Anticholinergic and muscarinic agonists for treatment of demodex-related disorders

Abstract

Provided herein are methods of treating a skin condition, an ophthalmic condition, or an autoimmune disease, the method comprising administering or applying an active agent to an individual having a skin condition in a dose sufficient to inactivate demodex brevis and / or demodex follicularis in hair follicles and / or skin of the individual, thereby improving or stopping the manifestation of anaphylaxis and / or vasosystolic response to demodex that causes conditions or disease symptoms and signs in an individual. Also provided herein are pharmaceutical formulations suitable for use in any method.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 334,085 filed on April 22, 2022 and U.S. Provisional Patent Application No. 63 / 422,839 filed on November 4, 2022, each of which is incorporated herein by reference to the extent not inconsistent herewith. Background Art

[0003] Provided herein are methods for treating a variety of human afflictions using topical or orally administered anticholinergics and / or muscarinic agonists to inactivate certain organisms associated with skin afflictions. For example, by reducing or eliminating Demodex organisms from affected skin areas, the methods reduce clinical symptoms of skin afflictions that are primarily due to the body's allergic and vasomotor responses to the organisms and the bacteria carried by the organisms.

[0004] Rosacea, originally known as acne rosacea, is a chronic inflammatory skin disorder that most commonly affects the face and eyelids of middle-aged adults. Clinical symptoms include erythema (redness), dryness, papules, pustules, and nodules, which may occur alone or together on the affected areas of skin. Eyelid involvement may present as mild conjunctival irritation or inflammation of the meiotic (oil) glands at the eyelid margins. Chronic eyelid irritation can lead to loss of eyelashes. Chronic nasal involvement in male rosacea can result in bulbous enlargements known as rhinophyma. Typically, this occurs in adults between 30 and 50 years of age. Although some lesions of rosacea may resemble those of acne vulgaris, the processes are separate and distinct, with the major differences being that comedones (whiteheads and blackheads) are only present in acne vulgaris and not in rosacea, the characteristic midface localization and flushing of rosacea are not seen in acne, and rosacea may involve the eyelids, which is never seen in acne.

[0005] Rosacea develops in four stages over the course of several years, with flare-ups being exacerbated by changes in temperature, alcohol, spices, exposure to sunlight, and mood. The disease's multiple stages include:

[0006] Stage 1: Erythematous phase. Due to the sudden dilation of the facial arterioles, the patient will experience spasms of reddening of the skin, which then presents a reddened appearance. These spasms are caused by emotions, diet, and temperature changes.

[0007] Stage 2: Couperosis, a permanent red patch with dilated capillaries. Some patients also have swelling on the cheeks and forehead.

[0008] Stage 3: Inflammatory stage, inflammatory papules and pustules appear, but the sebaceous follicles are not affected, so there are no cysts and comedones.

[0009] Stage 4: Rhinophyma stage. This advanced stage affects mainly men. The patient's nose becomes red, uneven, and enlarged, with sebaceous gland hyperplasia and fibrous reorganization of connective tissue.

[0010] The cause of rosacea is still not completely understood, although many theories have been proposed. It has been a frequently discussed topic in the medical community, but a complete consensus has not yet been reached. The prominent erythema (redness) and flushing of the face due to exacerbations caused by heat, sunlight, and alcohol have drawn attention to this aspect of the disease. A common hypothesis is based on the presence of the characteristic Demodex folliculorum parasite in patients with rosacea. Other factors that may contribute to the development of rosacea have been described, such as hormonal factors (especially endocrine factors), climatic and immune factors, as well as bacterial factors through the presence of Helicobacter pylori, a bacterium associated with gastrointestinal disease.

[0011] Treatments that use drugs to block these vasomotor flushes have no effect on other aspects of the disease, such as papules and pustules. Treatments with oral and topical antibiotics have been shown to be effective in stopping the progression of rosacea through unclear anti-inflammatory mechanisms or by destroying bacteria associated with Demodex folliculorum, but studies have shown that these drugs do not work by killing the Demodex folliculorum organisms in the affected skin. Antibiotics must be taken continuously and in many cases have little effect. Many times, patients cannot tolerate the side effects associated with oral antibiotics.

[0012] There is still no consensus on the many subtypes of rosacea and seborrheic dermatitis, demodex brevis and demodex folliculorum, although they are hypothesized to be the root cause of rosacea and there are no commercially available pharmaceutical solutions for their treatment. Demodex disease presents as either rosacea or seborrheic dermatitis, but is proven to be caused by demodex mites.

[0013] In a study funded by the National Rosacea Society, the National University of Ireland in Maynooth found that the bacterium Bacillus oleronius stimulated an immune system response, inducing high levels of T-cell proliferation in 79% of patients with type 2 rosacea, compared to only 29% of patients without this disorder. The T-cell proliferation induced an inflammatory response, evident in papules and pustules. This suggests that antigens produced by the bacillus found in Demodex mites may be responsible for the tissue inflammation associated with papulopustular rosacea. Many of the current antibiotic treatments for rosacea are theoretically effective based on their ability to effectively combat Bacillus oleronius.

[0014] Typically, rosacea is treated with oral or topical antibiotics (such as tetracyclines, erythromycin, or clindamycin), but also vitamin A, salicylic acid, antifungals, steroids, anti-infectives (such as benzoyl peroxide), or in severe cases isotretinoin, or most commonly, metronidazole (an antibacterial agent).

[0015] Metronidazole is known to have antiparasitic, antiprotozoal and antibacterial properties. It is particularly used to treat Helicobacter pylori infections. It is also used in the treatment of rosacea due to its favorable properties against inflammatory lesions of rosacea, especially papules and pustules. Metronidazole has selective toxicity against anaerobic microorganisms and anoxic cells. In the latter, metronidazole is reduced to various derivatives that are able to modify its DNA structure.

[0016] U.S. Patent Publication No. 2013 / 0095051 (published on April 18, 2013) describes a method of treating rosacea using avermectin / metronidazole in a topical application. U.S. Patent No. 5,952,372 describes a method of treating rosacea using ivermectin orally or topically to reduce and eliminate the presence of Demodex folliculorum parasites on the patient's skin.

[0017] Ivermectin belongs to the avermectin family, a group of macrolides produced by the bacterium Streptomyces avermitilis. Avermectins include in particular ivermectin, invermectin, avermectin, abamectin, doramectin, eprinomectin and selamectin. It is known in the prior art that ivermectin has antiparasitic and anthelmintic properties. The antiparasitic activity is believed to be due to the opening of chloride channels in the parasite neuronal membranes under the action of increased release of the neurotransmitter GABA (gamma aminobutyric acid), inducing neuromuscular paralysis, leading to the death of some parasites. Ivermectin also interacts with other chloride channels, especially those that are dependent on the neurotransmitter GABA (gamma aminobutyric acid).

[0018] Ivermectin is often administered in the treatment of skin diseases with clinical manifestations of internal parasites, such as onchocerciasis and myiasis. U.S. Patent No. 6,133,310 describes the use of ivermectin to treat rosacea to reduce and eliminate the presence of Demodex folliculorum parasites on the patient's skin. U.S. Patent No. 6,133,310 describes the use of ivermectin to treat rosacea to reduce and eliminate the presence of Demodex folliculorum parasites on the patient's skin.

[0019] However, these treatments and compounds all have disadvantages such as irritation and intolerance phenomena, especially when they are applied chronically. All current treatments for rosacea appear to be suppressive rather than curative, particularly acting on the pustular spasms that occur during the inflammatory phase.

[0020] The National Rosacea Society estimates that 16 million Americans have rosacea, but only a small percentage are receiving treatment. The etiology of rosacea is currently controversial in the dermatology community. Rosacea is a common skin condition that causes redness on the face and often produces small, red, pus-filled bumps. If not treated, rosacea tends to get worse over time. The signs and symptoms of rosacea may flare up for weeks to months and then subside until they flare up again. Rosacea may be mistaken for acne, an allergic reaction, or other skin problems. While there is currently no cure for rosacea, current treatments can only help control and reduce the signs and symptoms of the condition.

[0021] Rosacea is usually observed in individuals after the age of thirty and presents as redness on the cheeks, nose, chin, or forehead that may come and go. In some cases, rosacea may also occur on the neck, chest, scalp, or ears. Over time, the redness tends to become redder and more persistent, and visible blood vessels may appear. If left untreated, bumps and pimples tend to develop, and in severe cases, the nose may become swollen and bumpy with excess tissue. This condition is called rhinophyma, and is responsible for the late comedian W.C. Fields' iconic bulbous nose. Many people with rosacea also have their eyes affected, which can feel irritated and appear watery or bloodshot.

[0022] Although rosacea can affect all groups of people, fair-skinned individuals who easily flush or blush are thought to be at greatest risk. The disease is more common in women, but symptoms tend to be more severe in men. Rosacea can vary greatly from person to person, and in most cases, some but not all of the potential signs and symptoms will be present. According to a consensus committee and review panel of 17 medical experts from around the world, rosacea typically includes at least one of the following cardinal signs, and multiple secondary signs and symptoms may also be present.

[0023] The main signs of rosacea include: (1) Blushing: Many people with rosacea have a history of frequent blushing or flushing. This facial redness may come and go and is often the earliest sign of the disease. (2) Persistent redness: Persistent facial redness is the most common individual sign of rosacea and may resemble a blush that does not go away or a sunburn. (3) Bumps and papules: Small, solid, red bumps or pus-filled papules often appear. Although these may resemble acne, they do not have blackheads and may cause burning or stinging. (4) Visible blood vessels: In many people with rosacea, small blood vessels become visible in the skin. (6) Other possible signs and symptoms include: Eye irritation. Many people with rosacea may experience irritation in their eyes that appear watery or bloodshot, a condition known as ocular rosacea. The eyelids may also become red and swollen, and styes are common. Severe cases can lead to corneal damage and vision loss without medical attention. Burning or stinging: Burning or stinging may often occur on the face. Itching or tightness may also occur.Dry appearance: The skin in the center of the face may be rough and therefore appear very dry.Plaques: Raised, red patches, called plaques, may appear without changes in the surrounding skin.Thickened skin: The skin may thicken and enlarge due to extra tissue, most commonly on the nose. This condition is called rhinophyma, and men are more likely to be affected than women.Swelling: Swelling of the face, called edema, may occur with other signs of rosacea or on its own.Signs beyond the face: Signs and symptoms of rosacea may also develop beyond the face, most commonly on the neck, chest, scalp, or ears.

[0024] The various subtypes of rosacea include: subtype 1 (erythematoangiectatic rosacea), which is characterized by blushing and persistent redness that may include visible blood vessels; subtype 2 (papulopustular rosacea), which is characterized by persistent redness with transient bumps and papules; subtype 3 (phymatous rosacea), which is characterized by thickening of the skin, often leading to enlargement of the nose due to excess tissue; and subtype 4 (ocular rosacea), which is characterized by ocular manifestations such as dryness, tearing and burning of the eyes, eyelid swelling, recurrent styes, and corneal damage leading to potential vision loss.

[0025] Many patients will have features of more than one subtype at the same time, and these features may often progress in a continuum. Although rosacea may or may not progress from one subtype to another, each individual's signs or symptoms may progress from mild to moderate to severe. Early diagnosis and treatment are recommended.

[0026] Eyelid involvement may present as mild conjunctival irritation or inflammation of the meibomian (oil) glands at the eyelid margins. Chronic eyelid inflammation can lead to eyelash loss. Meibomian gland dysfunction due to irritation and inflammation can lead to chronic dry eyes and / or blepharitis.

[0027] Although Demodex folliculorum is postulated to be the underlying cause of dry eye associated with meibomian gland dysfunction, this is not a consensus, and there are no commercially available pharmaceutical solutions for the treatment of Demodex brevis and Demodex folliculorum in the eyelid. Responses to the presence or metabolic activity of Demodex in the eyelash follicles have been discussed as a cause of blepharitis, but prior studies using topical miticides other than anticholinergics and direct muscarinic agonists have shown inconsistent and marginal results. Demodex appears to play a role in meibomian gland dysfunction, which may ultimately cascade to the development of chronic dry eye.

[0028] The etiology of dry eye is becoming increasingly understood. Dry eye is progressive in nature and is fundamentally caused by inadequate tear coverage of the ocular surface. This poor tear coverage impedes healthy gas exchange and nutrient transport to the ocular surface, exacerbates cellular dryness, and produces a poor visual refractive surface. Poor tear coverage is often caused by: Excessive tear evaporation due to meibomian gland dysfunction. Meibomian glands at the eyelid margin within the tarsal plate are responsible for supplying meibum, an oily substance that prevents evaporation of the tear film from the eye. If the flow of meibum is blocked by inflammation caused by Demodex (or indirectly through an inflammatory response to Demodex), by cylindrical dandruff caused by Demodex, or by bacteria that proliferate from Demodex, then the lack of meibum can lead to a cascade effect that results in chronic dry eye. Low tear volume results in a hyperosmotic environment that induces a state of inflamed ocular surface. This inflammatory response induces apoptosis of surface cells, which in turn prevents proper distribution of the tear film over the ocular surface, making any given tear volume less effective. This sets off a vicious positive feedback loop where more inflammation leads to more surface cell damage, etc. Furthermore, because the sensory neurons on the ocular surface are damaged, the neural control circuits that control reflex tear activation are disrupted. As a result, fewer tears are secreted, and a second vicious cycle is created, leading to further progression of the disease, as fewer tears lead to nerve cell loss, which leads to even fewer tears, and so on.

[0029] There are multiple treatments for dry eye, however, none have substantial efficacy in the treatment of the condition. Treatment options include artificial tear substitutes, ointments, gels, warm compresses, environmental modification, topical cyclosporine, omega-3 fatty acid supplements, punctal plugs, and wet chamber lenses. Patients with severe disease may also be managed with punctal cautery, systemic cholinergic agonists, systemic anti-inflammatory agents, mucolytics, autologous serum tears, PROSE scleral contact lenses, and tarsorrhaphy. Despite these treatment options, dry eye is still considered one of the worst ophthalmic diseases. Therefore, a more effective treatment for dry eye is needed.

[0030] Typically, dry eyes are treated with frequent use of artificial tears or other lubricating eye drops. Artificial tears are usually the first step in dry eye treatment. Eye drops are the most commonly prescribed medication for dry eye syndrome ( (Cyclosporin ophthalmic emulsion) 0.05%) contains the immunosuppressant cyclosporin. Cyclosporin can increase tear secretion that is reduced due to eye inflammation. The eye drops are used to treat chronic dry eyes that may be caused by inflammation. Ocular rosacea is characterized by ocular manifestations such as dry eyes, tearing and burning sensations, eyelid swelling, recurrent styes, and potential vision loss due to corneal damage. There is a need in the art for a safe and effective treatment of ophthalmic conditions, particularly inflammatory eye diseases caused by Demodex, such as meibomian gland dysfunction, conjunctivitis, keratoconjunctivitis, congestion, dry eyes, and blepharitis.

[0031] Although Demodex folliculorum is assumed to be a benign human ectoparasite, it has been implicated in the autoimmune diseases lupus erythematosus, rheumatoid arthritis, juvenile idiopathic arthritis, systemic lupus erythematosus, systemic sclerosis, juvenile and adult dermatomyositis, Sjögren's syndrome ( syndrome) and porphyria cutanea tarda. There are no commercially available pharmacological treatment options for the treatment of Demodex brevis and Demodex folliculorum in autoimmune diseases. Responses to the presence or metabolic activity of Demodex in the human epidermis have been previously described in dermatological diseases such as rosacea and acne vulgaris, and in the ophthalmic condition meibomian gland dysfunction, which is considered a major cause of dry eye. All autoimmune diseases with a parasitic component, particularly those with a dermatological component, where the use of the drugs hydroxychloroquine and / or chloroquine as an effective intervention is considered to have an antiparasitic component, including a component that acts on Demodex in the etiology of the disease.

[0032] Repositioning anticholinergic and direct muscarinic agonist compounds to treat autoimmune diseases suspected of having a Demodex infestation provides a new treatment modality for patients with autoimmune diseases including systemic lupus erythematosus, rheumatoid arthritis, juvenile idiopathic arthritis, systemic lupus, systemic sclerosis, juvenile and adult dermatomyositis, Sjögren's syndrome and porphyria cutanea tarda, relapsing rheumatic disease, eosinophilic fasciitis, polymorphous light eruption, granuloma annulare , lichen planus, lupus panniculitis, discoid lupus, porphyria cutanea tarda, psoriatic arthritis, chronic ulcerative stomatitis, refractory chronic urticaria, sarcoidosis, frontal fibrosing alopecia, necrobiosis lipoidica, actinic reticulocytosis, actinic prurigo, epidermolysis bullosa, Kikuchi-Fujimoto disease, graft-versus-host disease, erythema nodosum chronicus, sleep-deprivation and systemic sclerosis, pemphigus vulgaris, pemphigus foliaceus and pemphigoid gestationis.

[0033] The causes of the following diseases are not fully understood: diseases Systemic lupus erythematosus, rheumatoid arthritis, juvenile idiopathic arthritis, systemic lupus, systemic sclerosis, juvenile and adult dermatomyositis, Sjögren's syndrome and porphyria cutanea tarda, relapsing rheumatic disease, eosinophilic fasciitis, polymorphic light eruption, granuloma annulare, lichen planus, lupus panniculitis, discoid lupus, porphyria cutanea tarda, psoriatic arthritis, chronic ulcerative stomatitis, refractory chronic urticaria, sarcoidosis, frontal fibrosing alopecia, necrobiosis lipoidica, actinic reticulocytosis, actinic prurigo, epidermolysis bullosa, Kikuchi-Fujimoto disease, graft-versus-host disease, chronic erythema nodosum, sleep-related and systemic sclerosis, pemphigus vulgaris, pemphigus foliaceus and pemphigoid gestationis. Autoimmune diseases are defined as those in which the body produces antibodies that attack its own tissues, leading to deterioration and, in some cases, destruction of such tissues. The immune system of some people may actually be upregulated by the presence of the ubiquitous human Demodex mite or pathogens carried on or in the mite.

[0034] Many of the triggers for rosacea flares overlap with those for autoimmune flares, especially lupus (SLE). Lupus rashes, much like the rash seen in patients with rosacea, tend to occur in the T and U zones of the face. In lupus, the rash is called a malar rash or butterfly rash. Antibiotics used to treat rosacea can induce lupus, especially tetracyclines. Antibiotics may act on the mites, leading to drug-induced lupus. The most effective treatment for drug-induced lupus is currently to stop the drug therapy that triggered the reaction. It should also be noted that tetracycline antibiotics are used to treat acne, rosacea, and meibomian gland dysfunction. Whether antibiotics act as antimicrobials and kill Demodex mites or act on the mite gut bacteria remains to be fully elucidated. We were able to show in our in vitro assays that doxycycline does have anti-mite activity.

[0035] The antimalarial drugs hydroxychloroquine and chloroquine treat autoimmune diseases by acting as antiparasitic agents against Demodex mites. Drugs used to treat dry eye, in which Demodex mites are now suspected of playing a pathogenic role, are treated with the same drugs used to treat many of these autoimmune diseases. Cyclosporine and Lifitegrast work by downregulating the patient's T cells or T lymphocytes. The immunosuppressant Tacrolimus has also been reported to treat dry eye. These immunosuppressants downregulate the host's immune response to Demodex mites in many different diseases that can be redefined as severe allergic or inflammatory reactions to Demodex mites or pathogens carried by the mites.

[0036] Demodex has been found in the lymph glands of dogs. Demodex canis may serve as an alternative model to better understand Demodex brevis and Demodex folliculorum in humans. It should also be noted that dogs have lupus, known as canine discoid lupus erythematosus, which may have a potential pathogenic role related to Demodex canis.

[0037] Intense pulsed light (IPL) has been used to treat rosacea and dry eyes. Demodex mites are believed to be a causative factor in both diseases. IPL treatment has been shown to kill Demodex mites. See Prieto V et al., “Effects of intense pulsed light on sun-damaged human skin, routine, and ultrastructural analysis.” (2002); Timothy Kirn, “Intense pulsed light eradicates Demodex mites.” Skin Allergy News (2002). In 2000, Dr. JL Levy described the treatment of chronic facial erythema of systemic lupus erythematosus in “Intense pulsed light treatment for chronic facial erythema of systemic lupus erythematosus: a case report.” In the report, a 33-year-old woman who had been diagnosed with systemic lupus erythematosus presented with chronic facial erythema and rosacea. The patient experienced redness and burning sensation on her facial skin and had sought treatment with antimalarial medications. After discussing multiple treatment options with the patient, she agreed to intense pulsed light therapy. Improvement was observed after the first treatment, and 75% clearance was observed 1 month after the second treatment. No treatment-related side effects occurred. One year later, the results of both treatments were observed to be maintained. Photosensitivity is one of the most prevalent symptoms of lupus, and ultraviolet light from the sun or artificial light is the most common trigger for lupus flare-ups. The use of IPL in patients with lupus is highly counterintuitive. The ultimate clearance in the case report is consistent with the mechanism of clearance of Demodex mites from the patient's facial skin.

[0038] US Patent Nos. 10,709,135 and 10,500,183 relate to treating skin disorders by inactivating Demodex brevis and / or Demodex folliculorum with organophosphates and acetylcholinesterase inhibitors. US Patent No. 11,446,241 relates to treating ophthalmic disorders by inactivating Demodex brevis and / or Demodex folliculorum with selected acetylcholinesterase inhibitors.

[0039] The art needs treatment methods with significant efficacy for these autoimmune diseases. Traditional treatment focuses on improving the quality of life by controlling symptoms and minimizing attacks. Therapeutic management includes lifestyle management and medications, such as anti-inflammatory drugs, steroids and immunosuppressants. Other treatments include acetaminophen or nonsteroidal anti-inflammatory drugs (NSAIDs), sometimes used in combination with antimalarials. Acetaminophen and nonsteroidal anti-inflammatory drugs are usually sufficient to relieve symptoms. Antimalarials include hydroxychloroquine (chloroquine) and chloroquine. Use low-dose corticosteroids and / or corticosteroid creams or ointments. Corticosteroids, including those such as prednisone. Immunosuppressive drugs such as azathioprine, belimumab, cyclophosphamide, methotrexate or mycophenolate mofetil can be used. Corticosteroids are the first-line treatment for muscle involvement in dermatomyositis, but skin lesions tend to be aggravated by reduction or discontinuation. There is a high unmet need for new treatment strategies that focus on skin lesions in systemic autoimmune diseases (including mites). Summary of the invention

[0040] There is a need in the art for new therapeutic strategies to treat a range of diseases associated with Demodex. Provided herein are methods of treatment for Demodex-induced inflammatory conditions (including skin conditions and / or eye conditions) comprising inactivating (e.g., by paralyzing, killing and / or eliminating) Demodex by administering or applying specific active agents, thereby alleviating, eliminating or otherwise slowing or stopping any one or more clinical symptoms associated with Demodex-induced inflammatory skin and / or eye conditions. Exemplary active agents include anticholinergic agents and / or muscarinic agonists that are effective in inactivating Demodex.

[0041] Based on the understanding of the role of Demodex in a range of diseases, a variety of treatment methods are provided herein that specifically target this Demodex-induced mechanism by applying an active agent. In an exemplary embodiment, the active agent applied can be one or more anticholinergic agents and / or muscarinic agonists applied topically to the hair follicles, skin, eyes, eyelids, eyelashes, or meibomian glands of an individual. For example, in one embodiment, the topically applied active agent penetrates the outer layer of the individual's skin, thereby exposing short Demodex and / or folliculorum Demodex present under the outer layer of the skin in the eyelid to the active agent. For example, in one embodiment, the topically applied active agent penetrates the subcutaneous area of ​​the individual's eyelid, thereby exposing short Demodex and / or folliculorum Demodex present in and around the meibomian glands to the active agent. Optional formulations of topical active agents that can be used in the methods provided herein include one or more compositions that increase skin permeability, such as formulations including an active agent and a skin permeation agent such as dimethyl sulfoxide (DMSO).

[0042] In an exemplary embodiment, the topically applied active agent is applied to the affected skin area at least once a day and no more than twice a day for about 2 to 12 weeks. In one embodiment, during a first application period, the topically applied active agent is applied to the affected area and / or unaffected area, thereby paralyzing, killing and / or eliminating Demodex mites, including adult Demodex brevis and / or Demodex folliculorum, in the hair follicles, skin, eyes, eyelids, eyelashes and / or meibomian glands. In one embodiment, during a second application period, the topically applied active agent is further applied to the affected area and / or unaffected area, thereby paralyzing, killing and / or eliminating individual Demodex brevis and / or Demodex folliculorum that matured from larval and / or egg forms present on and / or in the skin during the first application period from the hair follicles, skin, eyes, eyelids, eyelashes or meibomian glands. In one embodiment, during a third application period, a topically applied active agent is further applied to the affected area and / or unaffected area to paralyze, kill and / or eliminate from the hair follicles, skin, eyes, eyelids, eyelashes or meibomian glands individual Demodex brevis and / or Demodex folliculorum mites that matured from larval and / or egg forms present on and / or in the skin and / or on and / or in the hair follicles during the first application period and / or the second application period.

[0043] Provided herein are methods for treating a variety of autoimmune diseases, including inactivating certain organisms associated with autoimmune diseases in humans using topically applied, orally administered, or intravenously administered active agents (including anticholinergics and / or muscarinic agonists). For example, by reducing or eliminating Demodex organisms in a patient (including infected areas), the methods reduce clinical signs or symptoms of autoimmune diseases that are primarily due to the body's inflammatory and / or immune response to Demodex organisms and / or pathogens carried by the Demodex organisms.

[0044] Embodiments described herein relate to treating ophthalmic conditions by topical, intraocular, or oral administration of one or more active agents. Such treatment relieves clinical signs and symptoms of ophthalmic conditions more completely than any previously described method by effectively reducing or eliminating the Demodex population in infected areas and areas where Demodex may be present. Embodiments of the present invention may be used to treat ophthalmic conditions associated with Demodex-induced inflammatory eye diseases, including meibomian gland dysfunction, conjunctivitis, keratoconjunctivitis, hyperemia (excess blood flow to the eye), blepharitis, and dry eye.

[0045] Also provided herein are methods of treating an individual suffering from a Demodex-induced inflammatory eye disease. The method may include the steps of applying an active agent to an individual in need thereof, the active agent in a dose sufficient to inactivate Demodex brevis and / or Demodex folliculorum in an ocular region of the individual, wherein the ocular region includes one or more of hair follicles, eyelids, eyelashes, and meibomian glands in and around the eye region, including causing a sufficient amount of Demodex brevis and / or Demodex folliculorum to be inactivated to ameliorate or stop the manifestations of an allergic and / or inflammatory response to Demodex that causes the inflammatory eye disease. Depending on the sensitivity of the individual, the sufficient amount may correspond to at least 50%, 70%, 90%, or 95% elimination, including a reduction in and around the eye region and / or systemically.

[0046] The inflammatory eye disease may correspond to one or more of the following: meibomian gland dysfunction, dry eyes, blepharitis, conjunctivitis, keratoconjunctivitis, or hyperemia.

[0047] For any application described herein, the application may be topical application, including application to the affected skin area of ​​the individual and / or the periocular area of ​​the eye area, preferably avoiding direct application to the eye surface. The eye area may correspond to an area of ​​at least 1 cm to 10 cm around the outermost periphery defined by the eye, eyelid, eyebrow and eyelashes.

[0048] Provided herein are treatment methods for alleviating, eliminating or otherwise reducing or stopping any one or more of the above clinical symptoms by administering or applying anticholinergics and direct muscarinic agonists, naturally occurring anticholinergics and direct muscarinic agonists.

[0049] Embodiments of the invention described herein relate to treating skin disorders by topical or oral administration of one or more anticholinergic agents and direct muscarinic agonists, including naturally occurring anticholinergic agents or direct muscarinic agonists. Such treatment relieves the clinical signs and symptoms of the skin disorder more completely than any previously described method by effectively reducing or eliminating the demodex population in the affected skin area and in areas where demodex mites may be present. Embodiments of the invention can be used to treat skin disorders including acne vulgaris, seborrheic dermatitis, perioral dermatitis, acneiform eruptions, transient acantholytic dermatosis, acne necrotica milliaris, psoriasis, steroid-induced dermatitis, primary irritant dermatitis, rosacea, and hidradenitis suppurativa.

[0050] One exemplary method embodiment comprises the step of orally administering or topically applying to an individual with a skin disorder an anticholinergic agent and a direct muscarinic agonist in an amount sufficient to inactivate Demodex brevis and / or Demodex folliculorum in the hair follicles and / or skin of the individual, thereby alleviating or stopping the manifestation of an allergic and / or vasomotor response to the mites that causes the individual to develop symptoms and signs of the skin disorder. Optionally, the skin disorder affects the facial skin or the eyelids, or both the facial skin and the eyelids.

[0051] In an exemplary embodiment, the anticholinergic agent and the direct muscarinic agonist are applied topically. In one embodiment, for example, the topically applied anticholinergic agent and the direct muscarinic agonist are formulated into a carrier lotion, emulsion, cream, soap, wash, shampoo or gel. Optionally, the concentration of the anticholinergic agent and the direct muscarinic agonist in the topically applied lotion, cream, soap, wash, shampoo or gel is about 0.001% to 5% by weight or about 0.01% to 1% by weight. In an exemplary embodiment, the concentration of the anticholinergic agent and the direct muscarinic agonist in the topically applied lotion, cream, soap, wash, shampoo or gel is the minimum concentration that is effective in killing Demodex mites. In one embodiment, the dosage of the anticholinergic agent and the direct muscarinic agonist in the topically applied lotion, cream, soap, wash, shampoo or gel is less than about 150 mg / kg body weight or is about 0.01 mg / kg body weight to 50 mg / kg body weight. In an exemplary embodiment, the dosage of the anticholinergic agent and the direct muscarinic agonist in the topically applied lotion, cream, soap, wash, shampoo or gel is the minimum dosage effective to kill Demodex mites. Optionally, the topically applied anticholinergic agent and the direct muscarinic agonist are encapsulated in microliposomes before being formulated into a carrier lotion, cream, soap, wash, shampoo or gel.

[0052] In general, the methods of the present invention include those in which the topically applied anticholinergic agent and direct muscarinic agonist are applied to an area of ​​the skin affected by a skin disorder. However, in certain embodiments, the topically applied anticholinergic agent and direct muscarinic agonist are also applied to an area of ​​the skin not affected by a skin disorder. For example, in one embodiment, the topically applied anticholinergic agent and direct muscarinic agonist are applied to areas of the body skin where Demodex brevis and / or Demodex folliculorum are present. In an exemplary embodiment, the topically applied anticholinergic agent and direct muscarinic agonist are applied to all areas of the skin.

[0053] Optionally, the method of the present invention further comprises the step of applying the anticholinergic agent and the direct muscarinic agonist to the clothing, bedding, or both clothing and bedding of the individual. Such application is useful, for example, for preventing the clothing or bedding of the individual from becoming a source of demodex mites, thereby re-entering the skin of the individual. Similarly, the method of the present invention optionally further comprises the step of orally administering or topically applying the anticholinergic agent and the direct muscarinic agonist to other individuals in a dose sufficient to kill and eliminate the short demodex mites and / or the hair follicle demodex mites in the hair follicles and / or skin of other individuals in contact with the individual. For example, in an embodiment, the other individuals include cohabiting members, children, spouses, partners, family members, or domestic pets.

[0054] In an exemplary embodiment of the methods of the present invention, a topically applied anticholinergic agent and a direct muscarinic agonist are applied to the hair follicles and / or skin of an individual. For example, in one embodiment, the topically applied anticholinergic agent and direct muscarinic agonist penetrate the outer layer of the skin of the individual, thereby exposing Demodex brevis and / or Demodex folliculorum present under the outer layer of the skin to the anticholinergic agent and the direct muscarinic agonist. For example, in one embodiment, the topically applied anticholinergic agent and direct muscarinic agonist penetrate the subcutaneous region of the skin of the individual, thereby exposing Demodex brevis and / or Demodex folliculorum present in the subcutaneous region of the skin to the anticholinergic agent and the direct muscarinic agonist. Certain formulations of topical anticholinergic agents and direct muscarinic agonists useful in the methods of the present invention optionally include one or more compositions that increase skin permeability, such as dimethyl sulfoxide (DMSO).

[0055] In an exemplary embodiment, the topically applied anticholinergic agent and direct muscarinic agonist are applied to the affected skin area at least once a day, and no more than twice a day, for about two to six weeks. In one embodiment, during a first application period, the topically applied anticholinergic agent and direct muscarinic agonist are applied to the affected and / or unaffected skin area, thereby killing and eliminating adult Demodex brevis and / or Demodex folliculorum in individual skin hair follicles. In one embodiment, during a second application period, the topically applied anticholinergic agent and direct muscarinic agonist are further applied to the affected and / or unaffected skin area, thereby killing and eliminating individual Demodex brevis and / or Demodex folliculorum that have matured from larval forms and / or egg forms present on and / or in the skin during the first application period. In one embodiment, during the third application period, a topically applied anticholinergic agent and a direct muscarinic agonist are further applied to the affected and / or unaffected skin areas to kill and eliminate individual Demodex brevis and / or Demodex folliculorum mites that matured from larval and / or egg forms present on and / or in the skin and / or on and / or in hair follicles during the first application period and / or the second application period.

[0056] Optionally, the first application period and the second application period are separated by at least 5 days but no more than 10 days. Optionally, the first application period and the second application period are separated by at least seven days. In an exemplary embodiment, the time between the first application period and the second application period is sufficient to allow the larval form to mature into the adult form and / or allow the egg form to mature into the adult form.

[0057] Optionally, the second application period and the third application period are separated by at least 5 days but no more than 10 days. Optionally, the second application period and the third application period are separated by at least seven days. In an exemplary embodiment, the time between the second application period and the third application period is sufficient to allow the larval form to mature into the adult form and / or allow the egg form to mature into the adult form.

[0058] In an exemplary embodiment, the anticholinergic agent and the direct muscarinic agonist are administered orally or applied topically on a continuous intermittent regimen sufficient to prophylactically control the Demodex mite population in the hair follicles and / or skin of an individual.

[0059] In another embodiment, the active agent is administered orally. In a specific embodiment, for example, the orally administered anticholinergic agent and direct muscarinic agonist are administered at an oral dose of about 150 mg / kg body weight or less or about 0.01 mg / kg body weight to 50 mg / kg body weight of the anticholinergic agent and direct muscarinic agonist. In an exemplary embodiment, the orally administered anticholinergic agent and direct muscarinic agonist are administered at an oral dose of the anticholinergic agent and direct muscarinic agonist that is the lowest dose effective to kill Demodex mites. In certain embodiments, the orally administered anticholinergic agent and direct muscarinic agonist are formulated as a prodrug or a pharmaceutically acceptable salt.

[0060] Optionally, the orally administered anticholinergic agent and direct muscarinic agonist is administered at a daily dose of 10 mg / kg body weight. Optionally, the orally administered anticholinergic agent and direct muscarinic agonist is administered at a daily dose of 7.5 mg / kg body weight. Optionally, the orally administered anticholinergic agent and direct muscarinic agonist is administered at a dose of 5 mg / kg body weight three times a day. Optionally, the orally administered anticholinergic agent and direct muscarinic agonist is repeated about 2 to 4 times with an interval of 3 to 7 days between them.

[0061] In various embodiments, removing Demodex brevis and / or Demodex folliculosus from the hair follicles and / or skin of an individual can reduce the number of one or more bacteria in the hair follicles and / or skin of the individual. For example, in some embodiments, the allergic and / or vasomotor response to mites is caused by the presence of one or more bacteria associated with mites in the hair follicles and / or skin of the individual. In specific embodiments, the one or more bacteria include one or more bacteria from the genus Staphylococcus or Bacillus. For example, in one embodiment, the one or more bacteria include Bacillus oleronius bacteria. For example, in one embodiment, the one or more bacteria include Staphylococcus epidermidis bacteria. Optionally, the one or more bacteria are present in the digestive system of Demodex brevis and / or Demodex folliculosus.

[0062] Another exemplary method for treating a skin disorder includes the step of topically applying an active ingredient to an individual with a skin disorder, the active ingredient being in a dose sufficient to kill and eliminate Demodex brevis and / or Demodex folliculorum in the hair follicles and / or skin of the individual, thereby stopping the manifestation of an allergic and / or vasomotor response to the mites that causes the symptoms and signs of the individual's skin disorder, wherein the topically applied active ingredient is applied to an area of ​​the skin with the skin disorder and an area of ​​the skin that is not suffering from the skin disorder. In a specific embodiment, the topically applied active ingredient is applied to all of the skin of the individual, thereby killing and eliminating Demodex brevis and / or Demodex folliculorum on all of the skin of the individual. Likewise, the methods of the present invention can be used, for example, to treat skin disorders including acne vulgaris, seborrheic dermatitis, perioral dermatitis, acneiform eruptions, transient acantholytic dermatosis, miliary necrotizing acne, psoriasis, steroid-induced dermatitis, primary irritant dermatitis, rosacea, or hidradenitis suppurativa. In an exemplary embodiment, the skin disorder is caused by, exacerbated by, or otherwise comorbid with an infestation of the skin and / or hair follicles by Demodex mites.

[0063] In one aspect, anticholinergics and direct muscarinic agonists include the anticholinergics atropine (Atropen), belladonna alkaloids, benztropine mesylate (Cogentin), cyclamate, cyclopentolate (Cyclogyl), darifenacin (Enablex), dicylomine, fesoterodine (Toviaz), flavonoids (Urispas), glycopyrrolate, homatropine hydrobromide, scopolamine (Levsinex), ipratropium (Atrovent), orphenadrine, oxybutynin (Ditropan XL) propantheline (propantheline bromide), scopolamine, methscopolamine, solifenacin (VESIcare), tiotropium (Spiriva), tolterodine (Detrol), trihexyphenidyl, trospium, citalopram, fluoxetine, fluvoxamine, paroxetine, amitriptyline, clomipramine, desipramine, doxepin, imipramine mine), nortriptyline, trimipramine, desipramine, amoxapine, maprotiline, clozapine, quetiapine, biperiden, chlorpheniramine, dimenhydrinate, diphenhydramine, doxepin, doxelamine, glycopyrronium, oxitropium, promethazine, solifenacin, tropicamide, and umeclidinium. Direct muscarinic agonists: acetylcholine, carbachol, methacholine, bethanechol, muscarine, nicotine, pilocarpine, oxotremorine, bupropine, dextromethorphan, doxacurium, hexamethonium, mecamylamine, tubocurarine, cytisine, succinylcholine chloride (succinylcholine chloride), and varenicline.

[0064] In another embodiment, the anticholinergics and direct muscarinic agonists correspond to compounds currently used in medicine, including compounds with established safety in humans. These drugs include direct muscarinic agonists: atropine (Atropen), belladonna alkaloids, benztropine mesylate (Cogentin), cyclamate, cyclopentolate (Cyclogyl), darifenacin (Enablex), dicylomine, fesoterodine (Toviaz), flavonoids (Urispas), glycopyrrolate, homatropine hydrobromide, scopolamine (Levsinex), ipratropium (Atrovent), orphenadrine, oxybutynin (Ditropan XL) propantheline (propantheline bromide), scopolamine, methscopolamine, solifenacin (VESIcare), tiotropium (Spiriva), tolterodine (Detrol), trihexyphenidyl, trospium, citalopram, fluoxetine, fluvoxamine, paroxetine, amitriptyline, clomipramine, desipramine, doxepin, imipramine, Metriptyline, trimipramine, desipramine, amoxapine, maprotiline, clozapine, quetiapine, biperiden, chlorpheniramine, dimenhydrinate, diphenhydramine, doxepin, doxylamine, glycopyrronium, oxitropium, promethazine, solifenacin, tropicamide, and umeclidinium. Direct muscarinic agonists: acetylcholine, carbachol, methacholine, bethanechol, muscarine, nicotine, pilocarpine, oxotremorine.

[0065] Any of the methods provided herein may use an antinicotinic agent including, but not limited to, bupropine, dextromethorphan, docurium, hexamethonium, mecamylamine, tubocurarine, cytisine, succinylcholine chloride (succinylcholine chloride), and / or varenicline.

[0066] Also provided herein are pharmaceutical formulations suitable for use in any of the methods described herein. Also provided herein are uses of any of the compounds described herein for treating mite-induced conditions caused by the presence of one or both of Demodex brevis and Demodex folliculorum. Also provided herein are uses of the compounds described herein for the preparation of a medicament for treating a mite-induced skin condition, wherein the mite is one or both of Demodex brevis and Demodex folliculorum.

[0067] Provided herein is a method for treating acarid to at least alleviate clinical symptoms. For example, the method can be used for treating an individual suffering from Demodex, which causes the clinical symptoms that one or more acarids bring out, and the method comprises the step of applying an active agent to an individual suffering from acarid, the dosage of which is enough to inactivate the Demodex in the hair follicles, skin, eyes, eyelids, eyelashes and / or meibomian glands of the individual, thereby improving or stopping the clinical symptoms that acarids bring out. The active agent can be selected from: promethazine, benztropine, dicyclomine, fluvoxamine, imipramine, dextromethorphan, mecamylamine, bupropine, fesoterodine, paroxetine, amoxapine, trihexylfendi, nortriptyline, clozapine, dimethylhydrogen ester, chlorpheniramine, cyclopentyl diphenyl ester, quetiapine, its prodrug and its pharmaceutically acceptable salt. The individual needing treatment may suffer from skin disorders, ophthalmic disorders or autoimmune diseases.

[0068] Also provided herein is a method for treating skin disorders caused by Demodex. For example, the method can be used to treat an individual suffering from a skin disorder, comprising the step of topically applying or orally administering an active agent to an individual suffering from a skin disorder, wherein the dosage of the active agent is sufficient to inactivate Demodex in the hair follicles and / or skin of the individual, thereby improving or stopping the manifestation of allergic and / or vasomotor responses to mites that cause symptoms and signs of the individual's skin disorder. The active agent can be selected from: promethazine, benztropine, dicyclomine, fluvoxamine, imipramine, dextromethorphan, mecamylamine, bupropine, fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, cyclopentyl ester, quetiapine, its prodrugs, and its pharmaceutically acceptable salts.

[0069] Also provided herein is a method for treating an autoimmune disease caused by Demodex. For example, the method can be used to treat an individual suffering from an autoimmune disease, the method comprising the step of administering an active agent to the individual, the dosage of the active agent being sufficient to inactivate at least a portion of the Demodex in or on the body of the individual, wherein the inactivation results in an improvement or cessation of one or more symptoms associated with an inflammatory response and / or immune response to Demodex, the inflammatory response and / or immune response causing one or more symptoms associated with the autoimmune disease in the individual. The active agent can be selected from the group consisting of promethazine, benztropine, dicyclomine, fluvoxamine, imipramine, dextromethorphan, mecamylamine, bupropine, fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, cyclopentyl ester, quetiapine, prodrugs thereof, and pharmaceutically acceptable salts thereof.

[0070] Also provided herein is a method for treating an ophthalmic disorder in an individual. For example, the method can be used to treat an individual suffering from an ophthalmic disorder, the method comprising the step of topically applying or orally administering an active agent to an individual suffering from an ophthalmic disorder, the dosage of the active agent being sufficient to inactivate Demodex in the individual's hair follicles, skin, eyes, eyelids, eyelashes and / or meibomian glands, thereby improving or stopping the manifestation of allergic and / or inflammatory reactions to Demodex that cause symptoms and signs of the individual's ophthalmic disorder. The active agent can be selected from: promethazine, benztropine, dicyclomine, fluvoxamine, imipramine, dextromethorphan, mecamylamine, bupropine, fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, cyclopentyl ester, quetiapine, its prodrugs, and its pharmaceutically acceptable salts.

[0071] The active agent may be an anticholinergic and / or a muscarinic agonist, including a direct muscarinic agonist, so long as it is capable of inactivating the Demodex mite that is directly or indirectly responsible for the underlying condition.

[0072] The demodex mites may be Demodex brevis and Demodex folliculorum.

[0073] The step of topically applying or orally administering the active agent may kill the Demodex mites, paralyze the Demodex mites, and / or render at least a portion of the Demodex mites incapable of reproduction. Inactivation may be characterized as "substantially inactivating," meaning a reduction in the number of mites in a selected area of ​​at least 50%, at least 70%, at least 90%, at least 95%, at least 98%, at least 99%. Functionally, such reduction is considered to be sufficient to ameliorate, alleviate, and / or cause cessation of clinical symptoms. Of course, reinfestation of mites is generally to be expected, and therefore, treatment may be continued or repeated periodically as needed.

[0074] The active agent can be selected from the group of active agents that kill Demodex mites. For example, the active agent can be selected from: benztropine, dicyclomine, fluvoxamine, imipramine, promethazine, dextromethorphan, mecamylamine, bupropine, and pharmaceutically acceptable salts thereof.

[0075] The active agent can be selected from: promethazine hydrochloride, benztropine mesylate, dicyclomine hydrochloride, fluvoxamine, imipramine maleate, dextromethorphan hydrobromide, mecamylamine hydrochloride, and bupropine hydrochloride.

[0076] The active agent may be selected from the group of active agents that paralyze Demodex mites. The active agent may be selected from: fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, cyclopentyl, quetiapine, and pharmaceutically acceptable salts thereof.

[0077] The active agent may be selected from the group consisting of: fesoterodine fumarate, paroxetine hydrochloride, trihexyphenidyl hydrochloride, nortriptyline hydrochloride, chlorpheniramine maleate, clidinium bromide, and quetiapine fumarate.

[0078] The active agent may be Figure 1A-1E Any one or more compounds described in, including those screened or not screened by the in vitro Demodex test.

[0079] The skin disorder may be one or more of acne vulgaris, seborrheic dermatitis, perioral dermatitis, acneiform eruption, transient acantholytic dermatosis, miliary necrotizing acne, psoriasis, steroid-induced dermatitis, primary irritant dermatitis, rosacea or hidradenitis suppurativa.

[0080] The skin disorder may affect the facial skin, the eyelids, or both.

[0081] The active agent may be applied topically to the individual.

[0082] For topical application the active agent may be formulated as a carrier lotion, cream, soap, wash, shampoo or gel.

[0083] The active agent may be present in a concentration of 0.001% to 5% by weight in a topically applied lotion, cream, soap, wash, shampoo or gel.

[0084] The concentration of the active agent applied topically may be the lowest concentration effective to substantially inactivate (eg, kill or paralyze at least a substantial portion of) the Demodex mites.

[0085] The active agent is present in a dosage of about 0.01 mg / kg to 150 mg / kg body weight in a topically applied lotion, cream, soap, wash, shampoo or gel.

[0086] Active agents for topical application can be encapsulated within the microliposomes prior to formulation into a carrier lotion, cream, soap, wash, shampoo or gel.

[0087] The active agent may be topically applied to an area of ​​skin affected by the skin disorder, and optionally at least a portion of an area of ​​skin not affected by the skin disorder. The active agent may be topically applied to an area of ​​the body skin where Demodex brevis and / or Demodex folliculorum are present. The active agent may be topically applied to substantially all areas of the skin of the individual.

[0088] The ophthalmic disorder may be a Demodex-induced inflammatory eye disease, such as one or more of meibomian gland dysfunction, conjunctivitis, keratoconjunctivitis, congestion, dry eyes, and blepharitis.

[0089] The ophthalmic disorder may be one or more of meibomian gland dysfunction, blepharitis, dry eye. The ophthalmic disorder may affect the eye or the eyelid, or both the eye and the eyelid.

[0090] Demodex mites may be located in the hair follicles, skin, eyes, eyelids, eyelashes, and / or meibomian glands of an individual, requiring application of active agents accordingly.

[0091] The active agent may be applied topically and provided in a formulation effective to deliver the active ingredient of the active agent to the epidermal or subcutaneous area of ​​the subject.

[0092] The autoimmune disease may be one or more of lupus erythematosus, rheumatoid arthritis, juvenile idiopathic arthritis, systemic lupus erythematosus, systemic sclerosis, juvenile dermatomyositis, adult dermatomyositis, Sjögren's syndrome, or porphyria cutanea tarda.

[0093] The autoimmune disease may be one or more of systemic lupus erythematosus, rheumatoid arthritis, juvenile idiopathic arthritis, systemic lupus erythematosus, systemic sclerosis, juvenile and adult dermatomyositis, Sjögren's syndrome, porphyria cutanea tarda, relapsing rheumatic disease, eosinophilic fasciitis, polymorphic light eruption, granuloma annulare, lichen planus, lupus panniculitis, discoid lupus, porphyria cutanea tarda, psoriatic arthritis, chronic ulcerative stomatitis, refractory chronic urticaria, sarcoidosis, frontal fibrosing alopecia, necrobiosis lipoidica, actinic reticulocytosis, actinic prurigo, epidermolysis bullosa, Kikuchi-Fujimoto disease, graft-versus-host disease, chronic erythema nodosum, sleep-related and systemic sclerosis, pemphigus vulgaris, pemphigus foliaceus and pemphigoid gestationis.

[0094] Autoimmune diseases can affect an individual's skin, lymphatic system, muscles, joints, or internal organs.

[0095] Also provided herein are pharmaceutical formulations for practicing any of the disclosed methods. Also provided are active agents for treating any of the conditions described herein, including clinical symptoms caused by the presence of one or both of Demodex brevis and Demodex folliculorum, skin disorders, autoimmune diseases, and / or ophthalmic disorders.

[0096] Provided herein are pharmaceutical formulations for inactivating Demodex mites that cause one or more mite-induced clinical symptoms, the formulations comprising a therapeutically effective amount of an active agent selected from the group consisting of promethazine, benztropine, dicyclomine, fluvoxamine, imipramine, dextromethorphan, mecamylamine, bupropine, fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, cyclamate, quetiapine, prodrugs thereof, and pharmaceutically acceptable salts thereof.

[0097] The pharmaceutical formulation may further comprise DMSO or any additional skin penetration enhancer.

[0098] The pharmaceutical preparation may further comprise an ophthalmologically acceptable excipient selected from the group consisting of buffered saline, mineral oil, vegetable oil, petroleum jelly, Miglyol™ 182, ethanol solutions, liposomes, and liposome-like products.

[0099] The pharmaceutical preparation may further comprise one or more of a preservative, an antioxidant, an antibiotic and an immunosuppressant.

[0100] The pharmaceutical preparation can be formulated as a pad or a gel stick.

[0101] Without wishing to be bound by any particular theory, the present invention may discuss ideas or understandings of the underlying principles associated with the devices and methods disclosed herein. It should be recognized that regardless of whether any mechanistic explanation or hypothesis is ultimately correct, embodiments of the present invention remain viable and useful.

[0102] Statement on Compounds and Nomenclature

[0103] In one embodiment, the compositions or compounds used in the methods of the invention are isolated or purified. In one embodiment, the isolated or purified compounds are at least partially isolated or purified as understood in the art. In one embodiment, the compositions or compounds of the invention have a chemical purity of 95%, optionally 99% for some applications, optionally 99.9% for some applications, optionally 99.99% for some applications, and optionally 99.999% pure for some applications.

[0104] Many compounds used in the methods of the present invention contain one or more ionizable groups. Ionizable groups include groups from which protons can be removed (e.g., -COOH) or groups from which protons can be added (e.g., amines) and groups that can be quaternized (e.g., amines). All possible ionic forms of these molecules and their salts should be included in the disclosure herein alone. With regard to salts of the compounds herein, one of ordinary skill in the art can select from a variety of available counterions suitable for the preparation of salts of the present invention for a given application. In a specific application, the selection of a given anion or cation to prepare a salt can result in an increase or decrease in the solubility of the salt. BRIEF DESCRIPTION OF THE DRAWINGS

[0105] Figure 1A-1E This is a summary table of in vitro tests of various agents on survival time and paralysis of Demodex mites. DETAILED DESCRIPTION

[0106] In general, the terms and phrases used herein have their art-recognized meanings, which can be found by reference to standard texts, journal references and context known to those skilled in the art. The following definitions are provided to clarify their specific use in the context of the present invention.

[0107] "Active agent" refers to a composition capable of inactivating Demodex mites, particularly Demodex mites that cause one or more clinical symptoms ("mite-induced clinical symptoms"), including those associated with skin disorders, autoimmune diseases, or ophthalmic disorders.

[0108] "Inactivation" is used broadly herein to refer to the functional ability to reduce the effects of Demodex brevis and / or Demodex folliculorum. For example, inactivation may be achieved by applying an active agent that results in paralysis and / or death of the mite. Alternatively, inactivation may refer to rendering the mite unable to reproduce, so that the mite dies with aging and no longer reproduces. As long as the treatment results in an adverse effect on Demodex brevis and / or Demodex folliculorum that corresponds to an improved clinical outcome (e.g., improved symptoms), then the treatment is considered herein to have inactivated Demodex brevis and / or Demodex folliculorum.

[0109] "Anticholinergics" refers to agents that block the action of acetylcholine (ACh) in the mite nervous system. These agents compete with Ach on receptors in the nervous system, making it impossible for Ach to bind to the receptors and cause cholinergic effects. Many of these agents interact with muscarinic cholinergic receptors in the brain, secretory glands, heart, smooth muscle and eyes. "Muscarinic agonists" refers to agents that activate the activity of muscarinic acetylcholinergic receptors. The use of these classes of agents can be used to inactivate mites, including killing or paralyzing most of the mites, thereby reducing the total number of related mites, and correspondingly improving and / or stopping the clinical symptoms caused by the mites.

[0110] Demodex folliculorum and Demodex brevis may play a role in conditions including rosacea. Increased populations of Demodex mites have been observed in patients with rosacea. For most people, Demodex mites living in the skin are harmless, either by downregulating the host's immunity or simply evading the host's immune defenses. There is intense debate in the dermatology community as to whether they are causative agents in skin diseases such as rosacea and blepharitis (eyelid inflammation), a common problem in patients with rosacea.

[0111] Humans are the only host to this ubiquitous mite. In fact, both mites are considered the most common external parasites of humans. Women tend to have higher rates of Demodex infection. Infection rates also appear to correlate with age, with 84% of people over the age of 60 carrying the mite, increasing to 100% for people 70 and older. Whether people with compromised immune systems are more likely to have higher infection rates is unclear, although some studies have suggested that people with AIDs and leukemia may be more likely to have higher-than-average infection rates.

[0112] The mites are most commonly found on the scalp, face, and upper chest areas, with Demodex folliculorum preferring hair follicles and Demodex brevis preferring sebaceous ducts and meibomian glands at the margins of the eyelids (sebaceous ducts transfer the waxy sebum that lubricates the skin and hair from the sebaceous glands; the meibomian glands are a specialized type of this gland). Demodex folliculorum is gregarious and tends to congregate in the follicle area of ​​the hair or eyelashes, with their rear ends protruding from the follicle orifices. Demodex brevis, on the other hand, tends to be more solitary and will occupy sebaceous glands singly. Both species are small, less than 0.4mm, with an elongated, transparent body and four pairs of stout legs. Demodex brevis is usually slightly shorter than Demodex folliculorum, about 0.1mm. Both have ridged scales on their cephalothorax, with pointed, sharp teeth.

[0113] The mite is a short-lived organism with a life cycle of 14-18 days from egg to larva to adult. The adult mites emerge from the hair follicles and ducts, reproduce on the surface of the skin, and then the female lays eggs in the sebaceous glands. The larvae will mature through two nymphal stages in the glands until entering the hair follicles and ducts to begin a new cycle as adults. Both mites are presumed to feed on sebum as their primary food source, but may also chew on the epithelial cells of the hair follicles and glands. They are considered obligate ectoparasites and cannot survive outside of a human host.

[0114] Some studies have found that above-average mite densities, more than five mites per square centimeter, appear to play a role in both diseases in patients. Researchers believe that mites plugging the hair follicles and sebaceous ducts may cause epithelial hyperplasia, trigger a phagocytic granulomatous reaction, or cause an inflammatory response due to their waste products. The fact that treatment with certain antibiotics can reduce the severity of rosacea strongly suggests a microbial component to the mite-related disease. In fact, in 2007, researchers isolated a bacterium from Demodex folliculorum, Bacillus vegetative, which caused an inflammatory response in 73% of patients with rosacea, but only in 29% of subjects in a control group. These results suggest that patients with rosacea are sensitive to bacteria and may be immunosensitive to mites, bacteria, or both.

[0115] Specifically, two antigenic proteins found on the bacterial cell surface appear to cause an inflammatory response by stimulating the proliferation of peripheral blood mononuclear cells; an 83-kDa protein shows similarity to heat shock proteins, while the other 62-kDa protein has amino acid sequence homology to a protease found to be involved in signal transduction as well as carbohydrate metabolism. The sensitivity of B. vegetabilis to many antibiotics that have been shown to be effective in treating rosacea, particularly tetracycline, doxycycline, and minocycline, may also be further evidence of a pathogenic role for the bacterium.

[0116] In one exemplary embodiment, an anticholinergic and a direct muscarinic agonist are topically administered to a patient with an active skin condition whose underlying cause is Demodex. Because the target organisms (Demodex brevis and Demodex folliculorum) are ectoparasites of the family Acaridae, an effective treatment must be able to eradicate the entire life cycle of this tiny insect, including the egg, larval, and adult stages. For this reason, this embodiment uses several administrations to treat such patients. Such intervals allow time for the Demodex eggs to hatch into immature mites, which are killed before they develop into egg-laying adults. After the anticholinergic and direct muscarinic agonists exert their acaricidal activity against the skin Demodex brevis and Demodex folliculorum organisms, the inflammatory response to them begins to subside, but the remnants of the dead mites will still cause some flushing and lesion formation until the body's clearance process clears them, which takes 6 to 8 weeks. Conventional anti-rosacea drugs, such as oral tetracycline and topical metronidazole, may optionally be used during the initial phase of anticholinergic and direct muscarinic agonist administration to suppress early attacks and produce an early clinical response. After six to eight weeks, such drugs are no longer needed to treat rosacea symptoms. After a long period of absence of rosacea symptoms, if typical symptoms begin to reappear, treatment may be repeated. Optionally, such retreatment more than once or twice per year is not necessary. Anticholinergics and direct muscarinic agonists are formulated into cosmetically acceptable topical lotions, creams, shampoos or gels and applied specifically to skin affected by rosacea and any areas where Demodex brevis and Demodex folliculorum may be present. Due to the well-known barrier effect of the skin to the penetration of topical drugs, it is expected that this approach to treatment with anticholinergics and direct muscarinic agonists will require application once or twice daily for up to six weeks to achieve adequate follicular penetration and effective acaricidal activity. Topical formulations that achieve this effect may contain about 5% or less of anticholinergics and direct muscarinic agonists. Using smaller percentages of anticholinergics and direct muscarinic agonists while still achieving a miticide effect and successfully treating the skin condition is desirable to limit any possible side effects of the chemical. In addition, it is also useful to optionally employ systemic treatment to prevent re-entry of mites from other parts of the body, such as the facial skin.

[0117] Example 1: Representative Active Agents

[0118] A variety of compounds can be used as active agents in the present invention. As discussed, the active agent can be selected from: anticholinergics, selective serotonin reuptake inhibitor (SSRI) antidepressants, tricyclic antidepressants, antimuscarinics, antinicotinic agents, and direct muscarinic agonists.

[0119] The active agent can be an anticholinergic agent selected from the group consisting of atropine (Atropen), belladonna alkaloids, benztropine mesylate (Cogentin), cyclamate, cyclopentolate (Cyclogyl), darifenacin (Enablex), dicyclomine, fesoterodine (Toviaz), flavoxate (Urispas), glycopyrrolate, homatropine hydrobromide, hyoscyamine (Levsinex), ipratropium (Atrovent), orphenadrine, oxybutynin (Ditropan XL), propantheline (propantheline bromide), scopolamine, methscopolamine, solifenacin (VESIcare), tiotropium (Spiriva), tolterodine (Detrol), trihexyphenidyl, and tolotrapine.

[0120] The active agent may be a selective serotonin reuptake inhibitor (SSRI) antidepressant selected from the group consisting of: citalopram, fluoxetine, fluvoxamine, and paroxetine.

[0121] The active agent may be a tricyclic antidepressant selected from the group consisting of amitriptyline, clomipramine, desipramine, imipramine, nortriptyline, trimipramine, amoxapine, and maprotiline.

[0122] The active agent can be an antimuscarinic agent selected from the group consisting of clozapine, quetiapine, biperiden, chlorpheniramine, dimethylhydrochloride, diphenhydramine, doxepin, doxylamine, glycopyrrolate, oxitropium, promethazine, solifenacin, tropicamide, and umeclidinium.

[0123] The active agent may be an antinicotinic agent selected from the group consisting of: bupropine, dextromethorphan, docurium, hexamethonium, mecamylamine, tubocurarine, cytisine, succinylcholine chloride (succinylcholine chloride), and varenicline.

[0124] The active agent may be a muscarinic agonist selected from the group consisting of acetylcholine, methacholine, bethanechol, muscarine, oxotremorine, nicotine, pilocarpine, and carbachol.

[0125] Figure 1A-1E This is an overview of various agents tested in in vitro assays for Demodex paralysis and / or killing. The efficacy of various agents was evaluated in Demodex characterization and / or survival time experiments, including LT50 (time to kill 50% of Demodex mites) and / or related times (in hours: minutes or total minutes). LT50s of 12 hours or less were achieved for some actives. Interestingly, some of these actives are not generally considered cholinergic agents, although they all have cholinergic properties.

[0126] Anticholinergics are agents that reduce or block the effects of acetylcholine on parasympathetic nervous system receptors on smooth muscle cells, glands, and the central nervous system. Cholinergic receptors are generally classified as either nicotinic or muscarinic. Selective serotonin reuptake inhibitors (SSRIs) can modulate other neurotransmitter systems by inhibiting neuronal nicotinic acetylcholine receptors. Tricyclic antidepressants (TCAs) act primarily by blocking the reuptake of norepinephrine or serotonin, while also affecting cholinergic and histaminergic systems. They have a narrow therapeutic index and may be associated with severe central nervous system (CNS) toxicity. Interestingly, as Figure 1A-1E As shown, in any given class of the Demodex test, at least one agent worked and at least one did not.

[0127] In one embodiment, the active agent is characterized by having a LT50 of less than 12 hours, including as demonstrated in an in vitro Demodex assay, including as demonstrated in an in vitro Demodex assay. Figure 1A-1E In one embodiment, the active agent is characterized by having paralytic activity, including as demonstrated in an in vitro Demodex test, including as demonstrated in Figure 1A-1E As summarized in .

[0128] Of course, the invention provided herein is more broadly applicable to any antiparasitic compound that inactivates parasites, more specifically including Demodex, including Demodex brevis and Demodex folliculorum. With respect to paralytic compounds, paralysis of the mites prevents the mites from feeding and results in eventual death. Paralysis can also result in reduced reproduction of the mites, thereby reducing the mite load and mite population in an individual. The paralysis mechanism also results in the mites being displaced from the skin and cleared by the immune system of the individual.

[0129] Studies of oral paroxetine have revealed its effects on rosacea. The use of paroxetine was found to be effective and well tolerated, with 42.9% of patients experiencing improvement in clinical symptoms. Paroxetine is a selective serotonin reuptake inhibitor. See https: / / www.healio.com / news / dermatology / 20230221 / paroxetine-effective-in-refractory-erythema-of-rosacea-treatment and Wang et al. “Paroxetine is an effective treatment for refractory erythema of rosacea: Primary results from the Prospective Rosacea Refractory Erythema Randomized Clinical Trial.” JAm Acad Dermatol (2023Feb 15)S0190-9622(23)00197-4.doi:10.1016 / j.jaad.2023.01.044. This study further supports the trial results of 1% paroxetine in Figure 1, where demodex paralysis was noted within the first hour and persisted for at least 12 hours. Therefore, paralytics may be useful for treating mite-induced symptoms and illnesses, even for those that do not have a corresponding LT50 of 12 hours or less.

[0130] Example 2: Administration and Formulation

[0131] Salts and Prodrugs: The present invention contemplates pharmaceutically active compounds that are chemically synthesized or formed by in vivo biotransformation into the compounds described herein (eg, active agents).

[0132] Active agents of the present invention and compounds useful in the methods of the present invention include those compounds and formulae described herein, as well as pharmaceutically acceptable salts and esters of those compounds. In an embodiment, salts include any salts of acids and bases derived from the formulae herein that are acceptable for human or veterinary use. In an embodiment, the term "ester" refers to a hydrolyzable ester of a compound of the name and formulae herein. In an embodiment, salts and esters of compounds of the formulae herein may include salts and esters having the same or better general properties of treatment, diagnosis or medicine (for human or veterinary use) as the compounds of the formulae herein. In one embodiment, the composition of the present invention is a compound or its salt or ester suitable for pharmaceutical preparations.

[0133] The activating agents of the present invention and the activating agents used in the methods of the present invention may have a prodrug form. Prodrugs of the compounds of the present invention may be used in embodiments including compositions and methods. Any compound that is converted in vivo to provide a biological, pharmaceutical, diagnostic or therapeutically active form of the activating agent of the present invention is a prodrug. Multiple examples and forms of prodrugs are well known in the art. Examples of prodrugs can be found in, inter alia: Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985); Methods in Enzymology, Vol. 42, at pp. 309-396, edited by K. Widder et al. (Academic Press, 1985); A Textbook of Drug Design and Development, edited by Krosgaard-Larsen and H. Bundgaard, Chapter 5, "Design and Application of Prodrugs," by H. Bundgaard, at pp. 113-191 (1991); H. Bundgaard, Advanced Drug Delivery Reviews, Vol. 8, p. 1-38 (1992); H. Bundgaard, et al., Journal of Pharmaceutical Sciences, Vol. 77, p. 285 (1988); and Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392). Prodrugs, such as pharmaceutically acceptable prodrugs, can represent prodrugs of the compounds of the invention that are suitable for use in contact with tissues of humans and lower animals without excessive toxicity, irritation, allergic response, etc., within the scope of sound medical judgment, have a reasonable benefit / risk ratio, and are effective for their intended use. The prodrugs of the invention can be rapidly converted in vivo to the parent compound of the compounds described herein, for example, by hydrolysis in the blood or by other cellular, tissue, organ or systemic processes.For further discussion, see: T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, V. 14 of the ACSSymposium Series; and Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press (1987), edited by Edward B. Roche. For example, loxapine can be considered a prodrug of amoxapine because amoxapine is an N-demethylated metabolite of loxapine.

[0134] The active agents of the present invention may be formulated with pharmaceutically acceptable anions and / or cations. Pharmaceutically acceptable cations include alkali metal cations (e.g., Li + 、Na + , K + ), alkaline earth metal cations (such as Ca 2+ Mg 2+ ), non-toxic heavy metal cations and ammonium (NH 4+ ) and substituted ammonium (N(R')4 + etc., wherein R' is hydrogen, alkyl or substituted alkyl, i.e., including methyl, ethyl or hydroxyethyl, specifically, trimethylammonium cation, triethylammonium cation and triethanolammonium cation). Pharmaceutically acceptable anions include halides (e.g., F - , Cl - Br - 、At - ), sulfates, acetates (e.g., acetate, trifluoroacetate), ascorbates, aspartates, benzoates, citrates, and lactates, among others.

[0135] Pharmaceutically acceptable salts include pharmaceutically acceptable anions and / or cations. As used herein, the term "pharmaceutically acceptable salt" may refer to an acid addition salt or a base addition salt of a compound of the present disclosure. A pharmaceutically acceptable salt is any salt that retains at least a portion of the activity of the parent compound and does not produce a significant harmful or adverse effect on the subject when it is administered and under the circumstances in which it is administered. Pharmaceutically acceptable salts include metal complexes and salts of both inorganic and organic acids. Pharmaceutically acceptable salts include metal salts, such as aluminum salts, calcium salts, iron salts, magnesium salts, manganese salts, and complex salts. Pharmaceutically acceptable salts include, but are not limited to, acid salts such as acetate, aspartate, alkylsulfonate, arylsulfonate, axetil, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, butyrate, calcium edetate, camsylic, carbonate, chlorobenzoate, cilexetil, citrate, edetic, edisylic, estolic, esyl, esylic, formate, fumarate, gluceptic, gluconate, glutamate, glycolic, glycolylarsanilic, hexamethylenesulfonate. The pharmaceutically acceptable salts include, but are not limited to, hydroxynaphthoates, isethionates, lactates, lactobionates, maleates, malates, malonates, mandelates, methanesulfonates, methylnitrates, methylsulfates, mucates, muconics, napsylics, nitrates, oxalates, p-nitromethanesulfonates, pamoic, pantothenates, phosphates, monohydrogenphosphates, dihydrogenphosphates, phthalates, polygalacturonates, propionates, salicylates, stearates, succinates, sulfamics, sulfanics, sulfonates, sulfates, tannates, tartrates, teoclics, toluenesulfonates, and the like. Pharmaceutically acceptable salts may be derived from amino acids, including but not limited to cysteine. Other pharmaceutically acceptable salts can be found, for example, in Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Wiley-VCH, Verlag Helvetica Chimica Acta, Zürich, 2002. (ISBN 3-906390-26-8).

[0136] Efficacy: Generally, the compound of the present invention or a pharmaceutically acceptable salt thereof is administered to a subject in a diagnostically or therapeutically effective amount. Appropriate dosages can generally be determined by one skilled in the art.

[0137] Compositions for oral administration can be prepared, for example, in such a manner that a single dose in one or more oral formulations contains at least about 20 mg of the compound of the invention per square meter of subject's body surface area, or at least about 50 mg, 100 mg, 150 mg, 200 mg, 300 mg, 400 mg, or 500 mg of the compound of the invention per square meter of subject's body surface area (the average body surface area of ​​a person is, for example, 1.8 square meters). Specifically, a single dose of a composition for oral administration can contain about 20 mg to about 600 mg of the compound of the invention per square meter of subject's body surface area, and in some aspects about 20 mg to about 400 mg of the compound of the invention per square meter of subject's body surface area, on the other hand about 20 mg to about 300 mg of the compound of the invention per square meter of subject's body surface area, and on the other hand about 20 mg to about 200 mg of the compound of the invention per square meter of subject's body surface area. The composition for parenteral administration can be prepared in such a way that a single dose contains at least about 20 mg of the compound of the invention per square meter of subject's body surface area, or at least about 40 mg, 50 mg, 100 mg, 150 mg, 200 mg, 300 mg, 400 mg or 500 mg of the compound of the invention per square meter of subject's body surface area. Specifically, a single dose in one or more parenteral preparations comprises about 20 mg to about 500 mg of the compound of the invention per square meter of subject's body surface area, and comprises about 20 mg to about 400 mg of the compound of the invention per square meter of subject's body surface area in some aspects, and comprises about 20 mg to about 450 mg of the compound of the invention per square meter of subject's body surface area on the other hand, and comprises about 20 mg to about 350 mg of the compound of the invention per square meter of subject's body surface area on the other hand. It should be recognized that these oral and parenteral dosage ranges generally represent preferred dosage ranges, and are not intended to limit the present invention. The dosage regimen actually used can vary greatly, and therefore can deviate from the usual preferred dosage regimen. It is expected that those skilled in the art will adjust these ranges according to individual subjects.

[0138] The toxicity and efficacy of such compounds and bioconjugates can be determined by standard pharmaceutical procedures in cell cultures or experimental animals using the LD 50 (a dose lethal to 50% of the population) and ED 50 The dose ratio between toxicity and efficacy is the therapeutic index, which can be expressed as LD 50 / ED 50Compounds and bioconjugates that exhibit high therapeutic indices are preferred. Although compounds and bioconjugates that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets these compounds and bioconjugates to sites affected by the disease or condition to minimize potential damage to unaffected cells and reduce side effects.

[0139] The data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for use in humans and other mammals. Preferably, the dosage of such compounds and bioconjugates is within the range of ED 50 The therapeutically effective amount can be estimated initially from cell culture assays for any compound and bioconjugate of the invention. A dose can be formulated in animal models to achieve an ED assay including that determined in cell culture. 50 (the concentration of the test compound that achieves half-maximal inhibition of symptoms). Such information can be used to more accurately determine effective doses in humans and other mammals. Compound and bioconjugate levels in plasma can be measured, for example, by high performance liquid chromatography.

[0140] The amount of the compound or bioconjugate that can be combined with a pharmaceutically acceptable carrier to produce a single dosage form will vary depending on the patient being treated and the specific mode of administration. Those skilled in the art will appreciate that the unit content of the compound / bioconjugate contained in a single dose of each dosage form need not constitute a therapeutically effective amount in itself, as the desired therapeutically effective amount can be achieved by administering multiple single doses. The choice of dosage depends on the dosage form used, the condition being treated, and the specific purpose to be achieved according to the judgment of those skilled in the art.

[0141] The dosage and dosage regimen for treating a disease or condition can be selected based on a variety of factors, including the type, age, weight, sex, diet and / or medical condition of the patient, the route of administration, pharmacological considerations (e.g., the activity, efficacy, pharmacokinetic and / or toxicological profile of the specific compound / bioconjugate used, whether a compound / bioconjugate delivery system is used, and / or whether the compound / bioconjugate is administered as a prodrug or part of a drug combination). Therefore, the actual dosage regimen used can vary widely depending on the subject or disease, and different routes of administration can be used in different clinical settings.

[0142] The identified compounds / bioconjugates monitor, treat, inhibit, control and / or prevent or at least partially inhibit or partially prevent target diseases and conditions, and can be administered to a subject in a therapeutically effective amount and optionally a diagnostically effective amount. The compositions / formulations of the invention comprise a therapeutically effective amount (optionally, a diagnostically effective amount) of at least one compound or bioconjugate of the invention. Preferably, the subject receiving a treatment comprising a compound / bioconjugate of the invention is an animal (e.g., a mammal, a reptile and / or a bird), more preferably, the subject is a human, horse, cow, dog, cat, sheep, pig and / or chicken, and most preferably, the subject is a human.

[0143] Administration: Preferred compositions depend on the route of administration. Any route of administration can be used as long as the target of the active agent or pharmaceutically acceptable salt can be reached by the route. Suitable routes of administration include, for example, oral administration, intravenous administration, parenteral administration, inhalation administration, rectal administration, nasal administration, topical (e.g., transdermal and intraocular) administration, intravesical administration, intrathecal administration, gastrointestinal tract administration, pulmonary administration, intralymphatic administration, intracavity administration, vaginal administration, transurethral administration, intradermal administration, intraauricular administration, intramammary administration, buccal administration, in situ administration, intratracheal administration, intralesional administration, percutaneous administration, endoscopic administration, transmucosal administration, sublingual administration, and intestinal administration.

[0144] In one embodiment, the present invention provides a method for treating a medical condition, comprising administering a therapeutically effective amount of a composition of the present invention, such as an active agent, including anticholinergic agents and direct muscarinic agonist compositions, to a subject (e.g., a patient) in need thereof. In one embodiment, the present invention provides a method for diagnosing or assisting in the diagnosis of a medical condition, comprising administering a diagnostically effective amount of a composition of the present invention to a subject in need thereof. In one embodiment, the medical condition is a skin condition or a dermatological disease.

[0145] The diagnostic and therapeutic formulations of the invention may be administered alone or with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.

[0146] Any suitable administration form may be used in conjunction with the diagnostic and therapeutic formulations of the present invention. The diagnostic and therapeutic formulations of the present invention may be administered intravenously, in oral dosage forms, intraperitoneally, subcutaneously, or intramuscularly, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts.

[0147] Compositions, preparations and formulations of the present invention can be formulated into diagnostic or therapeutic compositions for gastrointestinal administration, parenteral administration, topical administration, aerosol administration, inhalation administration or cutaneous administration. Topical or cutaneous delivery of compositions, preparations and formulations can also include aerosol formulations, creams, gels, solutions, etc. Compositions, preparations and formulations of the present invention are administered in a dosage that effectively achieves the desired diagnostic and / or therapeutic effect. This dosage can vary widely depending on the specific composition used in the composition, the organ or tissue to be examined, the equipment used in the clinical process, the therapeutic effect achieved, etc. These compositions, preparations and formulations contain an effective amount of the composition and conventional pharmaceutical carriers and excipients suitable for the intended type of administration. Optionally, these compositions, preparations and formulations can also include stabilizers and skin penetration enhancers.

[0148] (i) Parenteral Administration: The compounds and bioconjugates of the present invention can be formulated for parenteral administration by injection (e.g., by rapid bolus injection or continuous infusion). Injectable formulations may be presented in unit dose form in ampoules or multi-dose containers, and optionally preservatives may be added. Parenteral formulations may be packaged in ampoules, disposable syringes or multi-dose vials made of glass, plastic, etc. The formulation may take the form of a suspension, solution or emulsion in an oily or aqueous vehicle, and may contain preparatants such as suspending agents, stabilizers and / or dispersants.

[0149] For example, parenteral formulations can be sterile injectable solutions or suspensions in nontoxic parenteral acceptable diluents or solvents (e.g., solutions in 1,3-butanediol). Acceptable carriers and solvents that can be used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are generally used as solvents or suspension media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can be used for parenteral formulations.

[0150] Alternatively, the compounds and bioconjugates of the present invention may be formulated in powder form for preparation with a suitable carrier (e.g., sterile pyrogen-free water) prior to use. For example, a compound / bioconjugate suitable for parenteral administration may include a sterile isotonic saline solution containing 0.1% to 90% by weight of the compound / bioconjugate per volume. For example, the solution may contain about 5% to about 20% by weight per volume, more preferably about 5% to about 17% by weight, more preferably about 8% to about 14% by weight, and still more preferably about 10% by weight of the compound / bioconjugate. The solution or powder formulation may also include a solubilizer and a local anesthetic such as lidocaine to relieve pain at the injection site. Other parenteral delivery methods of the compound / bioconjugate are known to technicians and are included within the scope of the present invention.

[0151] (ii) Oral Administration: For oral administration, the compounds / bioconjugates of the present invention may be formulated in the form of tablets or capsules, which are prepared by conventional methods together with one or more pharmaceutically acceptable carriers (e.g., excipients such as binders, fillers, lubricants and disintegrants).

[0152] (iii) Controlled-release administration: Controlled-release (or sustained-release) formulations can be formulated to prolong the activity of the compound / bioconjugate and reduce the frequency of dosing. Controlled-release formulations can also be used to affect the onset of action or other characteristics, such as blood levels of the compound / bioconjugate, thereby affecting the occurrence of side effects.

[0153] Controlled release formulations can be designed to initially release an amount of the compound / bioconjugate that produces the desired therapeutic effect, and gradually continue to release other amounts of the compound / bioconjugate to maintain the therapeutic level over a longer period of time. In order to maintain a near-constant level of the compound / bioconjugate in the body, the compound / bioconjugate can be released from the dosage form at a rate that replaces the amount of compound / bioconjugate that is metabolized and / or excreted from the body. Controlled release of the compound / bioconjugate can be stimulated by a variety of inducing factors, such as pH changes, temperature changes, enzymes, water, and / or other physiological conditions or molecules.

[0154] Controlled release systems can include, for example, infusion pumps that can be used to administer compounds / bioconjugates in a manner similar to that used to deliver insulin or chemotherapeutic drugs to the whole body or to a specific organ or tumor. Typically, such systems are used to administer compounds / bioconjugates in combination with biodegradable, biocompatible polymeric implants that release the compounds / bioconjugates at a selected site over a controlled period of time. Examples of polymeric materials include polyanhydrides, polyorthoesters, polyglycolic acid, polylactic acid, polyethylene vinyl acetate, and copolymers and combinations thereof. In addition, controlled release systems can be placed near a therapeutic target (e.g., an organ, tissue, or cell population) so that only a portion of the systemic dose is required.

[0155] The compounds / bioconjugates of the present invention can be administered by other controlled release means or delivery devices well known to those of ordinary skill in the art. These controlled release means or delivery devices include, for example, hydroxypropyl methylcellulose, other polymer matrices, gels, permeable membranes, permeable systems, multilayer coatings, microparticles, liposomes, microspheres, etc., or any combination of the above substances, to provide the desired release profile in different proportions. Other methods of controlled release delivery of compounds / bioconjugates are known to those of ordinary skill in the art and are included within the scope of the present invention.

[0156] (iv) Administration by Inhalation: The compounds / bioconjugates of the invention can be administered directly to the lungs of a patient / subject by inhalation. For administration by inhalation, the compounds / bioconjugates can be conveniently delivered to the lungs by a number of different devices. For example, a metered dose inhaler ("MDI") can be used to deliver the compounds / bioconjugates directly to the lungs, which utilizes a container containing a suitable low boiling point propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. MDI devices are available from a number of suppliers, such as 3M Company, Aventis, Boehringer Ingleheim, Forest Laboratories, GlaxoSmithKline, Merck & Co., and Vectura.

[0157] Alternatively, a dry powder inhaler (DPI) device can be used to administer the compound / bioconjugate to the lungs. The DPI device typically uses a mechanism such as a gas explosion to produce a dry powder cloud in a container, which can then be inhaled by the patient. The DPI device is also well known in the art and can be purchased from many suppliers, including, for example, GlaxoSmithKline, NektarTherapeutics, Innovata, and Vectura. A popular variant is a multi-dose DPI ("MDDPI") system that allows delivery of more than one therapeutic dose. The MDDPI device can be obtained from companies such as AstraZeneca, GlaxoSmithKline, TEVA, Merck&Co., SkyePharma, and Vectura. For example, gelatin cartridges and capsules for inhalers or insufflators can be formulated into a powder mixture containing the compound / bioconjugate and a suitable powder matrix such as lactose or starch for these systems.

[0158] Another type of device that can be used to deliver compounds / bioconjugates to the lungs is a liquid spray device, such as those provided by Aradigm Corporation. Liquid spray systems use extremely small nozzle holes to atomize liquid compound / bioconjugate formulations so that they can then be inhaled directly into the lungs. For example, a nebulizer device can be used to deliver compounds / bioconjugates to the lungs. Nebulizers generate aerosols from liquid compound / bioconjugate formulations by using, for example, ultrasonic energy to form microparticles that are easily inhaled. Examples of nebulizers include devices provided by Aventis and Battelle.

[0159] In another example, the compound / bioconjugate can be delivered to the lungs using an electrohydrodynamic ("EHD") aerosol device. An EHD aerosol device uses electrical energy to atomize a liquid compound / bioconjugate solution or suspension. When the compound / bioconjugate is delivered to the lungs using an EHD aerosol device, the electrochemical properties of the compound / bioconjugate formulation are important parameters that require optimization. Such optimization is typically performed by one skilled in the art. Other methods of intrapulmonary delivery of compounds / bioconjugates are known to those skilled in the art and are included within the scope of the present invention.

[0160] Liquid compound / bioconjugate formulations suitable for use with nebulizers and liquid spray devices and EHD aerosol devices generally include a compound / bioconjugate with a pharmaceutically acceptable carrier. In an exemplary embodiment, the pharmaceutically acceptable carrier is a liquid, such as ethanol, water, polyethylene glycol, or a perfluorocarbon. Optionally, another material can be added to change the aerosol properties of the solution or suspension of the compound / bioconjugate. For example, such a material can be a liquid, such as ethanol, a glycol, polyethylene glycol, or a fatty acid. Other methods of formulating liquid compound / bioconjugate solutions or suspensions suitable for aerosol devices are known to those skilled in the art.

[0161] (v) Depot administration: The compounds / bioconjugates of the present invention can be formulated as depot formulations. Such long-acting formulations can be administered by implantation (e.g., subcutaneous or intramuscular) or intramuscular injection. Thus, the compounds / bioconjugates can be formulated with suitable polymeric or hydrophobic materials such as emulsions in acceptable oils or ion exchange resins or sparingly soluble derivatives (e.g., sparingly soluble salts). Other depot delivery methods of compounds / bioconjugates are known to the skilled artisan and are included within the scope of the present invention.

[0162] (vi) Topical administration: For topical application, the compound / bioconjugate may be combined with a pharmaceutically acceptable carrier to deliver an effective dose based on the desired activity, e.g., an effective dose of 1.0 μM to 1.0 mM. In one aspect of the invention, a topical formulation of the compound / bioconjugate may be applied to the skin. The pharmaceutically acceptable carrier may be in the form of, for example, but not limited to, an ointment, cream, gel, paste, foam, aerosol, suppository, pad, or gel stick.

[0163] Topical preparations may include a therapeutically effective amount of a compound / bioconjugate in an ophthalmologically acceptable excipient, such as buffered saline, mineral oil, vegetable oil (such as corn oil or peanut oil), petroleum vaseline, Miglyol 182, ethanol solution, or liposomes or liposome-like products. Any of these preparations of these compounds / bioconjugates may include preservatives, antioxidants, antibiotics, immunosuppressants, and other biological or pharmaceutically effective agents that do not produce significant adverse effects on the compounds / bioconjugates. Other topical delivery methods of compounds / bioconjugates are known to technicians and are included within the scope of the present invention. Topical preparations of the present invention also include those containing one or more compositions that help penetrate the skin, such as dimethyl sulfoxide (DMSO).

[0164] (vii) Rectal Administration: The compounds / bioconjugates of the invention may be formulated into rectal preparations, such as suppositories or retention enemas, containing conventional suppository bases such as cocoa butter or other glycerides and / or binders and / or carriers such as triglycerides, microcrystalline cellulose, tragacanth or gelatin. Rectal preparations may contain, for example, 0.5% to 10% by weight of the compound / bioconjugate. Other rectal delivery methods of the compounds / bioconjugates are known to the skilled artisan and are included within the scope of the invention.

[0165] (viii) Other Administration Systems: A variety of other delivery systems are known in the art and can be used to administer the compounds / bioconjugates of the invention. In addition, these and other delivery systems can be combined and / or modified to facilitate optimizing administration of the compounds / bioconjugates of the invention. Exemplary formulations including the compounds / bioconjugates of the invention are described elsewhere herein (the compounds / bioconjugates of the invention are represented as active ingredients, but those skilled in the art will recognize that this term should also encompass prodrugs and compound combinations).

[0166] 5.d: Formulations: In one embodiment, the present invention provides a medicament comprising a therapeutically effective amount of one or more compositions of the present invention, such as an anticholinergic agent and a direct muscarinic agonist compound. In one embodiment, the present invention provides a medicament comprising a diagnostically effective amount of one or more compositions of the present invention. In one embodiment, the present invention provides a method for preparing a medicament for treating a condition described herein, such as a skin condition or a dermatological disease. In one embodiment, the present invention provides a method for preparing a medicament for diagnosing or aiding in the diagnosis of a condition described herein, such as a skin condition or a dermatological disease. In one embodiment, the present invention provides the use of one or more compositions described herein in the preparation of a medicament for treating a skin condition or a dermatological disease. In one embodiment, the present invention provides the use of one or more compositions described herein for treating a disease. In one embodiment, the present invention provides the use of one or more compositions described herein for diagnosing a disease. The compositions of the present invention include formulations and preparations, such as pharmaceutically acceptable formulations or preparations, comprising one or more anticholinergic agents and direct muscarinic agonists of the present invention provided in an aqueous solution. Optionally, the compositions of the present invention further comprise one or more pharmaceutically acceptable surfactants, buffers, electrolytes, salts, carriers, binders, coatings, preservatives and / or excipients.

[0167] In one embodiment, the present invention provides a pharmaceutical preparation having an active ingredient, the active ingredient comprising a composition of the present invention, such as an anticholinergic agent and a direct muscarinic agonist compound. In one embodiment, the present invention provides a method for synthesizing a composition of the present invention or a pharmaceutical preparation thereof (e.g., an anticholinergic agent and a direct muscarinic agonist compound). In one embodiment, the pharmaceutical preparation comprises one or more excipients, carriers, diluents, and / or other components known in the art. Preferably, these components conform to the standards of the National Formulary ("NF"), the United States Pharmacopeia ("USP"; United States Pharmacopeial Convention, Rockville, Maryland), or the Handbook of Pharmaceutical Formulations (Sarfaraz K. Niazi, all volumes, ISBN: 9780849317521, ISBN 10: 0849317525; CRC Press, 2004). See, e.g., United States Pharmacopoeia and National Formulary (USP 30-NF 25), Rockville, MD: United States Pharmacopeial Convention (2007 and 2008), and any earlier editions; Handbook of Pharmaceutical Excipients, a joint publication of the American Pharmacists Association and Pharmaceutical Press (Pharmaceutical Press (2005) (ISBN-10: 0853696187, ISBN-13: 978-0853696186)); Merck Index, Merck & Co., Rahway, NJ; and Gilman et al., (eds) (1996); Goodman and Gilman's: The Pharmacological Bases of Therapeutics, 8th Ed., Pergamon Press. In an embodiment, the formulation base of the formulation of the present invention comprises a physiologically acceptable excipient, i.e., at least one binder and optionally other physiologically acceptable excipients. Physiologically acceptable excipients are those excipients known for use in the field of pharmaceutical technology and adjacent fields, in particular those listed in the relevant pharmacopoeias (eg DAB, Ph. Eur., BP, NF, USP) and other excipients whose properties do not impair the physiological use.

[0168] The present invention also relates in part to pharmaceutical compositions comprising a therapeutically effective amount of the compounds of the present invention or salts, and methods for preparing such compositions. Such compositions typically include one or more pharmaceutically acceptable carriers (e.g., excipients, carriers, auxiliary agents, adjuvants, diluents), and may include other active ingredients. The formulation of these compositions can be achieved by a variety of methods known in the art. A general discussion of these methods can be found in, for example, Hoover, John E., Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, PA: 1975). See also Lachman, L., eds., Pharmaceutical Dosage Forms (Marcel Decker, New York, NY, 1980).

[0169] The diagnostic and therapeutic preparations of the present invention and the medicaments of the present invention may also comprise one or more pharmaceutically acceptable carriers, excipients, buffers, emulsifiers, surfactants, electrolytes or diluents. Such compositions and medicaments are prepared according to acceptable pharmaceutical methods, for example, those described in Remingtons Pharmaceutical Sciences, 17th edition, ed. Alfonoso R. Gennaro, Mack Publishing Company, Easton, Pa. (1985).

[0170] The compositions of the present invention include preparations and formulations comprising one or more compounds of the present invention provided in an aqueous solution, such as pharmaceutically acceptable preparations or formulations. Optionally, the compositions of the present invention further comprise one or more pharmaceutically acceptable surfactants, buffers, electrolytes, salts, carriers, binders, coatings, preservatives and / or excipients.

[0171] The compounds and bioconjugates of the present invention can be formulated by known methods for administration to a subject using several routes, including but not limited to parenteral, oral, topical, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and ocular routes. A single compound / bioconjugate can be co-administered with one or more additional compounds / bioconjugates of the present invention and / or with other biologically active agents or biologically inert agents. Such biologically active agents or inert agents can be in fluid or mechanical communication with the compound / bioconjugate, or attached to the compound / bioconjugate by ionic, covalent, van der Waals, hydrophobic, hydrophilic, or other physical forces. Local administration in the subject is preferred, but systemic administration is also possible.

[0172] The compounds and bioconjugates of the present invention can be formulated in any conventional manner using one or more pharmaceutically acceptable carriers. Therefore, the compounds / bioconjugates and pharmaceutically acceptable salts and solvates thereof can be specially formulated for administration, such as by inhalation or insufflation (through the mouth or nose) or oral, buccal, parenteral or rectal administration. The compounds / bioconjugates can take the form of charged, neutral and / or other pharmaceutically acceptable salts. Examples of pharmaceutically acceptable carriers include, but are not limited to, those described in REMINGTON'S PHARMACEUTICALSCIENCES (AR Gennaro, Ed.), 20th edition, Williams & Wilkins PA, USA (2000).

[0173] The compounds and bioconjugates of the present invention can be formulated into solutions, suspensions, emulsions, tablets, pills, capsules, powders, controlled-release or sustained-release formulations, etc. Such formulations will contain a therapeutically effective amount of the compound / bioconjugate (preferably in purified form) and a suitable amount of carrier to provide a form for appropriate administration to the patient. The formulation should be compatible with the mode of administration.

[0174] Pharmaceutically acceptable carriers that can be used in combination with the compounds of the present invention are well known to those of ordinary skill in the art. The carrier can be selected based on many factors, including, for example, the specific compound or pharmaceutically acceptable salt used; the concentration, stability and expected bioavailability of the compound; the condition being treated; the age, size and general condition of the subject; the route of administration; etc. A general discussion of carriers can be found, for example, in JG Nairn, Remington's Pharmaceutical Science, pp. 1492-1517 (A. Gennaro, ed., Mack Publishing Co., Easton, Pa. (1985)).

[0175] Solid dosage forms for oral administration include, for example, capsules, tablets, gel capsules, pills, dragees, troches, powders, granules and lozenges. In these solid dosage forms, the compound or its pharmaceutically acceptable salt can be combined with one or more pharmaceutically acceptable carriers. The compound and its pharmaceutically acceptable salt can be mixed with a carrier, and the carrier includes but is not limited to lactose, sucrose, starch powder, corn starch, potato starch, magnesium carbonate, microcrystalline cellulose, cellulose esters of alkanoic acid, cellulose alkyl esters, talcum powder, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric acid and sulfuric acid, sodium carbonate, agar, mannitol, sorbitol, saccharin sodium, gelatin, gum arabic, alginic acid, sodium alginate, tragacanth, colloidal silicon dioxide, cross-linked sodium carboxymethyl cellulose, polyvinyl pyrrolidone and / or polyvinyl alcohol, and then made into tablets or capsules for convenient administration. Such capsules or tablets can contain controlled release formulations, such as can provide in the form of a dispersion of a compound or salt in hydroxypropyl methylcellulose. In the case of capsules, tablets and pills, the dosage form can also include a buffer, such as sodium citrate, or a carbonate or bicarbonate of magnesium or calcium. Tablets and pills can also include, for example, a coating (such as an enteric coating) to delay disintegration and absorption. The concentration of the compounds of the present invention in a solid oral dosage form can be, for example, about 5% by weight to about 50% by weight based on the gross weight of the composition, about 8% by weight to about 40% by weight in some aspects, and about 10% by weight to about 30% by weight on the other hand.

[0176] Liquid dosage forms of the compounds of the present invention for oral administration include, for example, pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs containing inert diluents (e.g., water) commonly used in the art. Such compositions may also include adjuvants, such as wetting agents, emulsifiers, suspending agents, flavoring agents (e.g., sweeteners) and / or aromatics. The concentration of the compounds of the present invention in the liquid dosage form may be about 0.01 mg to about 5 mg per milliliter of composition, about 0.01 mg to about 1 mg in some aspects, and about 0.01 mg to about 0.5 mg on the other hand. In the case where the compounds of the present invention are more soluble at low concentrations, liquid dosage forms of the compounds of the present invention may be prepared at low concentrations. Techniques for preparing oral dosage forms that can be used in the present invention are generally described in, for example, Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors (1979)). Also see Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981). See also Ansel, Introduction to Pharmaceutical Dosage Forms, 2nd ed. (1976).

[0177] In some aspects of the invention, tablets or powders for oral administration can be prepared by dissolving the compound in a pharmaceutically acceptable solvent capable of dissolving the compound to form a solution, and then drying the solution under vacuum (the solvent is evaporated). A carrier can also be added to the solution before drying. The resulting solution can be dried under vacuum to form a glass. The glass can then be mixed with a binder to form a powder. This powder can be mixed with a filler or other conventional tableting agents and then processed into tablets. Alternatively, the powder can be added to a liquid carrier to form a solution, emulsion, suspension, etc.

[0178] In some aspects, a solution for oral administration is prepared by dissolving the compound in a pharmaceutically acceptable solvent capable of dissolving the compound to form a solution. An appropriate volume of the carrier is added to the solution while stirring to form a pharmaceutically acceptable solution for oral administration.

[0179] In some embodiments, liposomes or micelles can be used as carriers or vehicles for the composition. For example, in some embodiments, the compound can be part of a lipophilic bilayer or micelle, and the targeting ligand, if present, can be on the outer surface of the liposome or micelle. As another example, the targeting ligand can be attached externally to the liposome or micelle after formulation for targeting the liposome or micelle (which contains the anticholinergic agent and the direct muscarinic agonist) to a desired tissue, organ, or other part of the body.

[0180] Injectable preparations (e.g., sterile injectable aqueous or oily suspensions) can be prepared according to known techniques using suitable dispersants, wetting agents and / or suspending agents. Acceptable carriers for parenteral administration include aqueous and non-aqueous pharmaceutically acceptable solvents. Suitable pharmaceutically acceptable aqueous solvents include, for example, water, saline, glucose solutions (e.g., DW5), electrolyte solutions, etc.

[0181] In one embodiment, the compounds of the present invention are formulated into nanoparticles or microparticles. The use of such nanoparticle or microparticle preparations is advantageous for some applications, and can enhance the delivery, positioning, targeting specificity, administration, etc. of the compound. Potentially useful nanoparticles and microparticles include, but are not limited to, micelles, liposomes, microemulsions, nanoemulsions, vesicles, tubular micelles, cylindrical micelles, bilayers, folded sheet structures, spherical aggregates, swollen micelles, inclusion complexes, encapsulated droplets, microcapsules, nanocapsules, etc. As will be appreciated by those skilled in the art, the compounds of the present invention may be located inside nanoparticles or microparticles, inside the membrane or wall of nanoparticles or microparticles, or outside nanoparticles or microparticles (but in combination with or otherwise connected thereto). Agents formulated into nanoparticles or microparticles may be administered by any of the aforementioned routes. In preparations for topical application, the compound is slowly released over time. In injectable preparations, liposomes, micelles, capsules, etc. circulate in the bloodstream and are delivered to the desired site (e.g., target tissue).

[0182] The preparation and loading of nanoparticles and microparticles are well known in the art. As an example, liposomes can be prepared by dipalmitoylphosphatidylcholine (DPPC) or phosphatidylcholine (PC) because such lipids have low thermal transitions. Liposomes are prepared using standard methods known to those skilled in the art (e.g., Braun-Falco et al., (Eds.), Griesbach Conference, Liposome Dermatics, Springer-Verlag, Berlin (1992), pp.6981; 91 117). Polycaprolactone, poly (glycolic) acid, poly (lactic) acid, polyanhydride or lipid can be formulated into microspheres. As an illustrative example, the compound of the present invention can be mixed with polyvinyl alcohol (PVA), the mixture is then dried and coated with ethylene vinyl acetate, and then cooled again with PVA. In liposomes, the compound of the present invention can be in one or two lipid bilayers, in an aqueous solution between bilayers, or in the center or core. Liposomes can be modified with other molecules and lipids to form cationic liposomes. Liposomes can also be modified with lipids to make their surfaces more hydrophilic, thereby increasing their circulation time in the bloodstream. Such modified liposomes are referred to as "stealth" liposomes or long-lived liposomes, as described in U.S. Patent No. 6,258,378 and in Stealth Liposomes, Lasic and Martin (Eds.) 1995 CRC Press, London. Encapsulation methods include detergent dialysis, freeze drying, film formation, injection, as known to those skilled in the art and disclosed in, for example, U.S. Patent No. 6,406,713. Optionally, the compositions and methods of the present invention include micellar delivery systems, for example, including one or more PEG-based amphiphilic polymers developed for drug delivery, including: PEG-poly(ε-caprolactone), PEG-poly(amino acid), PEG-polylactide or PEG-phospholipid constructs; cross-linked poly(acrylic acid) polymer systems, phospholipid-based systems and / or block copolymer systems, the block copolymer systems comprising one or more of the following polymer blocks: poly(lactic acid) polymer blocks; poly(propylene glycol) polymer blocks; poly(amino acid) polymer blocks; poly(ester) polymer blocks; poly(ε-caprolactone) polymer blocks; poly(ethylene glycol) blocks, poly(acrylic acid) blocks; polylactide blocks; polyester blocks; polyamide blocks; polyanhydride blocks; polyurethane blocks; polyimine blocks; polyurea blocks; polyacetal blocks; polypolysaccharide blocks; and polysiloxane blocks.

[0183] Suitable pharmaceutically acceptable non-aqueous solvents include, but are not limited to, the following (and mixtures thereof):

[0184] (i) alcohols (these include, for example, sigma alcohol glycerol formal, beta aldehyde glycerol formal, 1,3-butanediol, aliphatic alcohols having 2 to about 30 carbon atoms or aromatic alcohols (e.g., methanol, ethanol, propanol, isopropanol, butanol, tert-butanol, hexanol, octanol, amyl hydrate, benzyl alcohol, glycerol (glycerine), glycol, hexylene glycol, glycol, tetrahydrofuran alcohol, hexadecanol and stearyl alcohol), fatty acid esters of fatty alcohols (e.g., polyalkylene glycols such as polypropylene glycol and polyethylene glycol), sorbitan, sucrose and cholesterol);

[0185] (ii) amides, which include, for example, dimethylacetamide (DMA), benzyl benzoate DMA, dimethylformamide, N-hydroxyethyl O-lactamide, N,N-dimethylacetamide-amide, 2-pyrrolidone, 1-methyl-2-pyrrolidone and polyvinylpyrrolidone;

[0186] (iii) Esters, which include, for example, acetates (e.g., monoacetin, diacetin, and triacetin), aliphatic and aromatic esters (e.g., ethyl caprylate or ethyl caprylate, alkyl oleates, benzyl benzoate or benzyl acetate), dimethyl sulfoxide (DMSO), glycerides (e.g., citric acid monoglycerides, diglycerides, and triglycerides and tartaric acid monoglycerides, diglycerides, and triglycerides), ethyl benzoate, ethyl acetate, ethyl carbonate, ethyl lactate, ethyl oleate, fatty acid esters of sorbitan, glyceryl monostearate, glyceryl esters (e.g., monoglycerides, diglycerides, and triglycerides), fatty acid esters (e.g., isopropyl myristate), fatty acid-derived PEG esters (e.g., PEG-hydroxyoleate and PEG-hydroxystearate), N-methylpyrrolidone, pluronic 60, polyoxyethylene sorbitan oleate (e.g., poly(ethoxylated) 30-60 Sorbitan poly(oleate) 2-4 , poly(ethylene oxide) 15-20 Monooleate, poly(ethylene oxide) 15-20 Mono 12-hydroxystearate and poly(ethylene oxide) 15-20polyoxyethylene sorbitan esters (e.g., polyoxyethylene-sorbitan monooleate, polyoxyethylene-sorbitan monopalmitate, polyoxyethylene-sorbitan monolaurate, polyoxyethylene sorbitan monostearate and POLYSORBATE 20, POLYSORBATE 40, POLYSORBATE 60 and POLYSORBATE 80 (from ICI Americas, Wilmington, DE)), polyvinylpyrrolidone, alkyleneoxy-modified fatty acid esters (e.g., polyoxyl 40 hydrogenated castor oil and polyoxyethylated castor oil, such as CREMOPHOR EL solution or CREMOPHOR RH 40 solution), sugar fatty acid esters (i.e., monosaccharides (e.g., pentoses, such as ribose, ribulose, arabinose, xylose, lyxose and xylulose; hexoses, such as glucose, fructose, galactose, mannose and sorbose; trioses; tetrasaccharides; heptoses; and octose), disaccharides (e.g., sucrose, maltose, lactose and trehalose), oligosaccharides or mixtures thereof with one or more C4-C 22 Fatty acids (e.g., condensates of saturated fatty acids such as caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, and stearic acid; and unsaturated fatty acids such as palmitoleic acid, oleic acid, elaidic acid, erucic acid, and linoleic acid), and steroidal esters;

[0187] (iv) ethers, such as alkyl, aryl and cyclic ethers having 2 to about 30 carbon atoms. Examples include diethyl ether, tetrahydrofuran, dimethyl isosorbide, diethylene glycol ethyl ether) and tetraethylene glycol ether (tetrahydrofuran alcohol polyethylene glycol ether);

[0188] (v) ketones, which typically have from about 3 to about 30 carbon atoms. Examples include acetone, methyl ethyl ketone, and methyl isobutyl ketone;

[0189] (vi) hydrocarbons, which are generally aliphatic, alicyclic or aromatic hydrocarbons having from about 4 to about 30 carbon atoms. Examples include benzene, cyclohexane, methylene chloride, dioxolane, hexane, n-decane, n-dodecane, n-hexane, sulfolane, tetramethylene sulfone, tetramethylene sulfoxide, toluene, dimethyl sulfoxide (DMSO); and tetramethylene sulfoxide;

[0190] (vii) Oils, including, for example, mineral oils, vegetable oils, animal oils, essential oils or synthetic oils. These include: mineral oils, such as aliphatic and wax-based hydrocarbons, aromatic hydrocarbons, mixed aliphatic and aromatic hydrocarbons and refined paraffin oils; vegetable oils, such as linseed oil, tung oil, safflower oil, soybean oil, castor oil, cottonseed oil, groundnut oil, rapeseed oil, coconut oil, palm oil, olive oil, corn oil, corn germ oil, sesame oil, almond oil (persic oil) and peanut oil; glycerides, such as monoglycerides, diglycerides and triglycerides; animal oils, such as fish oils, marine oils, spermaceti oils, cod liver oil, halibut liver oil, squalene oil, squalane oil and shark liver oil; oleic acid oils; and polyoxyethylated castor oils;

[0191] (viii) alkyl, alkenyl or aryl halides, including, for example, alkyl or aryl halides having 1 to about 30 carbon atoms and one or more halogen substituents. Examples include: methyl chloride; monoethanolamine; petroleum benzin; triethanolamine; ω-3 polyunsaturated fatty acids (e.g., α-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid or docosahexaenoic acid); polyethylene glycol esters of 12-hydroxystearic acid and polyethylene glycol (SOLUTOL HS-15, from BASF, Ludwigshafen, Germany); polyoxyethylene glycerol; sodium laurate; sodium oleate; and sorbitan monooleate.

[0192] Other pharmaceutically acceptable solvents useful in the present invention are well known to those of ordinary skill in the art. A general discussion of such solvents can be found, for example, in The Chemotherapy Source Book (Williams & Wilkens Publishing), The Handbook of Pharmaceutical Excipients, (American Pharmaceutical Association, Washington, DC, and The Pharmaceutical Society of Great Britain, London, England, 1968), Modern Pharmaceutics 3d ed., (G. Bankeret. al., eds., Marcel Dekker, Inc., New York, New York). York (1995)), The Pharmacological Basis of Therapeutics, (Goodman & Gilman, McGraw Hill Publishing), Pharmaceutical Dosage Forms, (H. Lieberman et.al., eds., Marcel Dekker, Inc., New York, New York (1980)), Remington'sPharmaceutical Sciences, 19thed., (A. Gennaro, ed., Mack Publishing, Easton, PA, (1995)), The United States Pharmacopeia 24, The National Formulary 19, (National Publishing, Philadelphia, PA (2000)); Spiegel, AJ, et al., "Use of Nonaqueous Solvents in Parenteral Products," J. Pharma. Sciences, Vol. 52, No. 10, pp. 917-927 (1963).

[0193] Solvents useful in the present invention include, but are not limited to, solvents known to stabilize the compounds of the present invention or their pharmaceutically acceptable salts. These may include, for example, triglyceride-rich oils such as safflower oil, soybean oil, and mixtures thereof; and alkyleneoxy-modified fatty acid esters such as polyoxyl 40 hydrogenated castor oil and polyoxyethylene castor oil (e.g., CREMOPHOREL solution or CREMOPHOR RH 40 solution). Commercially available triglycerides include INTRALIPID emulsified soybean oil (Kabi-Pharmacia Inc., Stockholm, Sweden), NUTRALIPID emulsion (McGaw, Irvine, California), LIPOSYN Il 20% emulsion (a 20% fat emulsion solution containing 100 mg safflower oil, 100 mg soybean oil, 12 mg lecithin, and 25 mg glycerol per ml of solution; Abbott Laboratories, Chicago, IL), LIPOSYN III 2% emulsion (a 2% fat emulsion solution containing 100 mg safflower oil, 100 mg soybean oil, 12 mg lecithin, and 25 mg glycerol per ml of solution; Abbott Laboratories, Chicago, IL), natural or synthetic glycerol derivatives containing from about 25% to about 100% (by weight based on total fatty acid content) of docosahexaenoic acid radicals (DHASCO, available from Martek Laboratories). Biosciences Corp., Columbia, MD; DHAMAGURO, from Daito Enterprises, Los Angeles, CA; SOYACAL and TRAVEMULSION). In particular, ethanol is a useful solvent for dissolving the compound or its pharmaceutically acceptable salt to form a solution, emulsion, etc.

[0194] Additional components may be included in the compositions of the present invention for a variety of purposes generally known in the pharmaceutical industry. These components tend to impart properties such as enhancing retention of the compounds of the present invention or their salts at the site of administration, protecting the stability of the composition, controlling pH, and facilitating processing of the compounds or their salts into pharmaceutical preparations. Specific examples of such components include cryoprotectants; agents for preventing reprecipitation of the surface of the compound or salt; active agents, wetting agents or emulsifiers (e.g., lecithin, polysorbate-80, Tween 80, pluronic 60, and polyoxyethylene stearate); preservatives (e.g., ethyl paraben); microbial preservatives (e.g., benzyl alcohol, phenol, m-cresol, chlorobutanol, sorbic acid, thimerosal, and parabens); pH adjusters or buffers (e.g., acids, bases, sodium acetate, sorbitan monolaurate, etc.); osmotic pressure regulators (e.g., glycerol); thickeners (e.g., aluminum monostearate, stearic acid, cetyl alcohol, stearyl alcohol, guar gum); , methylcellulose, hydroxypropylcellulose, tristearate, cetyl esters, polyethylene glycol, etc.); colorants; dyes; glidants; non-volatile silicones (e.g., cyclomethicone); clays (e.g., bentonite); binders; extenders; flavorings; sweeteners; adsorbents; fillers (e.g., sugars such as lactose, sucrose, mannitol, sorbitol, cellulose, calcium phosphate, etc.); diluents (e.g., water, saline, electrolyte solutions, etc.); adhesives (e.g., Gelatin; tragacanth gum; methylcellulose; hydroxypropyl methylcellulose; sodium carboxymethylcellulose; polyvinyl pyrrolidone; carbohydrates; polymers; gum arabic; starches such as corn starch, wheat starch, rice starch and potato starch; etc.); disintegrants (e.g., starches such as corn starch, wheat starch, rice starch, potato starch and carboxymethyl starch; cross-linked polyvinyl pyrrolidone; agar; alginic acid or its salts such as sodium alginate; cross-linked sodium carboxymethylcellulose; cross-linked polyvinyl pyrrolidone; etc.); lubricants (e.g., silicon dioxide; talc; stearic acid and its salts such as magnesium stearate; polyethylene glycol; etc.); coating agents (e.g., concentrated sugar solutions including gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, titanium dioxide, etc.); and antioxidants (e.g., sodium metabisulfite, sodium bisulfite, sodium sulfite, glucose, phenol, thiophenol, etc.).

[0195] The technology and composition for preparing parenteral dosage forms are generally known in the art. Preparations for parenteral administration can be prepared from one or more sterile powders and / or granules containing a compound or salt of the present invention and one or more carriers or diluents mentioned for oral administration preparations. Powders or granules are usually added to a suitable volume of solvent capable of dissolving the powder or granules (usually added while stirring (e.g., stirring) the solvent). Specific solvents that can be used in the present invention include, for example, water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride and / or a variety of buffers.

[0196] The preparation of emulsions for parenteral administration can be, for example, by dissolving the compound or salt of the present invention in any pharmaceutically acceptable solvent capable of dissolving the compound to form a solution; and adding an appropriate volume of a carrier to the solution under stirring to form an emulsion. The preparation of solutions for parenteral administration can be, for example, by dissolving the compound or salt of the present invention in any pharmaceutically acceptable solvent capable of dissolving the compound to form a solution; and adding an appropriate volume of a carrier to the solution under stirring to form a solution.

[0197] Suppositories for rectal administration can be prepared, for example, by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Suitable excipients include, for example, cocoa butter; synthetic mono-, di-, or triglycerides; fatty acids; and / or polyethylene glycols.

[0198] Unless otherwise stated, every formulation or combination of components described or exemplified herein can be used to practice the invention.

[0199] (i) Binders: Binders include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic gums such as gum arabic, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethylcellulose, cellulose acetate, carboxymethylcellulose calcium, sodium carboxymethylcellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropyl methylcellulose (e.g., No. 2208, No. 2906, No. 2910), microcrystalline cellulose, and mixtures thereof. Suitable forms of microcrystalline cellulose include, for example, materials sold as AVICEL-PH-101, AVICEL-PH-103, and AVICEL-PH-105 (available from FMC Corporation, American Viscose Division, Avicel Sales, Marcus Hook, Pennsylvania, USA). An exemplary suitable binder is a mixture of microcrystalline cellulose and sodium carboxymethylcellulose, sold as AVICEL RC-581 by FMC Corporation.

[0200] (ii) Fillers: Fillers include, but are not limited to, talc, calcium carbonate (eg, granules or powder), lactose, microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof.

[0201] (iii) Lubricants: Lubricants include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, electromagnetic radiation mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, and mixtures thereof. Other lubricants include, for example, syloid silica gel (AEROSIL 200, manufactured by WR Grace Co., Baltimore, Maryland, USA), condensed aerosols of synthetic silica (sold by Deaussa Co., Plano, Texas, USA), CAB-O-SIL (a pyrogenic silica product sold by Cabot Co., Boston, Massachusetts, USA), and mixtures thereof.

[0202] (iv) Disintegrants: Disintegrants include, but are not limited to, agar, alginic acid, calcium carbonate, microcrystalline cellulose, cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pregelatinized starches, other starches, clays, other algins, other celluloses, gums, and mixtures thereof.

[0203] Tablets or capsules may be optionally coated by methods well known in the art. If binders and / or fillers are used with the compounds / bioconjugates of the present invention, they are typically formulated as about 50% to about 99% by weight of the compound / bioconjugate. In one aspect, about 0.5% to about 15% by weight of disintegrants, particularly about 1% to about 5% by weight of disintegrants, may be used in combination with the compound. Optionally, a lubricant may be added, the amount of which is typically less than about 1% by weight of the compound / bioconjugate. The technology and pharmaceutically acceptable additives for preparing solid oral dosage forms are described in Marshall, SOLID ORAL DOSAGE FORMS, Modern Pharmaceutics (Banker and Rhodes, Eds.), 7: 359-427 (1979). Other formulations are known in the art.

[0204] Liquid formulations for oral administration may take the form of solutions, syrups or suspensions. Alternatively, liquid formulations may be presented as dry products for preparation with water or other suitable carriers before use. Such liquid formulations may be prepared by conventional methods using pharmaceutically acceptable additives, such as suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifiers (e.g., lecithin or gum arabic); non-aqueous carriers (e.g., almond oil, oily esters, ethanol or fractionated vegetable oils); and / or preservatives (e.g., methyl paraben or propyl paraben or sorbic acid). The formulation may also contain appropriate buffer salts, flavoring agents, colorants, fragrances (perfuming) and sweeteners. Formulations for oral administration may also be formulated to achieve controlled release of the compound / bioconjugate. Preferably, the oral formulation contains 10% to 95% of the compound / bioconjugate. In addition, the compound / bioconjugate of the present invention may be formulated into tablets or lozenges formulated in a conventional manner for buccal administration. Other methods of oral delivery of the compounds / bioconjugates of the invention are known to the skilled artisan and are within the scope of the invention.

[0205] Formulation 1: Prepare hard gelatin capsules using the following ingredients:

[0206] Table F1

[0207]

[0208] These ingredients are mixed and filled into hard gelatin capsules in an amount of 560 mg.

[0209] Formulation 2: Prepare the tablet formulation using the following ingredients:

[0210] Table F2

[0211]

[0212] The components were blended and compressed into tablets, 665 mg each.

[0213] Formulation 3: Prepare a dry powder inhaler formulation containing the following components:

[0214] Table F3

[0215]

[0216] The active ingredient is mixed with lactose and the mixture is added to a dry powder inhaler device.

[0217] Formulation 4: Tablets, each containing 60 mg of active ingredient, are prepared as follows:

[0218] Table F4

[0219]

[0220] The active ingredient, starch, and cellulose are passed through a No. 20 mesh U.S. sieve and mixed thoroughly. The polyvinyl pyrrolidone solution is mixed with the resultant powders and then passed through a No. 16 mesh U.S. sieve. The resulting granules are dried at 50-60°C and passed through a No. 16 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate, and talc, previously passed through a No. 30 mesh U.S. sieve, are then added to the granules and, after mixing, compressed on a tablet press to yield tablets weighing 150 mg each.

[0221] Formulation 5: Capsules, each containing 80 mg of active ingredient, are prepared as follows:

[0222] Table F5

[0223]

[0224] The active ingredient, cellulose, starch and magnesium stearate are blended, passed through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in amounts of 190 mg.

[0225] Formulation 6: Suppository, each containing 225 mg of active ingredient, prepared as follows:

[0226] Table F6

[0227]

[0228] The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides which have been previously melted using the minimum necessary heat. The mixture is then poured into suppository molds of nominal capacity of 2.0 g and allowed to cool.

[0229] Formulation 7: Suspension, containing 50 mg of active ingredient per 5.0 ml dose, prepared as follows:

[0230] Table F7

[0231]

[0232] The active ingredients, sucrose and xanthan gum are combined, passed through a No. 10 U.S. sieve, and mixed with the previously prepared aqueous solutions of microcrystalline cellulose and sodium carboxymethyl cellulose. The sodium benzoate, flavoring, and color are diluted with some of the water and added with stirring. Sufficient water is then added to produce the required volume.

[0233] Formulation 8: Capsules, each containing 150 mg of active ingredient, are prepared as follows:

[0234] Table F8

[0235]

[0236] The active ingredient, cellulose, starch and magnesium stearate are blended, passed through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in amounts of 560 mg.

[0237] Kits: Various embodiments of the present invention include kits. Such kits may include a compound / bioconjugate of the present invention, optionally one or more ingredients for preparing a pharmaceutically acceptable formulation of the compound / bioconjugate, and instructions for use (e.g., administration). The different components of the compound / bioconjugate formulation may be packaged in separate containers when provided as a kit and mixed immediately before use. If desired, such separate packaging of the components may be present in a packaging or dispensing device that may contain one or more unit dosage forms containing the compound / bioconjugate. The packaging may, for example, include metal or plastic foil, such as a blister pack. In some cases, such separate packaging of the components may also make long-term storage possible without losing the activity of the components. In addition, if more than one route of administration or more than one administration regimen is intended, the different components may be packaged separately and not mixed before use. In various embodiments, the different components may be packaged in a combination for administration together.

[0238] It is further contemplated that the compounds and salts of the present invention can be used in the form of a kit, which is suitable for use in practicing the methods described herein, packaged in a container. The kit may include one or more compounds, and optionally suitable diluents, devices or device components suitable for administration, and instructions for use according to the methods of the present invention. The device may include a parenteral injection device, such as a syringe or a transdermal patch, etc. The device component may include a barrel for an injection device, etc. In one aspect, the kit includes a first dosage form containing a compound or salt of the present invention and a second dosage form containing another active ingredient, in an amount sufficient to practice the methods of the present invention. The first dosage form and the second dosage form together may include a therapeutically effective amount of a compound for treating a target condition.

[0239] In certain embodiments, the kit may be provided with instructional materials. The instructions may be printed on paper or other substrates, and / or may be provided as an electronically readable medium, such as a floppy disk, mini CD-ROM, CD-ROM, DVD-ROM, Zip disk, videotape, audiotape, etc. The detailed instructions may not be physically associated with the kit; instead, the user may be directed to an Internet website designated by the manufacturer or distributor of the kit, or provided in the form of an e-mail.

[0240] If necessary, the above emulsions or solutions for oral or parenteral administration can be packaged in concentrated form in intravenous bags, vials or other conventional containers and then diluted with a pharmaceutically acceptable liquid (such as saline) to form an acceptable compound concentration before use.

[0241] The kit may include reagents in separate containers, for example, sterile water or saline to be added to the freeze-dried active components of a separate package. For example, a sealed glass ampoule may include freeze-dried superoxide dismutase mimics, and sterile water, sterile saline or sterile solution are provided in a separate ampoule, each of which is packaged under a neutral non-reactive gas such as nitrogen. The ampoule may be made up of any suitable material, such as glass, organic polymers (such as polycarbonate, polystyrene), ceramics, metal or any other material that is generally used to hold reagents. Other examples of suitable containers include bottles that may be made of materials similar to ampoule, and envelopes that may be composed of, for example, aluminum or alloys of foil linings. Other containers include test tubes, vials, flasks, bottles, syringes, etc. Containers may have sterile inlets, such as bottles with stoppers that may be pierced by hypodermic needles. Other containers may have two compartments that are separated by an easily removable film that allows component mixing when removed. Removable film may be glass, plastics, rubber, etc.

[0242] Statement Regarding Incorporation by Reference and Modifications

[0243] All references in this application, such as patent documents including issued or issued patents or equivalents, patent application publications, and non-patent literature documents or other source materials, are incorporated herein by reference in their entirety, as if individually incorporated by reference, to the extent that each reference is not at least partially inconsistent with the disclosure in this application (e.g., a partially inconsistent reference is incorporated herein by reference except for the portion of the reference that is partially inconsistent).

[0244] The terms and expressions used herein are used as descriptive terms only and not restrictive terms, and when such terms and expressions are used, it is not intended to exclude any equivalents of the features shown and described or parts thereof, but it should be recognized that various modifications can be made within the scope of the claimed invention. Therefore, it should be understood that although the present invention has been specifically disclosed through preferred embodiments, exemplary embodiments and optional features, those skilled in the art can still modify and change the concepts disclosed herein, and such modifications and changes are considered to be within the scope of the invention defined by the appended claims. The specific embodiments provided herein are examples of useful embodiments of the present invention, and it is obvious to those skilled in the art that a large number of variations of the devices, device components, and method steps described in this specification can be used to implement the present invention. It is obvious to those skilled in the art that the methods and devices used for the present method can include a large number of optional components and processing elements and steps.

[0245] When a group of substituents is disclosed herein, it is understood that all individual members of the group and all subgroups (including any isomers, enantiomers and diastereomers of the group members) are disclosed separately. When Markush groups or other groupings are used herein, all individual members of the group and all possible combinations and subcombinations of the group are intended to be included in the disclosure separately. When a compound is described herein without specifying the specific isomers, enantiomers and diastereomers of the compound, such as in a formula or chemical name, the description is intended to include each isomer and enantiomer of the compound described separately or any combination thereof. In addition, unless otherwise stated, all isotopic variants of the compounds disclosed herein are included in the disclosure. For example, it is understood that any one or more hydrogens in the disclosed molecules can be replaced by deuterium or tritium. Isotopic variants of molecules can generally be used as standards for the determination of molecules and chemical and biological studies related to molecules or their uses. Methods for preparing such isotopic variants are known in the art. Since it is known that those of ordinary skill in the art can name the same compound in different ways, the specific names of the compounds are exemplary.

[0246] Many molecules disclosed herein may contain one or more ionizable groups [groups from which protons can be removed (e.g., -COOH) or protons can be added (e.g., amines) or can be quaternized (e.g., amines)]. All possible ionic forms of these molecules and their salts are intended to be included in the disclosure herein individually. With regard to salts of the compounds described herein, one of ordinary skill in the art can select from a variety of available counterions those suitable for preparing salts of the invention for a given application. In a specific application, the selection of a given anion or cation to prepare a salt may result in an increase or decrease in the solubility of the salt.

[0247] Unless otherwise stated, every formulation or combination of components described or exemplified herein can be used to practice the invention.

[0248] Whenever a range is given in the specification, such as a temperature range, time range, or composition range or concentration range, all intermediate ranges and subranges and all individual values ​​included in the given range are intended to be included in the disclosure. It should be understood that any subrange or individual value within a range or subrange included in the description herein may be excluded from the claims herein.

[0249] All patents and publications mentioned in the specification are indicative of the technical level of those skilled in the art to which the invention belongs. The references cited herein are hereby incorporated by reference in their entirety to indicate the state of the art as of their publication or filing date, with the intent that this information may be used herein to exclude specific embodiments in the prior art when necessary. For example, when claiming a composition of matter, it should be understood that compounds known and available in the art prior to the applicant's invention, including compounds for which effective disclosure is provided in the references cited herein, are not intended to be included in the composition of matter claimed herein.

[0250] As used herein, "comprising" is synonymous with "including," "containing," or "characterized by," and is inclusive or open-ended and does not exclude additional, unlisted elements or method steps. As used herein, "consisting of" excludes any element, step, or ingredient not specified in the claim elements. As used herein, "consisting essentially of" does not exclude materials or steps that have no substantial effect on the basic and novel features of the claim. In each case herein, any one of the terms "comprising," "consisting essentially of," and "consisting of" may be replaced by any one of the other two terms. The invention illustratively described herein may be appropriately implemented in the absence of any one element or elements, limitation or limitations not specifically disclosed herein.

[0251] Those of ordinary skill in the art will appreciate that starting materials, biological materials, reagents, synthetic methods, purification methods, analytical methods, assay methods and biological methods other than those specifically exemplified can be used in the practice of the present invention without excessive experimentation. The present invention is intended to include all functional equivalents known in the art for any such materials and methods. The terms and expressions used are used as descriptive terms rather than restrictive terms, and when using these terms and expressions, there is no intention to exclude any equivalents of the features shown and described or parts thereof, but it should be recognized that various modifications can be made within the scope of the claimed invention. Therefore, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, those skilled in the art may modify and change the concepts disclosed herein, and such modifications and changes are considered to be within the scope of the present invention as defined in the appended claims.

Claims

1. A method for treating an individual suffering from Demodex mites, said Demodex mites causing one or more mite-induced clinical symptoms, said method comprising the step of applying to the individual suffering from said mite an active agent in an amount sufficient to inactivate Demodex mites in the hair follicles, skin, eyes, eyelids, eyelashes and / or meibomian glands of said individual, thereby improving or stopping said mite-induced clinical symptoms, wherein said active agent is selected from the group consisting of promethazine, benztropine, dicyclomine, fluvoxamine, imipramine, dextromethorphan, mecamylamine, bupropine, fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, prodrugs thereof, and pharmaceutically acceptable salts thereof.

2. A method of treating an individual suffering from a skin disorder, comprising the step of topically applying or orally administering to the individual suffering from the skin disorder an active agent in an amount sufficient to inactivate Demodex mites in the hair follicles and / or skin of the individual, thereby ameliorating or stopping the manifestation of an allergic reaction and / or vasomotor reaction to the mites that results in the symptoms and signs of the skin disorder in the individual, wherein the active agent is selected from the group consisting of promethazine, benztropine, dicyclomine, fluvoxamine, imipramine, dextromethorphan, mecamylamine, bupropine, fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, cyclamate, quetiapine, prodrugs thereof, and pharmaceutically acceptable salts thereof.

3. A method of treating an individual suffering from an autoimmune disease, the method comprising the step of administering to the individual an active agent in an amount sufficient to inactivate at least a portion of Demodex mites in or on the individual, wherein the inactivation results in an improvement or cessation of one or more symptoms associated with an inflammatory response and / or an immune response to the Demodex mites, the inflammatory response and / or immune response causing one or more symptoms associated with the autoimmune disease in the individual, wherein the active agent is selected from the group consisting of promethazine, benztropine, dicyclomine, fluvoxamine, imipramine, dextromethorphan, mecamylamine, bupropine, fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, cyclamate, quetiapine, prodrugs thereof, and pharmaceutically acceptable salts thereof.

4. A method of treating an individual suffering from an ophthalmic condition, the method comprising the step of topically applying or orally administering to the individual suffering from the ophthalmic condition an active agent in an amount sufficient to inactivate Demodex mites in the hair follicles, skin, eyes, eyelids, eyelashes and / or meibomian glands of the individual, thereby ameliorating or stopping the manifestation of an allergic and / or inflammatory response to the Demodex mites that results in the symptoms and signs of the individual's ophthalmic condition, wherein the active agent is selected from the group consisting of promethazine, benztropine, dicyclomine, fluvoxamine, imipramine, dextromethorphan, mecamylamine, bupropine, fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, cyclamate, quetiapine, prodrugs thereof, and pharmaceutically acceptable salts thereof.

5. The method according to any one of claims 1 to 4, wherein the Demodex mites are Demodex brevis and Demodex folliculorum.

6. The method of any one of claims 1-5, wherein the step of topically applying or orally administering the active agent kills, paralyzes, and / or renders at least a portion of the Demodex mites incapable of reproduction.

7. The method of any one of claims 1-6, wherein the active agent is selected from the group consisting of benztropine, dicyclomine, fluvoxamine, imipramine, promethazine, dextromethorphan, mecamylamine, bupropine, and pharmaceutically acceptable salts thereof.

8. The method according to claim 7, wherein the active agent is selected from the group consisting of promethazine hydrochloride, benztropine mesylate, dicyclomine hydrochloride, fluvoxamine, imipramine maleate, dextromethorphan hydrobromide, mecamylamine hydrochloride, and bupropine hydrochloride.

9. The method of any one of claims 1-6, wherein the active agent is selected from the group consisting of fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, cyclamate, quetiapine, and pharmaceutically acceptable salts thereof.

10. The method of claim 9, wherein the active agent is selected from the group consisting of: fesoterodine fumarate, paroxetine hydrochloride, trihexyphenidyl hydrochloride, nortriptyline hydrochloride, chlorpheniramine maleate, clidinium bromide, and quetiapine fumarate.

11. The method of any one of claims 2, 5-10, wherein the skin disorder is one or more of acne vulgaris, seborrheic dermatitis, perioral dermatitis, acneiform eruption, transient acantholytic dermatosis, miliary necrotizing acne, psoriasis, steroid-induced dermatitis, primary irritant dermatitis, rosacea, or hidradenitis suppurativa.

12. The method of any one of claims 2, 5-11, wherein the skin disorder affects the facial skin or the eyelids, or both the facial skin and the eyelids.

13. The method of any one of claims 1-12, wherein the active agent is applied topically to the individual.

14. The method of claim 13, wherein the topically applied active agent is formulated as a carrier lotion, cream, soap, wash, shampoo or gel.

15. The method of claim 14, wherein the active agent is present in a concentration of 0.001% to 5% by weight in a topically applied lotion, cream, soap, wash, shampoo or gel.

16. The method of claim 13, wherein the concentration of the active agent in the topically applied lotion, cream, soap, wash, shampoo or gel is the minimum concentration effective to kill or paralyze the Demodex mite.

17. The method of claim 13, wherein the active agent is present in a dosage of about 0.01 mg / kg to 150 mg / kg body weight in a topically applied lotion, cream, soap, wash, shampoo or gel.

18. The method of claim 13, wherein the topically applied active agent is encapsulated within microliposomes prior to formulation into the carrier lotion, cream, soap, wash, shampoo or gel.

19. The method of any one of claims 1-19, wherein the active agent is topically applied to an area of ​​skin affected by the skin disorder, and optionally to at least a portion of an area of ​​skin not affected by the skin disorder.

20. The method of any one of claims 1-19, wherein the active agent is topically applied to an area of ​​the body skin where Demodex brevis and / or Demodex folliculorum are present.

21. The method of any one of claims 1-20, wherein the active agent is topically applied to substantially all areas of the skin of the individual.

22. The method of any one of claims 4-21, wherein the ophthalmic disorder is Demodex-induced inflammatory eye disease.

23. The method of claim 22, wherein the Demodex-induced inflammatory eye disease is one or more of meibomian gland dysfunction, conjunctivitis, keratoconjunctivitis, congestion, dry eyes, and blepharitis.

24. The method according to any one of claims 4, 22-23, wherein the ophthalmic disorder is one or more of meibomian gland dysfunction, blepharitis, dry eye.

25. The method of any one of claims 4, 22-24, wherein the ophthalmic condition affects the eye or the eyelid, or both the eye and the eyelid.

26. The method of any one of claims 2-25, wherein the Demodex mite is located at the hair follicles, skin, eyes, eyelids, eyelashes, and / or meibomian glands of the individual.

27. The method of any one of claims 1-26, wherein the active agent is applied topically and is provided in a formulation effective to deliver the active ingredient of the active agent to the epidermis or subcutaneous region of the subject.

28. The method according to any one of claims 3, 5-26, wherein the autoimmune disease is one or more of lupus erythematosus, rheumatoid arthritis, juvenile idiopathic arthritis, systemic lupus erythematosus, systemic sclerosis, juvenile dermatomyositis, adult dermatomyositis, Sjögren's syndrome or porphyria cutanea tarda.

29. The method of any one of claims 3, 5-26, wherein the autoimmune disease is one or more of systemic lupus erythematosus, rheumatoid arthritis, juvenile idiopathic arthritis, systemic lupus erythematosus, systemic sclerosis, juvenile and adult dermatomyositis, Sjögren's syndrome, porphyria cutanea tarda, relapsing rheumatic disease, eosinophilic fasciitis, polymorphous light eruption, granuloma annulare, lichen planus, lupus panniculitis, discoid lupus, porphyria cutanea tarda, psoriatic arthritis, chronic ulcerative stomatitis, refractory chronic urticaria, sarcoidosis, frontal fibrosing alopecia, necrobiosis lipoidica, actinic reticulocytosis, actinic prurigo, epidermolysis bullosa, Kikuchi-Fujimoto disease, graft-versus-host disease, chronic erythema nodosum, sleep-related and systemic sclerosis, pemphigus vulgaris, pemphigus foliaceus, or pemphigoid gestationis.

30. The method of any one of claims 3, 5-29, wherein the autoimmune disease affects the epidermis, lymphatic system, muscles, joints, or internal organs of the individual.

31. The method of claim 1, wherein the subject suffers from a skin disorder, an ophthalmic disorder, or an autoimmune disease.

32. A pharmaceutical preparation for inactivating Demodex mites that cause one or more clinical symptoms caused by mites, the preparation comprising a therapeutically effective amount of an active agent selected from the group consisting of promethazine, benztropine, dicyclomine, fluvoxamine, imipramine, dextromethorphan, mecamylamine, bupropine, fesoterodine, paroxetine, amoxapine, trihexyphenidyl, nortriptyline, clozapine, dimethylhydrochloride, chlorpheniramine, cyclamate, quetiapine, prodrugs thereof, and pharmaceutically acceptable salts thereof.

33. The pharmaceutical formulation of claim 32, further comprising DMSO.

34. The pharmaceutical formulation according to claim 32 or 33, further comprising an ophthalmologically acceptable excipient selected from the group consisting of buffered saline, mineral oil, vegetable oil, petroleum vaseline, Miglyol™ 182, ethanolic solutions, liposomes, and liposome-like products.

35. The pharmaceutical formulation according to any one of claims 32-34, further comprising one or more of a preservative, an antioxidant, an antibiotic and an immunosuppressant.

36. The pharmaceutical formulation according to any one of claims 32 to 35, formulated as a pad or a gel stick.

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