprising:
a) Active fragment of bFGF in the range of 0.02 to 0.5 % w/w, as an active ingredient;
b) at least one penetration enhancing agent in the range of 10-40 % w/v;
c) at least one non-ionic surfactant in the range of 0.001% to 2.0 % w/v and acidulated medium in the range of 5 to 20 % v/v, using sodium acetate in solution having pH 4.00 to 5.00;
Wherein the pH of the final solution for topical administration is about 7.00 and said pharmaceutical composition does not contain a glycol.
2. The pharmaceutical composition as in claim 1, wherein the Active fragment of bFGF is selected from a group comprising of bFGF amino acid 106-115 (SEQ ID No 1), bFGF amino acid 106-120 (SEQ ID No 5), bFGF amino acid 1-24 (SEQ ID No 6),SEQ ID 2, SEQ ID 3, cyclic bFGF amino acid 106-115 (SEQ ID No 1) , cyclic bFGF amino acid 106-120 (SEQ ID No 5) , cyclic bFGF amino acid 1-24 (SEQ ID No 6), cyclic (SEQ ID 2) and cyclic (SEQ ID 3).
3. The pharmaceutical composition as in claim 1, wherein the penetrating enhancing agent is selected from propyl ethyl myristate, isopropyl ethyl myristate , propyl ethyl stearate, isopropyl ethyl stearate , propyl ethyl palmitate , isopropyl ethyl palmitate and mixtures thereof.
4. The pharmaceutical composition as in claim 1, wherein the non-ionic surfactant is selected from polyoxylated derivatives such as polysorbate 80, ethoxylated fatty alcohol and derivatives thereof such as Steareth-2 and Laureth, and mixtures thereof.
5. The pharmaceutical composition as in claim 1, wherein the aqueous acidulated medium incorporating acetate ions uses sodium acetate in solution having pH 4.5 to 5.00.

6. A process to prepare a stable pharmaceutical composition in the form of solution for
topical administration in the treatment of Vitiligo, said composition comprising;
a) Active fragment of bFGF in the range of 0.02 to 0.5 % w/w, as an active ingredient;
b) at least one penetration enhancing agent in the range of 10-40 % w/v;
c) at least one non-ionic surfactant in the range of 0.001% to 2.0 % w/v and acidulated medium in the range of 5 to 20 % v/v, using sodium acetate in solution having pH4.00 to 5.00;
Wherein the pH of the final solution for topical administration is about 7.00 and said pharmaceutical composition does not contain a glycol; Wherein the process comprises:
a) Dissolving non-ionic surfactant in acidulated water;
b) Adding active fragment of bFGF in step a) with continuous stirring to get dissolved and /or adding solvent with continuous stirring to make a clear solution;
c) Adding non-ionic surfactant in the solvent with continuous stirring to get dissolved;
d) Adding step c) to step b) with continuous stirring;
e) Adding penetration enhancing agent into step (d) with continuous stirring,
f) Making up the volume with solvent;
g) Filtering the solution with the nylon cloth.
7. The pharmaceutical composition as in claim 6, wherein the solvent(s) is selected from an alcohol such as isopropyl alcohol, propyl alcohol or ethyl alcohol, an acid such as caprylic acid and mixtures thereof.
8. The pharmaceutical composition as in claim 6, wherein the solvent(s) is used in the range of 10-90 % of total composition.