Anti-inflammatory compounds for the treatment of skin disorders - Patents.com

Abstract

The present invention relates to the use of compounds of Formula I for the treatment of wounds, poor wound healing, hair loss in and around wounds, scarring and / or wrinkles in and around wounds, psoriasis, and any combination thereof. The present invention also relates to methods of using such compounds, and compositions and kits containing the compounds. TIFF2023503886000027.tif34133

Description

[Technical field]

[0001] This application claims the benefit of Indian Patent Application No. 201911047906, filed on November 22, 2019, which is incorporated herein by reference.

[0002] The present invention relates to a compound of formula I (shown below), and topical compositions containing a compound of formula I and a pharmacologic acceptable excipient, for use in the treatment of one or more conditions selected from the group consisting of wounds, poor wound healing, hair loss in and around wounds, scarring and / or wrinkles in and around wounds, and psoriasis.

[0003] [ka] [Background technology]

[0004] Oxidative stress is one of the main events that impede tissue repair and regeneration processes. Removing excessively accumulated free radicals induces a faster healing process, possibly by promoting angiogenesis and neovascularization at the wound site, which are prerequisites.

[0005] Coumarins are a group of phenolic compounds that are widely distributed in natural plants and have a wide range of pharmacological activities.See, for example, Egan et al., Drug Metab. Rev., 22: 503-529, 1990.Of these, esculetin (6,7-dihydroxycoumarin) (compound 2) has been reported to reduce serum levels of liver enzyme markers ALT (alanine aminotransferase) and AST (aspartate aminotransferase) when administered intraperitoneally before treatment with t-butyl hydroperoxide.See, for example, Lin et al., Arch. Toxicol., 74, 467-72, 2000.

[0006] [ka]

[0007] However, their effectiveness remains limited because coumarins typically have poor in vivo bioavailability and do not significantly accumulate in mitochondria, so they must usually be used at higher concentrations to scavenge mitochondrial reactive oxygen species.

[0008] US Pat. No. 9,580,452 discloses triphenylphosphonium cation (TPP+)-conjugated esculetin of formula I (shown below) having anti-atherosclerotic effects.

[0009] [ka]

[0010] There is a need for treatment of wound-related disorders and psoriasis. Summary of the Invention

[0011] The present inventors have surprisingly found that compounds of formula I may be used to treat a variety of wound-related conditions and psoriasis. In one preferred embodiment, compounds of formula I are administered topically.

[0012] One embodiment is a compound of formula I for use in the treatment of one or more of wounds, poor wound healing, hair loss in and around wounds, scarring and / or wrinkles in and around wounds, and psoriasis.

[0013] [ka]

[0014] In one preferred embodiment, X - Br - or Cl - In one preferred embodiment, X - Br- It is.

[0015] Another embodiment is a method of treating wounds, poor wound healing, hair loss at and around the wound, scarring and / or wrinkles at and around the wound, psoriasis, and any combination thereof, in a subject in need thereof. The method comprises administering (preferably topically) to the subject a therapeutically effective amount of a compound of formula I:

[0016] [ka]

[0017] In one preferred embodiment, X - Br - or Cl - In one preferred embodiment, X - Br - In one preferred embodiment, the method comprises topically applying to the affected area of ​​the subject a therapeutically effective concentration of a compound of formula I.

[0018] In one embodiment, the compound of formula I is Compound 1:

[0019] [ka] [Brief description of the drawings]

[0020] [Figure 1] FIG. 1 is a set of photographs showing the effects of topical application of a cream solvent (vehicle) to wounds on the skin surface of diabetic (db / db) mice (vehicle control group) on days 1, 3, 6, 9, 10, and 26. [Diagram 2] FIG. 2 is a photograph showing the effect of compound 1 (0.625% w / w) prepared in solvent applied topically daily to superficial wounds on the skin of diabetic (db / db) mice on days 1, 3, 6, 9, and 10. [Diagram 3] FIG. 3 is a photograph showing the effect of compound 1 (2.5%) prepared in solvent applied topically daily to superficial wounds on the skin of diabetic (db / db) mice on days 1, 3, 6, 9, 10, and 26. [Figure 4a] FIG. 4a is a photograph showing the effects of topical application of a) compound 1 (0.625%, 1.25% and 2.5%) prepared in solvent, b) compound 2 (2.5%) prepared in solvent, and c) solvent to the wound area and surrounding wounds on the skin surface of diabetic (db / db) mice on days 0, 3, 6, 9, 12 and 20. [Figure 4b] FIG. 4b is a graph showing the percentage of wound closure (Y-axis) over time from day 0 to day 20 (X-axis) for the above groups. [Diagram 5] FIG. 5 shows photographs at days 0, 10, and 20 showing the effect of topical application of a) compound 1 prepared in solvent (at concentrations of 0.625%, 1.25%, and 2.5%), b) compound 2 prepared in solvent (2.5%), and c) solvent to wounds on the skin surface of non-diabetic (C57) mice and the effect on hair growth in and around the wound area. [Figure 6] FIG. 6 is a photograph showing the effect of a) vehicle and b) compound 1 (2.5%) prepared in vehicle applied topically to a surgical incision on the skin surface of a rabbit (incision depth 2 mm and incision length 3 cm) on days 0, 1-8, and 18. [Figure 7] FIG. 7 is a photograph showing the effect of topical application of a) vehicle and b) Compound 1 (2.5%) prepared in vehicle to wounds on the skin surface of rabbits 20 days after treatment. [Figure 8]FIG. 8 is a photograph showing the effect of topical administration of a) vehicle, b) compound 1 (0.625%) prepared in vehicle, c) a combination of compound 1 (0.625%) prepared in vehicle and EX-527 (6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide, also known as selisistat, a SIRT1 inhibitor) (0.025%), and d) EX-527 (0.025%) in vehicle on surgical incision healing in a diabetic (db / db) mouse model at days 0, 4, and 7. [Figure 9] FIG. 9 is a photograph showing measurements of tensile strength of skin tissue measured on day 8 after 7 days of treatment as described in Example 5 and Table 3. [Figure 10] FIG. 10 shows the Psoriasis Area and Severity Index (PASI) scoring for erythema, scaling, skin thickness and ear thickness, as well as the percentage of weight loss in the psoriasis model. [Figure 11] FIG. 11 shows phenotype images of the dorsal skin of representative mice from different treatment groups on days 0, 2, 4, 6, and 7 as described in Example 6. [Figure 12] FIG. 12 shows the levels of the pro-inflammatory cytokines IL-17 and IL23 (pg / mL) in skin tissue, measured using the enzyme-linked immunosorbent assay (ELISA) described in Example 6. [Figure 13] FIG. 13 shows hematoxylin and eosin (H&E) staining in the dorsal skin of representative mice from different treatment groups (10x and 40x), as described in Example 6. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] When utilized in accordance with this disclosure, unless otherwise indicated, all technical and scientific terms shall be understood to have the same meaning as commonly understood in the art. Unless otherwise required by context, singular terms shall include the plural and plural terms shall include the singular. The use of any and all examples or exemplary language provided herein is intended to better describe the embodiments and does not pose limitations on the scope of the claims unless otherwise stated.

[0022] The term "wound" refers to any trauma to the tissue of a subject's skin, which interrupts the continuity within the tissue and breaks, punctures or cuts the skin. Wounds typically include, for example, incisions, abrasions, lacerations, cuts, cuts, abrasions, puncture wounds, traumatic skin injuries, burns, and penetrating wounds. Wounds may be chronic, such as diabetic wounds and diabetic foot ulcers, bedsores or pressure ulcers, venous leg ulcers, or other chronic wounds that cause tissue damage, or acute, such as wounds caused by accidents, injuries, or surgery. Medical procedures may also cause wounds, such as dermatological or cosmetic procedures.

[0023] The term "scar" refers to a mark left on skin tissue when a wound does not heal completely and fibrous connective tissue develops on and around the wound. This includes the formation of such marks or fibrous connective tissue during or after wound healing. Symptoms of scarring include, for example, discoloration of the skin scar (including redness or changes in pigmentation), erythema, dryness, peeling, or itching of the skin, protrusion higher than the surrounding skin area, keloid formation, month thick bar, scar pain, scarring and / or reduced vascularity of the surrounding tissue, reduced flexibility, and poor aesthetic appearance (including the amount and texture of scar tissue). Scars caused by wounds can also be treated according to the present invention.

[0024] The term "wound healing" or "healing of a wound" refers to the restoration of tissue integrity, and the terms may be used interchangeably. Of course, these terms may refer to partial or complete restoration of tissue integrity. Thus, treating a wound refers to the enhancement, improvement, progression, facilitation, acceleration, or otherwise advancement of one or more steps or processes associated with the wound healing process. Poor wound healing for the purposes of the present invention includes any cause, procedure that may inhibit, inhibit, interfere with, or delay the enhancement, improvement, progression, facilitation, acceleration, or otherwise advancement of one or more steps or processes associated with wound healing. Poor wound healing as described herein includes that caused by chemotherapeutic agents or anti-angiogenic agents.

[0025] The terms "therapeutically effective amount" and "therapeutically effective concentration" herein refer to an amount or concentration of an active compound or pharmaceutical agent that elicits a biological or medicinal response in a tissue, system, animal, individual, or human, including one or more of: (1) prevention, suppression, or delay of disease, e.g., preventing, suppressing, or delaying a disease, condition, or disorder in an individual who is susceptible to the disease, condition, or disorder but who has not yet experienced or manifested the pathology or symptomology of the disease; (2) inhibition of disease, e.g., inhibiting the disease, condition, or disorder (i.e., preventing further development of the pathology and / or symptomology) in an individual who is experiencing or manifesting the pathology or symptomology of the disease; and (3) amelioration of disease, e.g., ameliorating the disease, condition, or disorder (i.e., reversing the pathology and / or symptomology) in an individual who is experiencing or manifesting the pathology or symptomology of the disease.

[0026] All concentrations of active ingredients, such as Compound 1, in topical compositions are weight percentages based on 100% total weight of the composition, unless otherwise specified.

[0027] Compounds of formula I can be prepared as described in US Pat. No. 9,580,452. Various embodiments of the invention are described below.

[0028] In one aspect, the present invention relates to a compound of formula I for use in treating one or more conditions selected from the group consisting of wounds, poor wound healing, hair loss in and around wounds, scarring and / or wrinkles in and around wounds, and psoriasis, by topical application (e.g., to the skin or wound area) to a subject in need thereof.

[0029] [ka]

[0030] In the formula, X - is any halogen (Br - or Cl - In one preferred embodiment, X - Br - or Cl - In one preferred embodiment, X - Br - It is.

[0031] In one embodiment, the present invention relates to a compound of formula I for use in the treatment of a wound.

[0032] In another embodiment, the present invention relates to a compound of formula I for use in the treatment of poor wound healing.

[0033] In a further embodiment, the present invention relates to a compound of formula I for use in the treatment of hair loss in and around a wound. The treatment of hair loss in and around a wound may be characterized by enhancing and / or promoting hair growth in and around the wound site.

[0034] In yet another embodiment, the present invention relates to a compound of formula I for use in the treatment of scarring and / or wrinkles at and around a wound. The treatment of scarring and / or wrinkles may be characterized by minimizing the appearance of scarring and / or wrinkles at and around the wound site.

[0035] The wound to be treated using a compound of formula I according to any one of the embodiments described herein may be a diabetic wound, an incision wound, a surgical wound, an accidental wound, a pressure or bedsore, a diabetic foot ulcer, pyoderma gangrenosum, a burn, a lesion, a cut, or any combination thereof.

[0036] In another embodiment, the present invention relates to a compound of formula I for use in the treatment of poor wound healing, wherein wound healing is impaired by the co-administration of one or more agents, such as, for example, immunosuppressants, chemotherapeutic agents, anticoagulants, NSAIDs, platelet aggregation inhibitors, antiangiogenic agents, and any combination thereof.

[0037] In another embodiment, the present invention relates to a compound of formula I for use in the treatment of psoriasis.

[0038] In a preferred embodiment of any of the uses or methods described herein, the compound of formula I is Compound 1:

[0039] [ka]

[0040] In another aspect, the present invention is a method for treating wounds, poor wound healing, hair loss at and around wounds, scars and / or wrinkles at and around wounds, psoriasis, or any combination thereof, in a subject in need thereof.The method comprises administering (e.g., topically applying to the affected area) a therapeutically effective amount of a compound of formula I (e.g., a compound of formula 1 where X is Br).In one preferred embodiment, the method comprises topically applying to the affected area of ​​the subject a therapeutically effective concentration of a compound of formula I.

[0041] In one embodiment, the invention relates to a method of treating a wound in a subject in need thereof, the method comprising administering (e.g., topically applying) to the subject a therapeutically effective amount of a compound of formula I, such as compound 1. In one preferred embodiment, the method comprises topically applying to the affected area of ​​the subject a therapeutically effective concentration of a compound of formula I.

[0042] In another embodiment, the present invention relates to a method of treating poor wound healing in a subject in need thereof, the method comprising administering to the subject (e.g., topically applying) a compound of formula I. In one preferred embodiment, the method comprises topically applying to the affected area of ​​the subject a therapeutically effective concentration of a compound of formula I.

[0043] In a further embodiment, the present invention relates to a method for treating hair loss at and around a wound in a subject in need thereof, the method comprising administering (e.g., topically applying) to the subject a compound of formula I. The treatment of hair loss at and around a wound may be characterized by enhancing and / or promoting hair growth at and around the wound site. In one preferred embodiment, the method comprises topically applying to the affected area of ​​the subject a therapeutically effective concentration of a compound of formula I.

[0044] In yet another embodiment, the present invention relates to a method of treating scarring and / or surrounding wrinkles over a wound in a subject in need thereof, the method comprising administering (e.g., topically applying) to the subject a compound of formula I. The treatment of scarring and / or wrinkles may be characterized by minimizing the appearance of scarring and / or wrinkles at and around the wound site. In one preferred embodiment, the method comprises topically applying to the affected area of ​​the subject a therapeutically effective concentration of a compound of formula I.

[0045] In any of the methods of treating a wound, poor wound healing, hair loss at and around a wound, or scarring and / or wrinkles at and around a wound described herein, the wound may be a diabetic wound, an incision wound, a surgical wound, an accidental wound, a pressure ulcer / bedsore, a diabetic foot ulcer, pyoderma gangrenosum, a burn, a lesion or a cut.

[0046] In another embodiment, the invention relates to a method of treating psoriasis, the method comprising administering (e.g., topically applying) to a subject in need thereof a compound of formula I. In one preferred embodiment, the method comprises topically applying to an affected area of ​​the subject a therapeutically effective concentration of a compound of formula I (such as compound 1).

[0047] In a preferred embodiment of any of the methods described herein, the compound of formula I is Compound 1:

[0048] [ka]

[0049] In any embodiment described herein, the wound is present on the surface of the skin of a subject. In one embodiment, the subject is a mammal. In a preferred embodiment, the subject is a human. The methods described herein are useful in both human therapeutics and veterinary applications. For veterinary purposes, the term "subject" includes, but is not limited to, livestock animals, including cows, sheep, pigs, horses, and goats; companion animals, such as dogs and cats; exotic and / or zoo animals; laboratory animals, including mice, rats, rabbits, guinea pigs, and hamsters; and poultry, such as chickens, turkeys, ducks, and geese.

[0050] Another embodiment relates to topical compositions and kits comprising a therapeutically effective concentration or amount of a compound of formula I, and optionally a pharma- ceutically acceptable excipient.The compositions described herein are useful for treating wounds, poor wound healing, hair loss in and around wounds, scars and / or wrinkles in and around wounds, psoriasis, or any combination thereof.

[0051] In one embodiment, the present invention relates to a topical composition comprising a therapeutically effective amount of a compound of formula I, and optionally a pharma- ceutically acceptable excipient, for the treatment of wounds.

[0052] In another embodiment, the present invention relates to a topical composition comprising a therapeutically effective amount of a compound of formula I, and optionally a pharma- ceutically acceptable excipient, for the treatment of poor wound healing.

[0053] In one embodiment, the present invention relates to a topical composition comprising a therapeutically effective amount of a compound of formula I, and optionally a pharma- ceutically acceptable excipient, for the treatment of hair loss in and around a wound. The treatment of hair loss in and around a wound may be characterized by enhancing and / or promoting hair growth in and around the wound site.

[0054] In yet another embodiment, the present invention relates to a topical composition comprising a therapeutically effective amount of a compound of formula I, and optionally a pharma- ceutically acceptable excipient, for the treatment of scarring and / or wrinkles at and around a wound. The treatment of scarring and / or wrinkles may be characterized by minimizing the appearance of scarring and / or wrinkles at and around the wound site.

[0055] In another embodiment, the present invention relates to a topical composition comprising a therapeutically effective amount of a compound of formula I, and optionally a pharma- ceutically acceptable excipient, for the treatment of psoriasis.

[0056] In a preferred embodiment of any of the compositions described herein, the compound of formula I is Compound 1.

[0057] Any topical composition described herein may further comprise one or more suitable pharma-ceutically acceptable excipients known to those skilled in the art.The composition may be in the form of a topical administration form, for example, a solution, a gel, an ointment, a cream, a lotion, a paste, a spray foam or an aerosol.The compound of formula I (such as compound 1) may be made into a topical composition by using a suitable pharma-ceutically acceptable carrier or diluent, and may be formulated into a semi-solid or liquid form.

[0058] In another aspect, the present invention relates to a kit comprising a therapeutically effective amount of a compound of formula I (e.g., in the form of a topical composition), and optionally instructions for using the kit.The kit described herein is useful for treating wounds, poor wound healing, hair loss at and around wounds, scars and / or wrinkles at and around wounds, psoriasis, or any combination thereof.

[0059] The following examples illustrate certain embodiments of certain aspects of the above disclosure, they are provided for illustrative purposes only and should not be construed as limiting the scope of the disclosure in any way.

[0060] [Example] General methods and procedures

[0061] Preparation of test compounds: For the purposes of animal testing in the following examples, topical formulations of test compounds were prepared in a solvent containing emulsifying ointment, a preservative (p-chlorocresol), and water. The solvent did not contain any test compounds. Test compounds (compounds 1 and 2) were incorporated into an aqueous cream preparation by trituration. The aqueous cream was prepared using emulsifying ointment BP, p-chlorocresol (preservative), and water according to the British Pharmacopoeia procedure. The control formulation did not contain any active ingredient.

[0062] Step 1: Preparation of emulsified ointment An emulsifying ointment was prepared using emulsifying wax, white soft paraffin, and liquid paraffin. These ingredients were melted and warmed to 60°C in a water bath, then stirred continuously until cooled.

[0063] Step 2: Preparation of anionic emulsion cream The emulsified ointment was melted and heated up to 60°C in a water bath, the required amount of parachlorocresol was dissolved in cold water (chlorocresol is soluble in cold water), the temperature was checked using a thermometer, purified water was added to the molten ointment and stirred continuously until it cooled down.

[0064] Step 3: Preparation of cream formulations of compounds 1 and 2 The required amount of Compound 1 or Compound 2 was added to the cream and mixed with a homogenizer. After proper mixing of the drugs, the formulations were transferred to suitable containers and stored at 4°C.

[0065] [Table 1]

[0066] [Table 2]

[0067] [Table 3]

[0068] [Table 4]

[0069] [Table 5]

[0070] [Table 6]

[0071] [Example 1] [Effects of Compound 1 on the wound healing process and hair growth in and around the wound area in a diabetic (db / db) mouse model]

[0072] Methods: In this study, mice of the db / db strain (age: 13–14 weeks) were used because this strain has a high similarity to diabetic wound healing in humans. 2 Wounds were created using punch biopsies on the dorsum of db / db mice under local anesthesia. Animals were divided into 5 groups with 5 animals per group as follows: Group 1 (n=5): treated with vehicle alone (diabetic control group). Group 2 (n=5): treated with Compound 2 (2.5%) prepared in solvent. Group 3 (n=5): treated with Compound 1 (0.625%) prepared in solvent. Group 4 (n=5): treated with Compound 1 (1.25%) prepared in solvent. Group 5 (n=5): treated with Compound 1 (2.5%) prepared in solvent.

[0073] Three concentrations of Compound 1 were tested for this study: 0.625%, 1.25% and 2.5%. Topical preparations were applied to the wound once a day for 12 days. Images of the wound area were visualized and measurements of the wound area were recorded on each day of the study.

[0074] Results: The diabetic control group and group 2 showed wound closure by day 26 (see Figures 1 and 3). In groups 3-5, complete closure of the wound was observed by day 10 (see Figures 2 and 4a and 4b (Percentage of wound closure over time)). Table 1 shows the wound healing rates observed in each group.

[0075] [Table 7]

[0076] [Example 2] [Effects of Compound 1 on wound healing and hair growth in and / or around the wound area in a non-diabetic C-57 mouse model]

[0077] Methods: In this study, C57BL / 6j mice strain (age: 12 weeks) were used to study the wound healing process. 2 Wounds were created using punch biopsies on the dorsum of C57 mice. Animals were divided into five groups of five animals each: Group 6 (n=5): treated with vehicle alone (non-diabetic control group). Group 7 (n=5): treated with Compound 2 (2.5%) prepared in solvent. Group 8 (n=5): treated with Compound 1 (0.625%) prepared in solvent. Group 9 (n=5): treated with Compound 1 (1.25%) prepared in solvent. Group 10 (n=5): treated with Compound 1 (2.5%) prepared in solvent.

[0078] Three concentrations of Compound 1 prepared in solvent, namely 0.625%, 1.25%, and 2.5%, were tested in this study. The topical formulation was applied to the wound once a day for 12 days, starting 12 hours after wound formation. Images of the wound area were visualized and area measurements were recorded on each day of the study.

[0079] Results: The non-diabetic control group and group 6 showed wound closure by day 14 (see Figure 5). In groups 8-10, complete closure of the wound was observed from days 9 to 12. Table 2 shows the wound healing rates observed in each group.

[0080] [Table 8]

[0081] [Example 3] [Effect of Compound 1 on wound healing in rabbits]

[0082] Methods: New Zealand rabbits weighing 2.0-2.5 kg were used in this study and were divided into two groups. Control group: treated with the solvent only. Test group: treated with Compound 1 (2.5%) prepared in solvent.

[0083] Paravertebral linear incisions, each 2 cm long, were made through the full thickness of the skin on either side of the spine using a sharp scalpel. After complete hemostasis, the wounds were closed with interrupted sutures at equidistant points spaced approximately 1 cm apart (see FIG. 6). Animals were treated once a day from the day of wounding, from day 0 to day 14. Images were taken daily throughout the study period. On day 7, the sutures were removed and wound closure was observed.

[0084] Results: The test rabbits were compared to the solvent-based control rabbits for wound closure at each time point during the study. Faster surgical wound healing was observed within 8 days in the test rabbits compared to the untreated control rabbits. No surgical scarring was observed in the test rabbits compared to the control rabbits. See Figure 7.

[0085] [Example 4] [Effect of Compound 1 on wound healing in diabetic mice (db / db)]

[0086] Methods: Mice of the db / db strain (age: 12 weeks) were used in this study. The animals were divided into four groups: Group 11: control mice treated with vehicle only. Group 12: Mice treated with Compound 1 (0.625%) prepared in solvent. Group 13: mice treated with Compound 1 (0.625%) and EX-527 (0.025%) in vehicle. Group 14: Mice treated with EX-527 (0.025%) in vehicle.

[0087] Paravertebral linear incisions, each 2 cm long and 1 mm deep, were made through the full thickness of the skin on either side of the spine using a sharp scalpel. After complete hemostasis, the wounds were closed with interrupted sutures at equidistant points spaced approximately 1 cm apart (see FIG. 8). All groups were treated with their respective test and control formulations once a day for 7 days. Images of the wound area were photographed and Doppler imaged on each day of the study. Wound area was measured using a transparent tracing sheet before and during treatment, and wound closure was measured at 4-day intervals. On the 7th day, the sutures were removed and the animals were observed for wound closure.

[0088] Results: Mice in group 12 showed complete healing of the incision wound within 7 days compared to mice in groups 11, 13, and 14. See Figure 8. Group 13 treated with EX-527 (0.025%), a SIRT1 inhibitor, in combination with Compound 1, disrupted the wound healing process induced by Compound 1. This study suggests that Compound 1 induces wound healing by enhancing SIRT1 activity. This result supports the hypothesis that Compound 1 enhances in vitro angiogenesis tube formation, which is perturbed in the presence of EX-527.

[0089] [Example 5] [Measurement of tensile strength of skin tissue on the 8th day after 7 days of treatment]

[0090] Methods: Fresh mouse skin was treated once a day and stretched between the two ends of a tensile tester with a load of 100 kg at a stretching rate of 5 mm / min (Figure 9). The results are shown in Table 3.

[0091] [Table 9]

[0092] [Example 6] [Effect of topical application of compound 1 in improving IMQ-induced psoriasis in Balb / c mice]

[0093] Methods: The efficacy of topical administration of Compound 1 for the treatment of psoriasis was tested in this animal study. In this study, typical features of psoriasis, namely erythema, scaling, and skin thickness, were induced by application of 5% w / w imiquimod (IMQ) cream to the dorsal skin of Balb / c mice for 6 consecutive days. Animals that developed psoriasis-like symptoms were used to test the efficacy of Compound 1, prepared in solvent (as described above). Male Balb / c mice (6-8 weeks old) were used in this study. The animals were divided into 8 groups of 10 animals in each group as follows:

[0094] Sham control group: Sham control: The backs of the mice were shaved and the animals received no treatment. This group served as the control. IMQ control group: this group of animals was treated topically with 62.5 mg of commercial imiquimod cream (5%) on the shaved back at a specific time in the morning for six consecutive days (study days 1 to 6). This negative control group represents a typical psoriasis model induced by imiquimod 5% (IMQ). IMQ+solvent-based group: This group represents mice following induction of psoriasis with imiquimod from day 1 to day 6 of the study, followed 12 hours later by treatment with the solvent used for the preparation of the compound of formula I on the shaved back of the animals. IMQ + clobetasol propionate cream 0.05%: This group received imiquimod induction of psoriasis on days 1 through 6 of the study, followed 12 hours later by the application of commercial clobetasol propionate (0.05%) cream (20 μg / 2 cm) on the shaved back of the animals. 2 The figures represent mice treated with 100 μg / mL IgG4 (region). IMQ+Compound 1 (0.313%) group: This group was treated with induction of psoriasis with imiquimod from day 1 to day 6 of the study, followed 12 hours later by administration of Compound 1 (0.313%, which is 82.6 mg / cm) prepared in a solvent on the shaved back of the animals. 2 Each figure represents mice treated with 100 μg / mL IgG4 (corresponding to the area of ​​the mouse). IMQ+Compound 1 (0.625%) group: This group was treated with induction of psoriasis with imiquimod from days 1 to 6 of the study, followed 12 hours later by administration of Compound 1 (0.625%, which is 82.6 mg / cm) prepared in a solvent on the shaved back of the animals. 2 Each figure represents mice treated with 100 μg / mL IgG4 (corresponding to the area of ​​the mouse). The activity schedule for this study is set out in Table 4 below.

[0095] [Table 10]

[0096] Histopathological studies: As part of the efficacy study, histopathological studies were performed to examine the morphological observations of the skin of mice representative of the different treatment groups. At the end of the experiment, mouse skin samples from each group were fixed in 4% paraformaldehyde and embedded in paraffin. From each paraffin block, sections of 4 μm thickness were made and mounted on glass slides. The sections were stained with hematoxylin and eosin, photographed and evaluated.

[0097] Results: Psoriasis Area and Severity Index (PASI) scoring was analyzed to determine the efficacy of compositions containing Compound 1 with appropriate controls as part of the study. The severity of skin inflammation was assessed by an objective scoring system based on clinical PASI scores. Erythema (0-4) and desquamation (0-4) were scored independently, where 0-none, 1-mild, 2-moderate, 3-severe, 4-very severe. The cumulative score (erythema + desquamation) served as a measure of the severity of inflammation (scale 0-8).

[0098] Figure 10 shows the PASI scoring including body weight, erythema, desquamation, and skin thickness determined during the study. Individual body weights of all mice were recorded daily. Weight loss was calculated relative to body weight on day 0. The negative control group, representing the general psoriasis condition, showed the highest PASI scores for all parameters. The treatment groups, including those treated with solvent-prepared compound 1 (0.313% and 0.625%), showed a decrease in PASI scores almost similar to the positive control during the treatment period. The IMQ+compound 1 (0.625%) group showed a greater decrease in PASI scores compared to the IMQ+compound 1 (0.313%) group and the positive control group. Based on the PASI scoring, the antipsoriasis effect visible from the treatment groups was comparable to the positive control group treated with commercially available clobetasol propionate cream.

[0099] FIG. 11 shows images of mice representing the different groups of the study, namely, sham group, negative control group, positive control group, and treatment groups treated with different concentrations of Compound 1 formulations from day 0 to day 7.

[0100] The IMQ-induced psoriasis model reproduces biochemical and histopathological parameters characteristic of human psoriatic lesions. Topical application of IMQ cream increased the levels of cytokines, including IL-23 and IL-17, in treated skin tissues. Animals were euthanized on day 7. Spleen, liver, and skin tissues were collected from all animals. Spleen and liver tissues were weighed. IL-17 and IL-23 levels were measured by ELISA using commercially available kits. For quantification of IL-17 and IL-23 levels, skin homogenates obtained from different groups of mice were analyzed using ELISA, since the selected interleukins are typical inflammatory markers of psoriasis. As shown in Figure 12, the IMQ group (p<0.01) and the IMQ+solvent-based (p<0.001) group showed a significant increase in IL-17 levels compared to the sham group. A decrease in IL-17 levels was seen in all treatment groups when compared to the negative control. The IMQ and IMQ + vehicle groups showed a significant increase in IL-23 levels compared to the sham control group (p<0.01). A decrease in IL-23 levels was seen in the treatment groups, including the IMQ + Compound 1 (0.313% and 0.625%) and IMQ + Clobetasol (p<0.01) groups, when compared to the negative control. Compared to the sham control group, the IMQ group showed a significant increase in interleukin levels reflecting the development of psoriatic inflammation (p<0.01).

[0101] The negative control group of the study (imiquimod-treated animals) displays typical psoriatic features of acanthosis, parakeratosis, hyperkeratosis, and skin infiltrates upon H&E staining. Similar staining of skin taken from mice in the treatment group showed reduced epidermal thickening, reduced thickening of the stratum corneum, and a reduced number of skin infiltrates. Histopathology is shown in Figure 13. Histopathology results from both skin and ear showed that IMQ-induced psoriasis was better improved by Compound 1 preparation (0.625%), comparable to the effect of clobetasol propionate cream.

Claims

1. Compounds of Formula I [In the formula, X － is any halogen.

1. A pharmaceutical composition for a method of treating wounds, poor wound healing, hair loss at and around wounds, scarring and / or wrinkles at and around wounds, psoriasis, or any combination thereof, in a subject in need thereof, said method comprising topically administering a therapeutically effective amount of a compound of formula I to the affected area of ​​said subject.

2. 2. The pharmaceutical composition of claim 1, wherein the wound is selected from a diabetic wound, an incision wound, a surgical wound, an accidental wound, a pressure / bedsore, a diabetic foot ulcer, pyoderma gangrenosum, a burn, a lesion, or a cut.

3. 3. The pharmaceutical composition of claim 1 or claim 2, wherein the method treats a wound.

4. 3. The pharmaceutical composition of claim 1 or claim 2, wherein the method treats poor wound healing.

5. 3. The pharmaceutical composition of claim 1 or claim 2, wherein the method treats hair loss in and around a wound.

6. 3. The pharmaceutical composition of claim 1 or claim 2, wherein the method treats scars and / or wrinkles in and around wounds.

7. 3. The pharmaceutical composition of claim 1 or claim 2, wherein the method treats psoriasis.

8. 7. The pharmaceutical composition of claim 6, wherein the treatment is characterized by reducing, minimizing or eliminating the appearance of scars and / or wrinkles at and around the wound site.

9. 6. The pharmaceutical composition of claim 5, wherein the treatment is characterized by increasing and / or promoting hair growth at and around the wound site.

10. X is Br － Or Cl － The pharmaceutical composition according to any one of claims 1 to 9,

11. The compound of formula I is The pharmaceutical composition according to any one of claims 1 to 9,

12. Use of a compound of formula I in the manufacture of a medicament for the treatment of one or more conditions selected from the group consisting of wounds, poor wound healing, hair loss at and around wounds, scarring and / or wrinkles at and around wounds, and psoriasis by topical administration to an affected area in a subject in need thereof. [In the formula, X － is any halogen.

13. 13. The use according to claim 12, wherein the wound is selected from diabetic wounds, incisional wounds, surgical wounds, accidental wounds, pressure / bedsores, diabetic foot ulcers, pyoderma gangrenosum, burns, lesions, and cuts.

14. 14. The use according to claim 12 or 13, wherein the condition is a wound.

15. 14. The use according to claim 12 or 13, wherein the condition is poor wound healing.

16. 14. The use according to claim 12 or 13, wherein the condition is hair loss in and around a wound.

17. 14. The use according to claim 12 or 13, wherein the condition is scarring and / or wrinkles in and around wounds.

18. 14. The use according to claim 12 or 13, wherein the condition is psoriasis.

19. 14. Use according to claim 12 or claim 13, wherein the treatment is characterised by reducing, minimising or eliminating the appearance of scars and / or wrinkles at and around the wound site.

20. 14. Use according to claim 12 or claim 13, wherein the treatment is characterised by increasing and / or promoting hair growth at and around the wound site.

21. X is Br － Or Cl － The use according to any one of claims 12 to 20, wherein

22. The use according to any one of claims 12 to 20, wherein said compound of formula I is compound 1.

23. 1. A topical composition for use in treating wounds, poor wound healing, hair loss in and around a wound, scarring and / or wrinkles in and around a wound, psoriasis, or any combination thereof, said composition comprising a compound of formula I, [In the formula, X － is any halogen.] and a pharma- ceutically acceptable excipient.

24. X － Br － Or Cl － 24. The topical composition of claim 23, wherein

25. X － Br － 24. The topical composition of claim 23, wherein

26. 24. The topical composition of claim 23, wherein the pharma- ceutically acceptable excipient is one or more of an emulsifier, a preservative, or any combination thereof.

27. 1. A kit for treating wounds, poor wound healing, hair loss at and around a wound, scarring and / or wrinkles at and around a wound, psoriasis, or any combination thereof, comprising a compound of formula I: [In the formula, X － is any halogen. Optionally, the kit includes instructions for using the kit.

28. X － Br － Or Cl － 28. The kit of claim 27,

29. X － Br － 28. The kit of claim 27,

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