WO2025037322A1

WO2025037322A1 - Pharmaceutical compositions for treatment of vitiligo and other skin disorders

Info

Publication number: WO2025037322A1
Application number: PCT/IN2024/050276
Authority: WO
Inventors: Pavan Kumar Kothapuvari
Original assignee: Individual
Current assignee: Individual
Priority date: 2023-08-12
Filing date: 2024-03-17
Publication date: 2025-02-20
Anticipated expiration: 2026-02-12

Abstract

: The present invention provides a topical pharmaceutical composition comprising Janus kinase (JAK) inhibitors like Tofacitinib, Immunosuppressive agent like Tacrolimus, Fingolimod and statins like atorvastatin, Simvastatin, Fluvastatin, Lovastatin, Pitavastatin, Pravastatin and Rosuvastatin dissolved or dispersed in a pharmaceutically acceptable base together with other pharmaceutically acceptable excipients or in a dermatologically acceptable carrier, useful in the treatment of vitiligo. The inventive concept provided herein are methods and pharmaceutical compositions by preparing oral dosage forms like Topical liquid spray or Topical cream or Topical gel dosage form for the management of vitiligo and its associated disorders and accordingly performing preclinical study to prove the efficacy of the selected drug substances from the class of JAK inhibitors, statins and immunosuppressants.

Description

PHARMACEUTICAL COMPOSITIONS FOR TREATMENT OF VITILIGO AND OTHER SKIN DISORDERS

FIELD OF THE INVENTION:

The present invention relates to pharmaceutical compositions containing Immunomodulators, JAK inhibitors and Statins for treatment of Vitiligo, Psoriasis, Atopic dermatitis, Alopecia, and other skin related disorders in suitable dosage form.

BACKGROUND OF THE INVENTION:

The etiology of vitiligo comprises genetic and autoimmune predispositions, as well as biochemical, neurochemical, and environmental factors.

Skin and blood of patients with an active form of vitiligo is abundant with autoreactive, melanocyte specific CD8+ T lymphocytes, which are critical and sufficient for the initiation of depigmentation Active CD8+ T lymphocytes produce interferon (IFN)-y which, accompanied by tumor necrosis factor (TNF)-a, represents the cytokine profile characteristic of a T-helper (Th)l cell response.

The stimulation of a Thl response plays a pivotal role in the pathogenesis of vitiligo. Additionally, the pathogenesis of vitiligo is associated with a decreased skin concentration of interleukin (IL)- 10, which belongs to the family of Th2 -dependent cytokines.

As a result of the actions of statins, a noticeable increase in anti-inflammatory cytokine (IL-4, IL-5, and IL-10) secretion can be observed. Moreover, a shift to a Th2 -dependent response occurs. IL-17, a cytokine that stimulates production of TNF-a, has been reported to be present in higher concentrations in the serum and tissue in vitiligous patients.

Vitiligo maintenance has also been proven to be associated with an increased IL- 17 concentration another cell population that plays an important role in pathogenesis of vitiligo is T regulatory lymphocytes (Treg).

It has been found that a decreased peripheral blood concentration of Tregs, as well as their impaired activity, are responsible for an enhanced destruction of melanocytes. As a result of statin use, inhibition of T cell differentiation into Th- 17 cells secreting IL- 17 can be observed. These phenomena result in inhibition of inflammatory processes and lead to Immunotolerance, Statins have also been found to cause lymphocyte anergy. According to the available data, the impairment of lymphocyte migration as well as a reduced influx to the inflammatory site underlie this condition.

Based on all the presented pathogenetic mechanisms of acquired vitiligo, we hypothesized that the use of statins may be beneficial for the development of vitiligous lesions and the appearance of re-pigmentation. Statins, acting directly on melanocyte -specific CD8+ T lymphocytes, lead to their limited proliferation as well as to decreased IFN-y production in murine vitiligo models.

These drugs are also known as immunosuppressants or immunomodulators & Examples of CD8+ T lymphocyte antagonist drugs include:

1. Cyclosporine: This drug blocks the activation of CD8+ T cells by inhibiting the production of interleukin-2, a cytokine that is required for T cell activation.

2. Tacrolimus: This drug also blocks T cell activation by inhibiting the production of interleukin-2.

3. Mycophenolate mofetil: This drug inhibits the proliferation of T cells, including CD8+ T cells, by blocking the production of nucleotides that are required for DNA synthesis.

4. Azathioprine: This drug also inhibits T cell proliferation by interfering with DNA synthesis.

Interferon (IFN)-y antagonist drugs are a class of medications that are used to block the activity of IFN-y, a cytokine that plays a key role in immune system function, in certain autoimmune disorders.

These drugs are also known as IFN-y inhibitors or IFN-y antagonists & Examples of IFN-y antagonist drugs include:

1. Tocilizumab: This drug is a monoclonal antibody that binds to the interleukin-6 (IL-6) receptor, which is involved in the production of IFN-and blocks its activity.

2. Anakinra: This drug is a recombinant protein that blocks the activity of IL-1, which is also involved in the production of IFN-y. 3. Ustekinumab: This drug is a monoclonal antibody that binds to the p40 subunit of interleukin- 12 (IL-12) and interleukin-23 (IL-23), both of which stimulate the production of IFN-y.

These drugs can have side effects such as increased risk of infection and injection site reactions. They are typically prescribed under close medical supervision and require regular monitoring of blood counts and organ function.

Anti-inflammatory cytokine antagonist drugs are a class of medications that are used to block the activity of interleukin-4 (IL-4), interleukin-5 (IL-5), and interleukin- 10 (IL-10), which are all involved in regulating immune responses and inflammation. These drugs are also known as IL-4, IL-5, and IL- 10 inhibitors or antagonists.

Examples of anti-inflammatory cytokine antagonist drugs include:

1. Dupilumab: This drug is a monoclonal antibody that binds to the receptor for IL-4 and blocks its activity. It is used to treat certain inflammatory conditions such as atopic dermatitis and asthma.

2. Mepolizumab: This drug is a monoclonal antibody that binds to IL-5 and blocks its activity. It is used to treat eosinophilic asthma and other eosinophilic disorders.

3. Nivolumab: This drug is a monoclonal antibody that blocks the activity of PD-1, a receptor that can inhibit the activity of immune cells, including those that produce IL- 10. It is used to treat certain types of cancer.

These drugs can have side effects such as increased risk of infection and allergic reactions. They are typically prescribed under close medical supervision and require regular monitoring of blood counts and organ function.

The exact cause of vitiligo is unknown, but it is thought to be an autoimmune disorder. There is no known cure for vitiligo, but there are several treatment options available to manage the symptoms.

1. Topical Corticosteroids: These are anti-inflammatory drugs that can help to reduce inflammation and stop the immune system from attacking the melanocytes (pigment cells). They are usually applied to the affected areas of the skin. 2. Topical Calcineurin Inhibitors: These are drugs that also help to reduce inflammation and stop the immune system from attacking the melanocytes. They are usually applied to the affected areas of the skin.

3. Psoralen Plus Ultraviolet A (PUVA) Therapy: This is a type of light therapy that involves taking a medication called psoralen and then exposing the skin to ultraviolet A (UVA) light. This helps to re-pigment the skin.

4. Narrowband Ultraviolet B (NB-UVB) Therapy: This is another type of light therapy that involves exposing the skin to narrowband UVB light. This helps to re-pigment the skin.

5. Excimer Baser: This is a type of laser therapy that uses a high-energy beam of UVB light to re-pigment the skin.

6. Depigmentation: In severe cases of vitiligo where more than 50% of the body is affected, depigmentation may be recommended. This involves using topical drugs to remove the remaining pigment from the skin, making the skin color uniform.

Chemokines are small glycoproteins that are activated by INF-y and act on a wide variety of cell types such as lymphocytes, fibroblasts, neutrophils, and endothelial cells. Chemokine receptors (CXCR3) and its ligand (CXCE10) are increased in vitiligo and other autoimmune diseases, leading to the induction of tissue inflammation and damage. High CXCR3 and CXCE10 reflects host immune response of Thl lymphocytes.

INF-y-specific Thl immune response provokes CXCE10 release and expression of CXCR3 on melanocyte-specific CD8+ T-cells that lead to melanocytes injury and depigmentation. Therefore, neutralization of CXCE10 reduces depigmentation and risk of vitiligo with a significant reversal effect on the depigmentation process.

Thus, CXCE10 is regarded as a novel target in the treatment of vitiligo, Statins therapy may produce significant inhibition of inflammatory reactions through the inhibition of chemokines and Veillard et al. illustrated that statins reduce chemokine and chemokine receptors in human macrophages and endothelial cells through suppression of geranyl-geranyl pyrophosphate pathway. Similarly, signal transducer and activator of transcription (STAT) protein family, mainly STAT-1 is required INF-y-signaling in vitiligo. Simvastatin downregulates JAK/STAT pathway in different inflammatory conditions. Furthermore, CXCR3 and its ligand CXCL10 induce the accumulation of cytotoxic autoreactive T-cells in the human epidermis leading to melanocyte degeneration and induction of vitiligo.

Atorvastatin inhibits epidermal cytotoxic autoreactive T-cells that may explain the potential role of this drug in the management of vitiligo. In addition, CXCR3/CXCL10 is an important pathway in the pathogenesis of vitiligo; serum level of CXCL10 is regarded as a novel biomarker in monitoring vitiligo activity and guiding treatment of progressive vitiligo.

The mechanism by which statins may be beneficial for vitiligo is not fully understood, but several theories have been proposed:

1. Anti-inflammatory effects: Statins have been shown to have anti-inflammatory effects by reducing the production of pro -inflammatory cytokines, such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF -alpha) . In vitiligo, it is thought that inflammation plays a role in the destruction of melanocytes, the cells that produce skin pigmentation.

2. Antioxidant effects: Statins have also been shown to have antioxidant effects by increasing the activity of the enzyme superoxide dismutase (SOD), which helps to neutralize free radicals that can damage cells. It is thought that oxidative stress may contribute to the development of vitiligo.

3. Promotion of melanocyte migration and proliferation: Some studies have suggested that statins may promote the migration and proliferation of melanocytes. This could potentially help to repopulate areas of the skin where melanocytes have been destroyed in vitiligo.

4. Statin inhibits the enzyme HMG-CoA reductase, which is responsible for the synthesis of cholesterol in the liver. By blocking this enzyme, Fluvastatin reduces the production of cholesterol.

In addition to lowering LDL cholesterol levels, Statin also has other beneficial effects on the cardiovascular system, such as improving endothelial function, reducing inflammation, and stabilizing atherosclerotic plaques. These effects are thought to be due to its ability to reduce the production of isoprenoids, which are lipid molecules that play a role in cell signaling pathways involved in inflammation and atherosclerosis.

It's important to note that the bioavailability of these drugs can be affected by various factors such as food, other medications, and individual differences in metabolism. Therefore, it is important to follow the instructions of your healthcare provider and pharmacist when taking these medications. The onset of action for these drugs can vary, but generally, it takes several weeks to see significant changes in cholesterol levels.

Atorvastatin typically starts to lower cholesterol levels within two weeks of starting treatment, but it may take up to four weeks to see the full effect. Simvastatin typically starts to lower cholesterol levels within four weeks of starting treatment, but it may take up to six weeks to see the full effect.

It's also important to follow the instructions of your healthcare provider and pharmacist when taking these medications to ensure optimal benefits and reduce the risk of side effects. Inflammation is a natural response of the body to injury or infection, but chronic inflammation can lead to various diseases, such as atherosclerosis, diabetes, and cancer. Lovastatin has been shown to inhibit the production of inflammatory cytokines, such as interleukin-6 and tumor necrosis factor-alpha, by immune cells, such as macrophages.

Lovastatin can also reduce the activation of a protein called nuclear factor-kappa B (NF-KB), which is a key regulator of inflammation. NF-KB controls the expression of various genes involved in inflammation, such as cytokines, chemokines, and adhesion molecules.

Moreover, lovastatin can modulate the activity of immune cells, such as T cells and B cells, which play a critical role in the development of chronic inflammation. Lovastatin can inhibit the proliferation and activation of these cells, which can prevent the progression of inflammation.

Overall, the anti-inflammatory effects of lovastatin suggest that this drug may have potential therapeutic applications in various inflammatory diseases. However, further studies are needed to fully understand the mechanisms of lovastatin's anti-inflammatory effects and to determine the optimal doses and treatment durations for different inflammatory conditions. Atorvastatin can inhibit the proliferation and activation of these cells, which can prevent the progression of inflammation. Overall, the anti-inflammatory effects of atorvastatin suggest that this drug may have potential therapeutic applications in various inflammatory diseases.

Statin medications are designed to be taken orally and absorbed through the gastrointestinal tract. While there are no topical formulations of statins approved for use in humans, some studies have investigated the potential for transdermal (skin) absorption of statins. Research has shown that some statins, such as atorvastatin and simvastatin, may have some degree of skin permeability and could potentially be absorbed through the skin.

However, the extent of absorption is generally considered to be low and insufficient to achieve therapeutic levels in the blood. It is important to note that the use of topical statins is not currently approved by regulatory agencies and should not be attempted without the guidance of a healthcare professional.

If you are interested in using a statin medication to manage cholesterol levels or another medical condition, it is best to speak with your doctor about the most appropriate treatment options for your individual needs. Skin re-pigmentation is the main goal of therapy regardless of its types; however, spontaneous re-pigmentation is occurring in about 1%— 25% of patients.

Topical corticosteroids are the first-line therapy and more effective for small vitiligo lesions and should not use more than 4 months due to the risk of skin atrophy.

Topical calcineurin inhibitors such as tacrolimus are effective as topical corticosteroids without risk of skin atrophy. Systemic corticosteroids, such as dexamethasone, prednisolone, and methylprednisolone are effective for generalized progressive vitiligo.

Besides, oral methotrexate is a useful therapy for vitiligo. On the other hand, physical therapy such as phototherapy with ultraviolet A(UVA), narrow-band UVB with psoralen and monochromatic excimer light, is safe and more effective than other therapeutic modalities.

Indeed, different previous studies illustrated the potential role of statins in the treatment of different types of vitiligo. Therefore, the objective of this study was to elucidate the mechanistic role of statins and/or molecular effects of different types of statins in the management of vitiligo.

Statins are conventionally used in the treatment of dyslipidemia, mainly hypercholesterolemia, which was previously evaluated in the treatment of vitiligo depending on its antioxidant, anti-inflammatory, and immune-modulating effects. Re-pigmentation and regression of vitiligo were initially reported in man with vitiligo who was on simvastatin therapy for hypercholesterolemia.

The animal model study showed that statins reverse and prevent melanocytes degeneration and depigmentation through inhibiting the proliferation of CD8-T-Cells. Statins are conventionally used in the treatment of dyslipidemia, mainly hypercholesterolemia, which was previously evaluated in the treatment of vitiligo depending on its antioxidant, anti-inflammatory, and immune -modulating effects.

Re -pigmentation and regression of vitiligo were initially reported in man with vitiligo who was on simvastatin therapy for hypercholesterolemia. The animal model study showed that statins reverse and prevent melanocytes degeneration and depigmentation through inhibiting the proliferation of CD8-T-Cells in the endothelial cells.

The dose-dependent effect of simvastatin leads to significant inhibition of INF-y-dependent MHC-II expressions with subsequent inhibition of activated T-lymphocytes in patients with active vitiligo.

This immune-modulating effect of statins may play an important role in the management of vitiligo. The immune-modulating effect of statins was previously reported since pravastatin prevents and reduces acute transplant rejections in human subjects.

Similarly, pravastatin prevents acute rejection of cardiac transplant due to the inhibition of proinflammatory mediators and expression of adhesion molecules, which are independent of its cholesterol-lowering effect.

It has been reported that different types of statins prohibit the expression of inflammatory and proinflammatory adhesion molecules such as lymphocyte function-associated antigen (LFA-1) and intercellular adhesion molecule- 1 (ICAM-1) on leukocytes.

Also, statins block LFA-1 on the lymphocytes and inhibit its interaction with ICAM-1 on antigen-presenting cells and by this way statins prevent the activation of lymphocytes and antigen presentation. In addition, statins inhibit chemokine release by endothelial cells, block chemokine receptors on T-cells, inhibition of natural killer cells, and attenuate the proliferation of stimulating leukocytes. Besides, lovastatin inhibits different mediators and cytokines such as inducible NOS, TNF-a, IL-6, and IL- 1 P leading to significant anti-inflammatory and immune-modulating activates in different modalities of vitiligo.

HMG-CoA reductase dependent pathway: inhibition of HMG-CoA reductase leads to the reduction of active inflammatory metabolites known as isoprenoids, HMG-CoA reductase independent pathway, Statins block LFA-1 so, prevent lymphocyte activations, Statins inhibit T-cell function through the inhibition of the second messenger phosphatidylinositol-30-kinase/ Akt transduction pathway. Moreover, IL- 17 serum level is increased in patients with vitiligo and statins have been found to be a potent inhibitor of IL- 17 through inhibition of T-cells proliferation and induction of immunotolerance.

OBJECTIVE OF THE INVENTION:

The present invention provides a topical pharmaceutical composition comprising Janus kinase (JAK) inhibitors like Tofacitinib, Immunosuppressive agent like Tacrolimus, Fingolimod and statins like atorvastatin, Simvastatin, Fluvastatin, Lovastatin, Pitavastatin, Pravastatin and Rosuvastatin dissolved or dispersed in a pharmaceutically acceptable base together with other pharmaceutically acceptable excipients or in a dermatologically acceptable carrier, useful in the treatment of vitiligo.

The inventive concept provided herein are methods and pharmaceutical compositions by preparing oral dosage forms like Topical liquid spray or Topical cream or Topical gel dosage form for the management of vitiligo and its associated disorders and accordingly performing preclinical study to prove the efficacy of the selected drug substances from the class of JAK inhibitors, statins and immunosuppressants.

SUMMARY OF THE INVENTION:

The present invention provides a topical pharmaceutical composition comprising Janus kinase (JAK) inhibitors like Tofacitinib, Immunosuppressive agent like Tacrolimus, Fingolimod and statins like Atorvastatin, Simvastatin, Fluvastatin, Lovastatin, Pitavastatin, Pravastatin and Rosuvastatin, dissolved or dispersed in a pharmaceutically acceptable base together with other pharmaceutically acceptable excipients or in a dermatologically acceptable carrier, useful in the treatment of vitiligo. The dosage form may be in the form of topical liquid spray or topical ointment, cream or topical gel dosage form for the management of vitiligo and its associated disorders.

The dermatologically acceptable carrier may be an oily carrier or an aqueous carrier. The carrier comprises water in combination with an excipient, such as benzalkonium chloride, sodium chloride, sodium phosphate dibasic, citric acid, or a mixture thereof. The carrier may further comprise liposomes.

In another aspect, the topical formulation is in the form of a cream, a gel (aqueous and/or alcoholic gel), a lotion, a spray, an ointment, an aqueous solution, a non-aqueous solution, or a transdermal patch. In yet another aspect, the topical formulation is used to treat a patient suffering from vitiligo in any of its forms.

In yet another aspect, the invention provides a method of treating vitiligo, by treating a depigmented skin surface with atopical formulation comprising drug substances from the class of JAK inhibitors, statins and immunosuppressants in a dermatologically acceptable carrier.

The method of treating vitiligo may comprise a animal study of treating the formerly depigmented skin surface with a topical formulation comprising a drug substances from the class of JAK inhibitors, statins and immunosuppressants in a dermatologically acceptable carrier.

In one embodiment, immunosuppressants like tacrolimus and fmgolimod are topically administrated as topical sprays comprising buffering agent, pH modifiers, Skin permeation agents, co-solvent and solvent. The Spray solution may be alcoholic or non-alcoholic in nature.

In one embodiment, the severity of vitiligo can be assessed by Vitiligo Assessment scale as shown in the table below.

1) 0 = No evidence of depigmentation

2) 1 = <10 % evidence of depigmentation

3) 2 = 10-25% evidence of depigmentation

4) 3 = >25-75% evidence of depigmentation

5) 4 = >75-99% evidence of depigmentation

6) 5 = 100% evidence of depigmentation As used herein, the term “human subject”, “individual” or “patient,” used interchangeably, refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, and most preferably humans. In some embodiments, the patient is a human. In some embodiments, the patient is a human aged 12 years or older.

In some embodiments, the pharmaceutical composition is a cream formulation. In some embodiments, the cream formulation is an oil-in-water emulsion.

In some embodiments, the oil-in-water emulsion comprises water, an oil component, an emulsifier, and Jak inhibitor like Tofacitinib, or statins like Atorvastatin or simvastatin the pharmaceutically acceptable salt thereof, on a free base basis. In some embodiments, the pH of the cream formulation is about 4.5 to about 5.5.

In some embodiments, the occlusive agent component comprises one or more substances selected from fatty acids (e.g., lanolin acid), fatty alcohols (e.g., lanolin alcohol), hydrocarbon oils & waxes (e.g., petrolatum), polyhydric alcohols (e.g., propylene glycol), silicones (e.g., dimethicone), sterols (e.g., cholesterol) vegetable or animal fat (e.g., cocoa butter), vegetable wax (e.g., carnauba wax), and wax ester (e.g., bees wax).

In some embodiments, the emollient component comprises one or more substances independently selected from light mineral oil, medium chain triglycerides, and dimethicone.

In some embodiments, the emulsifier component comprises one or more substances independently selected from glyceryl stearate, and polysorbate 20. In some embodiments, the stabilizing agent component like xanthan gum which improves the stability of the pharmaceutical formulation and/or the compatibility of the components in the formulation.

In some embodiments, the solvent component is present in an amount of about 10% to about 35% by weight of the formulation. In some embodiments, the solvent component is a liquid substance or mixture of liquid substances in which Jak inhibitor, or its pharmaceutically acceptable salt, has reasonable solubility.

Examples of the formulation compositions for topical composition having immunosuppressants are given below.

Table 1: Topical spray formulation containing Tacrolimus.

Table 2: Topical spray formulation containing Fingolimod.

Table 3: Topical spray formulation containing Tacrolimus and Fingolimod.

Table 4: Topical cream formulation containing Tofacitinib.

Table 5: Topical cream formulation containing combination of Tofacitinib and Atorvastatin

Table 6: Topical cream formulation containing combination of Tofacitinib and Simvastatin

Animal study data for vitiligo: Monobenzone induced vitiligo in female C57BL/6 mouse model: i) Species Details:

Species: Mice (C57BL/6). Animal age: 10 - 12 weeks iii) Table 7: Study Design

DOSE FORMULATION:

Formulation will be prepared as per the sponsor’s suggestions just before dosing or ready to use formulation.

II. EXPERIMENTAL PROCEDURES:

All mice were shaved to remove fur on the dorsal lateral region of back skin and 20 cm diameter shaved area is marked as the application area using a marker.

All animals except sham group were induced vitiligo like depigmentation using Monobenzone cream 40% w/w (Benoquin-40) daily once, topical application to application area using a spatula for fifty consecutive days.

The sham group animals will be treated with Vaseline daily once, topical application to application area using a spatula for fifty consecutive days. The disease induced mice were randomized into different treatment groups based on vitiligo observation scoring on day 50. The treatment groups will receive their respective test items according to the dose regimen i.e, topical spray applied b.i.d for 21 consecutive days i.e from Day 51 to Day 72. The sham group mice were kept aside without any induction or treatment which served as healthy group.

Vitiligo scoring:

7) 0 = No evidence of depigmentation

8) 1 = <10 % evidence of depigmentation

9) 2 = 10-25% evidence of depigmentation

10) 3 = >25-75% evidence of depigmentation

11) 4 = >75-99% evidence of depigmentation

12) 5 = 100% evidence of depigmentation

The following skin sites were evaluated and awarded a score between 0-5. 1) Tail

2) Ear

3) Nose

4) Footpad

5) Application area

During the study duration body weights and photographs of mice were recorded twice weekly. At the end of study plasma will be separated and archived at -80 degree Celsius. After euthanasia the skin samples were collected with one half snap freeze immediately and archived at -80 degree Celsius and remaining half stored in 10% formalin and evaluated the histopathological findings.

III. CLINICAL OBSERVATIONS

General clinical observations will be conducted once daily and detailed clinical observations will be conducted once and observations are.

1. Home cage observations

2. Handheld observations

3. Skin depigmentation scoring

IV. REPORT SUBMISSION:

The study report will be provided within one months after approval of study plan. The following information will be provided in the report.

Test Item

- Physical nature, purity and physicochemical properties.

- Identification data.

- Vehicle (if appropriate):

Test system:

- Species/strain used;

- Number, age and sex of animals;

- Source, housing conditions, diet, etc. - Individual weights of animals at the start of the test.

Test conditions:

- Rationale for dose level selection;

- Details of test substance formulation/achieved concentration;

- Details of the administration of the test substance;

- Details of food and water quality.

Results:

- Body weight/body weight changes;

- Clinical observations

- Vitiligo scoring and recordings

Table 8: Study design of treatment groups

RESULTS:

Disease induction: Vitiligo is induced by topical application of 40% Monobenzone ointment for 30 days. Confirmation of disease: By Observational vitiligo scoring based on patchy white spots appearance.

Treatment -21days: Each drug formulation dosed to the induced mice by topical application, twice daily for 21 days(n=10)

Samples collected: At the end of 21^st day treatment period all mice were sacrificed. Serum and back skin samples were collected and analyzed.

Claims

CLAIMS:

1. The present invention provides a topical pharmaceutical composition comprising Janus kinase (JAK) inhibitors like Tofacitinib, Immunosuppressive agent like Tacrolimus, Fingolimod and statins like Atorvastatin, Simvastatin, Fluvastatin, Lovastatin, Pitavastatin, Pravastatin and Rosuvastatin, dissolved or dispersed in a pharmaceutically acceptable base together with other pharmaceutically acceptable excipients or in a dermatologically acceptable carrier, useful in the treatment of vitiligo.

2. The dosage form may be in the form of topical liquid spray or topical ointment, cream or topical gel dosage form for the management of vitiligo and its associated disorders.

3. The dermatologically acceptable carrier may be an oily carrier or an aqueous carrier. The carrier comprises water in combination with an excipient, such as benzalkonium chloride, sodium chloride, sodium phosphate dibasic, citric acid, or a mixture thereof. The carrier may further comprise liposomes.

4. In another aspect, the topical formulation is in the form of a cream, a gel (aqueous and/or alcoholic gel), a lotion, a spray, an ointment, an aqueous solution, a non-aqueous solution, or a transdermal patch. In yet another aspect, the topical formulation is used to treat a patient suffering from vitiligo in any of its forms.

5. In yet another aspect, the invention provides a method of treating vitiligo, by treating a depigmented skin surface with a topical formulation comprising drug substances from the class of JAK inhibitors, statins and immunosuppressants in a dermatologically acceptable carrier.

6. The method of treating vitiligo may comprise a animal study of treating the formerly depigmented skin surface with atopical formulation comprising a drug substances from the class of JAK inhibitors, statins and immunosuppressants in a dermatologically acceptable carrier.

7. Immunosuppressants like tacrolimus and fingolimod are topically administrated as topical sprays comprising buffering agent, pH modifiers, Skin permeation agents, cosolvent and solvent. The Spray solution may be alcoholic or non-alcoholic in nature.

8. The severity of vitiligo can be assessed by Vitiligo Assessment scale as shown in the below. a. 0 = No evidence of depigmentation b. 1 = <10 % evidence of depigmentation c. 2 = 10-25% evidence of depigmentation d. 3 = >25-75% evidence of depigmentation e. 4 = >75-99% evidence of depigmentation f. 5 = 100% evidence of depigmentation

9. As used herein, the term “human subject”, “individual” or “patient,” used interchangeably, refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, and most preferably humans. In some embodiments, the patient is a human. In some embodiments, the patient is a human aged 12 years or older.

10. The pharmaceutical composition is a cream formulation. The cream formulation is an oil-in-water emulsion.

11. The oil-in-water emulsion comprises water, an oil component, an emulsifier, and Jak inhibitor like Tofacitinib, or statins like Atorvastatin or simvastatin the pharmaceutically acceptable salt thereof, on a free base basis. In some embodiments, the pH of the cream formulation is about 4.5 to about 5.5.

12. The occlusive agent component comprises one or more substances selected from fatty acids (e.g., lanolin acid), fatty alcohols (e.g., lanolin alcohol), hydrocarbon oils & waxes (e.g., petrolatum), polyhydric alcohols (e.g., propylene glycol), silicones (e.g., dimethicone), sterols (e.g., cholesterol) vegetable or animal fat (e.g., cocoa butter), vegetable wax (e.g., carnauba wax), and wax ester (e.g., bees wax).

13. The emollient component comprises one or more substances independently selected from light mineral oil, medium chain triglycerides, and dimethicone.

14. The emulsifier component comprises one or more substances independently selected from glyceryl stearate, and polysorbate 20. In some embodiments, the stabilizing agent component like xanthan gum which improves the stability of the pharmaceutical formulation and/or the compatibility of the components in the formulation.

15. The solvent component is present in an amount of about 10% to about 35% by weight of the formulation. In some embodiments, the solvent component is a liquid substance or mixture of liquid substances in which Jak inhibitor, or its pharmaceutically acceptable salt, has reasonable solubility.