RELATED APPLICATIONS

This application claims priority to U.S. Provisional application 60/656, 391, filed February 28, 2005.

BACKGROUND OF THE INVENTION

The present invention relates to in general to wound healing and, in particular to a process for improving the size and appearance of scar tissue. The ability to heal by forming scars is essential for mammalian systems to survive wounding after injury.

Although scar formation and remodeling are essential processes in skin wound healing, disorders of excess scar formation, such as hypertrophic scars and keloids, remain a common clinical problem. A hypertrophic scar is an excessive wound scar which is thick and raised, having grown in size beyond that required for normal wound healing. A hypertrophic scar stays essentially within the boundaries of the original injury. A keloid is a raised scar that exceeds the boundaries of the initial injury, and which is rarely corrected by surgical intervention.

The changing patterns of the connective tissue matrix during repair following injury require a delicate balance between synthesis and degradation of collagen and proteoglycans. Under normal circumstances this balance is maintained, while in many diseased states it is altered, leading to an excessive deposition of collagen, to a loss of functional tissue, or to disfigurement. With hypertrophic scars and keloids, the biosynthetic phase continues longer than necessary to repair the wound. In order to maintain nutrient supply in hypertrophic scars and keloids scars, vascular in-growth occurs, resulting in a large, highly vascularized scar which are unsightly and can be disabling.

Existing therapy for hypertrophic scars and keloids includes surgery, mechanical pressure, steroids, x-ray irradiation and cryotherapy. There are many

disadvantages associated with each of these methods. Surgical removal of the scar tissues is often incomplete and can result in further development of hypertrophic scars and keloids at the incision and suture points. Steroid treatments are unpredictable and often result in de-pigmentation of the skin. X-ray therapy is the only predictable effective treatment to date; however, because of its potential for causing cancer, it is not generally recommended or accepted. The most common approach to controlling scar, and in particular excessive scar formation, is to apply pressure, which appears to be somewhat effective in many instances. However, this treatment has limited application, generally based on the size and location of the scar tissue on the body. Other commonly used treatments are application of Vitamin E and corticosteroids. Each of these agents can interfere with collagen synthesis and promote collagen degradation.

Vitamin D is a name applied to two related fat soluble substances, cholecalciferol (vitamin D3) and ergocalciferol (vitamin D2), that have in common the ability to prevent or cure rickets (Reference - text book Godman & Gilman's -The pharmacological basis of Therapeutics, Ninth Edition , p 1529). These compounds require metabolic activation. The history of metabolic activation is primarily attributable to studies conducted in the labs of DeLuka in the USA and Kodiceck in England

Vitamin D as used herein, the term "activated vitamin D" or "active vitamin D" in reference to a compound is intended to include any biologically active vitamin D compound, including a pro-drug (or pro-hormone), a precursor, a metabolite or an analog, in any stage of its metabolism. It is contemplated that any of the biologically active forms of vitamin D can be used in the formulations in accordance with the present invention. Generally, an active vitamin D compound or analog is hydroxylated in at least the C-I, C-24 or C-25 position of the molecule, and either the compound itself or its metabolite binds to the vitamin D receptor (VDR).

Pro-drugs, for example, include vitamin D3 compounds that are, e.g., hydroxylated in the C-I. Such compounds undergo further hydroxylation in vivo, and their metabolites bind the VDR. Precursors include pre-vitamins such as Ia-,

25-hydroxyprevitamin D3 and Ia-, 25-dihydroxyprevitamin D3, which are thermal isomeric forms of the vitamins. Metabolites generally include compounds or analogs that have undergone further metabolic processing, e.g., hydroxylation.

Factors affecting the production of vitamin D3 synthesis are aging, sunscreens, latitude, time of day and season, and skin pigmentation. The amount of skin pigmentation has been directly correlated to vitamin D3 synthesis suggesting melanin competes with vitamin D for photons from the sun. It has been well established that the levels of provitamin D are similar in all ethnic populations but variations came when the skin was photolyzed. The amount of vitamin D3 produced was the function of the skin type (Matsuka L Y, Arch Dermatol., 1991; 127: 536-538). Clemens et al. (Lancet, 1982, 8263 (1), 74-76) published their results to show that increased skin pigmentation reduces the capacity of skin to synthesize Vitamin D3.

Vitamin D3 has been shown to act as an anti-inflammatory agent (Harant H, FEBS Lett. 1998; 436: 329-334; Arroyo CM et al., Pharmacol Toxicol. 2003; 92: 204-213; Inoue M et al., Eur J Dermatol. 1998; 8(1): 16-20). In dark skin individuals the amount of vitamin D3 production is lower thus increasing the susceptibility to inflammatory diseases. Elevated interleukin -6 expressions were exhibited in keloid fibroblasts (Xue H et al., J Surg Res. 2000; 89: 74-77). Vitamin D2 and vitamin D3 are generally considered equivalent in humans. Nevertheless, some differences between the two forms have been reported in the literature (Journal of Clinical Endocrinology and Metabolism, Vol. 89, no. 11, 5387-5391).

Hypertrophic scar are the major limiting consequence of injury from wound caused by laceration; a wound caused by avulsion; a wound caused by burn; a wound caused by radiation; a wound caused by chemical facial peel; and a wound caused by accident. There is a need for a safe and a cost effective therapy to reduce scarring conditions. One way to increase safety is to use an agent which is inexpensive and effective. One of them is vitamin D and its derivatives, analogues and active metabolites. It is possible to improve the size or appearance of a closed wound by administering to the healed wound, orally administering, externally administering, administering by injection or some combination thereof,

a therapeutically effective amount of a composition comprising Vitamin D or active Vitamin D analogues including a pro-drug (or pro-hormone), a precursor, a metabolite. A major problem is that vitamin D derivatives are unstable under heat and light and are toxic on long term use such as is desired here. They are chemically unstable to heat, light and oxygen and means such as refrigeration, light shading or replacement with an inert gas are required for storage of these vitamin D themselves. No formulation containing vitamin D and active vitamin D analogues as an active ingredient and having required stability, safety and desired performance has been reported to date for the treatment of hypertrophic scars and keloids and other related disorders such as cirrhosis of the liver, constrictive pericarditis, Duputryen's disease of the hand, plantar fibrosis of the foot and various other fibromastosis. The object of this invention is to solve this problem by providing active forms of vitamin D in the form of a stable, safe and efficacious preparation suitable for the treatment of excess scarring conditions.

The object of the invention is to provide methods and compositions for improving the size or appearance of a closed wound comprising administering to the healed wound, orally administering, externally administering, administering by injection or some combination thereof, a therapeutically effective amount of a composition comprising Vitamin D or active Vitamin D analogues including a pro-drug (or pro-hormone), a precursor, a metabolite. Also provided are novel hydrogels as thermal insulating material useful for treatment of excess scarring conditions.

Hydrogels and gels are preferred formulations for example due to their applicability in external as well as internal scarring conditions. Even though there are several hydrogels available, there is a need for a scar reducing hydrogel that incorporates the desired properties specifically tailored for this application. One criterion for commercial success of such a gel will be its ability to withstand washing with either water or water and mild soap. The scar reducing hydrogels needs to be applied for a long time (scar remodeling can last more than one year) thus safety requirements are high. Reusability is highly desirable especially in

external applications. A self-adhering gel able to withstand microbial infiltration is highly desirable.

There is a need for a scar reducing compositions that incorporates the desired properties adapted to cover and contact scar. Such has not been previously demonstrated in the prior art.

BRIEF SUMMARY OF THE INVENTION

The present invention provides a method for improving the size or appearance of a closed wound on a patient's skin comprising administering to the closed wound a gel or hydrogel and a therapeutically effective amount of a Vitamin D agent.

The present invention also provides a method for improving the size or appearance of a closed wound on a patient's skin comprising administering to the closed wound a gel or hydrogel containing a Vitamin D agent.

The present invention also provides a method for improving the size or appearance of a closed wound on a patient's skin comprising administering to the closed wound a gel or hydrogel and a composition containing a therapeutically effective amount of a Vitamin D agent.

DETAILED DESCRIPTION OF THE INVENTION

Conceptually, the wound healing process may be divided into three phases. The first phase is an intensely degradative phase called the inflammatory stage. It occurs immediately after injury and provides a means to remove the damaged tissues and foreign matter from the wound as well as regain immunological control over invading skin surface microbes. This phase lasts approximately one week when the wound is immediately closed, for example in a surgical incision. However, the level of the inflammatory response continues at elevated levels in open wounds until the wound surface is closed by regrowth of the epithelial barrier. If wound inflammation is prolonged or more intense, excessive scarring, called hypertrophic scars, usually appear.

Inflammation, or an "inflammatory response", is the net result of interconnected physiological events, including increased vascular permeability, fluid accumulation, and the migration of a changing population of inflammatory cells into an inflamed area. The clinical manifestations of inflammation include swelling, increased local temperature, erythema, and pain. The inflammatory response can be triggered by any of a number of causative factors, including certain bacteria, radiation, hypersensitivity to chemical agents, and the like. The inflammatory response is generally believed to be a primary defense mechanism in the body, but, unchecked, can become excessive resulting in functional impairment.

The second stage of wound healing typically occurs 2-3 days later and typically lasts about 3 weeks. This stage may be referred to as the proliferation and matrix synthesis stage. During this stage fibroblasts from the surrounding tissue invade the wound and proliferate. The fibroblasts in the wound proliferate and actively produce macromolecules, such as collagen and proteoglycans, which are secreted into the extracellular matrix. Fibroblast activity is driven by the chemical signals produced by inflammation. The newly-synthesized collagen fibrils are cross-linked by lysyl oxidase and provide structural integrity to the wound. During this stage, fibroblasts also contract the intact collagen in order to reduce the surface area of the wound.

In the third and final stage, called the remodeling stage, the previously constructed and randomly organized matrix is remodeled into an organized structure which is highly cross-linked and aligned to maximize mechanical strength. Natural skin wrinkles (relaxed skin tension lines) which align themselves in the direction of mechanical tension and become permanent on the face over time are a common manifestation of this control process.

This step can last for more than one year. The end result of mammalian wound healing is scar formation. Scars are not an exact replacement for undamaged tissue. Skin scars are generally less elastic, creating contour irregularities; color changes and maybe painful if they entrap nerves. Control of dermal scarring is one of the most important objectives in the management of

trauma particularly burn trauma. Minimizing dermal scarring and may lead to optimum post-traumatic functional and aesthetic recovery.

One such scar which can result from an overproduction of collagen and excess deposition of scar tissue is a hypertrophic scar. As used herein, the term "hypertrophic scar" includes a scar characterized by thick, raised scar tissue that stays essentially within the boundaries of the original injury. Hypertrophic scars contain characteristic nodules, and result from a full-thickness injury, such as a surgical incision on skin. These scars can cause problems such as aesthetic deformity and severe limitation of motion. For example, an excessive postoperative scar can develop as a result of "over-healing" or hypertrophic healing of a post-operative site.

Hypertrophic scars generally result from an over-production of cells, collagen and proteoglycan (Linares, H. A. et al., Plast. Reconst. Surg., 62:589 (1978); Linares, H. A., Plast. Reconstr. Surg., 818-820 (1983)). These scars more frequently occur among children and adolescents, suggesting that growth factors may influence the development of this type of scar.

Hypertrophic scars are especially common in patients who have burns or wounds that are allowed to remain open for more than a few weeks. These scars, by definition, exceed normal wound healing, causing problems that range from aesthetic deformity to severe limitation of motion. In these scars, the overproduction and compaction of collagen and proteoglycans exceeds the proliferation of cells. These histological observations suggest that the lesions result from loss of the normal control mechanisms which regulate the synthesis of extracellular matrix during wound healing.

Hypertrophic scars are more common on the anterior surfaces of the neck, the shoulder, the chest wall and, in general, the flexor surfaces of the extremities. While some hypertrophic scars will spontaneously resolve within a few years, in many instances, especially in the locations mentioned above, they persist indefinitely. Because these scars are so common, particularly in burns or wounds that heal by secondary intention, their management represents a major unsolved clinical problem.

Another type of scar in which there is an excess deposition of scar tissue is called a "reactive scar." As the term is used herein, a reactive scar is a normal, healed scar which, through mechanical disruption such as scratching or other irritation, is actively producing a hypertrophic tissue response.

Excessive scar deposition also occurs in a "fibrotic scar." As the term is used herein, a fibrotic scar is an accumulation of irritated fibrotic tissue at the site of a healed injury which may or may not have involved an observable wound.

Another type of scar that can result from an excess deposition of scar tissue is a keloid. As used herein, the term "keloid" includes a scar characterized by thick, raised scar tissue that exceeds the initial boundaries of the trauma and that lacks nodules. In contrast to hypertrophic scars, keloids proliferate beyond the wound edges, can result from superficial injuries, and are rarely treated successfully by surgery. Keloids frequently develop after burns, particularly where the skin is under tension, such as on the breastbone.

Although appearing similar, keloids and hypertrophic scars differ considerably. The former is genetically based with both autosomal dominant and autosomal recessive modes of transmission reported (Roseborough et al., J. Natl. Med. Assoc. 2004, 96(1): l-9).The basis for the genetic differences has been shown through several investigations describing the aberrant behavior of keloid fibroblasts. It has also resulted from reports of keloids exhibiting abnormal regulation of apoptosis to abnormally producing collagen, fϊbronectin, and proteoglycans with atypical responses to metabolic regulators.

Unlike hypertrophic scars, keloids may occur with only minor insults to the skin. Conversely, hypertrophic scars are usually the result of injury to the deep dermis. They also tend to be more pronounced in wounds with a prolonged inflammatory phase and may develop in areas with increased mechanical tension. Both types of scars may produce considerable cosmetic disfigurement and prompt many patients affected to seek treatment. Although keloids can be successfully treated in a single application, patients may require multiple modes of therapy (Shaffer JJ et al., J. Am. Acad. Dermatol. 2002; 46: 63-67).

The present invention is based, in part, on the discovery that the size and appearance of a healed wound can be improved, and the discomfort, itching, pain, and/or other symptoms caused by excessive tissue growth in a healed wound can be alleviated (partially or completely) by solving the problem of administering externally, orally or by injection or some combination thereof, a therapeutically effective amount of a composition comprising Vitamin D or active Vitamin D analogues including a pro-drug (or pro-hormone), a precursor, a metabolite or an analog.

The present invention relates to methods and compositions for improving the appearance and/or reducing the size of a closed wound, which may be a scar. For example, a closed wound may be a hypertrophic scar, keloid, Dupuytren's contracture, fϊbrotic scar, or a reactive scar. Accordingly, the present invention relates to methods that comprise administering to an individual having a closed wound or scar a therapeutically effective amount of a composition comprising Vitamin D or active Vitamin D analogues including a pro-drug (or pro-hormone), a precursor, a metabolite or an analogue.

As used herein, the terms "closed wound" or "scar" refer to a wound or a wound surface that is closed by regrowth of an epithelial barrier. A wound is "closed" after an open wound has been re-epithelialized. A wound is typically closed within 48-72 hours after injury. Closed wounds can result in the formation of a scar, which is never an exact replacement of the original tissue. Scar tissue is less elastic than the undamaged tissue and has surface and contour irregularities. As used herein, the term "affected area of skin" may also be used to refer to either a "closed wound" or a "scar."

In one embodiment, a closed wound is an area of skin that has pain, tingling, burning, and/or itching. In another embodiment, a closed wound is a scar. In another embodiment, a scar is an area of skin that has pain, tingling, burning, itching, discoloration, surface irregularities, and/or an erratic accumulation of fibrous tissue.

A closed wound may result from any of a number of types of skin traumas such as laceration, avulsion, burn, surgery, infection, chemical facial peel, and

accident. An open wound closes by regrowth of an epithelial barrier, the regrowth replacing some of the normal tissue which had been destroyed by trauma. Sometimes, in the closed wound or scar, excessive and disfiguring deposits of fibrous tissue having an erratic accumulation of collagen occur.

Normally, wound healing is a continuous process extending over a one-to- two-year period.

By the present invention vitamin D and active vitamin D compounds are formulated in gels or hydrogels to produce a stable and safe preparation with improved drug performance which is applied to inflamed scar tissue to relieve the inflamed condition and improve scar size and appearance.

In one embodiment, the method includes contacting a closed wound with a thermal insulating material that elevates the surface temperature of the closed wound, and that includes an effective amount of at least vitamin D or active vitamin D compound. The thermal insulating material may also include a deodorant agent to reduce surface bacteria and odor formation. The thermal insulating material is allowed to remain in contact with the closed wound for a period of time sufficient to allow a noticeable improvement in its size and appearance.

Keloids and hypertrophic scars can thus be treated by the present invention. Another type of scar tissue that may be treated by embodiments of the method and compositions of the present invention is Dupuytren's contracture. Dupuytren's contracture arises from unknown causes and is a progressive, scar-like shrinkage and thickening of the flexion contracture of the cusp-like extended palmar aponeurosis in the palm of the hand, whereby, as the curvature of the fingers increases, especially that of the fourth and fifth fingers, stretching of the fingers becomes ever more restricted. This ailment, which attacks men more frequently than women and can occur in one or both hands, begins with a dimple-like indentation in the palm of the hand and gradually but quite painlessly grows into nodules and fascicles. The flexor tendons of the fingers concerned are not in themselves diseased but their movement is impaired by the scar-fascicles of the palmar aponeurosis. A similar contracture concerning the toes is known.

Since the illness neither regresses spontaneously nor responds with any degree of long term success to conventional forms of treatment (without surgery) such as massage, heat treatment and the like, it can only be treated surgically, namely, by cutting away the proliferating atrophied tissue. In addition to ordinary risks and unpleasantness associated with any surgical operation, there exists a further risk that scars resulting from the operation can make a later recurrence of the ailment even worse. The related diseases treated by this invention are for example Peyronnie's Disease, a reactive scar, an excessive post-operative scar or a fibrotic scar.

The present invention also relates to pharmaceutical compositions which include a suitable pharmaceutical carrier containing Vitamin D or active Vitamin D analogues and a deodorant agent, aluminum hydroxide, and an anti-microbial substance such as sodium salicylate, parabens, aluminum zirconium trichlorohydrex, or other metallic anti-microbial. Any of the compositions of the present invention described herein may be administered with a suitable pharmaceutical carrier, the choice of which depends on the route of administration and the size of the scar. The terms "suitable pharmaceutical carrier," "pharmaceutically acceptable," and grammatical variations thereof, as they refer to compositions, carriers, diluents and reagents, are used herein interchangeably. As the terms are used herein, "suitable pharmaceutical carrier" and "pharmaceutically acceptable" refer to non-toxic materials that do not interfere with the effectiveness of the biological activity of active ingredients, and represent that the materials are capable of administration to or upon a vertebrate with a minimum of undesirable physiological effects such as nausea, dizziness, gastric upset and the like. The characteristics of the carrier will depend on the route of administration.

The compositions and methods according to embodiments of the invention can be used on any vertebrate with skin. Examples of such vertebrates include mammals (for example, human, bovine, porcine, canine, feline) and avian.

Hydrogels could be a good medium for reducing scars. They have been shown to be useful for the treatment of scarring conditions (for example US 5552162) and several types of hydrogels have been disclosed. It has been found by

us that gels with water and without water are preferred formulations for example due to their applicability in external as well as internal scarring conditions. Even though there are several hydrogels available/ theoretically possible, there is a need for a scar reducing hydrogel that incorporates the desired properties specifically tailored for this application. One criterion for the suitability of the gel will be washing the gel in water and cleaning with mild soap. The scar reducing hydrogels needs to be applied for a long time (scar remodeling can last more than one year) thus safety requirements are high. Reusability is highly desirable especially in external applications. Self Adhering nature of the gel along with ability to stand microbial infiltration is equally important.

In one embodiment of the invention, the thermal insulating material, when used to cover the affected area, causes an elevation in the surface temperature of the healed wound or scar of from about 0.5 ⁰ C to about 5 ⁰ C. In another embodiment, the thermal insulating material, when used to cover the affected area, causes an elevation in the surface temperature of the healed wound or scar of from about 1 °C to about 4 ⁰ C. In a preferred embodiment, the thermal insulating material, when used to cover the affected area, causes an elevation in the surface temperature of the healed wound or scar of from about 2 ⁰ C to about 3 ⁰ C.

The thermal insulating material may be a sponge. Examples of sponge materials suitable for use as a thermal insulating material in the present invention include collagen and cross-linked collagen. The term "cross-linked," as used herein, refers to covalent bonds formed among polymeric chains and to an interconnected structure wherein cross-links are formed between hydrophobic molecules, between hydrophilic molecules and between hydrophobic molecules and hydrophilic molecules.

The thermal insulating material may be a gel, a hydrogel, or a biodegradable hydrogel. Gels and hydrogels generally contain a very high concentration of water, e.g., about 60% to about 98% water and are held together by a variety of cellular groups. The water may be bound in the form of various hydrates, or unbound, entrapped in cellular pockets formed by the polymer network groups.

The term "hydrogel" is used herein to mean a polymeric material which can include a cross-linked macromolecular network, which exhibits the ability to swell in water and to retain a significant portion of water within its structure without dissolving.

A "biodegradable hydrogel," as the term is used herein, is a hydrogel formed from a hydrogel-forming system containing at least one biodegradable component, i.e., a component which is degraded by water and/or by enzymes found in nature. These hydrogels are particularly useful in preventing restenosis by coating implantation devices such as stents for implantation in human vessels for creating and sustaining openings and for preventing re-occlusion thereof after implantation.

Hydrogels could be a good medium for reducing scars. They have been shown to be useful for the treatment of scarring conditions (for example US 5552162) and several types of hydrogels have been disclosed. Even though there are several hydrogels available/ theoretically possible, there is a need for a scar reducing hydrogel that incorporates the desired properties specifically tailored for this application. For example it is not advisable to use the hydrogel patch on the skin as this may promote unwanted growth of bacteria. One criterion for the suitability of the gel will be the ability to with stand washing of the gel in water and cleaning with mild soap. The scar reducing hydrogels needs to be applied for a long time (scar remodeling can last more than one year) thus safety requirements are high. Reusability is highly desirable especially in external applications. Self Adhering nature of the gel along with ability to stand microbial infiltration is equally important.

A typical wound dressing hydrogel is applied on an open wound and should not stick to the open wound and absorb wound exudates and does not require repeated applications. On the other hand a typical scar reducing gel/ hydrogel is applied on the closed wound and does not have to absorb the exudates. The gel should actually stick to the skin without the aid of tapes and bandages and should have the ability to resist 'dry out' and retain its adhesion after repeated

applications. The gel should have a soft feel and gentle release from the skin as well as high safety index.

Particularly useful in this invention are polyacrylate polymer hydrogel containing from about 20 to 90 percent, and preferably from about 30 to 50 percent, by weight of a polymer prepared by the co polymerization of formula I (for example sodium 2-acrylamido-2-methylpropane sulphonate) or formula II (for example 2-acryloamino propyl trimethyl ammonium chloride compounds) with one or more monomers selected from monomers or the derivative of the monomers such as acrylic acid, water soluble acrylic functional monomers such as acrylamide, methacrylamide/N-vinyl2-pyrollidone, acrylic acidmethaacrylic acid, vinyl acetate, vinyl chloride, maleic anhydride. These hydrogels typically have a glycol component (can be selected from the group consisting of the formula OH- (C ₂ H ₄ O)-H, OH-(C ₃ H ₆ O) _n -H wherein n is in the range of about 1- 16). The balance can be water and or a pharmaceutical carrier.

Formula I

R-C(=CH ₂ )-C(=O)-NH-R -SO ₃ M

Formula II

R ¹

+ /

R-C(=CH ₂ )-C(=O)-NH-R ⁴ -N-R ²

\ R ³ wherein R is selected from the group consisting of hydrogen or alkyl radicals containing from 1 to 6 carbon atoms, R ¹ , R ² and R ³ are independently alkyl C ₁ - C ₈ . R ⁴ is selected from the group consisting of alkylene radicals containing from 1 to 6 carbon atoms or an arylene radical containing from 6 to 10 carbon atoms, and M is selected from the group consisting of hydrogen, ammonium, potassium, or sodium, of which 2-acrylamido-2-methylpropane sulphonic acid or sodium 2-

acrylamido-2-methylpropane sulphonate are the preferred examples of formula I. 2-Acrylamidopropyl trimethyl ammonium chloride is a preferred formula II.

Preferably, the monofunctional monomer component is predominantly acrylamidopropyl trimethyl ammonium chloride , 2-acryloamido-2-methyl propane sulphonic acid or a soluble salt. Further, it has been discovered that the gels containing glycerol along with water have exceedingly better applicability in reducing the scar. The glycerol content in the gel is preferred to be between 20- 60%.

Methods of preparing these polymers are well known and have been disclosed for example in US 4,822,842 and 5,037,930, both incorporated herein by reference. Hydrogels are known in prior art, for example, US 5,173,302, incorporated herein by reference. Hydrogel material based on polymerized 2- acryloamido-2 methyl propane sulphonate salts are well known as electrode material from US 4,391,278 and US patent 4,768,523, incorporated herein by reference. The polymerization of any of the above-described monomers and their resulting polymers are well known to those skilled in the art. There are numerous references which disclose methods of polymerizing the monomers and preparation of these types of gel. It is understood that various variations of these gels are possible and are covered under this invention.

Further, it has been discovered by us that the gels containing glycerol along with water have exceedingly better applicability in reducing the scar. The glycerol content in the gel is preferred to be between 20-60%. Glycerol and glycerine are two words for the same substance

Hydrogels materials based on polymerized 2- acryloamido- 2 methyl propane sulphonic acid are known from US pat numbers 4,391,278 an 4,768,523 for wound dressing. The polymerization of any of the above-described monomers and their resulting polymers are well known to those skilled in the art. There are numerous references which disclose methods of polymerizing the monomers and preparation of these types of gel. It is understood that various variations of these gels are possible and are covered under this invention.

Additionally, these hydrogels, due to exceptional self adherence properties can be used to cover an area on which vitamin D containing formulation have been applied. It is widely known that many vitamin D and active forms of vitamin D are chemically unstable to heat, light and oxygen and means such as refrigeration, light shading or replacement with an inert gas are required for storage of these vitamin D themselves. If a method to deliver vitamin D vitamin D and active forms of vitamin D can be produced. It will be practical to apply a hydrogel patch to cover the ointment which has been applied on the scar to help further in the treatment of the scarring conditions by further assisting the degradation of the scar.

Particularly, due to the intended long use of these gels, it has been found that it is highly desirable to have breathable top layer on the gel that control loss of moisture. Breathable film for example be of polyethylene or polyurethane. Our results showed that the gel's top layer has a great influence on gel's suitability for scar reduction. We experimented with polyurethane layers of varying MVTR (moisture vapor transmission rates) values on gel A-B (see table below). Our experiments indicated that the best MVTR values for a polyurethane top liner is between 200- 800 grams/sq. meters/ day. It needs to be recognized that the change of the gel will influence this parameter and MVTR values for example 100- 8000 grams/meters ² /day may be acceptable. Similarly, the thickness of the gel required for a particular gel is dependent on the hydrogel used.

When used as conductive adhesive hydrogels, high amounts of an electrical conductivity enhancer, such as potassium chloride, are generally added to the formulation. In the present invention, salt solutions of sodium or potassium chloride, at lower concentrations, may be present as polymerization enhancers in the formation of the hydrogels but are preferably restricted to 0.0% to 0.9%.

Ideally, the new hydrogel should be preserved and capable of use for intended wear period of 30 days without microbial growth. One way to ensure the sterility is to use antimicrobial gel or gamma irradiates the hydrogel. Unfortunately, the process of sterilization also changes the mechanical properties of the gel and may destroy integrity of vitamin D and active analogues. Several attempts to find a gel which would meet our product criteria were made but it was not possible to find a gel which after irradiation will retain desired mechanical properties. This resulted in a loss of lots of time.

Even if the gel does not allow the growth of microbes (for example disclosed gels in US patents 4,391,278 and 4,768,523) or is sterile before applying to the skin, once opened, sweat collection and bacterial growth starts. There have been reports of biofilm formation on the silicone sheets when worn for extended time period. Thus, it is preferable to add an antimicrobial agent in the hydrogel. This would extend the use time of the hydrogel, preferably to about 30 days or more.

The inclusion of antimicrobials in hydrogels are known in the literature for wound dressing hydrogels but no hydrogel for suitability for our application has been disclosed in the prior art. Methyl parabens and propyl parabens are most commonly used preservatives and are generally considered to be good

preservatives. In the new hydrogel they provide good protection against fungal and bacterial infections. However, parabens are not the perfect preservatives; several attempts have been made to combine parabens with other anti-microbial agents to overcome the shortcoming of parabens. Cocktails of preservatives have been prepared which contain additional anti-microbial agents. Any anti-microbial agent or combination of agents can be used. Preferably, a cocktail combination of 0.5% sodium salicylate, 0.2% methyl parabens and 0.1% propyl parabens is used.

Sodium salicylate is considered very safe and is commonly used as preservative (Cosmetic Ingredient Review committee refers to a review in International Journal of Toxicology; 2003 Supplement 3, Vol. 22 Issue s3, pl-108, 108p). It has been concluded in the review that 0.09 % to 2% of sodium salicylate is a safe widely used preservative.

Additionally, Staphylococcus epidermidis is a major cause of infections associated with medical devices. Biofilm production is an important virulence attribute in the pathogenesis of device-related infections. If biofilm builds up on our hydrogel that needs to be worn for extended time, it can become a source of irritation for the patient's scars. Therefore, elimination of any possibility of formation of biofilms is highly desirable.

Sodium salicylate has been demonstrated to have remarkable antibacterial activity, including the ability to enhance the activities of certain antibiotics. This drug inhibits adherence (55%), growth, and biofilm production of S. epidermidis (Farber, B. F. and A. G. Wolff, J. Infect. Dis., 1998, 166:861-865 (Medline); Muller, E., J. et al., J. Infect. Dis. (1998) 117:501-503).

It is noteworthy that the physical properties of the present gels were not affected by inclusion of these preservatives. Gels A and B were exceedingly good under washing procedures. Gels survived washing with water on daily basis for more than 30 days. Further, the expose mild soap did not destroy the gel. In fact washing was beneficial as it restored the water content just enough to make it adhere better to the skin.

The method of the present invention also may include the simultaneous administration of a substance such as an anti microbial agent or and anti-irritant,

for example, diphenhydramine, to reduce skin irritation, and/or the simultaneous administration of an ointment and/ or a gel/ hydrogel patch.

Vitamin D, a Vitamin D analogue, a Vitamin D precursor, a Vitamin D pro-drug or a Vitamin D metabolite or a combination thereof may be included in the present hydrogel either before or after the preparation of polymeric hydrogels. The appropriateness of the method is generally dependent on the stability of a particular Vitamin D, a Vitamin D analogue, a Vitamin D precursor, a Vitamin D pro-drug or a Vitamin D metabolite or a combination thereof.

On the other hand it has been discovered that waterless gels are exceedingly useful in the treatment of scarring conditions. One such gel can be made by using poly (alkylene) glycols. Several references on use of poly (ethylene glycol) in pharmaceutical preparations can be found in the literature. Polyethylene glycols are hygroscopic, i.e., they attract and retain moisture from the atmosphere. This property makes PEGs useful as water-soluble ointments and humectants, and as replacements for other hygroscopic materials such as glycerin and propylene glycol in certain applications. Hygroscopicity decreases as the molecular weight of the PEG increases. Above their melting/freezing temperatures, PEGs can be considered Newtonian fluids since their viscosities are nearly independent of shear. Kinematic viscosity measurements, therefore, are the most practical way of characterizing PEG viscosity. Viscosities of PEG materials decrease as temperature increases.

Polyethylene glycols may be liquids or solids at room temperature, depending on the average molecular weight of the PEG. Higher molecular weight PEGs form more rigid solids, while lower molecular weight PEGS tend to be viscous liquids at room temperature. For example, PEGs having average molecular weights from 200-600 are typically clear, viscous liquids at room temperature; PEGs having average molecular weights from 900-1500 are typically soft, opaque white solids at room temperatures; and PEGs having average molecular weights from 3350-8000 typically are hard, opaque white solids at room temperature. Increased molecular weight results in decreased solubility in water and other solvents, decreased hygroscopicity and vapor pressure, and increased

melting/freezing range and viscosity. Intermediate physical properties can be achieved by blending PEGs of differing molecular weights. It has been found by us that for example, blending two PEGs, for example a 400 MW PEG and a 3350 MW PEG can lead to a composition with the consistency of an ointment or salve, or a composition having a gel tendency.

Polyethylene glycols make excellent water-soluble ointment bases; they spread easily and evenly over the skin, even if the skin is moist. The good water solubility of PEGs makes it easy to incorporate aqueous ingredients in the formulation, and they do not become rancid or support microbial growth. Moreover, we also found that the vitamin D level in skin can be controlled over a wide range, from low to high levels, by incorporation of vitamin D by varying relative percentages of mixed polyethylene glycols.

Additionally, in practicing the present invention, it is advantageous if the PEG material creates a waxy gel on the surface of the closed wound or scar that can decrease UV light penetration and buffers superoxide radicals. PEG materials preferably limit the area of treatment to an appropriate level of moisture. Further, the structure of the PEG material provides a mechanical barrier for decreased penetration of bacteria and foreign materials. The PEG material also creates an evaporative barrier which augments transdermal delivery of the drugs into the closed wound or scar area. In one embodiment, the PEG material comprises an acidic composition, which is favorable for the skin and for longevity of the PEG material.

In one embodiment, one or more therapeutically effective substances may be applied to one surface of a thermal insulating material. The thermal insulating material is then applied to the closed wound in a manner such that the therapeutically effective substance is placed in contact with the closed wound.

In another embodiment, the therapeutically effective substance is dispersed within a hydrogel, a water-insoluble gel, a sponge or a PEG material. The hydrogel, water-insoluble gel, sponge or PEG material, within which the therapeutically effective substance is dispersed, is then placed in contact with the affected surface of the skin, and allowed to remain in place for a period of time

sufficient to bring about an improvement in the size and appearance of the closed wound.

As used herein, the term "dispersed" includes ionic, covalent, hydrophilic, or hydrophobic interactions between the therapeutically effective substance and the hydrogel, water-insoluble gel, sponge, or PEG material.

For example, a therapeutically effective substance containing a cationic moiety can be immobilized on a hydrogel polymer chain. As will be recognized by those skilled in the art, this cationic site may serve as a non-covalent, ionic binding site for anionic substances.

In another example, a hydrogel or sponge can be chosen which covalently bonds to the therapeutic substance used according to one embodiment. For example, through hydrophilic interactions with water in the hydrogel, any water soluble drug will dissolve in the hydrogel. A hydrophobic interaction between a non-water soluble therapeutic substance and a hydrogel can occur when the hydrogel selected includes a hydrophobic entity which is receptive to further interaction with a therapeutic substance having a hydrophobic moiety.

One skilled in the art will know, or will be able to ascertain with no more than routine experimentation, what hydrogels or PEG materials are suitable for dispersing a particular therapeutic substance.

A therapeutic substance which covalently bonds to the hydrogel, sponge or PEG material can form a drug delivery substance with controlled or sustained release. If a biodegradable hydrogel or sponge is used, delivery of the therapeutic substance to the closed wound or scar is also related to the rate of degradation of the hydrogel or sponge. The degradation rate of the hydrogel or sponge is usually slower than the diffusion rate of the therapeutic substance. It is well-known to those of skill in the art that the rate of delivery of the therapeutic substance can be controlled by choosing a particular concentration of each therapeutic substance used in a particular embodiment, and a particular hydrogel or sponge, one can control the rate of degradation or the rate of diffusion, and thus,.

The hydrogel, other thermal insulating material, or PEG material containing the therapeutically effective substance can remain in contact with the

surface of the affected area of skin for about between 0.5 to about one hour per day, from about one hour to about 8 hours per day, from about 12 hours to about 15 hours per day, from about 12 hours to about 18 hours per day, from about 18 hours to about 24 hours per day, or over a number of days, for a sufficient number of days to bring about an improvement in the size and appearance of the closed wound or scar. The hydrogel, other thermal insulating material, or PEG material can be removed periodically in order to cleanse the scar surface and to apply a fresh sample of therapeutically effective substance and hydrogel, other thermal insulating material, or PEG material.

In one embodiment, at least one cyclooxygenase inhibitor, and/or at least one NF-kB inhibitor, or combinations thereof, are administered topically with a suitable pharmaceutical carrier, including one or more substances that relieve skin irritation. In a particular embodiment of the invention wherein the method of administration is topical, the substance that relieves skin irritation includes at least one of the following substances: glyceryl monooleate, diphenhydramine, calamine, and a C3-C4 diol.

In one embodiment, a closed wound, such as a scar, is contacted with a hydrogel or PEG material comprising at least one Vitamin D or vitamin D derivative/ analogue or combinations thereof, and a deodorant agent to reduce surface bacteria and odor formation.

In one embodiment, a closed wound is treated by contacting the closed wound with a hydrogel or a PEG material comprising an effective amount of vitamin D and other Vitamin D active analogues or derivatives thereof in a pharmaceutically acceptable carrier. The hydrogel preferably elevates the surface temperature of the affected area of skin. The hydrogel or PEG material is allowed to remain in contact with the affected area of skin for a period of time sufficient to result in an improvement in the closed wound.

Examples of suitable patterns of use according to an embodiment of the invention include, among others: use of various hydrogel or PEG material combinations in sequence; use of various hydrogel or PEG material combinations simultaneously; use of various hydrogel or PEG material combinations in

systemic-topical co-administration, such as oral administration simultaneously with topical administration; use of combinations of active ingredients mixed by a pharmacist according to a prescription; and use of combinations of separate active ingredients available in kit form, mixed by the patient and self-administered according to physician instructions or directions provided with the kit.

Examples of various hydrogel or PEG combinations (i.e., hydrogel or PEG material combined with one or more active ingredients) that may be used according to an embodiment of the invention.

Hydrogel may apply an active ingredient with one or more of the following advantageous properties: efficacy, sustained delivery, consistency in, dosage, enhanced delivery, dosage control, efficiency, stability and bioavailability for scar reducing.

It is contemplated that any of the biologically active forms of vitamin D can be used in the formulations in accordance with the present invention. Generally, an active vitamin D compound or analog is hydroxylated in at least the C-I, C-24 or C-25 position of the molecule, and either the compound itself or its metabolite binds to the vitamin D receptor (VDR).

Pro-drugs, for example, include vitamin D compounds that are, e.g., hydroxylated in the C-I. Such compounds undergo further hydroxylation in vivo, and their metabolites bind the VDR. Precursors include pre vitamins, such as Ia-, 25-dihydroxyprevitamin D3, which are thermal isomeric forms of the vitamin forms. Metabolites generally include compounds or analogs that have undergone further metabolic processing, e.g., hydroxylation.

Examples of compounds suitable for formulations of the present invention include all biologically active forms of Vitamin D, without limitation vitamin D2,Viatmin D3, calcipotriene (a synthetic vitamin D3 derivative) lα ,25- dihydroxyvitamin D3 (calcitriol), lα-hydroxyvitamin D3 (α-calcidol), seocalcitol (EB- 1089), calcipotriol, 22-oxacalcitriol (maxacalcitol), fluorinated compounds such as falecalcitriol derivatives , and 19-nor compounds such as paricalcitol derivatives. Among those compounds that have a chiral center, e.g., in the sidechain, such as at C-24, it is understood that both epimers (e.g., R and S) and

the racemic mixture are within the scope of the present invention. Vitamin D2 and derivatives can also be used for the purpose of the invention (Armas LA et al., J Clin Endocrinol Metab. 2004 Nov; 89(11):5387-91)

One object of the invention was to produce a pharmaceutical preparation which is microbe free. Ideally, the preparation should be preserved and capable of use for long duration (long shelf life). Either raw materials are obtained in sterile conditions or / and formulations are produced aseptically.

It was found that even if the gel is sterile before applying to the skin, once opened sweat collection and bacterial growth can start and it may be more appropriate to add antimicrobial agent in the gel. This is particularly important for the products of this invention.

Based on consideration described above and after evaluation of existing preservatives, and compatibility consideration, it was decided that a combination of sodium salicylate, methyl parabens and propyl parabens will be used. Methyl parabens and propyl parabens are most commonly used preservatives and are generally considered to be good preservatives. In the new formulation they provide good protection against fungal and bacterial infections.

The invention includes an anti-irritant, an anti-microbial agent, an anti- prurient agent, a deodorant agent and combinations thereof. Amount of sodium salicylate (or related compounds) may be used in a range of 1-5 %, methyl- paraben in a range of 0.1-0.5%, and propyl-paraben in a range of 0.1-0.5% within a soluble PEG material. Other additives such as Vitamin E, aloe vera can be used, amount of salicylic acid in a range of 1-5%, aloe in a range of 0.2-2%, Vitamin E in a range of 0.2-2%, methyl-paraben in a range of 0.1-0.5%, and propyl-paraben in a range of 0.1-0.5% within a soluble PEG material that contained vitamin D3.

Oral and Parenteral Administration:

In addition to compositions suitable for topical or transdermal administration to the affected area of skin, in another embodiment of the invention, compositions may be those suitable for oral or parenteral (including intramuscular, sub-cutaneous and intravenous) administration, or those in a form

suitable for administration by inhalation. The therapeutically effective substance of the invention, together with a conventional adjuvant, carrier, or diluent, may thus be placed into the form of pharmaceutical compositions and unit dosages thereof, and in such form may be employed as solids, such as tablets or filled capsules, or liquids such as solutions, suspensions, emulsions, elixirs, or capsules filled with the same, all for oral use; or in the form of sterile injectable solutions for parenteral (including sub-cutaneous) use.

In one embodiment of the invention, aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizing and thickening agents, as desired. Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, or other well known suspending agents.

Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration. Such liquid forms include solutions, suspensions, and emulsions. These preparations may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like. Solid form preparations include, among others, powders, tablets, pills, capsules, and dispersible granules. A solid carrier can be one or more substances which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active component. In tablets, the active component is mixed with the carrier having the necessary binding capacity in suitable proportions and compacted in the shape and size desired.

According to an embodiment of the invention, powders and tablets preferably contain from five or ten to about seventy percent of the active compound. Suitable carriers are magnesium carbonate, magnesium stearate, sugar,

lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa buffer, and the like. The term "preparation" is intended to include the formulation of the active compound with encapsulating material as carrier providing a capsule in which the active component, with or without carriers, is surrounded by a carrier, which is thus in association with it. Similarly lozenges are included. Tablets, powders, capsules, pills and lozenges can be used as solid forms suitable for oral administration.

According to an embodiment of the invention, liquid preparations include solutions, suspensions, and emulsions, for example, sterile water or water- propylene glycol solutions. For example, parenteral injection liquid preparations can be formulated with PEG in aqueous solution. Other suitable pharmaceutical carriers for parenteral administration include, for example, physiological saline, bacteriostatic saline (saline containing about 0.9% mg/ml benzyl alcohol), phosphate-buffered saline, Hank's solution, Ringer's-lactate and the like. An embodiment of a therapeutically effective substance according to the present invention may thus be formulated for parenteral administration (by injection, for example, by bolus injection or continuous infusion) and may be presented in unit dose form in ampoules, pre-filled syringes, small volume infusion or in multi-dose containers with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulation agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution, for constitution with a suitable vehicle, e.g. sterile, pyrogen-free water, before use. Administration by Inhalation

According to an embodiment of the invention, administration may also be made to the respiratory tract by means of an aerosol formulation in which the active ingredient is provided in a pressurized pack with a suitable propellant such as a chlorofluorocarbon (CFC) for example dichlorodifluoromethane, trichlorofluoromethane, or dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. The aerosol may conveniently also contain a surfactant such as

lecithin. The dosage of the therapeutic substance may be controlled by provision of a metered valve. In compositions intended for administration to the respiratory tract, including intranasal compositions, compounds used in an embodiment will generally have a small particle size, for example of the order of 5 microns or less. Such a particle size may be obtained by means known in the art, for example by micronization.

Therapeutically Effective Amount and Dosage

As used herein, the terms "therapeutically effective amount" and "therapeutically effective dose" refer to the amount of an active agent, for example, a therapeutically effective substance, such as a, vitamin D and active vitamin D compounds or an anti-irritant, required to be administered in order to induce a desired result in the patient. That result may be alleviation or amelioration (complete or partial) of the symptoms or condition of irritation, pain, tingling, redness or other discoloration of a closed wound, an improvement in the appearance or reduction in the size of the closed wound, or any other desired improvement in the affected area of skin.

As used herein, the term "therapeutically effective amount" may also refer to the quantity of active agent or therapeutically effective substance, the administration of which results in improvement in the size, appearance, or condition of a closed wound, where little or no improvement would occur in the absence of the active agent. Typically, the active agent is administered for a sufficient period of time to achieve the desired therapeutic effect.

Therapeutic efficacy may be determined as described herein and by using standard pharmacological procedures in experimental animals.

The active ingredient of an embodiment of the invention, together with a conventional adjuvant, carrier, or diluent, may thus be placed into the form of pharmaceutical compositions and unit dosages thereof, and in such form may be employed as solids, such as tablets or filled capsules, or liquids such as solutions, suspensions, emulsions, elixirs, or capsules filled with the same, all for oral use, or in the form of sterile injectable solutions for parenteral (including subcutaneous)

use, or in the form of aerosol formulations for inhalation therapy. Such pharmaceutical compositions and unit dosage forms thereof may comprise conventional ingredients in conventional proportions, with or without additional active compounds or principles, and such unit dosage forms may contain any suitable effective amount of the active ingredient commensurate with the intended daily dosage range to be employed. Further details on techniques for formulation and administration may be found in the latest edition of Remington's Pharmaceutical Sciences (Merck Publishing Co., Easton, Pa.).

When desired, compositions adapted to give sustained release of the active ingredient may be employed.

The dose administered is adjusted to the size and severity of the closed wound or affected area of skin, the age, weight and condition of the individual being treated, as well as the route of administration, dosage form and regimen, and the result desired. The exact dosage should of course be determined by the practitioner.

The active ingredient can be administered in one or several doses per day. In one embodiment, it is presently contemplated that, for therapeutic treatments, at least one composition of the present invention, such as vitamin D and active vitamin D compounds and antimicrobial agent, can be administered in an amount comprising from about 0.1 microgram to about 3000 micrograms, from about 10 micrograms to about 2000 micrograms, from about 20 micrograms to about 1000 micrograms, or from about 40 micrograms to about 400 micrograms per square centimeter of treated tissue.

In another embodiment, for therapeutic treatments, at least one vitamin D and active vitamin D compounds is administered in an amount comprising from about less than 0.1 microgram to about 2000 micrograms, from about 10 micrograms to about 1000 micrograms, or from about 40 micrograms to about 400 micrograms per square centimeter of treated tissue. The amount of composition of the present invention can be administered by any suitable method of administration, including, but not limited to, topical application, subcutaneous or

parenteral administration, oral administration, administration by inhalation, and by combinations of these methods.

It is to be understood that the present invention is not limited to the particular compositions, methodologies or protocols described herein. Further, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. 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 limit the scope of the present invention, which will be limited only by the claims.

EXAMPLES

Example 1

Ointment Formulation:

In one example, the PEG material of the present invention may be prepared by combining approximately 63% PEG 400 (e.g. Carbowax® available from The Dow Chemical Company® ) with approximately 37 % of PEG 3350 (e.g. Carbowax® available from The Dow Chemical Company®) in a reaction vessel made with glass. The combined PEG materials are heated until the temperature reaches 65 ⁰ C. The combined PEG materials are mixed until they are completely in liquid form.

Once the combined PEG materials are liquid, the liquid PEG is maintained at 65 ⁰ C and methyl parabenzene, propyl parabenzene and BHT are added. Then the mixture is cooled. Vitamin D is dissolved in ethanol and added. PEG materials were formed into a shape for use in the present invention.

It has been found that PEGs can also be combined in a stainless steel kettle for the purpose of commercialization and scaling up. Preferably, the kettle has been clean and sanitized prior to combining the PEG materials.

Example 2

Ointment Formulation: Ointment Formulation:

In one example, the PEG material of the present invention may be prepared by combining approximately 63% ( w/w) PEG 400 (e.g. Carbowax® available from The Dow Chemical Company® ) with approximately 37% ( w/w) of PEG 3350 (e.g. Carbowax® available from The Dow Chemical Company®) in a reaction vessel made with glass. The combined PEG materials are heated until the temperature reaches 65 ⁰ C. The combined PEG materials are mixed until they are completely in liquid form.

Once the combined PEG materials are liquid, the liquid PEG is maintained at 65 ⁰ C and methyl parabenzene, propyl parabenzene and BHT are added. Then the mixture is cooled. Vitamin D is dissolved in ethanol and added during stirring. PEG materials were formed into a shape for use in the present invention.

It has been found that PEGs can also be combined in a stainless steel kettle for the purpose of commercialization and scaling up. Preferably, the kettle has been clean and sanitized prior to combining the PEG materials.

Example 3

Ointment Formulation:

In one example, the PEG material of the present invention may be prepared by first dissolving 0.05 gm BHT by stirring in 16 gm PEG 400 (e.g. Carbowax® available from The Dow Chemical Company® ) with approximately 9 gm of PEG 3350 (e.g. Carbowax® available from The Dow Chemical Company®) in a reaction vessel made with glass. The combined PEG materials are heated until the temperature reaches 65°C. The combined PEG materials are mixed until they are completely in liquid form.

Once the combined PEG materials are liquid, the liquid PEG is maintained at 65°C and methyl parabenzene and propyl parabenzene are added. Then the mixture is cooled. Vitamin D3 is dissolved in minimum amount of ethanol and added. PEG materials were formed into a shape for use in the present invention.

The following formulations were prepared as described in example 2, unless otherwise indicated. Example 4

Example 14 Ointment Formulation:

Example 15 Lotion formulation:

Example 16

Lotion formulation:

Example 17

13.8 mg vitamin D3 was dissolved by stirring it in 4 gm PEG 400 at room temperature.

Example 18

13.8 mg vitamin D3 was dissolved in 1 gm PEG 400 by warming in a water bath maintained at 65 degree centigrade.

Example 19

Patches made up of Gel A and B were worn by four patients continuously for at least 14 days. The hydrogel patches were washed in plain water or with mild soap almost daily.

Aerobic count CFU/g for hydrogel was less than 10. Both mold and yeast counts were also less than 10.

Example 20

This sample passed microbial contamination test using specially validated procedure for this product. Aerobic count CFU/g was less than 10. Both mold and yeast counts were also less than 10.

It has also passed antimicrobial preservative effectiveness test (USP) where this formulation was inoculated with a microorganism suspension to achieve a inoculation level of Ix 10 (raise to the power 5) to Ix 10 ( raise to the power 6) with P. aeuginosa, E. coli, S. aureus, C. albicans, A.niger. There was 2 log reduction of bacterial organisms at day 14 with no increase at day 28. The sample passed the USP category 2 test.

Example 21

1.00 % w/w vitamin D3 was added to the preparation of example 20 while stirring at room temperature.

Example 22

This formulation passed ISO skin irritation study. Under the conditions of this study, no erythmia and no edema were observed on the skin of the rabbits. The primary irritation index for the test was calculated to be 0.0.

It also passed microbial contamination test using specially validated procedures for this product. Aerobic count CFU/g was less than 10. Both mold and yeast counts were also less than 10.

Example 23

1.00 % w/w vitamin D3 was added to the preparation of example 22 while stirring at room temperature.