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WO2011031990A1 - Traitements intermittents et ponctuels faisant appel au lithium pour moduler la pousse des poils et des cheveux - Google Patents

Traitements intermittents et ponctuels faisant appel au lithium pour moduler la pousse des poils et des cheveux Download PDF

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Publication number
WO2011031990A1
WO2011031990A1 PCT/US2010/048457 US2010048457W WO2011031990A1 WO 2011031990 A1 WO2011031990 A1 WO 2011031990A1 US 2010048457 W US2010048457 W US 2010048457W WO 2011031990 A1 WO2011031990 A1 WO 2011031990A1
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Prior art keywords
lithium
hair
treatment
skin
follicles
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Inventor
William D. Ju
Stephen M. Prouty
Shikha P. Barman
Scott C. Kellogg
Eric Schweiger
Seth Lederman
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Follica Inc
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Follica Inc
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/19—Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/63—Steroids; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00—Preparations for affecting hair growth
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/74—Biological properties of particular ingredients
    • A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00—Preparations for affecting hair growth
    • A61Q7/02—Preparations for inhibiting or slowing hair growth

Definitions

  • the invention relates to intermittent lithium treatments, or a single pulse lithium treatment for modulating hair growth in human subjects.
  • Uses of compositions containing compounds that liberate lithium ions are described, including adjuvants and devices for administration.
  • the intermittent treatment protocol involves multiple courses of lithium treatment interrupted by lithium treatment "holidays.”
  • a dose of lithium is administered over a short period of time.
  • the lithium treatment s) can be used in combination with other treatments for the enhancement or inhibition of hair growth.
  • Such combination treatments may involve mechanical or physical treatments that cause integumental perturbation (e.g.
  • the combination treatment(s) may be any treatment that causes integumental perturbation or that enhance or inhibit hair growth.
  • the combination treatment(s) may be any treatment that causes integumental perturbation or that enhance or inhibit hair growth.
  • the combination treatment(s) may be any treatment that causes integumental perturbation or that enhance or inhibit hair growth.
  • the combination treatment(s) may be any treatment that causes integumental perturbation or that enhance or inhibit hair growth.
  • the combination treatment(s) may be
  • the skin of an adult human is essentially covered with hair follicles and contains approximately five million hair follicles, with approximately 100,000 - 150,000 covering the scalp.
  • the portions of human skin that lack visible hair contain, for the most part, hair follicles that produce "vellus hair” while certain other hair follicles may contain or produce no hair (see Figure 1).
  • Essentially, only the glaborous skin on palmar and plantar aspects of hands and feet, respectively, and the lips and labia lack hair follicles.
  • Only a minority of human hair follicles produce a hair fiber that can be appreciated visibly (a "terminal hair") and these specialized follicles are localized on specific regions of skin. Accordingly, both the presence and absence of visible hair on human non-glaborous skin is mediated by regulation of activity of specialized follicles.
  • Hair follicles and particularly human hair follicles, are crypt structures comprised of distinct components, each comprised of several different specialized cells (see Figures 2 and 3).
  • the vast majority of hair follicles contain units called sebaceous glands (which produce sebum).
  • Some hair follicles have apocrine glands attached to them, and are located in the axilla and other specific areas of the body.
  • the structures of the hair follicle include the follicular papilla (FP) and the germinative epithelium (GE) (together, the bulb).
  • the FP is comprised of mesenchymal cells (and connective tissue).
  • the other cells of the follicle are epithelial and include at least 8 cellular lineages including the outer root sheath (ORS), the companion layer (CL), the internal root sheath Henle's layer (He), internal root sheath Huxley's layer (Hu). the cuticle of the internal root sheath (Csth), the cuticle of the hair shaft (Csft), the cortex of the hair shaft, and the medulla of the shaft (Med).
  • ORS outer root sheath
  • CL the companion layer
  • He internal root sheath Henle's layer
  • Huxley's layer Hu
  • the cuticle of the internal root sheath Csth
  • the cuticle of the hair shaft Csft
  • a follicular unit of scalp hair is typically composed of one to four terminal hair follicles; one to two vellus hair follicles; their associated sebaceous glands, neurovascular plexus, an erector pilorum muscle and a circumferential band of adventitial collagen, termed the
  • Hair follicles are believed to produce approximately 20 individual hair shafts over the life of the follicle as the follicle progresses through cycles of hair production, shedding (ejection), involution and new growth.
  • mouse follicles While the mouse has certain specialized follicles (e.g., whiskers, guard, awl, suchene, and zigzag hair), mouse follicles are generally not subject to developmental and gender-specific hair patterning. In contrast, a significant number of human follicles are individual participants in choreographed hair patterning that affects the type, length and color of shaft produced at different times in development and aging and in a gender specific manner.
  • follicles e.g., whiskers, guard, awl, suchene, and zigzag hair
  • mouse follicles are generally not subject to developmental and gender-specific hair patterning.
  • a significant number of human follicles are individual participants in choreographed hair patterning that affects the type, length and color of shaft produced at different times in development and aging and in a gender specific manner.
  • follicle formation occurs but once in a lifetime (in utero), so that a mammal, and particularly a human, is born with a fixed number of follicles, which does not normally increase thereafter. It has been proposed, however, that follicle neogenesis can be associated with wound healing in animals (e.g., rabbits, mice). See, Stenn & Paus, 2001 , Physiol. Revs. 81 :449-494. Nevertheless, severe wounds and burns are usually associated with cutaneous repair that results in scar tissue, no hair follicles, and the loss of regenerative capability that hair follicles may provide (see, Fathke et al.. 2006, BMC Cell Biol. 7:4).
  • Hair growth in each follicle occurs in a cycle that includes the following principal phases: anagen (growth phase), catagen (involuting/regressing stage), telogen (the quiescent phase), exogen (shedding phase), and re-entry into anagen (sometimes referred to herein as the "Follicle Cycle”).
  • anagen growth phase
  • catagen involuting/regressing stage
  • telogen the quiescent phase
  • exogen shedding phase
  • re-entry into anagen sometimes referred to herein as the "Follicle Cycle”
  • kenogen a latent phase
  • the Follicle Cycle has been relied on to explain many phenomena associated with hair growth and hair loss.
  • Human hair follicles are relatively unique among mammals (and particularly non- primates) since hair follicles in a region of skin are not synchronized. On an adult human scalp, at any particular time approximately 90% are in anagen; 10-14% in telogen and 1 -2% in catagen.
  • each hair follicle belongs to a class of follicles that accounts for the distinctive length, temporal appearance, regulation by sex hormones, etc., of the hair shaft it produces.
  • the duration of the Follicle Cycle length is believed to be a characteristic of each distinct specialized human hair follicle that accounts for the length of hair produced and other aspects of the follicle function.
  • the anagen stage for eyebrow hair follicles is approximately 4 months and for scalp hair follicles is approximately 3-4 years. It has been proposed that eyebrow hairs have a shorter length compared to scalp hairs because the former have a shorter anagen phase.
  • the Follicle Cycle has also been used to distinguish two different types of pathologic hair loss (effluvium): Anagen effluvium and Telogen effluvium.
  • Anagen is associated with intense mitotic activity; therefore, follicles in anagen arc sensitive to cancer chemotherapeutic agents.
  • Anagen effluvium is believed to be the process in which certain hair follicles undergo hair loss and involution during chemotherapy because these agents typically target cells with high metabolic or mitotic activity (e.g. , 5- fluorouracil, methotrexate, cyclophosphamide, vincristine), in addition to chemotherapy drugs, Anagen effluvium can be caused by other toxins, radiation exposure (e.g. , radiation overdose), endocrine diseases, trauma, pressure, and certain diseases like alopecia areata (an autoimmune disease that attacks anagen follicles.)
  • Telogen effluvium is a premature interruption of anagen and early entry of anagen follicles into the Telogen (or resting) phase. The proportion of telogen hairs on the scalp increases to 25-50% and telogen follicles remain in telogen for more than the usual 3-6 months. Telogen effluvium is caused frequently by drugs like lithium and other drugs like valproic acid and carbamazepine and numerous other drugs including the commonly used beta blockers and oral contraceptives. In addition to psychiatric drugs, telogen effluvium can be induced by childbirth, traction, febrile illnesses, surgery, stress, or poor nutrition (see Mercke et al, 2000, Ann. Clin. Psych. 12:35-42).
  • human skin is essentially covered with hair follicles.
  • the portions of human skin that lack visible hair contain, for the most part, hair follicles that produce "vellus hair” which is thin and short (i.e., less than 2 cm in length) and often colorless. Certain other hair follicles may contain or produce no hair. Only a minority of human hair follicles produce a hair fiber that can be appreciated visibly (a "terminal hair") and these specialized follicles are localized on specific regions of skin.
  • Another follicle type is the "sebaceous" follicle, which has a large sebaceous gland and a vellus-like hair shaft localized in the acne-prone areas. Accordingly, both the presence and absence of visible hair on human skin is mediated by regulation of the activity of specialized hair follicles.
  • follicle types By the time of birth, distinct specialized follicle types are positioned in specific areas of the skin where they will each play a programmed role in hair patterning over the life of the human individual, producing various hair types (lanugo, vellus or terminal hair) either constitutively or depending on certain signals, such as sex hormones or other factors (e.g., lanugo hair can reappear in starvation or in eating disorders such as anorexia nervosa and bulimia).
  • Gender is associated with specific patterning of human hair.
  • the growth and loss of visible hair in specific areas of the skin, in stereotypical gender dimorphic patterns, are regarded as “Secondary Sexual Characteristics/'
  • This terminology relates "secondary” features such as hair patterning to the genitals and reproductive organs, which are termed "Primary Sexual Characteristics.”
  • Primary Sexual Characteristics The distinctive genitals and reproductive organs of males and females acquired during embryonic development undergo further changes in puberty and menopause/andropause. In addition to hair growth and loss, breasts in females are also considered Secondary Sexual Characteristics.
  • Certain human hair follicles are targeted to specific skin areas and develop specialized characteristics during embryogenesis under the influence of sex hormones such as testosterone and dihydrotestosterone ("androgens") and/or estrogens. Further, certain human hair follicles are driven to change activity by sex hormones during puberty and in
  • the appearance and intensity of secondary sex characteristics can be described as being regulated by ratios of androgens and estrogens, since to a certain extent either of these groups of hormones (androgens and estrogens) can act to induce certain activities or to inhibit the effect of the other group (i.e.. androgens inhibit estrogen effects and estrogens inhibit androgen effects).
  • androgens induce male characteristics and suppress female characteristics while estrogens induce female characteristics and suppress male
  • Male and female refer to the extremes of genetic gender dimorphism and include by reference the various conditions and states that represent a spectrum of male and female features (such as XO syndromes or conditions that result from exogenous sex steroid administration).
  • J Specialized human hair follicles have quantitative variation in activity as well as qualitative variation.
  • sex steroids have qualitative effects on hair patterning either in embryogenesis or in adult life or both (e.g., males have beard hair follicles that produce terminal hair after puberty whereas females do not).
  • Males and females also vary in the amount of gender-specific hair patterning (e.g., a higher density of leg hair follicles produce terminal hair on male rather than female legs).
  • individuals of the same gender exhibit quantitative variation. For example, male chest and back hair presents in different individuals as a spectrum from almost hairless to dense hair and from small regions of follicles producing terminal hair to large regions.
  • Gender specific human hair patterning highlights the distinct biological programming of specific hair follicles. Distinct hair follicles in relative proximity on the male scalp and face respond to high androgen/estrogen ratios in diametrically opposite ways: high androgen/estrogen ratios induce vellus to terminal hair transformation in male moustache/beard skin (particularly during puberty), but induce terminal to vellus follicle transformation change in male frontal/temporal scalp (progressively post puberty) in MPHL.
  • Hair follicles in the axillary and pubic regions appear to be more sensitive to androgen than moustache/beard follicles; since terminal hair in axillac/pubis grows: (a) in females with relatively low levels of androgen; (b) early in male puberty before beard/moustache; and (c) in patients with genetic 5-alpha-Reductase Type II deficiency.
  • MHL Male pattern hair loss
  • Androgenetic alopecia is a genetically-mediated disorder that occurs in approximately 50% of men by the age of 50 years (see review, Stough ct al, 2005). in women, the histological features of the condition are the same as in men, but susceptibility, age at onset, rate of progression and pattern of hair loss differ between genders (Dinh and Sinclair, 2007).
  • the loss of scalp hair in men is known to be a process driven by the androgen, DHT, which can be inhibited and to some extent reversed by finasteride, which inhibits 5-alpha-reductase II (which converts testosterone to DHT).
  • DHT androgen
  • finasteride which inhibits 5-alpha-reductase II (which converts testosterone to DHT).
  • DHT androgen
  • finasteride which inhibits 5-alpha-reductase II (which converts testosterone to DHT).
  • the affected hair follicles on the bald vertex or temples are considered to be atrophied, or perhaps involuted irreparably ("senescent").
  • senescent irreparably
  • Telogen hairs are more loosely anchored and prone to shedding or being pulled out (for example, by combing or brushing hair).
  • a club hair is produced that is a fully keratinized hair.
  • the hair follicles on MPHL affected areas also undergo follicular miniaturization in which a growing proportion of terminal follicles become vcllus follicles.
  • androgenetic alopecia is thought to involve the progressive conversion of hair follicle units with 3 or more terminal hairs to follicular units having fewer terminal hairs (e.g., units with 2 terminal hairs progress to units with 1 terminal hair).
  • MPHL is associated with specific polymorphisms of the androgen receptor, the EDA2R gene. Men who are genetically deficient in Type II 5-alpha-reductase do not experience MPHL (see Jenkins et al, 1992, J Clin Invest 89:293-300).
  • Hic-5/ARA55 Inui, 2007, J Invest Dermatol 127:2302-2306
  • Hic-5/ARA55 mRNA expression was high in dermal papilla cells from the beard and bald frontal scalp but low in cells from the occipital scalp.
  • ARA70/ELE 1 had decreased expression of a splice variant form (ARA70beta/ELE 1 beta) in the dermal papilla of balding recipient areas than non- balding areas (Lee et al, 2005, J Cutan Pathol 32:567-571 ).
  • FPHL is thought to share some features with MPHL in terms of progressive reduction in the duration of anagen and progressive follicular miniaturization, although recent studies have found a prolongation of kenogen.
  • transplanted which has been refen-ed to as "donor dominance” (Orentreich N, 1959, Ann NY Acad Sci. 83:463-479).
  • donor dominance Orentreich N, 1959, Ann NY Acad Sci. 83:463-479.
  • This principle is evidenced by the results of the commonly performed procedure of transplanting scalp hair (skin, follicles or follicle units) in males from areas that are not subject to androgen-triggered, MPHL (e.g. occipital scalp) to areas in which specialized follicles have begun producing vellus hair or have stopped producing hair under the influence of androgens (e.g. frontal/temporal; crown or vertex scalp.
  • the transplanted follicles retain the programmed terminal hair producing features from their original location.
  • more recent studies suggest that the recipient site may affect some characteristics of transplanted hairs. See Hwang et al , 2002, Dermatol. Surg. 28:795-799.
  • Human hair loss can be categorized as ( 1 ) gender specific hair patterning, (2) pathological hair loss, or (3) hair loss after wounding, all which can be associated with effects on self-esteem and self-image, and many individuals explore whether their hair loss process can be treated.
  • Current treatments offered involve a limited selection of agents and regimens, such as chemical and surgical approaches that either stimulate or transplant pre-existing hair - none are associated with true follicular neogenesis.
  • Chemical treatments involve the use of drugs for the treatment of certain MPHL. These include, for example, minoxidil (an antihypertensive drug that opens the + channel); and antiandrogens such as finasteride, dutasteride or ketoconazole. While these types of treatments are reasonably effective in preventing or delaying MPHL, they are less effective in stimulating the growth of significant terminal hair in scalp of MPHL after baldness has been present for 6 months or more. Moreover, minoxidil and finasteride require continuous treatment for lasting effects. Consequently, patients with advanced MPHL may express dissatisfaction with even statistically significant, but cosmetically insignificant increase in hair counts and such frustration may contribute to poor compliance and further unsatisfactory outcomes.
  • minoxidil an antihypertensive drug that opens the + channel
  • antiandrogens such as finasteride, dutasteride or ketoconazole.
  • bimatoprost a prostaglandin analog used to control the progression of glaucoma in the management of ocular hypertension
  • Latisse® a prostaglandin analog used to control the progression of glaucoma in the management of ocular hypertension
  • a device that uses low level light energy directly on the scalp has received FDA clearance as a 51 OK device. Although the device is advertised as a "Laser,” it operates by applying low level monochromatic light energy directly to the scalp, which is thought to stimulate hair growth through "photo-biostimulation " of hair follicles.
  • Various types of devices operating on similar principles were referenced as the predicate for HairMax (see, Lolis et ai, 2006, J. Cosmetic Dermatol. 5:274-276).
  • hair transplantation in which scalp strips, hair follicles or follicular units from the occipital scalp (which are resistant to the effects of androgens in inducing MPHL type alopecia) are excised and transplanted to a person's balding or thinning areas.
  • Another surgical method that has been used is scalp reduction; in this procedure, the skin in the balding area of the scalp is surgically excised and the surrounding skin (with hair) is pulled together and sutured.
  • Surgical methods are best for focal hair loss, and are less effective for diffuse hair loss, are less effective for women and younger patients are not ideal candidates because the pattern and extent of hair loss is variable, and may be inconvenient because of the expense of the surgery, duration of time to show a cosmetic effect, creation of scarring.
  • cosmetic coverage is constrained by the area of and the number of hairs in a patient's donor sites.
  • Depilation affects the part of the hair above the surface of the skin.
  • the most common form of depilation is shaving.
  • Another popular option is the use of chemical depilatories (e.g., Nair®), which work by breaking the disulfide bonds that link the protein chains that give hair its strength, making the hair disintegrate.
  • Epilation is removal of the entire hair, including the part below the skin, and is believed to be longer-lasting.
  • Methods include plucking with tweezers, waxing, sugaring, epilation devices, threading, home pulsed light, and can include the use of hair growth retardants (e.g., Vaniqa® (eflornithine)).
  • Electrology electrolysis
  • laser and intense pulsed light are used for permanent hair removal.
  • permanent hair removal is an imperfect process.
  • laser hair removal does not work well on light hair and/or on dark skin.
  • multiple sessions with trained medical personnel are required.
  • Intermittent lithium treatments or a single pulse lithium treatment are used to modulate the growth of hair in human subjects.
  • Any pharmaceutically acceptable compound that releases the lithium ion also referred to herein as lithium cation, Li+, ionized lithium
  • lithium treatment can be used for the lithium treatment; such compounds include, but are not limited to lithium gluconate, lithium succinate, and other organic salts/acids; and lithium chloride and other inorganic salts/acids, as described in Section 5.1, infra.
  • the intermittent lithium treatment protocol involves multiple courses of lithium treatment interrupted by lithium treatment "holidays" (periods during which no lithium treatment is administered).
  • a lithium treatment holiday is a period of time during which the patient stops the lithium treatment with the intent of resuming treatment.
  • a dose of lithium is administered over a short period of time.
  • the lithium treatment can be administered topically, transdermally, intradermal ly, cutaneously, subcutaneously, intramuscularly, intravenously, orally, sublingually, or can be bucchal.
  • Adjuvants that target the lithium to the desired hair follicles may be included in the formulations used.
  • Topical lithium treatment is a preferred embodiment because high local concentrations can be achieved while minimizing systemic exposure.
  • lithium gluconate 8% weight/weight (w/w) gel e.g. , Lithioderm 8% gel
  • Dreno B 2007, Ann Dermatol Venereol.
  • lithium formulations including various modified release forms, may be delivered topically as additives to shampoos and other hair products, as a lotion, cream, or ointment, may be delivered using devices such as iontophoresis, micro-needle injection arrays, or auto-injector devices.
  • Embodiments of the invention include combination therapies, involving the addition of other treatment(s) concurrently with, or during the breaks between, the cycles of intermittent lithium treatments; or the addition of other treatments ) concurrently with, or before and/or after the pulse lithium treatment.
  • Such combination therapies can include, but are not limited to, the concurrent or sequential use of other chemical agents, or mechanical or physical treatments including but not limited to electrology, laser, intense pulsed light, dermabrasion, or surgical treatments (e.g., hair transplant, strip harvesting, follicular unit extraction (FUE), scalp reduction, etc. ) that either promote or inhibit the growth of hair.
  • intermittent lithium treatment or a pulse lithium treatment can be in combination with perturbation (e.g. , debriding, peeling, or wounding) of the skin and/or other tissues of the integumentary system by methods such as dermabrasion, microneedles, laser treatment, electromagnetic disruption, electroporation, or sonoporation, chemically (e.g., to induce inflammation), or by any other method described herein or known in the art, prior to or concurrent with administration of the lithium formulation.
  • the procedure can be controlled to limit perturbation to the epidermis, or extend deeper into the dermis and/or hypodermis.
  • the occurrence of pinpoint bleeding would indicate removal of the epidermis and portions of the upper layer of the dermis.
  • the occurrence of increased bleeding would indicate deeper penetration (and thus perturbation) into the dermis layer.
  • intermittent lithium treatment or a pulse lithium treatment can be used concurrently or in sequential/alternating combination with other agents or treatments that stimulate hair growth to increase overall hair density in a human subject.
  • intermittent lithium treatments or a pulse lithium treatment administered concurrently or alternating sequentially with one or more of the following agents can be used to promote hair growth: minoxidil, finasteride, bimatoprost (Latisse), CaCl 2 , or adenosine.
  • the lithium treatment, with or without an additional agent for promoting hair growth can be used in combination with integumental perturbation to promote hair growth, such as by, e.g.
  • a low-level laser therapy treatment e.g., HairMax
  • a low-level laser therapy treatment can be applied concurrently with the intermittent lithium treatment or pulsed lithium treatment to stimulate hair growth.
  • intermittent lithium treatments or a pulse lithium treatment administered concurrently or alternating sequentially with one or more agents that prevent hair follicle cells from sencscing can be used to promote hair growth, for example, anti-oxidants such as glutathione, ascorbic acid, tocopherol, uric acid, or polyphenol antioxidants); inhibitors of reactive oxygen species (ROS) generation, such as superoxide dismutase inhibitors; stimulators of ROS breakdown, such as selenium; mTOR inhibitors, such as rapamycin; or sirtuins or activators thereof, such as resveratrol, or other SIRT1 , SIRT3 activators, or nicotinamide inhibitors.
  • anti-oxidants such as glutathione, ascorbic acid, tocopherol, uric acid, or polyphenol antioxidants
  • ROS reactive oxygen species
  • stimulators of ROS breakdown such as selenium
  • mTOR inhibitors such as rapamycin
  • Success of a pulse or intermittent lithium treatment can be measured by:
  • Human subjects who are candidates for such treatments include any subject for whom increased hair growth is desired including, but not limited to, subjects with
  • nonscarring (noncicatricial) alopecia such as androgenetic alopecia (AGA), including MPHL or FPHL, or any other form of hair loss caused by androgens, toxic alopecia, alopecia areata (including alopecia universalis), scarring (cicatricial) alopecia, pathologic alopecia (caused by, e.g. , medication, chemotherapy, trauma, wounds, burns, stress, autoimmune diseases), senescence (age-related hair loss), malnutrition, or endocnne dysfunction), or hypotrichosis, or any other disease, disorder, or form of hair loss as discussed infra and/or known in the art.
  • AGA androgenetic alopecia
  • intermittent lithium treatments or a pulse lithium treatment is used concurrently or in sequential combination with a cytotoxic drug, a hair growth retardant, epilation or depilation methods to reduce unwanted hair growth.
  • intermittent lithium treatments or a pulse lithium treatment alternating sequentially with one or more of the following agents or treatments can be used to inhibit unwanted hair growth: etlornithine HC1 (Vaniqa), 5-fluorouracil (5-FU) (e.g., Efudex 5% cream), and/or epilation. Success of treatment can be measured by:
  • Human subjects who are candidates for such treatment include any subject for whom elimination of unwanted visible hair is desired including, but not limited to, those afflicted with hypertrichosis, excess hair in androgen-dependent areas of the skin, idiopathic hirsutism, female post-menopausal facial hair, axillary hair, leg hair, back hair, ear hair, nares or nose hair, or any other disease, disorder, or form of unwanted hair or excessive hair as discussed infra and/or known in the art.
  • the invention is based in part on the realization that human hair follicles and follicular units are relatively unique (among other mammals, particularly non-primates) in that they enter and progress through different stages of the hair Follicle Cycle relatively independently of each other, even independently of neighboring follicles or follicular units. Consequently, the normal biology of human hair patterning is based on a probability distribution of the hair cycle stage that follicles will be in. generated by a stochastic (random) process by which follicles cycle independently.
  • the object of the invention is to synchronize hair follicle growth in the treated area to more effectively promote the growth of terminal hair (in preference to vellus hair); and/or promote the branching of pre-existing hair follicles (seen as an increased number of hair shafts per pore); and/or increase the width of hair follicles (thereby promoting growth of an increased shaft width); and /or promote regeneration of hair follicles or generation of new hair follicles ("follicle neogenesis"); and/or delay or prevent follicle senescence.
  • the object is to inhibit the growth of unwanted hair (as measured by, e.g., decreased terminal hair formation or inhibition of follicle regeneration or generation of new follicles).
  • the intermittent lithium treatments or pulse lithium treatments may achieve these results by:
  • the invention is based, in part, on the principle that the lithium ion (Li+) is an inhibitor of the polyphosphoinositide cycle that can reversibly arrest cells in cell cycle. In plant cells which demonstrate remarkable precision in the timing of mitotic events, the lithium ion has been shown to cause metaphase arrest that can be reversed by the addition of CaCl 2 or myo-inositol.
  • the lithium ion has also been shown to arrest cancer cell lines at certain stages of the cell cycle (see, e.g., Wang JS, 2008, World J. Gastroenterol. 14:3982-3989).
  • the invention is based, in part, on the inventors' recognition that the lithium ion can be used in a pulse or intermittent treatment regimen to synchronize groups of hair follicle cells that are in various stages of cell cycle (cycling asynchronously). More particularly, a pulse lithium treatment or intermittent lithium treatments can be used to reversibly arrest mitosis in hair follicle cells.
  • Restarting the cell cycle at the termination of a pulse lithium treatment, or during the "holidays" between intermittent lithium treatments should restart cell cycle synchronously.
  • the synchronization phenomenon can be described by analogy to traffic lights: periodically arresting the motion of individual cars generates synchronization because cars pile up behind stop lights. Similarly, by introducing a signal that periodically arrests cell division, synchronization is generated because when the "stop" signal is removed, cells initiate division at the same time.
  • Such synchronization of cell cycles in the hair follicle cells results in relative synchronization of hair follicle cycle stage in groups of follicles that otherwise have a stochastic distribution of stages of follicle cycle (asynchronous follicle cycle), as is the case with the human scalp.
  • the invention is based in part on the recognition that the timing of the administration of lithium is important for it to function as an effective modulator of hair growth in human subjects.
  • lithium treatment results, indirectly, in increasing Wnt signaling, but agents that increase Wnt signaling have had conflicting effects on follicle development.
  • they stimulate follicle morphogenesis but also induce hair follicle tumors (Gat et ai, 1998, Cell 95: 605-614), leading to decreased hair growth (Millar et ai , 1999, Dev. Biol. 207: 133-149).
  • it has been shown to arrest mitosis (Wolniak, 1987, Eur. J. Cell Biol.
  • the invention is also based, in part, on the principle that human skin is replenished by bone-marrow derived and tissue-derived stem cells throughout life.
  • the lithium treatment(s) is used in combination with methods that mobilize tissue stem cells (e.g., using integumental perturbation); and/or methods that mobilize bone marrow-derived stem cells (e.g., growth factors such as G-CSF and/or chemical agents such as plerixafor (Mozobil®)); and/or methods that regulate the differentiation of these stem cells into gender-specific specialized human hair follicles (e.g., using agents such as finasteride, fluconazole, spironolactone, flutamide, diazoxide, 1 l -alpha-hydroxyprogesterone, ketoconazole, U58841 , dutasteride, fluridil, or QLT-7704, an antiandrogen oligonucleotide, cyoctol, topical proge
  • agents such as fin
  • any other antiestrogen, an estrogen, or estrogen-like drug (alone or in combination with agents that increase stem cell plasticity; e.g., such as valproate), etc. , known in the art), that can result in the appearance of specialized follicles having features that arc different from natural follicles in the target location of skin.
  • Such combination treatments can further include the use of agents that enhance the growth of hair (e.g., minoxidil, finasteride, bimatoprost (Latisse), CaCl 2 , adenosine, and others described herein) or aid in the removal of hair (e.g.
  • the Follicle Stem Cells involved can be derived from (1 ) other Follicle Stem Cells, (2) from other tissue stem cells, termed "pre-Follicle Stem Cells" (from the interfollicular skin), (3) from bone marrow- derived stem cells (“BMST”). and/or (4) from mesenchymal stem cells such as adipocyte stem cells.
  • pre-Follicle Stem Cells from the interfollicular skin
  • BMST bone marrow- derived stem cells
  • mesenchymal stem cells such as adipocyte stem cells.
  • BMST bone marrow derived stem cells
  • their differentiation into Follicle Stem Cells requires intact follicles, whose cells can play the role of "nurse cells” and provide appropriate signals to guide the differentiation of bone marrow derived stem cells into Follicle Stem Cells.
  • Integumental perturbation (for example, by the induction of inflammation, wounding, or laser treatment) (1 ) provides signals for Follicle Stem Cells to divide symmetrically to begin the process of forming new follicles; (2) mobilizes tissue stem cells ("pre-Follicle Stem Cells") from interfollicular skin to differentiate into stem cells and (3) increases the trafficking of bone marrow derived stem cells to affected areas of skin and promotes their differentiation into Follicle Stem Cells by nurse cells in existing follicles.
  • intermittent or pulse lithium treatment organizes the normally asynchronous state of human hair follicle cells in Cell Cycle and human hair follicles in Follicle Cycle into relatively more synchronous states of human hair follicle cells in Cell Cycle and human hair follicles in Follicle Cycle.
  • Alopecia Abnormal hair loss:
  • Alopecia areata Hair loss in patches, thought to be caused by an autoimmune response to hair follicles in the anagen stage; extensive forms of the disorder are called alopecia areata totalis (hair loss over the entire scalp) and alopecia areata universalis (hair loss over the entire body).
  • Anagen Growth stage of the hair-Follicle Cycle.
  • Anagen effluvium Abrupt shedding of hair caused by interruption of active hair- follicle growth (e.g., in patients undergoing chemotherapy).
  • AGA Androgenetic alopecia
  • Bulb Lowermost portion of the hair follicle, containing rapidly proliferating matrix cells that produce the hair.
  • Catagen Stage of the hair cycle characterized by regression and involution of the follicle.
  • Club hair Fully keratinized, dead hair—the final product of a follicle in the telogen stage; 50 to 150 club hairs are shed daily from a normal scalp.
  • Female Pattern Hair Loss (FPHL): form of gender specific hair patterning in females (also sometimes referred to as female pattern alopecia).
  • Follicle cycle Hair growth in each follicle occurs in a cycle that includes the following phases: anagen (growth phase), catagen (involuting/regressing stage), telogen (the quiescent phase), exogen (shedding phase), and re-entry into anagen.
  • Kenogen Latent phase of hair cycle after hair shaft has been shed and growth is suspended in follicle.
  • Hirsutism Excessive hair growth in androgen-dependent areas in women.
  • Hypertrichosis Excessive hair growth (usually diffuse) beyond that considered normal according to age, race, sex, and skin region.
  • Integumental Pertaining to the integumentary system, which comprises the skin (epidermis, dermis, hypodermis (or subcutanea)) and all cells contained therein regardless of origin, and its appendages (including, e.g., hair and nails).
  • Lanugo hair Fine hair on the body of the fetus, usually shed in utero or within weeks after birth.
  • MHL Male Pattern Hair Loss
  • niM millimolar units
  • Miniaturization Primary pathological process in androgenetic alopecia, resulting in conversion of large (terminal) hairs into small (vellus) hairs.
  • Permanent alopecia Caused by destruction of hair follicles as a result of inflammation, trauma, fibrosis, or unknown causes; examples include lichen planopilaris and discoid lupus erythematosus. Include diseases referred to as scarring alopecia.
  • Telogen effluvium Excessive shedding of hair caused by an increased proportion of follicles entering the telogen stage; common causes include drugs and fever.
  • Terminal hair Large, usually pigmented hairs on scalp and body.
  • Vellus hair Very short, nonpigmented hairs (e.g. , those found diffusely over nonbeard area of face and bald scalp as a result of miniaturization of terminal hairs ). 4. DESCRIPTION OF THE FIGURES
  • Figure 1 Types of human hair follicles.
  • Figure 2 Architecture of the skin.
  • Figure 5 Hamilton Norwood classification of male pattern hair loss (MPHL).
  • FIG. 6 Permeation of lithium ions (also referred to as "Li ions”) through the dermis (y-axis) from Formulation 35A' (lithium chloride emulsion cream; see Table 2) is plotted over time, in hours (x-axis).
  • Cadaver skin was dermabraded with a standard dermabrader to remove the stratum corneum and epidermis prior to administration of the lithium compound.
  • Formulation 35 A' lithium chloride emulsion cream; see Table 2 is plotted over time, in hours (x-axis).
  • Figure 8 Release of Li ions through dermis (y-axis) from Formulation BX (lithium chloride gel; see Table 2) is plotted over time, in hours (x-axis).
  • Cadaver skin was dermabraded, with a standard dermabrader to remove the stratum corneum and epidermis prior to administration of the lithium compound.
  • Figure 9 Release of Li ions through dermis (y-axis) from Formulation BV-001 - 003A (lithium chloride hydrogei; see Table 2) is plotted over time, in hours (x-axis).
  • Cadaver skin was dermabraded with a standard dermabrader to remove the stratum corneum and epidermis prior to administration of the lithium compound.
  • Figure 10 Release of Li ions through dermis (y-axis) from Formulation 28A (lithium chloride topical dispersion cream; see Table 2) is plotted over time, in hours (x-axis).
  • Cadaver skin was dermabraded, with a standard dermabrader to remove the stratum corneum and epidermis prior to administration of the lithium compound.
  • FIG. 13 Skin lithium concentrations calculated in mM, as a function of increasing doses of a formulation of lithium chloride dissolved in isotonic saline in mg kg administered subcutaneously to mice dermabraded prior to dosing. "Peak samples" were taken 1 h post last dosing.
  • Figure 14 Comparison of Peak lithium concentrations in plasma and skin upon subcutaneous administration of a formulation of lithium chloride dissolved in isotonic saline following DA.
  • FIG. 15 Lithium concentrations calculated in mM, in total blood (red blood cells (RBC) + plasma), as a function of increasing doses of a formulation of lithium chloride dissolved in isotonic saline in mg kg, administered subcutaneously to mice dermabraded prior to dosing.
  • B Skin lithium concentrations calculated in g/kg, as a function of increasing doses in mg/kg. In the wounded groups, skin was dermabraded prior to administration of the formulation of lithium chloride dissolved in isotonic saline. Non-wounded comparisons are shown (square, diamond) with dermabrasion wounded groups (cross, triangle). * It is noted that in this experiment, dermabrasion was accomplished using a microdermabrasion device.
  • FIG. 16 Skin lithium concentrations calculated in mM, as a function of increasing doses of a formulation of lithium chloride dissolved in isotonic saline in mg/kg.
  • the lithium formulation was administered subcutaneously following full thickness excision (FTE) of skin. Dosing was started on the day of scab detachment (-1 1 -14 days post-FTE). Lithium ion concentrations were measured by a validated bioanalytical ICP method (see Section 29 infra),
  • FIG. Plasma lithium Concentrations calculated in mM, as a function of increasing doses of a formulation of lithium chloride dissolved in isotonic saline in mg/kg.
  • the lithium formulation was administered subcutaneously following FTE. Dosing was started on the day of scab detachment (-1 1- 14 days post-FTE). Lithium concentrations were measured by a validated bioanalytical ICP method (see Section 29 infra).
  • Figure 18 Comparison of Peak lithium concentrations in plasma and skin upon subcutaneous administration of a formulation of lithium chloride dissolved in isotonic saline following FTE.
  • ICP/MS/MS using a validated method (see Section 20 infra).
  • N 2 per time point— error bars denote range
  • Figure 20 Pharmacokinetic analysis of lithium concentrations in skin and plasma with twice daily topical dosing of lithium gluconate, 1%; lithium gluconate, 8%; and lithium gluconate, 16% following DA.
  • FIG. 21 Topical lithium 8% increases the proportion of mature neogenic hair follicles in healed FTE wounds, based on histologic examination.
  • A Diagrams of selected stages of hair follicle development.
  • B Percentage of stageable neogenic hair follicles at stage 5 or greater following administration of topical lithium gluconate hydrogel (""lithium gluconate”). 1 %, 8%, or 16% or lithium chloride hydrogel (“LiCr " ), 8%. Numbers in the bars indicate the number of mice per group that were used for quantitation. Ratios above the bars indicate the number of NHF (neogenic hair follicles) > stage 5 divided by the total number of stageable NHF.
  • lithium gluconate hydrogel lithium gluconate hydrogel
  • LiCl lithium chloride hydrogel
  • Topical lithium 8% increases both the number and maturation of neogenic hair follicles in FTE wounds, as measured following administration of topical lithium gluconate hydrogel ("lithium gluconate' " ), 1%, 8%, or 16% or lithium chloride hydrogel (“LiCl”), 8%.
  • FIG 24 No adverse systemic effects of topical lithium as indicated by equal weight gain. Weight gain profile of mice administered topical lithium gluconate hydrogel ("lithium gluconate”), 1 %, 8%, or 16%, or lithium chloride hydrogel (“LiCl”), 8%, following FTE.
  • Figure 25 Topical lithium 8% increases shaft thickness of regenerated hair follicles following DA. Tissues analyzed following administration of topical lithium gluconate hydrogel ("lithium gluconate " ), 1 %, 8%, or 16% or lithium chloride hydrogel (“LiCl”), 8%.
  • FIG. 26 Topical lithium gluconate 8% results in a 16% increase thickness of regenerated hair shafts following DA.
  • LiCF * lithium chloride hydrogel
  • Graph on right side shows simultaneous 90% confidence intervals, corrected for 4 comparisons by the Treatment Groups Bonferroni method.
  • FIG. 27 Healed FTE wounds treated with topical LiCT 8% have increased numbers of neogenic hair follicles, as assessed by in vivo scanning laser microscopy, imaging the wounded area approximately 60-80 ⁇ beneath the skin surface.
  • FIG. 29 Topical LiCl 8% increases the total number of neogenic hair follicles (also referred to as "HF") per FTE wound by 3-fold, based on histology of tissue sections.
  • Topical LiCl 8% increase the percentage of FTE wound covered with neogenic hair follicles, based on in vivo scanning confocal microscopy.
  • FIG 31 Topical LiCl 8% results in a 1.57 fold increase over placebo in coverage of the FTE wound with neogenic hair follicles.
  • Figure 32 Following FTE, topical LiCl 8%, as compared to placebo, does not affect the density of neogenic hair follicles in the region where follicles are forming.
  • Figure 34 Complexed Lithium Gluconate encapsulated within biodegradable poly (D,L-lactide-co-glycolide) ("PLG”) microspheres.
  • PLG biodegradable poly
  • Figure 36 Synthetic Biodegradable Matrices from PLA PLG Blends. A:
  • any compound or composition that can release a lithium ion (also referred to herein as lithium cation, Li+, or ionized lithium) is suitable for use in the compositions and methods.
  • Such compounds include but are not limited to a pharmaceutically acceptable prodrug, salt or solvate (e.g., a hydrate) of lithium (sometimes referred to herein as "'lithium compounds").
  • the lithium compounds can be formulated with a pharmaceutically acceptable vehicle, carrier, diluent, or excipient, or a mixture thereof.
  • lithium- polymer complexes can be utilized to developed various sustained release lithium matrices.
  • lithium is best known as a mood stabilizing drug, primarily in the treatment of bipolar disorder, for which lithium carbonate (L12CO3), sold under several trade names, is the most commonly used.
  • Other commonly used lithium salts include lithium citrate (L13C6H5O7), lithium sulfate (L12SO 4 ), lithium aspartate, and lithium orotate.
  • a lithium formulation well-suited for use in the methods disclosed herein is lithium gluconate, for example, a topical ointment of 8% lithium gluconate (LithiodermTM), is approved for the treatment of seborrheic dermatitis.
  • lithium succinate for example, an ointment comprising 8% lithium succinate, which is also used to treat seborrheic dermatitis. See, e.g., Langtry et al..
  • the lithium formulation is an ointment comprising 8% lithium succinate and 0.05% zinc sulfate (marketed in the U.K. as Efalith). See, e.g., Efalith Mullicenter Trial Group, 1992, J Am Acad Dermatol 26:452-457, which is incorporated by reference herein in its entirety.
  • Examples of lithium succinate formulations and other lithium formulations for use in the intermittent lithium treatments or pulse lithium treatment described herein are also described in U.S. Patent No. 5,594,03 1 , issued January 14, 1997. which is incorporated herein by reference in its entirety.
  • any pharmaceutically acceptable lithium salt may be used as a source of lithium ions in the intermittent lithium treatments or a pulse lithium treatment. It will be understood by one of ordinary skill in the art that pharmaceutically acceptable lithium salts are preferred. See, e.g. , Berge et al., J. Pharm. Sci. 1977, 66: 1- 19; Stahl & Wermuth, eds., 2002, Handbook of Pharmaceutical Salts, Properties, and Use. Zurich, Switzerland: Wiley- VCH and VHCA; Remington 's Pharmaceutical Sciences, 1990, 18 th eds., Easton, PA: Mack Publishing;
  • the compositions used for intermittent lithium treatment or a pulse lithium treatment comprise mixtures of one or more lithium salts.
  • a mixture of a fast-dissolving lithium salt can be mixed with a slow dissolving lithium salt proportionately to achieve the release profile.
  • the lithium salts do not comprise lithium chloride.
  • the lithium salt can be the salt form of anionic amino acids or poly(amino) acids. Examples of these are glutamic acid, aspartic acid, polyglutamic acid, polyaspartic acid.
  • lithium salts of the acids set forth above, applicants do not mean only the lithium salts prepared directly from the specifically recited acids. In contrast, applicants mean to encompass the lithium salts of the acids made by any method known to one of ordinary skill in the art, including but not limited to acid-base chemistry and cation-exchange chemistry.
  • lithium salts of anionic drugs that positively affect hair growth such as prostaglandins can be administered.
  • a large anion or multianionic polymer such as polyacrylic acid can be complexed with lithium, then complexed with a cationic compound, such as finasteride, to achieve a slow release formulation of both lithium ion and finasteride.
  • a lithium complex with a polyanion can be complexed further with the amines of minoxidil, at pHs greater than 5.
  • Lithium compounds for use in the methods provided herein may contain an acidic or basic moiety, which may also be provided as a pharmaceutically acceptable salt. See, Berge et al, J. Pharm. Sci. 1977, 66: 1-19; Stahl & Wermuth, eds., 2002, Handbook of Pharmaceutical Salts, Properties, and Use Zurich, Switzerland: Wiley-VCH and VHCA.
  • the lithium salts are organic lithium salts.
  • Organic lithium salts for use in these embodiments include lithium 2,2-dichloroacetate, lithium salts of acylated amino acids (e.g. , lithium N-acetylcysteinate or lithium N-stearoylcysteinate), a lithium salt of poly(lactic acid), a lithium salt of a polysaccharides or derivative thereof, lithium acetylsalicylate, lithium adipate, lithium hyaluronate and derivatives thereof, lithium polyacrylate and derivatives thereof, lithium chondroitin sulfate and derivatives thereof, lithium stearate, lithium linoleate, lithium oleate, lithium taurocholate, lithium cholate, lithium glycocholate, lithium deoxycholate, lithium alginate and derivatives thereof, lithium ascorbate, lithium L-aspartate, lithium benzenesulfonate, lithium benzoate, lithium 4- acetamidobenzoate, lithium
  • lithium laurate lithium (-)-L-malate, lithium maleate, lithium malonate, lithium ( ⁇ )-DL-mandelate, lithium methanesulfonate, lithium naphthalene- 2-sulfonate, lithium naphthalene- 1 ,5-disulfonate, lithium 1 -hydroxy-2-naphthoate, lithium nicotinate, lithium oleate, lithium orotate, lithium oxalate, lithium palmitate, lithium pamoate, lithium L-pyroglutamate, lithium saccharate, lithium salicylate, lithium 4-amino-salicylate, sebacic acid, lithium stearate, lithium succinate, lithium tannate, lithium (+)-L-tartarate, lithium thiocvanate, lithium p-toluenesulfonate, lithium undecylenate, or lithium valerate.
  • the organic lithium salt for use in these embodiments is lithium (S)-2- alkylthio-2-phenylacetate or lithium (R)-2-alkylthio-2-phenylacetate (e.g., wherein the alkyl is C2-C22 straight chain alkyl, preferably C8- 16). See, e.g., International Patent Application Publication No. WO 2009/019385, published February 12, 2009, which is incorporated herein by reference in its entirety.
  • the organic lithium salts used for intermittent lithium treatment or a pulse lithium treatment comprise the lithium salts of acetic acid. 2,2- dichloroacetic acid, acetylsalicylic acid, acylated amino acids, adipic acid, hyaluronic acid and derivatives thereof, polyacrylic acid and derivatives thereof, chondroitin sulfate and derivatives thereof, poly(lactic acid-co-glycolic acid), poly(lactic acid), poly(glycolic acid), pegylated lactic acid, stearic acid, linoleic acid, oleic acid, taurocholic acid, cholic acid, glycocholic acid, deoxycholic acid, alginic acid and derivatives thereof, anionic derivatives of polysaccharides, poly(sebacic anhydride)s and derivatives thereof, ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid
  • organic lithium salts for use in these embodiments is the lithium salt of (S)-2-alkylthio-2-phenylacetic acid or the lithium salt of (R)-2-alkylthio-2-phenylacetic acid (e.g., wherein the alkyl is C2- C22 straight chain alkyl, preferably C8-16). See, e.g., International Patent Application Publication No. WO 2009/019385, published February 12, 2009, which is incorporated herein by reference in its entirety.
  • the organic lithium salt can be modified to create sustained release lithium salts. Due to the size of the lithium ion, it is possible that the residence time of ion at the treatment site will be short. In efforts to generate sustained release lithium salts, the hydrophobicity of the salt can be enhanced and made "lipid-like,' " to, for example, lower the rate of ionization of the salt into lithium ions. For example, lithium chloride has a much faster rate of ionizing into lithium ions, than lithium stearate or lithium orotate. In that regard, the lithium salt can be that of a cholesterol derivative, or a long chain fatty acids or alcohols. Lipid complexed lithium salts of size less than 10 microns can also be effectively targeted to the hair follicles and "tethered" to the sebaceous glands, by hydrophobic-hydrophobic interactions.
  • the organic lithium salt can be in the form of complexes with anionic compounds or anionic poly(amino acids) and other polymers.
  • the complexes can be neutral, wherein all of the negative charges of the complexation agent are balanced by equimolar concentrations of Li ions.
  • the complexes can be negatively charged, with Lithium ions bound to an anionic polymer.
  • the complexes can be in the form of nano-complexes, or micro-complexes, small enough to be targeted to the hair follicles. If the complexes are targeted to the dermis, the charged nature of the complexes will "tether" the complexes to the positively charged collagen.
  • This mode of tethering holds the Li ions at the site of delivery, thereby hindering fast in-vivo clearance.
  • negatively charged polymers that can be used in this application are poly(acrylates) and its copolymers and derivatives thereof, hyaluronic acid and its derivatives, alginate and its derivatives, etc.
  • the anionic lithium complexes formed as described above can be further complexed with a cationic polymer such as chitosan, or polyethylimine form cell-permeable delivery systems.
  • the salt can be that of a fatty acid, e.g. , lithium stearate, thereby promoting absorption through skin tissues and extraction into the lipid compartments of the skin.
  • the lithium salt of sebacic acid can be administered to the skin for higher absorption and targeting into structures of the skin, such as hair follicles.
  • the lithium salts are inorganic lithium salts.
  • Inorganic lithium salts for use in these embodiments include halide salts, such as lithium bromide, lithium chloride, lithium fluoride, or lithium iodide.
  • the inorganic lithium salt is lithium fluoride.
  • the inorganic lithium salt is lithium iodide.
  • the lithium salts do not comprise lithium chloride.
  • Other inorganic lithium salts for use in these embodiments include lithium borate, lithium nitrate, lithium perchlorate, lithium phosphate, or lithium sulfate.
  • the inorganic lithium salts used for intermittent lithium treatment or a pulse lithium treatment comprise the lithium salts of boric acid, hydrobromic acid, hydrochloric acid, hydrofluoric acid, hydroiodic acid, nitric acid, perchloric acid, phosphoric acid, or sulfuric acid.
  • the lithium compounds used for intermittent lithium treatment or a pulse lithium treatment may be formulated with a pharmaceutically acceptable carrier (also referred to as a pharmaceutically acceptable excipients), i.e., a pharmaceutically-acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, an encapsulating material, or a complexation agent.
  • a pharmaceutically acceptable carrier also referred to as a pharmaceutically acceptable excipients
  • each component is "pharmaceutically acceptable” in the sense of being chemically compatible with the other ingredients of a pharmaceutical formulation, and biocompatible, when in contact with the biological tissues or organs of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity, or other problems or complications, commensurate with a reasonable benefit/risk ratio. See.
  • Suitable excipients are well known to those skilled in the art, and non-limiting examples of suitable excipients are provided herein. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form depends on a variety of factors well known in the art, including, but not limited to, the method of administration. For example, forms for topical administration such as a cream may contain excipients not suited for use in transdennal or intravenous administration. The suitability of a particular excipient depends on the specific active ingredients in the dosage form.
  • Exemplary, non-limiting, pharmaceutically acceptable carriers for use in the lithium formulations described herein are the cosmetically acceptable vehicles provided in
  • the lithium compounds suitable for use in intermittent lithium treatments or a pulse lithium treatment may be formulated to include an appropriate aqueous vehicle, including, but not limited to, water, saline, physiological saline or buffered saline ⁇ e.g., phosphate buffered saline (PBS)), sodium chloride for injection, Ringers for injection, isotonic dextrose for injection, sterile water for injection, dextrose lactated Ringers for injection, sodium bicarbonate, or albumin for injection.
  • PBS phosphate buffered saline
  • Suitable non-aqueous vehicles include, but are not limited to, fixed oils of vegetable origin, castor oil, corn oil, cottonseed oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, hydrogenated vegetable oils, hydrogenated soybean oil, and medium-chain triglycerides of coconut oil, lanolin oil, lanolin alcohol, linoleic acid, linolenic acid and palm seed oil.
  • Suitable water- miscible vehicles include, but are not limited to, ethanol, wool alcohol, 1 ,3-butanediol, liquid polyethylene glycol ⁇ e.g., polyethylene glycol 300 and polyethylene glycol 400), propylene glycol, glycerin, N-methyl-2-pyrrolidone ( ⁇ ), NN-dimefhylacetamide (DMA), and dimethyl sulfoxide (DMSO).
  • the water-miscible vehicle is not DMSO.
  • the lithium compounds for use in the methods disclosed herein may also be formulated with one or more of the following additional agents.
  • Suitable antimicrobial agents or preservatives include, but are not limited to, alkyl esters of p-hydroxybenzoic acid, hydantoins derivatives, propionate salts, phenols, cresols, mercurials, phenyoxyethanol, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoates, thimerosal, benzalkonium chloride ⁇ e.g., benzethonium chloride), butyl, methyl- and propyl-parabens, sorbic acid, and any of a variety of quarternary ammonium compounds.
  • Suitable isotonic agents include, but are not limited to, sodium chloride, glycerin, and dextrose.
  • Suitable buffering agents include, but are not limited to, phosphate, glutamate and citrate.
  • Suitable antioxidants are those as described herein, including ascorbate, bisulfite and sodium metabi sulfite.
  • Suitable local anesthetics include, but are not limited to, procaine
  • Suitable suspending and dispersing agents include but are not limited to sodium carboxymethylcelluose (CMC), hydroxypropyl methylcellulose (HPMC), polyvinyl alcohol (PVA), and polyvinylpyrrolidone (PVP).
  • Suitable emulsifying agents include but are not limited to, including polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate 80. and triethanolamine oleate.
  • Suitable sequestering or chelating agents include, but are not limited to, EDTA.
  • Suitable pH adjusting agents include, but are not limited to, sodium hydroxide, hydrochloric acid, citric acid, and lactic acid.
  • Suitable complexing agents include, but are not limited to, cyclodextrins, including a-cyclodextrin, ⁇ -cyclodextrin, hydroxypropyl-P-cyclodextrin, sulfobutylether-p-cyclodextrin, and sulfobutylether 7- ⁇ - cyclodextrin (CAPTISOL ® , CyDex, Lenexa, KS).
  • Soothing preparations may contain sodium bicarbonate (baking soda), and coal tar based products.
  • Formulations may also optionally contain a sunscreen or other skin protectant, or a waterproofing agent.
  • a product for application to the scalp or face may additionally be formulated so that it has easy rinsing, minimal skin/eye irritation, no damage to existing hair, has a thick and/or creamy feel, pleasant fragrance, low toxicity, good biodegradability, and a slightly acidic pi I (pi I less than 7), since a basic environment weakens the hair by breaking the disulfide bonds in hair keratin.
  • lithium gluconate - for example, 8% lithium gluconate (LithiodermTM), which is approved for the treatment of seborrheic dermatitis (see, e.g., Dreno and Moyse, 2002, Eur J Dermatol 12:549-552; Dreno et al, 2007, Ann Dermatol Venereol 134:347-351 (abstract); and Ballanger et al, 2008, Arch Dermatol Res 300:215-223, each of which is incorporated by reference herein in its entirety); 8% lithium succinate (see, e.g.
  • a preparation of lithium or lithium salt comprises an anionic polymer (such as, e.g., crosslinked polyacrylic acid), which may form a gel.
  • an anionic polymer such as, e.g., crosslinked polyacrylic acid
  • a preparation provided in the Examples of Sections 27, 30, 33, 34, or 37 below may be used.
  • the intermittent lithium treatments or a pulse lithium treatment can be provided by administration of the lithium compound (or combination treatments, discussed in Section 5.3 infra) in forms suitable for topical (e.g. , applied directly to the skin, transdermal, or intradermal), subcutaneous, intramuscular, intravenous or by other parenteral means, oral
  • the topical (e.g., applied directly to the skin, transdermal, or intradermal) administration is accomplished with the use of a mechanical device, such as, e.g., an iontophoretic device.
  • the lithium compounds (or combination treatment) can also be formulated as modified release dosage forms, including delayed-, extended-, prolonged-, sustained-, pulsatile-, controlled-, accelerated-, fast-, targeted-, programmed-release, and gastric retention dosage forms.
  • the intermittent lithium treatments or a pulse lithium treatment can be administered by a health care practitioner or by the subject. In some embodiments, the subject administers the intermittent lithium treatments or a pulse lithium treatment to him or herself.
  • topical administration is to the skin, either to the skin surface, transdermally, or intradermally. Topical administration can be with or without occlusion with a bandage or other type of dressing. In some embodiments, topical administration is to orifices or mucosa, or conjunctival, intracorneal, intraocular, ophthalmic, auricular, nasal, vaginal, urethral, respiratory, and rectal administration. The formulation used for topical administration can be designed to retain the lithium in the skin or to deliver a dose of lithium systematically. In some embodiments, topical administration of a lithium compound is combined with another treatment described herein, such as, but not limited to, a technique of integumental perturbation described in Section 5.3.3 infra.
  • Dosage forms that are suitable for topical administration for preferably local but also possible systemic effect, include emulsions, solutions, suspensions, creams, gels, hydrogels, ointments, dusting powders, dressings, elixirs, lotions, suspensions, tinctures, pastes, powders, crystals, foams, films, aerosols, irrigations, sprays, suppositories, sticks, bars, ointments, bandages, wound dressings, microdermabrasion or dermabrasion particles, drops, and transdermal or dermal patches.
  • the topical formulations can also comprise micro- and nano-sized capsules, liposomes, micelles, microspheres, microparticles, nanosystems, e.g., nanoparticles, nano-coacervates and mixtures thereof. See, e.g., International Patent Application Publication Nos. WO 2005/107710, published November 17, 2005, and WO 2005/020940, published March 10, 2005, each of which is incorporated herein by reference in its entirety.
  • the nano-sized delivery matrix is fabricated through a well- defined process, such as a process to produce lithium encapsulated in a polymer.
  • the lithium-releasing compound is spontaneously assembled in aqueous solutions, such as in liposomes and micelles.
  • the formulation for topical administration is a shampoo product, hair conditioner, hair dye, hair styling product, or skin lotion or cosmetic.
  • the selected formulation will penetrate into the skin and reach the hair follicle.
  • the stratum corneum and/or epidermis are removed by a method of integumental perturbation described herein (or by microdcrmabrasion, a less vigorous form of dermabrasion), permitting application of the dosage form for topical administration directly into the exposed dermis.
  • the formulation for topical administration will be lipid-based, so that it will penetrate the stratum corneum.
  • the formulation for topical administration will contain a skin penetrant substance, such as, e.g., propylene glycol or transcutol. See, e.g., International Patent Application Publication No.
  • a formulation in ointment form comprises one or more of the following ingredients: wool alcohol (acetylated lanolin alcohol), hard paraffin, white soft paraffin, liquid paraffin, and water. See, e.g., Langtry et ai, supra.
  • the selected formulation is inconspicuous when applied to the skin, for example, is colorless, odorless, quickly-absorbing, etc.
  • the selected formulation is applied on the skin surface as a solution, which can crosslink into a hydrogel within a few minutes, thus creating a biocompatible dressing.
  • the hydrogel may be biodegradable.
  • the solution will absorb into the skin and crosslink into depots releasing drug.
  • the lithium ion will be used to crosslink the polymer, with release of the lithium ion controlled by the rate of degradation of the hydrogel.
  • Pharmaceutically acceptable earners and excipients suitable for use in topical formulations include, but are not limited to, aqueous vehicles, water-miscible vehicles, nonaqueous vehicles, antimicrobial agents or preservatives against the growth of
  • microorganisms such as, stabilizers, solubility enhancers, isotonic agents, buffering agents, antioxidants, local anesthetics, suspending and dispersing agents, wetting or emulsifying agents, complexing agents, sequestering or chelating agents, penetration enhancers, cryoprotectants, lyoprotectants, thickening agents, and inert gases.
  • Forms for topical administration can also be in the form of ointments, creams, and gels.
  • Suitable ointment vehicles include oleaginous or hydrocarbon vehicles, including lard, benzoinated lard, olive oil, cottonseed oil, mineral oil and other oils, white petrolatum, paraffins; emulsifiable or absorption vehicles, such as hydrophilic petrolatum, hydroxysteann sulfate, and anhydrous lanolin; water-removable vehicles, such as hydrophilic ointment; water-soluble ointment vehicles, including polyethylene glycols of varying molecular weight; emulsion vehicles, either water-in-oil (W/O) emulsions or oil-in-water (O/W) emulsions, including cetyl alcohol, glyceryl monostearate, lanolin, wool alcohol (acetylated lanolin alcohol), and stearic acid (see, Remington: The Science and
  • Suitable cream base can be oil-in-water or water-in-oil.
  • Suitable cream vehicles may be water-washable, and contain an oil phase, an emulsifier, and an aqueous phase.
  • the oil phase is also called the "internal" phase, which is generally comprised of petrolatum and a fatty alcohol such as cetyl or stearyl alcohol.
  • the aqueous phase usually, although not necessarily, exceeds the oil phase in volume, and generally contains a humectant.
  • the emulsifier in a cream formulation may be a nonionic, anionic, cationic, or amphoteric surfactant.
  • Gels are semisolid, suspension-type systems. Single-phase gels contain organic macromolecules distributed substantially uniformly throughout the liquid carrier. Suitable gelling agents include, but are not limited to, crosslinked acrylic acid polymers, such as carbomers, carboxypolyalkylenes, and CARBOPOL ® ; hydrophilic polymers, such as polyethylene oxides, polyoxyethylene-polyoxypropylene copolymers, and polyvinylalcohol; cellulosic polymers, such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and methylcellulose; gums, such as tragacanth and xanthan gum; sodium alginate; and gelatin.
  • dispersing agents such as alcohol or glycerin can be added, or the gelling agent can be dispersed by trituration, mechanical mixing, and/or stirring.
  • lithium gluconate e.g., 8% lithium gluconate (LithiodermTM)
  • LithiodermTM lithium gluconate
  • topical administration is by electrical current, ultrasound, laser light, or mechanical disruption or integumental perturbation. These include
  • electroporation electroporation, RF ablation, laserporation, laser ablation (fractional or non-fractional), non- ablative use of a laser, iontophoresis, phonophoresis, sonophoresis, ultrasound poration, or using a device that accomplishes skin abrasion, or microneedle or needle-free injection, such as topical spray or POWDERJECTTM (Chiron Corp., Emeryville, CA), BIOJECTTM (Bioject Medical Technologies Inc., Tualatin, OR), or JetPeelTM (from TavTech, Tel Aviv, Israel), which uses supersonically accelerated saline to remove epidermis.
  • POWDERJECTTM Chiron Corp., Emeryville, CA
  • BIOJECTTM Bioject Medical Technologies Inc., Tualatin, OR
  • JetPeelTM from TavTech, Tel Aviv, Israel
  • the device for topical administration of lithium compounds is an automatic injection device worn continuously but delivers lithium intermittently.
  • the device for topical administration of lithium compounds is an automatic injection device that is inconspicuous, for example, can be worn without undue discomfort under clothes, in the hair, under a hairpiece, etc.
  • a device for administration of the intermittent lithium treatment or a pulse lithium treatment delivers the lithium at a controlled depth in the skin so that it reaches hair follicles, but entry into the circulation is minimized.
  • Other methods for administration of the lithium compounds described herein, used alone or in combination with other treatments described in Section 5.3.3 e.g..
  • integumental perturbation methods such as dermabrasion, laser treatment, or partial thickness or full thickness excision
  • a transdermal particle injection system such as, e.g., a "gene gun. ' '
  • Such systems typically accelerate drug or drug particles to supersonic velocities and "shoot" a narrow stream of drug through the stratum corneum.
  • the stratum corneum and epidermis is previously removed using a method of integumental perturbation described herein, and thus the required delivery pressures and velocities can be reduced. This reduction reduces the required complexity of the firing mechanisms.
  • a narrow firing stream is used, particularly to accomplish systemic delivery.
  • the particle injection system administers the lithium compound over a broad area of skin.
  • An exemplary particle delivery device compatible with broad-based skin delivery includes a low pressure / low velocity firing mechanism with a spray nozzle designed to deliver to a broad area.
  • a single- shot device that delivers to a 25-cm 2 area could be fired or used multiple times on the scalp or other skin surface until the entire area is treated.
  • a dry particle spraying mechanism similar to an airbrush or miniature grit-blaster can be used to "paint" drug or drug particles onto the perturbed area.
  • the stratum corneum and epidermis are already removed, e.g., by a method of integumental perturbation described herein, and thus permits effective use of the mechanism using lowered pressure and velocity requirements to achieve dermal delivery.
  • the lithium compound (and/or additional drug) is present in an aqueous suspension, permitting use of standard aerosol spray can technology to deliver the lithium compound to the desired skin area.
  • dermabrasion e.g., using a mechanical device, including microdermabrasion devices that can be used to dermabrade, or alumina-, silica- or ice-based dermabrasion (as described by Webber, U.S. 6,764,493; U.S. 6,726,693; and U.S. 6,306, 1 19
  • a mechanical device including microdermabrasion devices that can be used to dermabrade, or alumina-, silica- or ice-based dermabrasion (as described by Webber, U.S. 6,764,493; U.S. 6,726,693; and U.S. 6,306, 1 19
  • the device tires ablation particles at the skin, it could also fire smaller drug particles that would simultaneously embed in the exposed dermis.
  • via an internal valve control ihe device could switch over to firing dmg particles once it is determined that adequate skin disruption has occurred. See, International Patent Application Publication No. WO 2009/061349, which is incorporated herein by
  • a standard dermabrasion device can be modified to incorporate any of the devices described above, e.g., a spraying painting device.
  • a spray nozzle is located behind the dermabrasion wheel such that drug is sprayed into the dermis as it is exposed by the wheel.
  • the dermabrasion device via internal controls, could turn off the abrasion wheel once it is determined that adequate skin disruption has occurred, and switch on the drug spray to convert to drug painting mode.
  • a non-fractional C0 2 or Erbium- YAG laser is combined with drug spraying either without skin disruption, in conjunction with skin disruption, or following skin disruption.
  • a fractional non-ablative laser e.g. , an Erbium- YAG laser used at 1540-1550 nm
  • a fractional ablative laser e.g. , an Erbium- YAG laser used at 2940 nm or a C0 2 laser used at 10,600 nm
  • drug spraying either without skin perturbation, in conjunction with skin perturbation, or following skin perturbation.
  • fractional ablative laser treatment of the skin e.g., an Erbium- YAG laser used at 2940 nm or a C0 2 laser used at 10.600 nm
  • lithium compound delivery e.g., lithium compound delivery.
  • a fractional laser could be combined with a precise delivery means such that as the laser forms a hole in the skin, the inkjet-like delivery component could fill that same hole with drug.
  • adequate integrated hardware and software controls are required such that the laser ablation and drug delivery are properly timed resulting in each newly formed hole being properly filled with drug.
  • fractional ablative laser treatment of the skin e.g., an Erbium- YAG laser used at 2940 nm or a CO 2 laser used at 10,600 nm
  • lithium compound delivery e.g., lithium compound delivery.
  • a non-ablative, fractional laser could be combined with a precise delivery means such that as the laser forms a hole in the skin, the inkjet-like delivery component could fill that same hole with drug.
  • adequate integrated hardware and software controls are required such that the laser treatment and drug delivery arc properly timed resulting in each newly formed hole being properly filled with drug.
  • topical administration comprises administration of lithium- containing particles.
  • the particles can be delivered to the skin in combination with any of the means above and described elsewhere infra. Additionally, the particles can be designed for intermittent or pulse delivery of lithium. In one embodiment, particles with different release properties are be delivered simultaneously to achieve pulse delivery.
  • topical administration comprises administration of a lithium-containing formulation that is delivered through channels that are created by the use of micro-needle technology.
  • the formulation can be, e.g. , a liquid, a gel or a dry spray.
  • topical administration may be through delivery of a lithium-containing formulation through hollow needles.
  • topical administration comprises administration of a lithium-containing formulation that is delivered into the skin by an iontophoretic patch.
  • a patch can be developed in which the lithium-containing formulation is incorporated.
  • topical administration comprises administration of a lithium-containing formulation that is incorporated into micro-needle shaped biodegradable polymers.
  • the biodegradable microneedles penetrate the targeted skin tissue, and are optionally left in place to deliver the lithium ions in a sustained fashion over time.
  • Administration can be parenterally by injection, infusion, or implantation, for local or systemic administration.
  • Parenteral administration includes intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, intrasynovial, intravesical, and subcutaneous administration.
  • Compositions for parenteral administration can be formulated in any dosage forms that are suitable for parenteral administration, including solutions, suspensions, emulsions, micelles, liposomes, microspheres, nanosystems, and solid forms suitable for solutions or suspensions in liquid prior to injection.
  • compositions intended for parenteral administration can include one or more pharmaceutically acceptable carriers and excipients, including, but not limited to, aqueous vehicles, vater-miscible vehicles, non-aqueous vehicles, antimicrobial agents or preservatives against the growth of microorganisms, stabilizers, solubility enhancers, isotonic agents, buffering agents, antioxidants, local anesthetics, suspending and dispersing agents, wetting or emulsifying agents, complexing agents, sequestering or chelating agents, cryoprotectants, lyoprotectants, thickening agents, pH adjusting agents, and inert gases.
  • aqueous vehicles vater-miscible vehicles
  • non-aqueous vehicles non-aqueous vehicles
  • antimicrobial agents or preservatives against the growth of microorganisms stabilizers, solubility enhancers
  • isotonic agents buffering agents, antioxidants, local anesthetics, suspending and dispersing agents, wetting or e
  • compositions for parenteral administration can be formulated as a suspension, solid, semi-solid, or thixotropic liquid, for administration as an implanted depot.
  • the compositions are dispersed in a solid inner matrix, which is surrounded by an outer polymeric membrane that is insoluble in body fluids but allows the active ingredient in the pharmaceutical compositions diffuse through.
  • Suitable inner matrixes include, but are not limited to, polymethylmethacrylate, polybutyl-methacrylate, plasticized or unplasticized polyvinylchloride, plasticized nylon, plasticized polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene-vinyl acetate copolymers, silicone rubbers, polydimethyl siloxanes, silicone carbonate copolymers, hydrophilic polymers, such as hydrogels of esters of acrylic and methacrylic acid, collagen, cross-linked polyvinyl alcohol, and cross-linked partially hydrolyzed polyvinyl acetate.
  • Suitable outer polymeric membranes include but are not limited to, polyethylene, polypropylene, ethylene/propylene copolymers, ethylene/ethyl acrylate copolymers, ethylene/vinyl acetate copolymers, silicone rubbers, polydimethyl siloxanes, neoprene rubber, chlorinated polyethylene, polyvinylchloride, vinyl chloride copolymers with vinyl acetate, vinylidene chloride, ethylene and propylene, ionomer polyethylene terephthalate, butyl rubber epichlorohydrin rubbers, ethylene/vinyl alcohol copolymer, ethylene/vinyl acetate/vinyl alcohol terpolymer, and ethylcne/vinyloxyethanol copolymer.
  • compositions comprising lithium compounds for oral
  • oral administration can be provided in solid, semisolid, or liquid dosage forms for oral administration.
  • oral administration also includes buccal, lingual, and sublingual administration.
  • Suitable oral dosage forms include, but are not limited to, tablets, fastmelts, chewablc tablets, capsules, pills, strips, troches, lozenges, pastilles, cachets, pellets, medicated chewing gum, bulk powders, effervescent or non-effervescent powders or granules, oral mists, solutions, emulsions, suspensions, wafers, sprinkles, elixirs, and syrups.
  • the pharmaceutical compositions can contain one or more pharmaceutically acceptable carriers or excipients, including, but not limited to, binders, fillers, diluents, disintegrants, wetting agents, lubricants, glidants, coloring agents, dye-migration inhibitors, sweetening agents, flavoring agents, emulsifying agents, suspending and dispersing agents, preservatives, solvents, non-aqueous liquids, organic acids, and sources of carbon dioxide.
  • Compositions for oral administration can be also provided in the forms of liposomes, micelles, microspheres, or nanosystems. Micellar dosage forms can be prepared as described in U.S. Pat. No. 6,350,458.
  • oral formulations approved for treating mood disorders e.g., lithium carbonate (L12CO3), sold under several trade names, lithium citrate L13C H5O7), lithium sulfate (L1 2 SO 4 ), lithium aspartate, or lithium orotate, may be administered in accordance with the methods described herein.
  • lithium carbonate L12CO3
  • lithium citrate L13C H5O7 lithium citrate L13C H5O7
  • lithium sulfate L1 2 SO 4
  • lithium aspartate lithium orotate
  • modified release refers to a dosage form in which the rate or place of release of the lithium or other active ingredient(s) is different from that of an immediate dosage form when administered by the same route.
  • Modified release dosage forms include, but are not limited to, delayed-, extended-, prolonged-, sustained-, pulsatile-, controlled-, accelerated- and fast-, targeted-, programmed-release, and gastric retention dosage forms.
  • compositions in modified release dosage forms can be prepared using a variety of modified release devices and methods known to those skilled in the art, including, but not limited to, matrix controlled release devices, osmotic controlled release devices, multiparticulate controlled release devices, ion-exchange resins, enteric coatings, multilayered coatings, microspheres, liposomes, and combinations thereof.
  • the release rate of the active ingredient(s) can also be modified by varying the particle sizes and polymorphism of the active ingredient(s).
  • the controlled release is achieved by using an adjuvant that causes a depot effect, i.e., that causes an active agent or antigen to be released slowly, leading to prolonged exposure to a target cell or tissue (e.g., cells of the follicle, or, in the case of
  • Examples of formulations for modified release to skin or hair include those described in International Patent Application Publication No. WO 2008/1 15961 , published September 25, 2008, which is incorporated herein by reference in its entirety.
  • Other examples of modified release include, but are not limited to, those described in U.S. Pat. Nos.: 3.845.770; 3,916,899; 3.536,809; 3.598, 123: 4,008,719; 5.674,533; 5,059.595;
  • the modified release dosage form can be fabricated using a matrix controlled release device known to those skilled in the art. See, Takada et al, 1999, in Encyclopedia of Controlled Drug Delivery, Mathiowitz E, ed., Vol. 2, Wiley.
  • the modified release dosage form is formulated using an erodible matrix device, which is water-swellable, erodible, or soluble polymers, including, but not limited to, synthetic polymers, and naturally occurring polymers and derivatives, such as polysaccharides and proteins.
  • Materials useful in forming an erodible matrix include, but are not limited to. chitin, chitosan, dextran.
  • gum agar gum arabic, gum karaya, locust bean gum, gum tragacanth, carrageenans, gum ghatti, guar gum, xanthan gum, and scleroglucan; starches, such as dextrin and maltodextrin; hydrophilic colloids, such as pectin; phosphatides, such as lecithin; alginates; propylene glycol alginate; gelatin; collagen;
  • cellulosics such as ethyl cellulose (EC), methylethyl cellulose (MEC). carboxymethyl cellulose (CMC), CMEC, hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), cellulose acetate (CA), cellulose propionate (CP), cellulose butyrate (CB), cellulose acetate butyrate (CAB), CAP, CAT, hydroxypropyl methyl cellulose (HPMC), HPMCP, HPMCAS, hydroxypropyl methyl cellulose acetate trimellitate (HPMCAT), and ethyl hydroxyethyl cellulose (EHEC); polyvinyl pyrrolidone; polyvinyl alcohol; polyvinyl acetate; glycerol fatty acid esters; polyacrylamide; polyacryiic acid; copolymers of ethacrylic acid or methacrylic acid (EUDRAGIT ® , Rohm America, Inc., Piscataway
  • degradable lactic acid-glycolic acid copolymers poly-D-(-)-3-hydroxybutyric acid; and other acrylic acid derivatives, such as homopolymers and copolymers of butylmethacrylate, methyl methacrylate, ethyl methacrylate, ethylacrylate, (2-dimethylaminoethyi)methacrylate, and (trimethylaminoethyl)methacrylate chloride.
  • the compositions are formulated with a non-erodible matrix device.
  • the active ingredient(s) is dissolved or dispersed in an inert matrix and is released primarily by diffusion through the inert matrix once administered.
  • Materials suitable for use as a non-erodible matrix device include, but are not limited to, insoluble plastics, such as polyethylene, polypropylene, polyisoprene, polyisobutylene, polybutadiene,
  • the desired release kinetics can be controlled, for example, via the polymer type employed, the polymer viscosity, the particle sizes of the polymer and/or the active ingredient(s), the ratio of the active ingredient(s) versus the polymer, and other excipients or carriers in the compositions.
  • modified release dosage forms can be prepared by methods known to those skilled in the art, including direct compression, dry or wet granulation followed by compression, and melt-granulation followed by compression. 5.2.3.2 OSMOTIC CONTROLLED RELEASE DEVICES
  • the modified release dosage form can be fabricated using an osmotic controlled release device, including, but not limited to, one-chamber system, two-chamber system, asymmetric membrane technology (AMT), and extruding core system (ECS).
  • AMT asymmetric membrane technology
  • ECS extruding core system
  • such devices have at least two components: (a) a core which contains an active ingredient; and (b) a semipermeable membrane with at least one delivery port, which encapsulates the core.
  • the semipermeable membrane controls the influx of water to the core from an aqueous environment of use so as to cause drug release by extrusion through the delivery port(s).
  • the core of the osmotic device optionally includes an osmotic agent, which creates a driving force for transport of water from the environment of use into the core of the device.
  • an osmotic agent which creates a driving force for transport of water from the environment of use into the core of the device.
  • osmotic agents water- swellable hydrophilic polymers, which are also referred to as "osmopolymers" and
  • hydrogels Suitable water-swellable hydrophilic polymers as osmotic agents include, but are not limited to, hydrophilic vinyl and acrylic polymers, polysaccharides such as calcium alginate, polyethylene oxide (PEO), polyethylene glycol (PEG), polypropylene glycol (PPG), poly(2-hydroxyethyl methacrylate), poly(acrylic) acid, poly(methacrylic) acid,
  • hydrophilic vinyl and acrylic polymers polysaccharides such as calcium alginate, polyethylene oxide (PEO), polyethylene glycol (PEG), polypropylene glycol (PPG), poly(2-hydroxyethyl methacrylate), poly(acrylic) acid, poly(methacrylic) acid,
  • PVP polyvinylpyrrolidone
  • PVA polyvinyl alcohol
  • PVA/PVP copolymers PVA/PVP copolymers with hydrophobic monomers such as methyl methacrylate and vinyl acetate, hydrophilic polyurethanes containing large PEO blocks, sodium
  • croscarmellose hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose (HPMC), carboxymethyl cellulose (CMC) and carboxvethyi, cellulose (CEC), sodium alginate, polycarbophil, gelatin, xanthan gum, and sodium starch glycolate.
  • HEC hydroxyethyl cellulose
  • HPMC hydroxypropyl cellulose
  • HPMC hydroxypropyl methyl cellulose
  • CMC carboxymethyl cellulose
  • CEC carboxvethyi
  • sodium alginate sodium alginate
  • polycarbophil gelatin
  • gelatin xanthan gum
  • sodium starch glycolate sodium starch glycolate
  • the other class of osmotic agents is osmogens, which are capable of imbibing water to affect an osmotic pressure gradient across the barrier of the surrounding coating.
  • Suitable osmogens include, but are not limited to, inorganic salts, such as magnesium sulfate, magnesium chloride, calcium chloride, sodium chloride, lithium chloride, potassium sulfate, potassium phosphates, sodium carbonate, sodium sulfite, lithium sulfate, potassium chloride, and sodium sulfate; sugars, such as dextrose, fructose, glucose, inositol, lactose, maltose, mannitol, raffinose, sorbitol, sucrose, trehalose, and xylitol; organic acids, such as ascorbic acid, benzoic acid, fumaric acid, citric acid, maleic acid, sebacic acid, sorbic acid, adipic acid, edetic
  • Osmotic agents of different dissolution rates can be employed to influence how rapidly the active ingredient(s) is initially delivered from the dosage form.
  • amorphous sugars such as MANNOGEM EZ (SPI Pharma, Lewes, DE) can be used to provide faster delivery during the first couple of hours to promptly produce the desired therapeutic effect, and gradually and continually release of the remaining amount to maintain the desired level of therapeutic or prophylactic effect over an extended period of time.
  • the active ingredients is released at such a rate to replace the amount of the active ingredient metabolized and excreted.
  • the core can also include a wide variety of other excipients and carriers as described herein to enhance the performance of the dosage form or to promote stability or processing.
  • Materials useful in forming the semipermeable membrane include various grades of acrylics, vinyls, ethers, polyamides, polyesters, and cellulosic derivatives that are water- permeable and water-insoluble at physiologically relevant pHs, or are susceptible to being rendered water-insoluble by chemical alteration, such as crosslinking.
  • Suitable polymers useful in forming the coating include plasticized, unplasticized, and reinforced cellulose acetate (CA), cellulose diacetate, cellulose triacetate, CA propionate, cellulose nitrate, cellulose acetate butyrate (CAB), CA ethyl carbamate, CAP, CA methyl carbamate, CA succinate, cellulose acetate trimellitate (CAT), CA dimethylaminoacetate, CA ethyl carbonate, CA chloroacetate, CA ethyl oxalate, CA methyl sulfonate, CA butyl sulfonate, CA p-toluene sulfonate, agar acetate, amylose triacetate, beta glucan acetate, beta glucan triacetate, acetaldehyde dimethyl acetate, triacetate of locust bean gum, hydroxylated ethylene-vinylacetate, EC, PEG, PPG, PEG/PPG copo
  • a semipermeable membrane can also be a hydrophobic microporous membrane, wherein the pores are substantially filled with a gas and are not wetted by the aqueous medium but arc permeable to water vapor, as disclosed in U.S. Pat. No. 5,798,1 1 .
  • Such hydrophobic but water-vapor permeable membrane are typically composed of hydrophobic polymers such as polyalkenes, polyethylene, polypropylene, polytetrafluoroethylene, polyacrylic acid derivatives, polyethers, polysulfones, polyethersulfones, polystyrenes, polyvinyl halides, polyvinylidene fluoride, polyvinyl esters and ethers, natural waxes, and synthetic waxes.
  • hydrophobic polymers such as polyalkenes, polyethylene, polypropylene, polytetrafluoroethylene, polyacrylic acid derivatives, polyethers, polysulfones, polyethersulfones, polystyrenes, polyvinyl halides, polyvinylidene fluoride, polyvinyl esters and ethers, natural waxes, and synthetic waxes.
  • the deliver port(s) on the semipermeable membrane can be formed post-coating by mechanical or laser drilling. Delivery port( s) can also be formed in situ by erosion of a plug of water-soluble material or by rupture of a thinner portion of the membrane over an indentation in the core. In addition, delivery ports can be formed during coating process, as in the case of asymmetric membrane coatings of the type disclosed in U.S. Pat. Nos.
  • the total amount of the active ingredient(s) released and the release rate can substantially by modulated via the thickness and porosity of the semipermeable membrane, the composition of the core, and the number, size, and position of the delivery ports.
  • An osmotic controlled-release dosage form can further comprise additional conventional excipients or carriers as described herein to promote performance or processing of the formulation.
  • the osmotic controlled-release dosage forms can be prepared according to conventional methods and techniques known to those skilled in the art. See Remington: The Science and Practice of Pharmacy, supra; Santus and Baker, J. Controlled Release 1995, 35, 1 -21 ; Verma et ⁇ , Drug Development and Industrial Pharmacy 2000, 26, 695-708; and Verma et al. , ,1. Controlled Release 2002, 79, 7-27.
  • compositions are formulated as AMT controlled- release dosage form, which comprises an asymmetric osmotic membrane that coats a core comprising the active ingredient(s) and other pharmaceutically acceptable excipients or carriers.
  • AMT controlled-release dosage forms can be prepared according to conventional methods and techniques known to those skilled in the art, including direct compression, dry- granulation, wet granulation, and a dip-coating method.
  • compositions are formulated as ESC controlled-release dosage form, which comprises an osmotic membrane that coats a core comprising the active ingredient(s), a hydroxylethyl cellulose, and other pharmaceutically acceptable excipients or carriers.
  • the lithium-containing compound can be loaded into a polymeric solution that consists of a water-soluble polymer that is a solution at room temperature (20-25°C) and below, but gels at physiological temperatures of 32-37°C.
  • the lithium-containing solution can be cooled to 2-8°C to impart a soothing effect, while being sprayed as a liquid spray on the tissue surface. Once sprayed on, the lithium- loaded solution will thicken into a gel, releasing the lithium-containing compound slowly over time.
  • the lithium-loaded solution can be injected as a liquid, to form an in-situ depot within the tissue.
  • the lithium-loaded solution can be delivered as a solution, which can flow into orifices of the tissue, such as hair follicles and then, form a gel to release lithium for follicle-associated conditions, such as MPHL, folliculitis, or another condition described herein.
  • the temperature and time of gelation can be correlated to the concentration of the polymers and the length of the polymer blocks that constitute the polymers.
  • the a modified release dosage form can be fabricated as a multiparticulate controlled release device, which comprises a multiplicity of particles, granules, or pellets, ranging from about 10 ⁇ to about 3 mm, about 50 ⁇ to about 2.5 mm, or from about 100 ⁇ to about 1 mm in diameter.
  • Such multiparticulates can be made by the processes known to those skilled in the art, including microfluidization, membrane-controlled emulsification, oil-in-water, water-oil-water and oil-in oil emulsification and homogenization processes, complex coacervation, wet-and dry-granulation, extrusion/spheronization, roller-compaction, melt-congealing, and by spray-coating seed cores.
  • microfluidization membrane-controlled emulsification
  • compositions can be blended with the compositions to aid in processing and forming the multiparticulates.
  • the resulting particles can themselves constitute the multiparticulate device or can be coated by various film- forming materials, such as enteric polymers, water-swellable, and water-soluble polymers.
  • the multiparticulates can be further processed as a capsule or a tablet.
  • the lithium compounds for use herein may be formulated with a carrier that delivers the lithium to the site of action, for example, a follicle in a particular tissue. Such targeted delivery may be preferable in formulations for systemic administration, in order to reduce side effects associated with lithium therapy and/or ensure that the lithium reaches only follicles of particular tissues.
  • the carrier may be an aptamer targeted to a particular protein or cell type in the follicle, an antibody or antigen-binding fragment thereof, a virus, virus-like particle, virosome, liposome, micelle, microsphere, nanoparticle, or any other suitable compound.
  • compositions for use in the methods provided herein can also be formulated to be targeted to a particular tissue, follicle, or other area of the body of the subject to be treated, including liposome-, resealed erythrocyte-, and antibody-based delivery systems. Examples include, but are not limited to, those disclosed in U.S. Pat. Nos. 5,709,874; 5,759,542;
  • targeting is accomplished by the attachment of specific targeting moieties to the delivery systems containing the drug.
  • Targeting moieties can be in the form of antibodies, aptamers or small molecules that bind to specific proteins expressed in specific tissues.
  • Specific or guided targeting can "channel" the drug only to the specific tissue type, thus minimizing distribution to all tissues. This concept is especially useful if the drug causes side effects.
  • microspheres and nanospheres have been utilized, to deliver drugs into the hair follicle. Entry into the hair follicle is governed by the size of the drug-containing spheres, with microspheres of size 0.5-0.7 microns of ideal size for entry.
  • the surface of the microspheres can be functionalized with moieties that bind to specific surfaces in the follicular orifice to "retain " them at the site.
  • These moieties can be non-specific, such as hydrophobic coatings, or cationic coatings, in order to be bioadhesive to cells within the follicle.
  • the moieties can be specific and targeted to certain proteins that are expressed specifically on specific cell membranes. For example, proteins over-expressed on the follicular lymphoma cell surfaces can be targeted by delivery systems that have antibodies or aptamers designed to bind to these proteins.
  • the surface of the delivery systems can also be functionalized with cell-penetrating moieties such as cell-permeable peptides, positively charged polymers that bind to anionic cell surfaces. 5.2.3.6 LOCA DELIVERY
  • the intermittent lithium treatments or a pulse lithium treatment described herein may be delivered locally to any part of the subject in which modulation of hair growth is desired, including, e.g., the head (e.g. , the scalp, cheek, chin, upper lip, lower lip, ears, nose, eyelashes, or eyebrow), neck, abdomen, chest, breast, e.g., the nipples, back, arms, armpits (axillary hair), stomach, genital area, buttocks, legs, hands, or feet of a subject.
  • the intermittent lithium treatment or a pulse lithium treatment is applied to wounded or scarred skin.
  • Such local delivery of the intermittent lithium treatment or a pulse lithium treatment can be achieved by topical administration, transdermal, intradermal, subcutaneous (depot effect), or by intramuscular, intravenous and oral routes of delivery in formulations for targeting systemically delivered lithium to desired follicles. Such modes of delivery are discussed supra.
  • a lithium treatment described herein is delivered in combination with a pre-designed biomaterial dressing that may serve as a substrate to encourage a step-wise attachment of keratinocytes and epithelial cells to it, such that formation of an organized extra-cellular matrix (ECM) is enhanced.
  • Delivery of lithium in combination with, or via, such a "scaffold" may be of particular benefit when the lithium is administered in connection with integumental perturbation (see, e.g. , Section 5.3.4 infra), including but not limited to techniques that involve skin excision (such as full thickness excision), surgical techniques for hair transplantation, or other techniques in which the skin is wounded as described herein.
  • the scaffold for use in combination with lithium treatment may be comprised of a mesh of a biocompatible, bioabsorbable material that cells recognize and attach to, preferably with ease.
  • these materials can be collagen type I/III, hyaluronic acid, chitosan, alginates, or combinations and derivatives thereof or any other such material described herein or known in the art.
  • the mesh scaffold may be neutral, or charged. If the mesh is positively charged, it may permit cells (which are negatively charged) to adhere to it more effectively. If the mesh scaffold is negatively charged, it may contain signaling moieties that the cells will recognize and attach to. For example, polymers such as hyaluronic acid are present already in skin, and thus a mesh comprised of this material is thought to be compatible with cells.
  • the scaffold is pre-fabricated with a fine microstructure that is of the dimension of cells, for example, red blood cells that will initially diffuse throughout the scaffold, or epithelial cells and keratinocytes from surrounding tissue.
  • the mesh scaffold has an "open-cell " structure, with the pores interconnected, much like an open-celled foam. The open, interconnecting nature of the scaffold may allow free diffusion of oxygen and cells.
  • the mesh scaffold has the capacity to hydrate and remain hydrated throughout a wound healing period.
  • the mesh scaffold has moieties that act as molecular signals to the cells, for example, to aid their proliferation. These moieties include, but are not limited to,
  • peptidoglycans and RGD integrin recognition sequences that encourage cell attachment and subsequent proliferation.
  • the mesh scaffold has incorporated within it one or more additional active agents, for example, a small molecule, or a nucleic acid, or a protein, such as described in Section 5.3.4 infra.
  • the additional active agent is a protein, such as noggin or WNT, or is a nucleic acid that encodes noggin or NT.
  • a small molecule is incorporated into the scaffold, such as, e.g., a GS inhibitor. BMP inhibitor, or PPAR antagonist.
  • the compound incorporated in the mesh scaffold is a compound considered for use in the combination therapies described herein, for example, in Section 5.3.4.
  • the compounds aid in hair follicle migration or hair follicle neogenesis in the wound site.
  • the scaffold may incorporate superoxide dismutase, a free radical quenching molecule that functions in the reduction of inflammation. (00209J)
  • the lithium compound itself is incorporated within the mesh scaffold.
  • the lithium compound is incorporated within one or more layers of a multilayered mesh scaffold.
  • the mesh scaffold contains the lithium compound in alternating layers, which may achieve a pulsatile delivery of lithium.
  • the lithium compound in incorporated in microspheres in the scaffold, enabling a controlled release of lithium from the scaffold.
  • the mesh scaffold can be fibrin gels that additionally contain lithium.
  • the fibrin delivery matrix may contain lithium, fibrinogen and thrombin, that '"gels" in situ.
  • One issue that may be encountered when lithium is administered to integumental perturbed skin is the ability of lithium to diffuse through the fibrin "scab" - making the drug part of the scab may help solve this issue.
  • the mesh scaffold is a synthetic biodegradable dressing and lithium delivery system that also acts as a "sponge" and absorbs the exudates/bloods from integumental perturbed skin.
  • These exudates intercalating with the synthetic scaffold may contain an abundance of fibrinogen, thrombin, fibronectin, cell adhesion proteins, growth factors and hyaluronic acid, all of which create an integrated staicture that is an attractive matrix for cell attachment /differentiation and delivery of lithium.
  • the release rate of lithium can be modulated by varying the composition of polymers that comprise the synthetic scaffold, or sponge.
  • a synthetic scaffold fabricated out of poly(lactide)-co-(glycolide) (PLG) and poly(lactide) (PLA) can be developed to have varied release profiles of lithium. Changing the ratio of PLA to PLG will change the release profile of the lithium from the scaffold.
  • polymers that can utilized to generate synthetic scaffolds are chitosan, carregenan, alginate, poly(vinyl alcohol), poly(ethylcne oxide) (PEO), poIy(ethylene oxide)-co-poly( propylene oxide)-co-poly(ethylene oxide) (PEO-PPO-PEO), poly(acrylates) and polyvinyl pyrrolidone) (PVP).
  • the rate of lithium release from the formulation e.g., scaffold or sponge
  • the rate of lithium release from the formulation e.g., scaffold or sponge
  • the mesh scaffold releases the aforementioned compounds in a timed release manner, acting as a controlled release formulation such as described in Section 5.2.3 above.
  • the compounds may be bound to the mesh scaffold, and are then released at a sustained release manner as a result of de-binding kinetics from the mesh.
  • the compound may be bound to a polymer, which is then incorporated to the mesh scaffold, and which may allow the compound to diffuse from the mesh at a slow rate, resulting in sustained release.
  • the mesh scaffold is extruded as a gel, with certain components of the gel precipitating out to form a mesh in situ.
  • the in situ mesh can be sprayed on the integumentally perturbed surface. A large area can be covered in this manner.
  • the mesh scaffold is pre- abricated as a dressing or a wrap, to cover large areas of skin.
  • the mesh scaffold can be cut to size to fit the size of the area of the integumental perturbed skin.
  • the scaffold is prepared by melt spinning, electrospinning, micromachining, weaving, or other methods known in the art in which open cell foams are fabricated.
  • the mesh scaffold can be fabricated by these methods, with the optional incorporation of additional compound(s) (which are optionally sterilized), then sterilized by gentle ethylene oxide sterilization.
  • the additional compounds are sterilized, and then added to the sterile mesh scaffold.
  • a combinatorial strategy that uses a biodegradable scaffold combined with administration of a lithium formulation described herein (alone or in combination with another treatment, such as described in Section 5.3.4), is applied.
  • This approach may be used together with a form of integumental perturbation described in Section 5.3.4 (e.g. , dermabrasion accomplished by a standard dermabrader or a laser, or full-thickness excision accomplished by a bulk ablative laser) or integumental perturbation by wounding.
  • the scaffold is biodegradable. Placement of a 3-dimensional biodegradable scaffold on the integumentally perturbed site assists the attachment, growth and differentiation of cells.
  • tissue repair has been by autologous cell/tissue transplantation— however, autografts are associated with donor site morbidity and limited availability.
  • An alternative is allografts, but these are susceptible to immune responses and also carry the risk of disease transfer.
  • tissue engineering has emerged as an
  • tissue engineering strategy generally involves isolation of healthy cells from a patient, followed by their expansion in vitro. These expanded cells are then seeded onto three-dimensional biodegradable frameworks that provide structural support for the cells and allow cellular infiltration, attachment, proliferation and growth ultimately- leading to new tissue.
  • the biodegradability of the scaffold is modulated.
  • the biodegradability of the scaffold should be matched to the formation of the new epithelium as the integumentally perturbed skin heals.
  • One skilled in the art would know how to measure whether a synthetic matrix is biodegradable.
  • biodegradability can be measured ex vivo in implants or using rats or another animal model, by histological and HPLC analysis.
  • biodegradability by hydrolysis can be assessed.
  • the scaffold structure of choice is incubated in phosphate buffered saline, pH 7.4 and 37 °C.
  • the incubation buffer includes enzymes.
  • the scaffolds are weighed prior to incubation.
  • the scaffolds are retrieved two-at-a-time at predetermined time points and dried in a vacuum oven.
  • the scaffolds are weighed at each time point and a plot of weight versus time is generated to develop the rate of
  • the biodegradability of the scaffold matrix is modulated to coincide with the healing process, and can be modulated by changing the composition of polymers utilized to fabricate the mesh.
  • a percentage of polyethylene glycol (PEG) can be included in a composition with PLG (e.g. , described in the example in Section 37) to increase biodegradation (for example, see ASTM El 279 - 89, 2008, Standard Test Method for Biodegradation By a Shake-Flask Die-Away Method).
  • Biodegradable synthetic matrices can be created to mimic the extra-cellular micro- environment for the enhanced cellular attachment necessary for tissue regeneration.
  • cell-recognition motifs such as RGD peptides may be incorporated to encourage cells to attach themselves to the scaffold.
  • biomirnetic nature of the scaffold is judged on the basis of the content of the mesh and resultant intercalating fibrin.
  • the properties of the synthetic scaffold are dependent upon the three-dimensional geometry, matching of the modulus of the matrix with the tissue type and the porosity. It has been shown that the differentiation process can be modulated if the modulus of the tissue type is matched with the modulus of the scaffold.
  • the modulus of the scaffold is matched with the modulus of the tissue ty pe.
  • the compressive modulus of a scaffold or hydrogel can be measured by a standard Instron instrument (e.g. , using the TA Instruments DMA Q800).
  • the micro-environment created by the cells is optimally highly biocompatible to the cells present at the site, namely keratinocytes and stem cells derived from the dermal papilla.
  • this can be accomplished through the use of hydrophilic components that can absorb water.
  • hydrophobic components such as petrolatum is likely to be occlusive and prevent rapid cell proliferation.
  • the scaffold is incubated with human foreskin fibroblasts (HFF) in vitro and the scaffold is considered to be biocompatible if the cells maintain their shape and attach appropriately.
  • HFF human foreskin fibroblasts
  • the biodegradable scaffold is permeable to water, nutrients, oxygen and growth factors, enabling easy exchange of nutrients between tissues and cells (see, e.g., ASTM D39857). In some embodiments, a non-occlusive, non-permeable barrier is avoided. 5.2.4.6 COMBINED BIOLOGICAL/SYNTHETIC MESH
  • a loose, dry, highly porous network or scaffold or mesh is placed in a bleeding site of integumentally perturbed skin to gently absorb the blood and the cell adhesion proteins released at the site.
  • This will result in creation of a highly rich environment that consists of a combination of a 3-dimensional scaffold combined with fibrinogen and thrombin, which will ultimately result in a highly biocompatible hydrogel suitable for cell attachment and growth.
  • inclusion of blood components and cell adhesion proteins into the network is critical for establishment of the ECM (extracellular matrix) necessary to form continuous tissue in-growth.
  • a dry scaffold has the added advantage of absorbing the blood at the perturbed site.
  • a person's own blood components can be used to create a combined
  • the scaffold has an added advantage of serving as a blood absorbing gauze.
  • the scaffold has cell-recognition motifs, such as RGD peptides, to recruit cells to the site and attachment thereof. Once attached, cells will proliferate. Without being bound by any theory, it is hypothesized that the primary attachment of cells to the scaffold is a critical step to prevent premature cell death.
  • a dry, sterile biodegradable scaffold is placed onto the freshly integumentally perturbed (e.g. , wounded) skin.
  • the properties of the scaffold will be such that it will transform into an adherent hydrogel upon water absorption.
  • Methods that may be employed to fabricate the scaffold are known in the art, and include electrospinning, micromachining, and others. Nano-fiber meshes fabricated by electrospinning and hydrogel imprint technologies have been utilized to create three- dimensional microstructures that match the supramolecular architecture of the tissue type. In situ forming scaffolds are also contemplated.
  • the active agents are administered using an active agent-containing spray-on hydrogel.
  • the active agent after placement of the biodegradable scaffold, the active agent is sprayed on the tissue.
  • the active agent (or combination of active agents, e.g., lithium and another stem cell signaling agent) may be incorporated into a spray-on hydrogel that will be sprayed on as a liquid, but which transforms into a hydrogel after it is sprayed on the tissue. This will be especially useful if the area of the integumentally perturbed site is large and uniform coverage is needed.
  • the active agent-containing spray-on hydrogel is applied on the integumentally perturbed site, forming a cross-linked hydrogel that releases active agent over the time period of healing or a shorter or longer time period, as necessary.
  • the active agent will either be incorporated in micro-encapsulates or nano-encapsulates and suspended into the pre-hydrogel solution.
  • the active agent can also be dissolved into the pre-hydrogel solution.
  • the "pre-hydrogel' " solution is defined as the solution that will be sprayed on the tissue and which also contains the active agent.
  • the active agent is contained within microspheres that can be positively charged to rapidly bind themselves to the negatively charged collagen present in the dermis. Binding the microspheres to the dermis renders the active agent-releasing moiety immobile at the site.
  • the integumentally perturbed site may be covered with a breathable, non-occlusive spray-on hydrogel to cover the site during healing.
  • the lithium compound or formulation thereof can be administered topically, subcutaneously, orally, etc.
  • the dosing regimen should be adjusted to achieve peak concentrations of lithium in the target skin area of at least about 0.1 mM to 10 mM, and/or peak concentrations of lithium in the blood (serum or plasma samples) of at least about 1 mM (these values are sometimes referred to herein as the ''target concentration").
  • concentrations of lithium including its concentration in formulations, in tissue, in serum, etc. , and as a salt form, as an ionized atom in solution, etc.
  • ionized lithium is a monovalent cation
  • the peak concentration of lithium can be established by taking samples when peak concentrations are achieved and assaying them for lithium content using techniques well known to those skilled in the art (see. e.g.
  • samples can be taken when peak blood concentrations are typically achieved - for example, within 1 to 2 hours for standard release formulations, and 4-5 hours for sustained release formulations.
  • the peak concentration times for other formulations, including topical preparations, can be determined for the particular formulation used, and sampling can be adjusted accordingly.
  • the target concentration of lithium should be maintained in the skin and/or blood for at least 1 day; at least 2 days; at least 3 days; at least 5 days; at least 14 days; or at least 21 days; and, in certain embodiments, not more than 21 days.
  • This can be accomplished using, e.g., repeated applications of the lithium compound or a single application of a sustained release or extended release lithium formulation.
  • Either the single pulse protocol or the intermittent treatments can be used to achieve the target concentration of lithium for the shorter maintenance periods (i.e., for at least 1, 2 or 3 days). Maintenance periods longer than 3 days may require repeated application of the intermittent lithium treatments or the single pulse protocol.
  • topical administration of a lithium compound is preferred over oral or subcutaneous administration.
  • a topically administered lithium compound may achieve a higher concentration of lithium in skin than in the blood, thereby reducing the risk of toxicity associated with elevated blood levels of lithium.
  • a subcutaneously or orally administered lithium compound may be preferred in order to achieve a controlled release of lithium from the blood to the skin.
  • lithium doses should be adjusted on the basis of the blood concentration (serum or plasma) drawn (by convention) 12 or 24 hours after the last dose of the lithium compound; this trough blood concentration should be maintained below 2 mM Li+ and preferably, below about 1.5 mM Li+. In some embodiments, the steady state blood concentration of lithium should not exceed a maximum of 1.5 mM to 2 mM.
  • a trough concentration of lithium in the skin of no less than 0.01 mM to 0.05 mM is preferred. In some embodiments, a trough concentration of lithium in the skin of 0.05 mM to 0.1 mM is preferred. In some embodiments, a trough concentration of lithium in the skin of less than 1 mM is preferred. In some embodiments, a trough concentration of lithium in the skin of less than 3 mM is preferred. In some embodiments, lithium concentrations at trough can be increased by twice daily dosing, or more frequent dosing. In such embodiments, topical administration of a lithium compound is preferred.
  • a pulsatile effect is achieved by the multiple dosing, but the trough concentrations do not decline as much as when once daily dosing is used.
  • a trough skin concentration of lithium is maintained at 0.25 mM or higher, for example from 0.25 mM to 0.5 mM or 0.5 mM to 0.75 mM.
  • the trough concentration is maintained at approximately 0.6 mM to 1.4 mM lithium.
  • a trough skin concentration is maintained at 1 mM to 3 mM lithium. In some such
  • the trough skin concentration is maintained at less than 0.5 mM, or less than 0.75 mM, or less than 1 mM, or less than 2 mM, or less than 3 mM of lithium.
  • an effective amount of a lithium compound is administered such that the target concentration of lithium ions in plasma or serum, as measured 30 minutes to 1 hour after the lithium treatment, is 0.10-0.20 ⁇ , 0.20-0.50 ⁇ , 0.50- 1.0 ⁇ , 1 .0-5.0 ⁇ , 5.0-10 ⁇ , 10-20 ⁇ , 20-50 ⁇ , 50- 100 ⁇ , 100-500 ⁇ , 0.1- 0.5 mM, 0.5- 1 .0 mM. 1.0 mM-2.0 mM, 2.0-2.5 mM, 2.5-3.0 mM, 3.0-4.0 mM. 4.0 mM-5.0 mM, 5.0-7.0 mM, or 7.0 mM or greater.
  • an effective amount of lithium is administered such that the plasma or serum lithium ion concentration measured either 8 hours, 16 hours, 1 day, 1 week, 2 weeks, or 1 month after the lithium treatment, is 0.1 to 0.5 ⁇ , 0.1 to 1.0 ⁇ , 0.5 to 1 .0 ⁇ , 0.5 to 1.5 ⁇ , 1 to 10 ⁇ , 10 to 50 ⁇ , 50 to 100 ⁇ , 100 to 150 ⁇ , 150 to 200 ⁇ , 250 to 300 ⁇ , 100 to 250 ⁇ , 100 to 500 ⁇ , 200 to 400 ⁇ , 500 to 1000 ⁇ ; or 1000 to less than 1500 ⁇ .
  • the plasma or serum lithium concentration reaches at least 1 ⁇ . In one embodiment, the plasma or serum lithium concentration reaches at least 100 ⁇ .
  • the plasma or serum lithium concentration reaches at least 1 mM. In one embodiment, the plasma or serum lithium concentration does not exceed 1 mM. In another embodiments, the plasma or serum concentration of lithium does not exceed 1.5 mM. Serum lithium concentration may be measured using any technique known in the art, such as described in Sampson et al, 1992, Trace Elements in Medicine 9:7-8.
  • an amount of a lithium compound is administered such that the target concentration of lithium in the skin is 0.01 to 0.05 ⁇ , 0.05 to 0.1 ⁇ , 0.1 to 0.5 ⁇ , 0.1 to 1 ⁇ .
  • the concentration of lithium achieved in the skin is greater than 0.1 mM. In some embodiments, the concentration of lithium achieved in the skin is greater than 1.0 mM. In some embodiments, the concentration of lithium achieved in the skin is greater than 1.5 mM. In one embodiment, the amount of lithium achieved in the skin is approximately 1 mM to 5 mM. In one embodiment, the amount of lithium achieved in the skin is approximately 5 mM to 10 mM. In one embodiment, the amount of lithium achieved in the skin is approximately 100 to 200 mM. In one embodiment, the amount of lithium achieved in the skin does not exceed 5 mM. In one embodiment, the amount of lithium achieved in the skin does not exceed 10 mM.
  • the amount of lithium achieved in the skin does not exceed 50 mM.
  • an amount of lithium is administered such that the concentration of lithium delivered to the stratum comeum is 0.1 to 0.5 mM, 0.5 to 1 mM, 1 to 10 mM, 10 to 100 mM, 100 to 200 mM, or 500 to 1000 mM.
  • the concentration of lithium delivered to the stratum corneum is greater than 1.5 mM.
  • the amount of lithium achieved in the stratum corneum is approximately 100 to 200 mM.
  • the amount of lithium achieved in the stratum corneum does not exceed 5 mM.
  • the amount of lithium achieved in the stratum corneum does not exceed 10 mM.
  • lithium concentrations in skin using techniques known in the art, for example, mass spectroscopy, e.g. , inductively coupled plasma mass spectroscopy (ICP-MS).
  • ICP-MS inductively coupled plasma mass spectroscopy
  • the concentration of lithium in skin can be measured using the method provided in the example of Section 29.2 below or equivalent methods.
  • the lithium concentration is measured in the hair shaft using techniques known in the art, e.g., Tsanaclis & Wicks, 2007. Forensic Science Intl. 176: 19-22, which is incorporated by reference herein in its entirety.
  • lithium can be applied topically, e.g. , as a cream, gel, ointment, or other form for topical administration as described in Section 5.2.2.1 and 5.3 supra.
  • Topical lithium may be administered to wounded or unwounded skin.
  • the lithium formulation for topical administration (e.g. , gel, cream, ointment, salve, etc.) comprises lithium (or monovalent lithium salt) at a concentration of 50 mM, 75 mM, 100 raM, 125 mM, 150 mM, 175 mM, 200 mM, 250 mM, 300 niM, 350 mM, 400 mM, 450 mM, 500 mM, 550 mM, 600 mM, 650 mM, 700 mM, 750 mM, 800 mM, 900 mM, 1 M, 1.1 M. or 1.2 M, or more.
  • a monovalent lithium salt e.g. , lithium gluconate, lithium chloride, lithium stearate, lithium orotate, etc.
  • a divalent lithium salt e.g. , in some embodiments, lithium succinate, lithium carbonate
  • a trivalent lithium salt e.g., in some embodiments, lithium citrate
  • a lithium formulation comprising lithium (or monovalent lithium salt) at a concentration in the range of 50 mM to 200 mM is chosen for use in the embodiments described herein.
  • a lithium formulation comprising lithium (or monovalent lithium salt) at a concentration in the range of 200 mM to 400 mM is used.
  • a lithium formulation comprising lithium (or monovalent lithium salt) at a concentration in the range of 400 mM to 600 mM is used.
  • a lithium formulation comprising lithium (or monovalent lithium salt) at a concentration in the range of 600 mM to 800 mM is used.
  • concentration of lithium in a particular topical lithium formulation to deliver the intended dose of lithium will depend on the release properties of the lithium ion, the hydrophobicity of the lithium salt form, the partition coefficient of the lithium salt form, etc.
  • Lithium formulations comprising the foregoing lithium (or monovalent lithium salt) concentrations may be achieved using, for example, a formulation comprising, w/w. lithium ions at a concentration of 0. 10% lithium, 0.15% lithium, 0.20% lithium, 0.25% lithium. 0.30% lithium. 0.35% lithium, 0.40% lithium, 0.45% lithium, 0.50% lithium, 0.55% lithium, 0.60% lithium, 0.65% lithium, 0.70% lithium, 0.75% lithium, 0.80% lithium, 0.85% lithium, 0.90% lithium, 0.95% lithium.
  • the form of lithium for topical administration comprises, w/w, 0.1% to 0.5% lithium ions, 0.2% to 0.5% lithium ions, 0.5% to 1 % lithium ions, or more.
  • the amount of a salt form of lithium to generate a topical lithium formulation with one of the aforementioned concentrations of lithium ion is readily deducible by one of ordinary skill in the art, and depends upon several factors including, e.g., the valency of the salt form, the stability of the salt form, the ability of the salt form to release the lithium ion, the hydrophobicity or hydrophilicity, etc.
  • Lithioderm (Labcatal) comprises 8% lithium gluconate, which corresponds to 0.275% lithium ion (i.e. , 274.8 mg Li+/100 g gel).
  • a formulation for topical administration comprises a salt form of lithium (e.g. , lithium gluconate or other form described in Section 5.1 above) at a concentration, w/w, of 1 %, 2%, 3%, 4%, 5%, 6%. 7%, 8%. 9%, 10%, 12%. 15%, 16%, 18%. 20%, or more.
  • a salt form of lithium e.g. , lithium gluconate or other form described in Section 5.1 above
  • a salt form of lithium for topical administration comprises, w/w, 1 % to 2% lithium salt (e.g., lithium gluconate or other form described in Section 5.1 above). 2% to 5% lithium salt, 5% to 10% lithium salt, 10% to 15% lithium salt, 15% to 20% lithium salt, 20% to 25% lithium salt, or 25% to 50% lithium salt. In one embodiment, the form of lithium for topical administration is 1 % to 20% w/w lithium salt.
  • a topical formulation of lithium comprises l%-4% lithium gluconate (w/w). In some embodiments, a topical formulation of lithium comprises 4%-8% lithium gluconate (w/w). In some embodiments, a topical formulation of lithium comprises 8%- 16% or more lithium gluconate (w/w). In some embodiments, a topical formulation of lithium comprises 0.2%- 1%, or l %-5%, or more lithium chloride (w/w). In some
  • a topical formulation of lithium comprises 0.5%-2%, or 2%-4%, or 4%-8%, or 8%-16, or more lithium succinate (w/w). In some embodiments, a topical formulation of lithium comprises 0.5%-6%, 6%-12%, or 12%-25%, or more lithium stearate (w/w). In some embodiments, a topical formulation of lithium comprises l %-4%, 4%-8%, or 8%- 16%, or more lithium orotate (w/w). In some embodiments, a topical formulation of lithium comprises 0.25%-0.75%, 0.75%- 1.5%, or 1.5%-3%, or more lithium carbonate (w/w). In some embodiments, a topical formulation of lithium comprises 0.25%- 1.5%, 1.5%-3.0%, or 3%-6%, or more 8% lithium citrate (w/w).
  • a 50 kg patient is administered a single droplet - approximately 0.1 ml - of 8% (w/w) lithium gluconate at 3 sites, twice daily. This corresponds to approximately 8 mg lithium gluconate (0.274 mg Li+) per site, i.e., 0.16 mg/kg lithium gluconate (0.005 mg/kg Li+) per site. Over three sites twice daily, this corresponds to approximately 0.96 mg/kg lithium gluconate (0.033 mg/kg Li+) per day.
  • a patient e.g., a 50 kg patient
  • a topical lithium formulation is administered once daily.
  • a topical lithium formulation is administered twice daily.
  • doses are administered 6 hours apart, or 7 hours apart, or 8 hours apart, or 9 hours apart, or 10 hours apart, or 1 1 hours apart, or 12 hours apart. In a particular embodiment, the doses are administered 7 to 8 hours apart.
  • an amount of lithium is administered such that the peak lithium concentration in skin is between 0.01 mM and 0.05 mM, 0.05 mM and 0.1 mM, 0.1 mM and 0.5 mM or between 0.5 mM and 10 mM, for example, between 0.1 and 0.5 mM, 0.5 mM and 1 mM, 1 mM and 2 mM, between 2 mM and 5 mM, 5 mM to 10 mM, or 10 mM to 50 mM.
  • the peak lithium concentration in blood may be one or more orders of magnitude lower than the peak concentration in skin (for example, 0.001 mM to 0.01 mM, 0.01 mM to 0.1 mM, or 0.1 mM to 0.5 mM, 0.5 mM to 1.0 mM, or 1.0 mM to 10 mM).
  • the steady state blood concentration of lithium should not exceed a maximum of 1.5 mM to 2 mM.
  • a formulation of lithium described herein (by non-limiting e.g. , lithium gluconate, lithium chloride, lithium succinate, lithium carbonate, lithium citrate, lithium stearate, lithium orotate, etc. ) is administered subcutaneously, to either wounded or unwounded skin.
  • the form of lithium for subcutaneous administration is administered at a dose comprising 0.001 mg lithium ion per kg of patient weight.
  • the dose is 0.001 mg/kg, 0.002 mg kg, 0.003 mg/kg, 0.004 mg/kg, 0.005 mg/kg, 0.006 mg kg, 0.007 mg/kg, 0.008 mg/kg, 0.009 mg/kg, 0.010 mg kg, 0.020 mg kg, 0.025 mg/kg, 0.050 mg/kg, 0.075 mg/kg, 0.10 mg/kg, 0.15 mg/kg, 0.20 mg/kg, 0.25 mg/kg.
  • the lower ranges of dosages may be preferably used for bolus dosing.
  • a controlled release e.g., a delayed release or a sustained release
  • the maximum dosage that may be administered at any one time may vary depending on the release kinetics of the lithium and the concentration of efficacy of the formulation.
  • concentration of a salt form of lithium required to generate a subcutaneously administered formulation that delivers lithium ions at one of the aforementioned dosages is readily deducible by one of ordinary skill in the art, and depends upon several factors including, e.g. , the valency of the salt form, the stability of the salt form, the ability of the salt form to release the lithium ion, the hydrophobicity or hydrophilicity, etc.
  • a formulation comprising lithium gluconate may be subcutaneously administered at a dosage of approximately 10 mg lithium gluconate per kg of patient weight (mg/kg), 20 mg/kg, 25 mg/kg, 30 mg/kg, 35 mg/kg, 40 mg kg, 50 mg/kg, 75 mg/kg, 100 mg/kg, 125 mg/kg, 150 mg/kg, 175 mg/kg, 200 mg/kg.
  • the formulation for subcutaneous administration contains a dose of 10 mg/kg to 50 mg/kg, 50 mg/kg to 100 mg/kg, 100 mg/kg to 200 mg/kg, 200 mg/kg to 400 mg kg, 400 mg/kg to 600 mg/kg. or 100 mg/kg to 600 mg / kg of lithium gluconate.
  • the formulation for subcutaneous administration contains a dose in the range of 30 mg/kg to 150 mg/kg lithium gluconate. In one embodiment, the formulation for subcutaneous administration contains a dose in the range of about 30 mg/kg to 300 mg/kg lithium gluconate. In one embodiment, the dose for subcutaneous administration does not exceed 300 mg/kg lithium gluconate. In another embodiment, the dose for subcutaneous administration does not exceed 600 mg kg lithium gluconate.
  • the lower ranges of dosages may be preferably used for bolus dosing.
  • the maximum dosage that may be administered at any one time may vary depending on the release kinetics of the lithium and the concentration of efficacy of the formulation.
  • the lithium formulation is administered subcutaneously once daily. In some embodiments, the lithium formulation is administered subcutaneously twice daily, in some embodiments of a twice daily treatment regimen, doses are administered 6 hours apart, or 7 hours apart, or 8 hours apart, or 9 hours apart, or 10 hours apart, or 1 1 hours apart, or 12 hours apart. In a particular embodiment, the doses are administered 7 to 8 hours apart.
  • an amount of lithium is administered such that the peak lithium concentration in skin is between 0.1 ⁇ and 0.2 ⁇ , 0.2 ⁇ and 0.5 ⁇ , 0.5 and 1 ⁇ , 1 ⁇ and 2 ⁇ , 2 ⁇ to 10 ⁇ , 10 ⁇ to 100 ⁇ , 100 ⁇ to 500 ⁇ , 500 ⁇ to 1000 ⁇ .
  • peak values will depend on the lithium release properties of the formulation, the hydrophobicity of the lithium salt form, the partition coefficient of the lithium salt form, etc.
  • the concentration in skin is 0.2 ⁇ to 1.5 ⁇ lithium. In some embodiments, the peak concentration in skin should not exceed 1 ⁇ or 1.5 ⁇ lithium. In some embodiments, the peak concentration in skin is 10 ⁇ to 100 ⁇ lithium. In some embodiments, the peak concentration in skin is 100 ⁇ to 1000 ⁇ lithium. In some such embodiments, the peak lithium concentration in blood may be several orders of magnitude higher, for example, 0.1 mM to 0.5 mM, or 0.5 mM to 1.1 mM, 1.1 to 1.5 mM, 1.5 mM to 5 mM, 5 mM to 10 mM, 10 mM to 50 mM, or 50 mM to 100 mM.
  • the steady state blood concentration of lithium should not exceed a maximum of 1.5 mM to 2 mM.
  • a formulation of lithium described herein (by non-limiting e.g. , lithium gluconate, lithium chloride, lithium succinate, lithium carbonate, lithium citrate, lithium stearate, lithium orotate, etc. ) is administered orally, for example, once daily, or twice daily as determined by the medical practitioner and in accordance with Section 5.3.1 above.
  • an oral formulation comprising of 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 1.1 mM, 1.2 mM, 1.3 mM, 1.4 mM, 1.5 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, or more, but preferably less than 10 mM, of lithium ions (or monovalent lithium salt) is administered.
  • an oral formulation comprising lithium ions or a monovalent lithium salt in the range of 0.1 to 0.5 mM, 0.4 to 0.6 mM. 0.5 to 1 mM, 0.6 to 1.2 mM, or 1 to 1 .5 mM, is administered.
  • Administration of the foregoing amounts of lithium may be achieved by oral administration of a lithium formulation at a dosage comprising 0.001 mg lithium ion per kg of patient weight.
  • the dose is 0.001 mg/kg, 0.002 mg/kg, 0.003 mg/kg, 0.004 mg kg, 0.005 mg/kg, 0.006 mg/kg, 0.007 mg/kg, 0.008 mg kg, 0.009 mg/kg, 0.010 mg/ kg, 0.020 mg/kg, 0.025 mg/kg, 0.050 mg/kg, 0.075 mg/kg, 0.10 mg/kg, 0.15 mg/kg, 0.20 mg/kg, 0.25 mg/kg, 0.30 mg kg, 0.40 mg/kg, 0.50 mg/kg, 0.75 mg/kg, 1 mg/kg, 1 .5 mg/kg, 2 mg kg, 2.5 mg/kg, 3 mg/kg, 3.5 mg/kg, 4 mg/kg, 4.5 mg/kg, 5 mg/kg, 5.5 mg/kg, 6 mg/kg
  • the dose does not exceed 50 mg/kg Li+.
  • the maximum dosage that may be administered at any one time may vary depending on the release kinetics of the lithium and the concentration of efficacy of the formulation.
  • the concentration of a salt form of lithium required to generate an orally administered formulation that delivers lithium ions at one of the aforementioned dosages is readily deducible by one of ordinary skill in the art, and depends upon several factors including, e.g. , the valency of the salt form, the stability of the salt form, the ability of the salt form to release the lithium ion, the hydrophobicity or hydrophilicity, etc.
  • a formulation comprising lithium carbonate, which is a divalent lithium salt e.g.
  • Ekalith CR trade names Eskalith CR, Eskalith, Lithobid
  • Eskalith CR trade names Eskalith CR, Eskalith, Lithobid
  • the oral formulation contains a dose of 2 mg/kg to 10 mg/kg, 10 mg/kg to 25 mg/kg, 25 mg/kg to 50 mg/kg, 50 mg/kg to 100 mg/kg, 100 mg kg to 200 mg/kg, or 200 mg/kg to 500 mg/kg of lithium carbonate. In one embodiment, the oral formulation contains a dose in the range of 5 mg/kg to 100 mg/kg lithium carbonate. In one embodiment, the oral formulation contains a dose in the range of about 5 mg/kg to 50 mg/kg lithium carbonate. In one embodiment, the oral formulation contains a dose in the range of about 10 mg/kg to 100 mg/kg lithium carbonate. In one embodiment, the oral formulation contains a dose that does not exceed 300 mg/kg lithium carbonate.
  • the maximum dosage that may be administered at any one time may vary depending on the release kinetics of the lithium and the concentration of efficacy of the formulation.
  • an amount of lithium compound is administered such that the peak lithium concentration in skin is between 0.1 ⁇ and 0.2 ⁇ , 0.2 ⁇ and 0.5 ⁇ , 0.5 and l ⁇ , 1 ⁇ and 2 ⁇ , 2 ⁇ to 10 ⁇ , 10 ⁇ to 100 ⁇ , 100 ⁇ to 500 ⁇ , 500 ⁇ to 1000 ⁇ .
  • peak values will depend on the lithium release properties of the formulation, the hydrophobicity of the lithium salt form, the partition coefficient of the lithium salt form, etc.
  • the peak concentration in skin is 0.2 ⁇ to 1.5 ⁇ lithium.
  • the peak concentration in skin should not exceed 1 ⁇ or 1 .5 ⁇ lithium. In some embodiments, the peak concentration in skin is 10 ⁇ to 100 ⁇ lithium. In some embodiments, the peak concentration in skin is 100 ⁇ to 1000 ⁇ lithium. In some such embodiments, the peak lithium concentration in blood may be several orders of magnitude higher, for example, 0.1 mM to 0.5 mM, or 0.5 mM to 1.1 mM, 1.1 to 1.5 mM, 1.5 mM to 5 mM. 5 mM to 10 mM, 10 mM to 50 mM, or 50 mM to 100 mM.
  • the steady state blood concentration of lithium should not exceed a maximum of 1.5 mM to 2 mM.
  • the pulse lithium treatment can be administered one time, or multiple times at intervals of time. It is understood that the precise dosage and duration of treatment may v ary with the age, weight, and condition of the patient being treated, and may be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test or diagnostic data. It is further understood that for any particular individual, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the formulations.
  • a pulse lithium treatment is administered at the time of integumental perturbation.
  • a pulse lithium treatment is administered following integumental perturbation.
  • the pulse lithium treatment is administered before scab formation.
  • a pulse lithium treatment is administered following an integumental perturbation that leads to formation of a scab
  • the pulse lithium treatment is administered during scab formation.
  • the pulse lithium treatment is administered periscab detachment.
  • the pulse lithium treatment is administered immediately after scab detachment.
  • the pulse lithium treatment is administered 1 hour after scab detachment.
  • the pulse lithium treatment in which a pulse lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the pulse lithium treatment is administered up to 6 hours after scab detachment. In one embodiment, in which a pulse lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the pulse lithium treatment is administered 6- 12 hours after scab detachment. In one embodiment, in which a pulse lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the pulse lithium treatment is administered 12- 18 hours after scab detachment.
  • the pulse lithium treatment in which a pulse lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the pulse lithium treatment is administered 18- 24 hours after scab detachment. In one embodiment, in which a pulse lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the pulse lithium treatment is administered 1 day after scab detachment. In one embodiment, in which a pulse lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the pulse lithium treatment is administered 2 days after scab detachment.
  • the pulse lithium treatment in which a pulse lithium treatment is administered following an integumental perturbation that leads to formation of a scab, is administered 3 days after scab detachment. In some embodiments, in which a pulse lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the pulse lithium treatment is administered within 3 days. 5 days, 7 days, 10 days, 2 weeks, or 3 weeks after integumental perturbation.
  • the pulse lithium treatment is administered at the time of integumental perturbation and then maintained for 3 or 4 or 5 days thereafter (in some embodiments, a scab forms during this time). In some embodiments, a pulse lithium treatment is administered as soon as the scab falls of and maintained for 3 or 4 or 5 days. In some embodiments, the pulse lithium treatment is administered in order to modulate the neoepidermis that forms underneath the scab. In some such embodiments, the pulse lithium treatment is administered at the time of integumental perturbation and is maintained up to some time after scab falls off, for example, between 5 - 14 days following integumental perturbation.
  • the course of treatment with lithium is short, for example, limited to a few days just following scab detachment, or even continued only for as long as the scab is still attached.
  • the timing of integumental perturbation and lithium administration is preferably monitored and adjusted so that optimal results are achieved.
  • a pulse treatment is combined with a form of integumental perturbation that does not lead to formation of a scab.
  • the pulse lithium treatment is administered at the time of integumental perturbation.
  • a pulse lithium treatment is administered following integumental perturbation.
  • the pulse lithium treatment is administered within 15 minutes of, or 15 minutes, 30 minutes, 45 minutes, 1 hour. 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, 18 hours, 1 day, 2 days, 3 days, 5 days, 7 days. 10 days, 2 weeks, or 3 weeks after integumental perturbation.
  • the intermittent lithium treatment can be administered one time (e.g. , using a controlled release formulation), or multiple times at intervals of time. It is understood that the precise dosage and duration of treatment may vary with the age, weight, and condition of the patient being treated, and may be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test or diagnostic data. It is further understood that for any particular individual, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the formulations.
  • lithium can be administered daily (e.g. , once, twice or three times daily) for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 7 days: and in some embodiments not more than 14 days.
  • Holidays can be interspersed for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 7 days; and in some embodiments not more than 14 days.
  • an intermittent lithium treatment is begun at the time of integumental perturbation. In some embodiments, an intermittent lithium treatment is begun following integumental perturbation. In one embodiment, in which an intermittent lithium treatment is begun following an integumental perturbation that leads to formation of a scab, the intermittent lithium treatment is begun before scab formation. In one embodiment, in which an intermittent lithium treatment is begun following an integumental perturbation that leads to formation of a scab, the intermittent lithium treatment is begun during scab formation. In one embodiment, in which an intermittent lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the first
  • administration of lithium in the intermittent lithium treatment is periscab detacliment.
  • the intermittent lithium treatment is administered following an integumental perturbation that leads to fonnation of a scab
  • the first administration of lithium is immediately after scab detachment.
  • the intermittent lithium treatment is administered following an integumental perturbation that leads to formation of a scab
  • the first administration of lithium is up to 6 hours after scab detachment.
  • the intermittent lithium treatment is administered following an integumental perturbation that leads to formation of a scab
  • the first administration of lithium is 6- 12 hours after scab detachment.
  • the first administration of lithium is 12- 18 hours after scab detachment. In one embodiment, in which the intermittent lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the first administration of lithium is 1 8-24 hours after scab detachment. In one embodiment, in which the intermittent lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the first administration of lithium is I day after scab detachment.
  • the first administration of lithium is 2 days after scab detachment. In one embodiment, in which the intermittent lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the first administration of lithium is 3 days after scab detachment. In one embodiment, in which the intermittent lithium treatment is administered following an integumental perturbation that leads to formation of a scab, the first administration of lithium is administered immediately after scab detachment, followed by another administration each day for several days to 1 week.
  • the pulse lithium treatment is begun within 3 days, 5 days, 7 days, 10 days, 2 weeks, or 3 weeks after integumental perturbation.
  • the intermittent lithium treatment is begun at the time of integumental perturbation and then administered daily (or twice daily) for 5 days thereafter (in some embodiments, a scab forms during this time). In some embodiments, the intermittent lithium treatment is begun as soon as the scab falls off, and administered daily for 5 days. In some embodiments, the intermittent lithium treatment is to modulate the neoepidermis that forms underneath the scab. In some such embodiments, the intermittent lithium treatment is begun at the time of integumental perturbation and is continued with daily dosing up to some time after scab falls off, for example, between 5 - 14 days following integumental perturbation.
  • the course of treatment with lithium is short, for example, limited to daily doses for a few days just following scab detachment, or even continued only for as long as the scab is still attached.
  • the timing of integumental perturbation and lithium administration is preferably monitored and adjusted so that optimal results are achieved.
  • an intermittent lithium treatment is combined with a form of integumental perturbation that does not lead to formation of a scab.
  • the intermittent lithium treatment is begun at the time of integumental perturbation.
  • an intermittent lithium treatment is begun following integumental perturbation.
  • the intermittent lithium treatment is begun within 15 minutes of, or 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, 18 hours, 1 day, 2 days, 3 days, 5 days, 7 days, 10 days, 2 weeks, or 3 weeks after integumental perturbation.
  • intermittent lithium treatment or a pulse lithium treatment in combination with conventional methods for enhancing hair growth or removal of unwanted hair enhances the effectiveness of these methods.
  • the effect that each drug offers could be an additive or synergistic improvement, or a combination of two different pharmacologically defined effects, to achieve the desired end result.
  • Most drugs for hair loss aim to retain the existing hair follicles in their active cycling states, or to rejuvenate telogen hair follicles to actively cycling anagen states.
  • a dosage form that encourages the growth of "new" hair follicles, combined with one that retains the hair follicles in their actively cycling states offer significant value to the individual who is balding.
  • the combined modality of treatment could involve alternating treatment of each dosage form or concurrent or simultaneous treatment.
  • a reason for this enhancing effect, or synergism is that intermittent lithium treatment or a pulse lithium treatment synchronizes the hair/Follicle Cycle. Since all the follicles are at the same stage in their growth cycle, a treatment intended to enhance hair growth or remove hair will be more effective, efficient, cost-effective, and user friendly. For example, fewer treatments may be required.
  • the intermittent lithium treatments or the pulse lithium treatment described herein may be in combination with any additional treatment(s) described or incorporated by reference herein or determined to be appropriate by the medical practitioner.
  • the amount of an additional treatment(s) will depend on the desired effect and the additional compound that is selected. Dosages and regimens for administering such additional treatment(s) are the dosages and regimens commonly in use, which can be easily determined by consulting, for example, product labels or physicians' guides, such as the Physicians' Desk Reference (“PDR”) (e.g., 63rd edition, 2009, Montvale, NJ: Physicians' Desk Reference).
  • PDR Physicians' Desk Reference
  • the combination treatment comprises lithium and an additional compound(s) formulated together.
  • the lithium in such formulations may be released concurrently with or separately from the additional compound(s), or may be released and/or delivered to the tissue site with different pharmacokinetics.
  • one or more of the compounds in the formulation undergoes controlled release, whereas one or more of the other compounds does not.
  • one or more of the compounds in the formulation undergoes sustained release whereas one or more of the other compounds undergoes delayed release.
  • the combination treatment comprises lithium and an additional compound(s) formulated separately.
  • the separate formulations may be administered concurrently, sequentially, or in alternating sequence.
  • the lithium compound may be administered sequentially, or concurrently with another compound such as finasteride or minoxidil, to achieve the desired effect of hair retention and growth.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more treatments selected from, e.g., cell therapy (such as a stem cell), a formulation for gene therapy (such as, e.g., a virus, vims-like particle, virosome). an antibody or antigen-binding fragment thereof, an herb, a vitamin (e.g., a form of vitamin E, a vitamin A derivative, such as, e.g., all-trans retinoic acid (ATRA), a B vitamin, such as. e.g.
  • cell therapy such as a stem cell
  • a formulation for gene therapy such as, e.g., a virus, vims-like particle, virosome
  • an antibody or antigen-binding fragment thereof an herb
  • a vitamin e.g., a form of vitamin E, a vitamin A derivative, such as, e.g., all-trans retinoic acid (ATRA)
  • ATRA all-trans retinoic acid
  • a shampoo ingredient e.g. , ammonium chloride, ammonium lauryl sulfate, glycol, sodium laureth sulfate, sodium lauryl sulfate, ketoconazole, zinc pyrithione, selenium sulfide, coal tar, a salicylate derivative, dimethicone, or plant extracts or oils
  • a conditioning agent e.g. , a soap product, a moisturizer, a sunscreen, a waterproofing agent, a powder, talc, or silica, an oil-control agent, alpha-hydroxy acids, beta-hydroxy acids (e.g.
  • astringent salts e.g., zinc salts, such as zinc pyrithione, inorganic or organic salts of aluminum, zirconium, zinc, and mixtures thereof, aluminum chloride, aluminum
  • chlorohydrate aluminum chlorohydrex, aluminum chlorohydrex PEG, aluminum
  • chlorohydrex PG aluminum dichlorohydrate, aluminum dichlorohydrex PEG.
  • aluminum dichlorohydrex PG aluminum sesquichlorohydrate, aluminum sesquichlorohydrex PEG, aluminum sesquichlorohydrex PG, aluminum sulfate, aluminum zirconium
  • octachlorohydrate aluminum zirconium octachlorohydrex GLY (abbreviation for glycine), aluminum zirconium pentachlorohydrate, aluminum zirconium pentachlorohydrex GLY, aluminum zirconium tetrachlorohydrate, aluminum zirconium trichlorohydrate, aluminum zirconium tetrachlorohydrate GLY, and aluminum zirconium trichlorohydrate GLY, potassium aluminum sulphate, (also known as alum ( A1(S04) 2 12H20)), aluminum undecylenoyl collagen amino acid, sodium aluminum Iactate+ aluminum sulphate
  • octyl methoxycinnamate ParsolTM MCX
  • 2-hydroxy-4-methoxy benzophenone also known as oxybenzone and available as BenzophenoneTM , and preservatives
  • an anti-age cream e.g., carboxyalkylates of branched alcohols and/or alkoxylates thereof, e.g., tridecyl carboxy alkylates, cerulenin or a cerulenin analog, including pharmaceutically acceptable salts or solvates thereof, another fatty acid synthase inhibitor, such as triclosan or analogs thereof, a polyphenol extracted from green tea (EGCG), available from Sigma Corporation (St.
  • EGCG green tea
  • a massage agent e.g., a massage agent, an exfoliant, an anti-itch agent, a pro-inflammatory agent, an immunostimulant (e.g., cytokines, agonists or antagonists of various ligands, receptors and signal transduction molecules of the immune system, immunostimulatory nucleic acids, an adjuvant that stimulates the immune response and/or which causes a depot effect), a cell cycle regulator, a hormonal agonist, hormonal antagonist (e.g., flutamide, bicalutamide, tamoxifen, raloxifene, leuprolide acetate
  • an immunostimulant e.g., cytokines, agonists or antagonists of various ligands, receptors and signal transduction molecules of the immune system, immunostimulatory nucleic acids, an adjuvant that stimulates the immune response and/or which causes a depot effect
  • a cell cycle regulator e.g., a hormonal agonist, hormonal antagonist (e.g., flu
  • LUPRON LH-RH antagonists
  • an inhibitor of hormone biosynthesis and processing a steroid (e.g., dexamethasone, retinoids, deltoids, betamethasone, Cortisol, cortisone, prednisone, dehydrotestosterone, glucocorticoids, mineralocorticoids, estrogen, testosterone, progestins), antigestagens (e.g., mifepristone, onapristone), an antiandrogen (e.g., cyproterone acetate), an antiestrogen, an antihistamine (e.g., niepyramine, diphenhydramine, and antazoline), an anti-inflammatory (e.g., corticosteroids, NTHEs, and COX-2 inhibitors), a retinoid (e.g., 13-cis-retinoic acid, adapalene, all-trans-retinoic acid, and etretinate), an immunosuppressant (
  • an antibiotic such as, e.g., fluorouracil (5-FU or f5U) or other pyrimidine analogs, methotrexate, cyclophosphamide, vincristine
  • an anti-cancer agent such as, e.g., fluorouracil (5-FU or f5U) or other pyrimidine analogs, methotrexate, cyclophosphamide, vincristine
  • a mood stabilizer e.g., valproic acid or carbamazepine
  • an antimetabolite an anti-viral agent
  • an antimicrobial e.g., benzyl benzoate, benzalkonium chloride, benzoic acid, benzyl alcohol, butylparaben, ethylparaben, methylparaben, propylparaben, camphorated metacresol, camphorated phenol, hexylresorcinol, methylbenzethonium chloride, cetrim
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with integumental perturbation and, optionally, also comprises another treatment known in the art or described herein.
  • Integumental perturbation can be achieved by any means known in the art or described herein, such as, for example, using chemical or mechanical means.
  • integumental perturbation comprises disrupting the skin of the subject (for example, resulting in the induction of re-epithelialization of the skin of the subject ).
  • a certain area of the epithelium is partially or wholly disrupted.
  • a certain area of both the epithelium and stratum corneum are partially or wholly disrupted.
  • Integumental perturbation can be used to induce, for example, a burn, excision,
  • dermabrasion dermabrasion, full-thickness excision, or other form of abrasion or wound.
  • Mechanical means of integumental perturbation include, for example, use of sandpaper, a felt wheel, ultrasound, supersonically accelerated mixture of saline and oxygen, tape-stripping, spiky patch, or peels.
  • Chemical means of integumental perturbation can be achieved, for example, using phenol, trichloroacetic acid, or ascorbic acid.
  • Electromagnetic means of integumental perturbation include, for example, use of a laser (e.g., using lasers, such as those that deliver ablative, non-ablative, fractional, non-fractional, superficial or deep treatment, and/or are C02-based, or Erbium- Y AG-based, etc.).
  • Integumental perturbation can also be achieved through, for example, the use of visible, infrared, ultraviolet, radio, or X-ray irradiation.
  • integumental perturbation is by light energy, such as described in Leavitt et al , 2009, Clin. Drug. Invest. 29:283-292.
  • Electrical or magnetic means of disruption of the epidermis can be achieved, for example, through the application of an electrical current, or through electroporation or RF ablation.
  • Electric or magnetic means can also include the induction of an electric or a magnetic field, or an electromagnetic field. For example, an electrical current can be induced in the skin by application of an alternating magnetic field.
  • a radiofrequency power source can be coupled to a conducting element, and the currents that are induced will heat the skin, resulting in an alteration or disruption of the skin. Integumental perturbation can also be achieved through surgery, for example, a biopsy, a skin transplant, hair transplant, cosmetic surgery, etc.
  • integumental perturbation is by laser treatment.
  • Exemplary laser treatments for integumental perturbation include Erbium- YAG laser, Ultrapulse C0 2 fractional laser. Ultrapulse C0 2 ablative laser, Smooth Peel Full-ablation Erbium laser (Candela), as described, for example, in the examples of Sections 7, 9, 1 1 , 13, 15, 17, 1 , 21. 23, and 24, below.
  • a laser treatment is chosen in which the integumental perturbation achieved most resembles that achieved by dermabrasion (for example, a dermabrasion method described herein).
  • integumental perturbation by laser treatment is by a fractional laser. See, e.g., the laser treatments described in U.S. Provisional Application Nos.
  • a fractional laser treatment is treatment with an Erbium- YAG laser at around 1540 nm or around 1550 nm (for example, using a Fraxel® laser (Solta Medical)). Treatment with an Erbium- YAG laser at 1540 or 1550 nm is typically non-ablative, and pinpoint bleeding typical of laser treatment is not observed since the stratum corneum is left in tact.
  • integumental perturbation by laser treatment is by a fractional laser, using, e.g., a CO 2 laser at 10,600 nm. Treatment with a C0 2 laser at 10,600 nm is typically ablative, and typically leads to the appearance of pinpoint bleeding.
  • a standard C0 2 or Erbium-YAG laser can be used to create superficial and, optionally, broad based, integumental perturbation similar to dermabrasion (discussed below). Although the pinpoint bleeding clinical endpoint may not be achieved due to the coagulation properties of (particularly non-ablative) lasers, use of a laser has an advantage making it possible to select the specific depth of skin disruption to effectively remove the stratum corneum and epidermis, or portions thereof.
  • the laser treatment is ablative.
  • full ablation of tissue is generated by the targeting of tissue water at wavelengths of 10,600 nm by a C0 2 laser or 2940 nm by an Erbium-YAG laser.
  • the epidermis is removed entirely and the dermis receives thermal tissue damage.
  • the depth of tissue ablation may be a full ablation of the epidermis, or a partial ablation of the epidermis, with both modes causing adequate wounding to the skin to induce the inflammatory cascade requisite for regeneration.
  • the depth of ablation may extend partially into the dermis, to generate a deep wound.
  • a lithium composition described herein is delivered by a sustained release depot that is comprised of biocompatible, bioabsorbable polymers that are compatible to tissue.
  • the standard full thickness excision model is created using scissors or with a scalpel in animal models (see, also, the examples of Section 28-30, and 32 infra).
  • This procedure while contemplated for use herein (see, e.g., the example of Section 36 infra), carries with it the risk of scarring.
  • various fractional laser modalities could be used to achieve a similarly deep disruption on a grid pattern.
  • a fractional laser can be use to "drill,” for example, 1-mm diameter holes with a 1-mm hole spacing (the fractional laser can make holes of smaller dimensions).
  • the laser treatment is ablative and fractional.
  • fractional tissue ablation can be achieved using a C0 2 laser at 10,600 nm or an Erbium- YAG laser at 2940 nm (e.g., the Lux 2940 laser, Pixel laser, or Profractional laser).
  • the lasing beam creates micro-columns of thermal injury into the skin, at depths up to 4 mm and vaporizes the tissue in the process.
  • Ablative treatment with a fractional laser leads to ablation of a fraction of the skin leaving intervening regions of normal skin intact to rapidly repopulate the epidermis. Approximately 15%-25% of the skin is treated per session.
  • micro thermal zones can be varied to create a dense "'grid" of injury columns surrounded by intact skin and viable cells.
  • the density of the grid on the treatment area plays an important role. The denser the grid, the more the thermal injury and the type of injury begins to approximate full ablation. Therefore, it is appreciated that there may be an "optimum" MTZ density that is appropriate for use in the methods disclosed herein.
  • a lithium composition described herein is delivered into the dermis immediately after wounding, or after initial re-epithelialization has occurred.
  • the mode of laser treatment is non-ablative, wherein the stratum corneum and the epidermis are intact after treatment, with the dermis selected for the deep thermal treatment required for the requisite injury to tissue.
  • This can be accomplished by cooling the epidermis during the laser treatment.
  • the depth of treatment may be 1 mm to 3 mm into the skin.
  • contact cooling such as a copper or sapphire tip.
  • Lasers that are non- ablative have emission wavelengths between 1000- 1600 nm, with energy fluences that will cause thermal injury, but do not vaporize the tissue.
  • the non-ablative lasers can be bulk. wherein a single spot beam can be used to treat a homogenous section of tissue. In some embodiments, multiple treatments are required to achieve the desired effect.
  • a lithium composition described herein is delivered deep into the dermis in polymeric micro-depots and released in a sustained fashion.
  • Lasers that are non-ablative include the pulsed dye laser (vascular), the 1064 Nd:YAG laser, or the Erbium- YAG laser at 1540 nm or 1550 nm (e.g., the Fraxel® laser).
  • the mode of laser treatment is fractional and non-ablative. Treatment with a fractional, non-ablative laser leads to perturbation of a fraction of the skin, leaving intervening regions of normal skin intact to rapidly repopulate the epidermis.
  • Approximately 15%— 25% of the skin is treated per session.
  • the skin barrier function is maintained, while deep thermal heating of dermis can occur.
  • zones of dermis and epidermis are coagulated and the stratum corneum is left essentially intact.
  • This process has been coined "fractional photothermolysis" and can be accomplished, e.g., using the Erbium- YAG laser with an emission at or around 1540 nm or 1550 nm.
  • a lithium composition described herein is delivered immediately after the tissue injury, deep into the dermis.
  • a combination of bulk and fractional ablation modes of tissue injury are used.
  • the mode of laser treatment for, e.g., a Caucasian male 30-50 years old is fractional and non-ablative using an Erbium- YAG laser at 1550 nm, with the following settings: 50-70 J/cm 2 , treatment level 8-10 (density of the "dots"), with 8 passes.
  • the laser device can be equipped with a touch pad screen that offers the operator a menu of options for setting the parameters for operating the laser to promote hair growth.
  • the device can be programmed to offer the operator selections for hair growth vs. removal, choice of skin color, hair follicle density, power settings, etc.
  • a combination treatment comprising use of a laser includes administration to the skin of a compound absorbing light at wavelengths between 1000-1600 nm for the purpose of efficient channeling of light to heat energy.
  • This method of channeling energy may cause micro-zones of thermal injury within the skin.
  • the compound may be delivered to the skin homogenously in the treatment zone, then subsequently irradiated with a non-ablative laser to efficiently capture the vibrational energy of the infrared beam. This method may result in evenly distributed and deep thermal injury, without causing tissue vaporization.
  • a combination treatment comprising use of a laser includes administration of a lithium compound formulation that is encapsulated in matrices that are highly hydrophilic and charged, for example, linked to the dermis by covalent or ionic bonding to prevent the matrices from being cleared by phagocytosis, as part of the wound healing process.
  • a combination treatment comprising use of a laser includes the step of placing a biocompatible, synthetic skin substitute on the newly created wound, especially if the wound is deep, covers large area and is bulk ablated. This process can help minimize or prevent the rapid wound contraction that occurs after loss of a large area of tissue, frequently culminating in scar tissue formation and loss of skin function.
  • the biocompatible synthetic skin substitute is be impregnated with depots of a slow releasing lithium formulation described herein. This method of treatment may enable treating a large bald area on the scalp in one session at the treatment clinic.
  • other molecules are also co-eluted at the site through the skin substitute, such as, e.g., anesthetics and antibiotics, to prevent further pain and minimization of infection, or any other compound described herein.
  • the skin substitute in the presence or absence of a lithium compound and/or other compounds described herein, may also be pre-cooled and applied to the wound to provide a feeling of comfort to the patient. This mode of lithium or other compound application may prevent the lithium or other compound from being cleared away from the wound site as the wound heals.
  • a fractional like hole pattern (similar to that achieved with a fractional laser or full thickness excision) is achieved with using an array of punch biopsy needles.
  • 1 -mm punch biopsies can be arranged with 1-mm hole spacing.
  • the cored skin samples can be removed and, thus, an effect approximating the full thickness excision model is invoked within each hole.
  • microneedles e.g., 19 or 21 gauge needles
  • micro-coring needles could be used.
  • integumental perturbation is by dermabrasion (also referred to herein as "DA"), a well-established dermatological procedure that has been used for decades as a skin resurfacing technique (Grimes, 2005, Microdermabrasion. Dermatol Surg 31 : 1351 - 1 3 4). While the popularity of mechanical DA has decreased in recent years with the advent of laser-based procedures, DA is still used for removing facial scars resulting from acne and other trauma. Small, portable mechanical dermabrasion equipment uses interchangeable diamond fraises operated at different rotation speeds to remove the epidermis and dermis to differing skin depths levels.
  • Dermabrasion may be carried out using any technique known in the art or as described herein, e.g. , in the examples of Sections 25, 26, and 28-36 (see, e.g. , Sections 35 and 36 for protocols for dermabrasion in human subjects), infra.
  • dermabrasion may be carried out using standard DA with aluminum oxide crystals using the Aseptico Econo-Denmabrader, Advance Microderm DX system, or M2-T system; standard DA with Bell Hand Engine with diamond fraize; etc.
  • DA is carried out using an abrasive wheel.
  • DA with an abrasive wheel is used in order to achieve pinpoint bleeding.
  • DA may be carried out using an abrasive wheel to achieve larger globules of bleeding and frayed collagen.
  • non-powered devices such as abrasive cloths can also be used to achieve the DA, with the optional achievement of the same endpoint(s).
  • DA is accomplished using a device typically used for microdermabrasion.
  • a microdermabrasion device is used to remove a greater depth and/or area of skin than is typical for microdermabrasion (also referred to herein as "MDA"').
  • MDA microdermabrasion
  • the microdermabrasion device is used under sterile conditions.
  • DA is achieved by using a device typically used for microdermabrasion to the point where treatment is stopped upon the observation of pinpoint bleeding, which signals the removal of the stratum corneum and epidermis into the papillary dermis.
  • DA is achieved by using a device for
  • this extended use is reduced by using a microdermabrasion device with increased output pressure and increased abrasion particle size, which may accelerate the skin removal process.
  • DA is accomplished by removal of surface skin by particle bombardment (also referred to herein as "particle mediated dermabrasion" (“PMDA”)), for example, with alumina-, ice- or silica-based particles.
  • particle bombardment also referred to herein as "particle mediated dermabrasion” (“PMDA)
  • PMDA particle mediated dermabrasion
  • micron- sized particles are propelled toward the surface of the skin via short strokes of a handpiece, such as a particle gun, as known in the art.
  • the velocity of particles is controlled through positive or negative pressure.
  • the depth of skin removed by particle bombardment DA (e.g. , PMDA) is a function of the volume of particles impacting the skin, the suction or positive pressure, the speed of movement of the handpiece, and the number of passes per area of the skin.
  • integumental perturbation by one or more of the aforementioned methods achieves removal of part or all of the epidermis. In some embodiments, integumental perturbation removes the entire epidermis. In some
  • integumental perturbation disrupts the papillary dermis.
  • integumental perturbation removes the papillary dermis.
  • integumental perturbation removes the reticular dermis.
  • the depth of integumental perturbation depends on the thickness of the skin at a particular treatment area. For example, the skin of the eyelid is significantly thinner than that of the scalp.
  • the occurrence of pinpoint bleeding indicates that the epidermis and portions of the dermis have been removed. Deeper penetration can results in much more bleeding, and the perturbation can go as deeps as the hypodermis.
  • integumental perturbation by one or more of the aforementioned methods is to a skin depth of 60 ⁇ . In some embodiments, integumental perturbation is to a skin depth of 60- 100 ⁇ . In some embodiments, integumental perturbation is to a skin depth of 100 ⁇ . In some embodiments, integumental perturbation is to a skin depth of 150 pm. In some embodiments, integumental perturbation is to a skin depth of 100-500 ⁇ . In some embodiments, integumental perturbation is to a skin depth of less than 500 ⁇ . In some embodiments, integumental perturbation is to a skin depth of 500- 1000 ⁇ .
  • integumental perturbation is to a skin depth of 1 mm or more. In some embodiments, integumental perturbation is to a skin depth of 1 mm to 3 mm. In some embodiments, integumental perturbation is to a skin depth of 1 mm to 5 mm.
  • the pulse or intermittent lithium treatments described herein potentiate the formation of new hair follicles.
  • Integumental perturbation produces in the affected skin tissue an increase in the number of hair follicle stem cells and in the plasticity of hair follicle cells, such that resident hair follicles may be reprogrammed. Accordingly, and without being bound by any theory for how the invention works, integumental perturbation in combination with a pulse or intermittent lithium treatment provides an environment for the formation of a large number of follicles with desired properties.
  • New hair follicles originate from Hair Follicle Stem Cells (FSCs), oligopotent cells whose progeny can differentiate into the highly differentiated specialized cells of the hair follicle (see Amoh Y. et al. Human hair follicle pluripotent stem (hfPS) cells promote regeneration of peripheral-nerve injury: an advantageous alternative to ES and iPS cells. J Cell Biochem, 2009, 107: 1016-1020; and Amoh Y, et al. Nascent blood vessels in the skin arise from nestin-expressing hair-follicle cells. Proc Natl Acad Sci U S A. 2004 Sep.
  • FSCs originate from one or more of the following: (i) existing follicles ("follicle derived follicle stem cells” or “FDFSC”) (see, e.g. , Toscani et al., 2009, Dermatol Surg. 2009; (ii) the skin (“tissue derived follicle stem cells” or “TDFSC”) (see, e.g. , Ito M, 2007, Nature 447:316-320); (iii) bone marrow (“bone marrow derived follicle stem cells” or “BMDFSC”) (see, e.g.
  • FSCs generate new hair follicles that preserve the type of hair follicle that is typical for each location of skin or scalp.
  • FSCs from the coronal scalp of a male with MPHL typically generate atrophic follicles with vellus or club hairs.
  • FSCs from the occipital scalp of the same male typically generate follicles with terminal hair that are not subject to involution in response to DHT.
  • FSCs responsible for follicle formation may be reprogrammed.
  • FSCs in the process of asymmetric division and subsequent differentiation are susceptible to signals (such as estrogen or testosterone) that alter the determinism of their differentiation program.
  • signals such as estrogen or testosterone
  • Such follicles have characteristics usually associated with: (i) pre-alopecia follicles in the coronal scalp; (ii) female-type follicles on the coronal scalp; or (iii) occipital scalp type follicles.
  • pre-alopecia follicles in the coronal scalp e.g., pre-alopecia follicles in the coronal scalp
  • female-type follicles on the coronal scalp e.g., occipital scalp type follicles.
  • a pulse or intermittent lithium treatment in combination with integumental perturbation provides a window during which a third treatment that alters the follicle development program may be administered in order to significantly change the number and quality of follicles in a particular area of skin.
  • the third treatment e.g. , estrogen or testosterone modulator, such as those described in Poulos & Mirmirani, 2005, Expert Opin. Investig. Drugs 14: 177-184 (incorporated herein by reference
  • the third treatment is administered simultaneously with integumentai perturbation.
  • the third treatment is administered after integumentai perturbation.
  • the third treatment is administered 1 day, 2 days.
  • the third treatment is administered at the time of integumentai perturbation and then daily for 5 days thereafter (in some embodiments, a scab forms during this time). In some embodiments, the third treatment is administered daily for 5 days beginning as soon as the scab falls off. In some embodiments, the third treatment is administered in order to modulate the neoepidermis that forms underneath the scab. In some such embodiments, the third treatment is administered at the time of integumentai perturbation and up to some time after scab falls off , for example, between 5 - 14 days following integumentai perturbation.
  • the course of treatment with the third treatment is short, for example, limited to a few days just following scab detachment, or even continued only for as long as the scab is still attached.
  • the timing of the integumentai perturbation, lithium administration, and the third treatment is preferably monitored and adjusted so that optimal results are achieved.
  • the following sections contain non-limiting examples of combination treatments that are specific for, respectively, (i) enhancing hair growth and/or treating conditions associated with hair loss and (ii) removal of unwanted hair and/or treating conditions associated with excessive hair growth.
  • the treatments described in these sections may optionally be combined with the aforementioned treatments and/or with one another.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with a treatment that enhances hair growth and/or treats a disease or condition associated with excessive hair loss. Any treatment that enhances hair growth and/or treats a disease or condition associated with excessive hair loss that is known in the art or yet to be developed is contemplated for use in such combination treatments.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more antiandrogens, such as. e.g., finasteride (e.g., marketed as Propecia or Proscar), fluconazole, spironolactone, flutamide, diazoxide, 17-alpha-hydroxyprogesterone, 1 1-alpha-hydroxyprogesterone, ketoconazole, RU58841 , dutasteride (marketed as Avodart), fluridil, or QLT-7704, an antiandrogen oligonucleotide, or others described in Poulos & Mirmirani. 2005, Expert Opin. Investig. Drugs 14: 177-1 84, the contents of which is incorporated herein by reference.
  • finasteride e.g., marketed as Propecia or Proscar
  • fluconazole e.g., marketed as Propecia or Proscar
  • fluconazole spironolactone
  • Commonly used dosage forms of finasteride that may be used in such combination therapies are, for example, oral finasteride at 1 mg/day. See, e.g. , Physicians ' Desk Reference, 2009, 63rd ed., Montvale, NJ: Physicians' Desk Reference Inc., entries for Propecia® and Proscar® at pages 2095-2099 and 2102-2106, respectively, which are incorporated herein by reference in their entireties.
  • the regular dosages may be increased or decreased as directed by the physician. For example, a lower dosage may be used over a shorter duration owing to the synergistic effect of the combination with the intermittent lithium treatment or a pulse lithium treatment.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more channel openers, such as, e.g., minoxidil (e.g., marketed as Rogaine or Regaine), diazoxide, or phenytoin.
  • channel openers such as, e.g., minoxidil (e.g., marketed as Rogaine or Regaine), diazoxide, or phenytoin.
  • Commonly used dosage forms of minoxidil that may be used in such combination therapies are topical solutions comprising 2% minoxidil or 5% minoxidil, for example, topical minoxidil foam 5%.
  • the regular dosages may be increased or decreased as directed by the physician. For example, a lower dosage may be used over a shorter duration owing to the sy nergistic effect of the combination with the intermittent lithium treatment or a pulse lithium treatment.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with an antiandrogen (e.g., finasteride, 5-alpha reduction inhibitors) and a channel opener (e.g. , minoxidil).
  • an antiandrogen e.g., finasteride, 5-alpha reduction inhibitors
  • a channel opener e.g. , minoxidil
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more antiandrogens, such as ketoconazole, herbs (such as, e.g. , saw palmetto, glycine soja, Panax ginseng, Castanea Sativa, Arnica Montana, Hedera Helix Geranium Maculatum), triamcinolone acetonide (e.g., suspension of 2.5 to 5 mg/ml for injection), a topical irritant (e.g. , anthralin) or sensitizer (e.g. , squaric acid dibutyl ester [SADBE] or diphenyl cyclopropenone [DPCPJ),
  • antiandrogens such as ketoconazole, herbs (such as, e.g. , saw palmetto, glycine soja, Panax ginseng, Castanea Sativa, Arnica Montana, Hedera Helix Geranium Maculatum), triamcinol
  • the combination therapy comprises intermittent lithium treatment or a pulse lithium treatment in combination with nitroxide spin labels (e.g., TEMPO and TEMPOL). See United States Patent 5,714,482. which is incorporated herein by reference.
  • the combination treatment comprising intermittent lithium treatment or a pulse lithium treatment to enhance hair growth in a female subject is not finasteride or ketoconazole.
  • the combination therapy comprising intermittent lithium treatment or a pulse lithium treatment to enhance hair growth in a pregnant, female subject is not finasteride or ketoconazole.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with a copper peptide(s), preferably applied topically, or another compound with superoxide dismutation activity.
  • the combination therapy comprises intermittent lithium treatment or a pulse lithium treatment in combination with an agent that increases nitric oxide production (e.g. , arginine. citrulline, nitroglycerin, amyl nitrite, or sildenafil (Viagra)).
  • an agent that increases nitric oxide production e.g. , arginine. citrulline, nitroglycerin, amyl nitrite, or sildenafil (Viagra)
  • such compounds are administered further in combination with a catalase or catalase mimetic, or other antioxidant or free radical scavenger.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more agents that counteract hair follicle cell senescence (also referred to herein as "anti-senescence agents"), for example, anti-oxidants such as glutathione, ascorbic acid, tocopherol, uric acid, or polyphenol antioxidants); inhibitors of reactive oxygen species (ROS) generation, such as superoxide dismutase inhibitors; stimulators of ROS breakdown, such as selenium; mTOR inhibitors, such as rapamycin; or sirtuins or activators thereof, such as resveratrol, or other SIRT1 , SIRT3 activators, or nicotinamide inhibitors.
  • anti-oxidants such as glutathione, ascorbic acid, tocopherol, uric acid, or polyphenol antioxidants
  • ROS reactive oxygen species
  • stimulators of ROS breakdown such as selenium
  • mTOR inhibitors such as rapamycin
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more agents that induce an immune response or cause inflammation, such as, e.g. , tetanus toxoid, topical non-specific irritants (anthralin), or sensitizers (squaric acid dibutyl ester [SADBE] and diphenyl cyclopropenone [DPCP]). While not intending to be bound by any theory, it is thought that by contacting these agents to the skin, lymphocytes and hair follicle stem cells may be recruited to skin.
  • agents that induce an immune response or cause inflammation such as, e.g. , tetanus toxoid, topical non-specific irritants (anthralin), or sensitizers (squaric acid dibutyl ester [SADBE] and diphenyl cyclopropenone [DPCP]). While not intending to be bound by any theory, it is thought that by contacting these agents to the skin, lymph
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with a chemical or mechanical (such as those discussed infra) treatment that induces an inflammatory process in the skin. While not intending to be bound by any theory, inducing intlammation in the site where hair growth is desired helps to recruit stem cells to the tissues that drive the formation of new follicles.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with an antiapoptotic compound.
  • the antiapoptotic compound is not a Wnt or a Wnt agonist.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more of stem cell therapy, hair cloning, hair transplantation, scalp massage, a skin graft, hair plugs, or any surgical procedure aimed at hair restoration.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with use of a laser device or other mode of accomplishing "photo-biostimulation" of the hair follicles.
  • a laser device or other mode of accomplishing "photo-biostimulation" of the hair follicles.
  • the Hairmax Lasercomb or the Leimo laser are non-limiting examples of devices that can be used to stimulate growth of hair and can be used in combination with the methods described herein.
  • intermittent lithium treatment or a pulse lithium treatment in combination with the aforementioned methods for enhancing hair growth prevents, delays, or reverses scalp hair loss in MPHL and/or diffuse hair thinning associated with aging.
  • intermittent lithium treatment or a pulse lithium treatment alone or in combination with the aforementioned methods for enhancing hair growth, synchronizes hair follicle cells in the cell cycle.
  • lithium is administered to arrest hair follicle cells in G2/M phase, which synchronizes them; then the lithium treatment is removed; and then their re-entry into the cell cycle and mitotic division is stimulated with other drugs (which leads to anagen follicles and an increased number of follicles).
  • other drugs which leads to anagen follicles and an increased number of follicles.
  • the lithium treatment arrests hair follicle cells in late prophase or metaphase, which synchronizes them; the lithium treatment is removed; and then their re-entry into the cell cycle and mitotic division is stimulated with other drugs (which leads to anagen follicles and an increased number of follicles).
  • the lithium treatment arrests hair follicle stem cells in G2/M phase, which synchronizes them; then the lithium treatment is removed: and then their re-entry in to the cell cycle and mitotic division is stimulated with other drugs (which leads to anagen follicles and an increased number of follicles).
  • the lithium treatment arrests hair follicle stem cells in late prophase or metaphase, which synchronizes them; the lithium treatment is removed; and then their re- entry into the cell cycle and mitotic division is stimulated with other drugsfwhich leads to anagen follicles, and an increased number of follicles).
  • intermittent lithium treatment or a pulse lithium treatment alone or in combination with the aforementioned methods for enhancing hair growth, synchronizes hair follicle cells in the Follicle Cycle.
  • the treatment regimen induces follicles to enter anagen.
  • the treatment regimen prevents follicles from entering catagen.
  • the treatment regimen induces follicles in telogen to enter exogen. or induces follicles in exogen to enter anagen.
  • intermittent lithium treatment or a pulse lithium treatment in combination with the aforementioned methods for enhancing hair growth improves the effectiveness of these methods, making the treatment more effective, efficient, cost-effective, and/or user friendly. For example, fewer treatments may be required.
  • intermittent lithium treatment or a pulse lithium treatment in combination with the aforementioned methods for enhancing hair growth improves the effectiveness of these methods, making the treatment more effective, efficient, cost-effective, and/or user friendly. For example, fewer treatments may be required.
  • one of the previously described hair growth enhancement treatments on its own is not cosmetically satisfactory, the benefits are too short-lived, or the hair that results from the treatment is vcllus hair, or other thin or patchy hair, or has inadequate pigmentation.
  • the hair that results may be more cosmetically satisfactory, longer lasting, terminal hair or head hair (depending on the type of hair intended as opposed to vellus hair, and/or thicker, more uniform, and properly pigmented hair.
  • more than one hair will emerge from each follicle, leading to the appearance of thicker hair.
  • one of the aforementioned treatments to enhance hair growth is administered following integumental perturbation (as described herein) and lithium treatment.
  • integumental perturbation is followed by lithium treatment, which is then followed by one of the aforementioned treatments to enhance hair growth.
  • integumental perturbation accompanies lithium treatment, which is then followed by one of the aforementioned treatments to enhance hair growth.
  • integumental perturbation is prior to lithium treatment administered together with one of the aforementioned treatments to enhance hair growth.
  • Integumental perturbation is achieved by either treatment with a fractional Erbium- YAG laser to epidermal or dermal depth, a fractional C0 2 laser to epidermal or dermal depth, or dermabrasion as described herein. This is followed by the stimulation of follicle formation by a pulse lithium treatment.
  • the follicles are reprogrammcd, e.g., a miniaturizing male temporal scalp follicle (or Follicle Stem Cell) is changed to a non-miniaturizing female-type temporal scalp follicle (or Follicle Stem Cell) using one or more of the following drugs: estrogen, finasteride, dutasteride (AvodartTM).
  • the follicle type can be reprogrammed, e.g.
  • a miniaturizing male temporal scalp follicle can be changed to a non- miniaturizing male occipital scalp-type follicle, using a drug such as valproate.) Then, terminal hair growth is stimulated by the application of low energy light (using, e.g. , LaserMax or IPL) or minoxidil.
  • enhancement of hair growth is accomplished by a combination of integumental perturbation, as described herein, and lithium treatment without one of the aforementioned treatments for enhancing hair growth.
  • the combination of integumental perturbation and lithium treatment of an area of skin that already contains hair-producing follicles (preferably, terminal hair) increases production of hair in that area of skin.
  • the combination of integumental perturbation and lithium treatment is administered to skin that has been damaged and which no longer contains follicles.
  • the combination of integumental perturbation and lithium treatment may restore follicle production in that area of skin.
  • an area of skin containing a wound that has not healed correctly such as a scar (e.g., a keloid scar) is administered a combination treatment of integumental perturbation and lithium in order to restore hair follicles and/or growth to that area of skin.
  • a scar e.g., a keloid scar
  • enhancement of hair growth is accomplished by lithium treatment alone.
  • lithium treatment of an area of skin that already contains hair-producing follicles increases production of hair in that area of skin.
  • the lithium treatment is administered to skin that has been damaged and which no longer contains follicles.
  • the lithium treatment may restore follicle production in that area of skin.
  • an area of skin containing a wound that has not healed correctly such as a scar (e.g. , a keloid scar), is administered a lithium treatment in order to restore hair follicles and/or hair growth to that area of skin.
  • a scar e.g. , a keloid scar
  • Synergism occurs when the combination has an effect that is more than would be expected from merely the additive effect of each element in the combination, for example, if branched hair follicles or multiple shafts per pore were produced by the combination and not by either alone.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more techniques of depilation (removal of the part of the hair above the surface of the skin) or epilation (removal of the entire hair, including the part below the skin) of a part of the skin affected by unwanted hair.
  • Intermittent lithium treatment or a pulse lithium treatment can be used in combination with any form of epilation or depilation known in the art. Any treatment that inhibits the growth of unwanted hair, removes unwanted hair and/or treats a disease or condition associated with unwanted hair that is known in the art or yet to be developed is contemplated for use in such combination treatments. For example, see U.S. Patent No. 6,050,990, issued April 18, 2000, which is incorporated herein by reference in its entirety.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more prescription drugs such as, e.g., eflornithine (trade name: Vaniqa, which is formulated as eflornithine hydrochloride, 13.9%) or another ornithine decarboxylase inhibitor, and/or 5-fluorouracil (5- FU, Efudex 5% cream).
  • prescription drugs such as, e.g., eflornithine (trade name: Vaniqa, which is formulated as eflornithine hydrochloride, 13.9%) or another ornithine decarboxylase inhibitor, and/or 5-fluorouracil (5- FU, Efudex 5% cream).
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with epidermal growth factor ( EGF) or a mimetic thereof.
  • EGF epidermal growth factor
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more polyamine derivatives and/or analogs including pharmaceutically acceptable salts and solvates thereof; inhibitors of ornithine decarboxylase, such as difluoryl methyl ornithine (DFMO),
  • DFMO difluoryl methyl ornithine
  • MGBG methylglyoxalbisguanylhydrazone
  • HAVA hydrozino ornithine
  • AC N-acetyl cysteines
  • AC neutralized salts of a non-hydroxy C 2 -C 40 dicarboxylic acids, preferably malonate salts; and mixtures thereof.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with Stryphnodendron adstringens (Martius) Coville bark extract, which suppresses terminal hair. See Vicente et al., 2009. J Eur Acad Dermatol Venereol 23:410-414.
  • the combination therapy comprises the use of fennel. See Javidnia et al., 2003, Phytomedicine 10:455-458.
  • the combination treatment for hair removal comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more antiandrogen drugs, such as cyproterone acetate, ketoconazole, or spironolactone (the latter marketed under the trade names Aldactone, Novo-Spiroton, Aldactazide, Spiractin,
  • antiandrogen drugs such as cyproterone acetate, ketoconazole, or spironolactone (the latter marketed under the trade names Aldactone, Novo-Spiroton, Aldactazide, Spiractin,
  • such therapy is for use in female subjects affected by idiopathic hirsutism.
  • such anti-androgen combination therapy is not for use in female subjects predisposed to, at risk for, or suffering from, cancer.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with a pro-apoptotic compound or a cytotoxic agent (such as, e.g. , those described in U.S. Patent No. 6,050,990, which is incorporated herein by reference).
  • a pro-apoptotic compound or a cytotoxic agent (such as, e.g. , those described in U.S. Patent No. 6,050,990, which is incorporated herein by reference).
  • the pro-apoptotic compound is not a Wnt antagonist.
  • the combination treatment for hair removal comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more of hair bleaching, shaving, waxing, sugaring, threading, plucking, use of an abrasive material, laser, electrolysis or electrology, use of an epilation device (e.g., commercially available devices such as Emjoi®, Epilady®, Duet, Legend, and EpiGirl, While Philips Norelco's Satinelle Epilator, and similar products by Panasonic, Braun, and Norelco), use of friction, exfoliation, burning, intense pulsed light ("IPL"; e.g., Flashlamp or F.piLight), use of a mechanical device, or use of a chemical depilatory such as thioglycolic acid (e.g., Nair®), use of turmeric optionally with other ingredients like besan powder and milk, or enzymes such as the Epiladerm-Complex.
  • an epilation device e.g., commercially available devices such as Emjoi
  • intermittent lithium treatment or a pulse lithium treatment in combination with the aforementioned methods for hair removal or inhibiting hair growth improves the effectiveness of these methods, making the treatment more effective, efficient, cost-effective, and/or user friendly. For example, fewer treatments may be required.
  • one of the previously described hair removal or growth inhibition treatments on its own is not cosmetically satisfactory, the benefits are too short-lived, or the hair that remains or grows back after the treatment is too pigmented, thick or course.
  • intermittent lithium treatment or a pulse lithium treatment the results may be more cosmetically satisfactory, longer lasting, and the hair that remains or grows back are fewer in number, thinner (e.g., vellus hair only), less pigmented, and/or shorter.
  • a subject previously had one or more hair follicles with numerous hairs emerging from it, and after the combination therapy, only one hair, preferably a vellus hair, emerges from each follicle, leading to the appearance of thinner hair.
  • intermittent lithium treatment or a pulse lithium treatment in combination with the aforementioned methods for hair removal or inhibiting hair growth prevents, delays, or reverses terminal hair growth on female axilla, female face, female legs, male ears, male nose, or male back.
  • intermittent lithium treatment or a pulse lithium treatment in combination with the aforementioned methods for hair removal or inhibiting hair growth synchronizes hair follicle cells in the Cell Cycle.
  • the lithium treatment arrests hair follicle cells in G2/M phase, which synchronizes them; the lithium treatment is removed; and then their entry into the cell cycle and mitotic division is stimulated with other drugs that expose their DNA and renders them susceptible to cytotoxic drugs or apoptosis-inducing drugs that cause them to enter catagen and involute.
  • the lithium treatment arrests hair follicle cells in late prophase or metaphase, which synchronizes them; the lithium treatment is removed; and then their entry into the cell cycle and mitotic division is stimulated with other drugs, which exposes their DNA and renders them susceptible to cytotoxic drugs or apoptosis-inducing drugs, causing them to enter catagen and involute.
  • the lithium treatment arrests hair follicle stem cells in G2/M phase, which synchronizes them; the lithium treatment is removed; and then their entiy into the cell cycle and mitotic division is stimulated with other drugs which exposes their DNA and renders them susceptible to cytotoxic drugs or apoptosis-inducing drugs, causing the stem cells to enter catagen and involute.
  • the lithium treatment arrests hair follicle stem cells in late prophase or metaphase, which synchronizes them; the lithium treatment is removed and then their entry into the cell cycle and mitotic division is stimulated with other drugs which expose their DNA and renders them susceptible to cytotoxic drugs or apoptosis-inducing drugs, and thus they enter catagen and involute.
  • intermittent lithium treatment or a pulse lithium treatment in combination with the aforementioned methods for hair removal or inhibiting hair growth synchronizes hair follicles in the Follicle Cycle.
  • the lithium treatment may induce follicles to leave anagen, enter catagen and involute, or enter exogen.
  • the lithium treatment may inhibit follicles from entering anagen, or maintain follicles in telogen, or maintain follicles in exogen.
  • one of the aforementioned treatments to inhibit hair growth or remove unwanted hair is administered following integumental perturbation (as described herein) and lithium treatment.
  • integumental perturbation is followed by lithium treatment, which is then followed by one of the aforementioned treatments to inhibit hair growth or remove unwanted hair.
  • integumental perturbation accompanies lithium treatment, which is then followed by one of the aforementioned treatments to inhibit hair growth or remove unwanted hair.
  • integumental perturbation is prior to lithium treatment administered together with one of the aforementioned treatments to inhibit hair growth or remove unwanted hair.
  • inhibition of hair growth or removal of unwanted hair is accomplished by a combination of integumental perturbation, as described herein, and lithium treatment w ithout one of the aforementioned treatments for inhibition of hair growth or removal of unwanted hair.
  • the combination of integumental perturbation and lithium treatment of an area of skin that already contains follicles that do not produce hair or that produce only vellus hair further reduces hair in that area of skin.
  • inhibition of hair growth or removal of unwanted hair is accomplished by lithium treatment alone.
  • lithium treatment of an area of skin that already contains follicles that do not produce hair or that produce only vellus hair further reduces hair in that area of skin. These effects may be accomplished by modulating the dosage of lithium.
  • the intermittent lithium treatment or a pulse lithium treatment can be administered prior to, concurrently with, or subsequent to the administration of a second (or third, or more) treatment.
  • the intermittent lithium treatment or a pulse lithium treatment is administered to a subject at reasonably the same time as the other treatment. This method provides that the two administrations are performed within a time frame of less than one minute to about five minutes, or up to about sixty minutes from each other, for example, at the same doctor's visit. [00342J In another embodiment, the intermittent lithium treatment or a pulse lithium treatment and other treatment are administered at exactly the same time.
  • the intermittent lithium treatment or a pulse lithium treatment and the other treatment are administered in a sequence and within a time interval such that the intermittent lithium treatment or a pulse lithium treatment and the other treatment can act together to provide an increased benefit than if they were administered alone.
  • the intermittent lithium treatment or a pulse lithium treatment and other treatment are administered sufficiently close in time so as to provide the desired outcome.
  • Each can be administered simultaneously or separately, in any appropriate form and by any suitable route.
  • the intermittent lithium treatment or a pulse lithium treatment and the other treatment are administered by different routes of
  • each is administered by the same route of administration.
  • the intermittent lithium treatment or a pulse lithium treatment and the other treatment can be administered at the same or different sites of the subject's body.
  • the intermittent lithium treatment or a pulse lithium treatment and the other treatment may or may not be administered in admixture or at the same site of administration by the same route of administration.
  • the intermittent lithium treatment or a pulse lithium treatment and the other treatment are administered less than 1 hour apart, at about 1 hour apart, 1 hour to 2 hours apart, 2 hours to 3 hours apart, 3 hours to 4 hours apart, 4 hours to 5 hours apart, 5 hours to 6 hours apart, 6 hours to 7 hours apart, 7 hours to 8 hours apart, 8 hours to 9 hours apart, 9 hours to 10 hours apart, 10 hours to 1 1 hours apart, 1 1 hours to 12 hours apart, no more than 24 hours apart or no more than 48 hours apart.
  • the intermittent lithium treatment or a pulse lithium treatment and other treatment are administered 2 to 4 days apart, 4 to 6 days apart, 1 week a part, 1 to 2 weeks apart, 2 to 4 weeks apart, one month apart, 1 to 2 months apart, 2 to 3 months apart, 3 to 4 months apart, 6 months apart, or one year or more apart.
  • the intermittent lithium treatment or a pulse lithium treatment and the other treatment are administered in a time frame where both are still active. One skilled in the art would be able to determine such a time frame by determining the half life of each administered component.
  • the intermittent lithium treatment or a pulse lithium treatment and the other treatment are administered within the same patient visit. In one embodiment, the intermittent lithium treatment or a pulse lithium treatment is administered prior to the administration of the other treatment. In an alternate embodiment, the intermittent lithium treatment or a pulse lithium treatment is administered subsequent to the administration of the other treatment.
  • the intermittent lithium treatment or a pulse lithium treatment and the other treatment are cyclically administered to a subject. Cycling treatment involves the administration of the intermittent lithium treatment or a pulse lithium treatment for a period of time, followed by the administration of the other treatment for a period of time and repeating this sequential administration.
  • the first treatment may be with the intermittent lithium treatment or a pulse lithium treatment or with the other treatment, depending on the subject's prior treatment history and the intended outcome.
  • cycling treatment can also reduce the development of resistance to one or more of the treatments, avoid or reduce the side effects of one of the treatments, and/or improve the efficacy of the treatment.
  • alternating administration of the intermittent lithium treatment or a pulse lithium treatment may be followed by the administration of another treatment (or vice versa) 1 year later, 6 months later, 3 months later, 1 month later, 3 weeks later, 2 weeks later, 1 week later, 4 to 6 days later, 2 to 4 days later, or 1 to 2 days later, wherein such a cycle may be repeated as many times as desired.
  • the intermittent lithium treatment or a pulse lithium treatment and the other treatment are alternately administered in a cycle of 3 weeks or less, once every two weeks, once every 10 days or once every week.
  • Such time frames can be extended or reduced depending on whether a controlled release formulation of either the lithium compound or the other treatment formulation is used, and/or depending on the progress of the treatment course.
  • subjects discontinue their current treatment (e.g., topical minoxidil, finasteride, eflornithine), and the lithium compound is applied for one week (to synchronize the hair follicle cells in G2/M phase arrest). After one week, treatment with the lithium compound is discontinued and treatment with the current treatment is re-started.
  • the subject is treated with 10 cycles of the protocol: alternating topical lithium (e.g., for 1 week) with the other treatment (e.g. , for 3 weeks).
  • treatment with lithium at the wounds(s) from which transplanted tissue was obtained and/or the site of implantation is initiated for one week, and then discontinued and followed by treatment with, e.g. , minoxidil or finasteride for three weeks.
  • Human hair patterning consists of gender specific changes that occur over the life of subjects and vary in degree between individuals and more generally between humans of different racial and ethnic backgrounds.
  • males and females Before puberty, males and females have similar patterns of scalp hair and the rest of their bodies are covered with largely invisible vellus hair.
  • the forearms and legs grow thin, fine terminal hair gradually even before puberty.
  • puberty In males and females puberty is associated with terminal hair growth in the axilla, and anogenital regions.
  • both males and females grow terminal hair over forearms and legs, but males have quantitatively more growth in these regions.
  • Males after puberty grow terminal hair over the moustache/beard, chest, and back regions. Later, males manifest varying degrees of loss of terminal hair on the scalp (vertex/corona and frontal/
  • Sex hormones are synthesized by the skin locally where they exert intracrine or paracrine actions. (Reviewed in, Zouboulis CC, Chen VVC.
  • Estrogens act on Estrogen Receptor alpha (ERalpha) and Estrogen Receptor alpha (ERbeta) in human skin are expressed in site specific localizations. In addition to regulating hair patterning and growth, androgens have effects in sebaceous gland growth and differentiation, epidermal barrier homeostasis and wound healing; and estrogens regulate skin aging, pigmentation, hair growth, sebum production and skin cancer. (Ohnemus et al., 2006, Endocr Rev. 27(6):677-706, "The hair follicle as an estrogen target and source.")
  • a candidate subject for intermittent lithium treatment i.e., alternating lithium treatment with "vacationed iday" periods
  • a pulse lithium treatment for promoting hair growth is any subject suffering from hair loss, hair thinning, balding, or who has or has had a disease or condition associated therewith, or who wishes to enhance the growth or thickness of hair.
  • the subject may be any subject, preferably a human subject, including male, female, intermediate/ambiguous (e.g. , XO), and transsexual subjects.
  • a human subject including male, female, intermediate/ambiguous (e.g. , XO), and transsexual subjects.
  • the subject is a human adolescent. In certain embodiments, the subject is undergoing puberty. In certain embodiments, the subject is a middle-aged adult. In certain embodiments, the subject is a premenopausal adult. In certain embodiments, the subject is undergoing menopause. In certain embodiments, the subject is elderly. In certain embodiments, the subject is a human of 1 year old or less, 2 years old or less, 2 years old, 5 years old.
  • the subject is a male 20 to 50 years old.
  • the subject is a male 20 to 60 years old. In some embodiments, the subject is a male 30 to 60 years old. In some embodiments, the subject is a male 40 to 60 years old. In some embodiments, the subject is a male or female 12 to 40 years old. In some embodiments, the subject is not a female subject. In some embodiments, the subject is not pregnant or expecting to become pregnant. In some embodiments, the subject is not a pregnant female in the first trimester of pregnancy. In some embodiments, the subject is not breastfeeding.
  • the intermittent lithium treatment or a pulse lithium treatment is delivered to an area in which hair growth is desired, for example, the scalp or face (e.g. , the eyebrow, eyelashes, upper lip, lower lip, chin, cheeks, beard area, or mustache area) or another part of the body, such as, e.g. , the chest, abdomen, arms, armpits (site of axillary hair), legs, or genitals.
  • hair restoration to a wounded or scarred part of the skin is desired.
  • the scar is caused by surgery, such as a face lift, skin graft, or hair transplant.
  • the subject may have a disease or disorder of balding or hair loss (including hair thinning), such as forms of nonscarring (noncicatricial) alopecia, such as androgenetic alopecia (AGA), including MPHL or FPHL (e.g. , thinning of the hair, i.e., diffuse hair loss in the frontal/parietal scalp), or any other form of hair loss caused by androgens, toxic alopecia, alopecia areata (including alopecia universalis), scarring (cicatricial) alopecia, pathologic alopecia (caused by, e.g.
  • a disease or disorder of balding or hair loss including hair thinning
  • forms of nonscarring (noncicatricial) alopecia such as androgenetic alopecia (AGA)
  • AGA androgenetic alopecia
  • MPHL or FPHL e.g. , thinning of the hair, i.e
  • trichotillomania a form of hypotrichosis, such as congenital hypotrichosis, or lichen planopilaris, or any other condition of hair loss or balding known in the art or described infra.
  • the subject has hair loss caused by a genetic or hereditary disease or disorder, such as androgenetic alopecia.
  • the subject has hair loss caused by anagen effluvium, such as occurs during chemotherapy (with, e.g., 5-fluorouracil, methotrexate, cyclophosphamide, vincristine).
  • anagen effluvium such as occurs during chemotherapy (with, e.g., 5-fluorouracil, methotrexate, cyclophosphamide, vincristine).
  • Anagen effluvium can be caused by other toxins, radiation exposure (such as radiation overdose), endocrine diseases, trauma, pressure, and certain diseases, such as alopecia areata (an autoimmune disease that attacks anagen follicles.)
  • the subject has hair loss caused by telogen effluvium.
  • telogen effluvium is caused frequently by drugs like lithium and other drugs like valproic acid and carbamazepinc. In addition to psychiatric drugs, telogen effluvium can be induced by childbirth, traction, febrile illnesses, surgery, stress, or poor nutrition. (See, Mercke et al., 2000, Ann. Clin. Psych. 12:35-42).
  • the subject has hair loss caused by or associated with medication, such as chemotherapy (e.g. , anti-cancer therapy or cytotoxic drugs), thallium compounds, vitamins (e.g. , vitamin A), retinoids, anti-viral therapy, or psychological therapy, radiation (e.g.
  • an autoimmune disease or disorder malnutrition
  • an infection such as, e.g., a fungal, viral, or bacterial infection, including chronic deep bacterial or fungal infections
  • dermatitis psoriasis
  • eczema pregnancy, allergy, a severe illness (e.g.
  • the subject has hair thinning, or "shock loss,” or a bald patch caused by prior use as a source of tissue or follicles for hair transplantation or follicular unit transplantation.
  • a candidate subject is a subject who wishes to enhance hair growth, for example, to have more hair, faster-growing hair, longer hair, and/or thicker hair.
  • the candidate is a subject who wishes to increase hair pigmentation.
  • the subject is not affected by a condition of excessive hair loss.
  • Minoxidil is FDA approved for both males and females. Finasteride, while not approved for females, does provide a benefit. See, PDR entry for Propecia. which is incorporated herein by reference.
  • Intermittent lithium treatment or a pulse lithium treatment can be used in combination with minoxidil or other channel openers, finasteride, dutasteride, flutamide, or other antiandrogens, laser therapy or other mode of photo-stimulation of hair follicles, dermabrasion, hair transplantation or other surgical treatment for treatment, or any other treatment provided in Section 5.3 supra. These treatments can be administered during the lithium treatment "holidays.” Alternatively, these treatments can be administered prior to or subsequent to a pulse lithium treatment.
  • Minoxidil and finasteride treatment are more effective at delaying the progression of MPHL than in reversing it. These agents are generally not effective a year or more after hair loss has occurred, consistent with some kind of terminal senescence or involution of the follicle (as will be discussed below). These treatments can be administered during the lithium treatment "holidays.” Alternatively, these treatments can be administered prior to or subsequent to a pulse lithium treatment.
  • Most drugs for hair loss aim to retain the existing hair follicles in their active cycling states, or to rejuvenate telogen hair follicles to actively cycling anagen states.
  • a treatment that encourages the growth of "new" hair follicles combined with one that retains the hair follicles in their actively cycling states offer significant value to the individual who is balding.
  • intermittent lithium treatment or a pulse lithium treatment synchronizes the hair/Follicle Cycle
  • a treatment intended to enhance hair growth or remove hair will be more effective, efficient, cost-effective, and user friendly. For example, fewer treatments may be required.
  • the hair that results may be more cosmetically satisfactory, longer lasting, more terminal hair and/or thicker, more uniform, and properly pigmented hair.
  • the combination treatments disclosed herein for age-related hair loss comprise a combination of intermittent lithium treatment or pulse lithium treatment and estrogen replacement therapy or androgen inhibition therapy.
  • Intermittent lithium treatment or pulse lithium treatment may be used in combination with any of the treatments described in Section 5.3 in order to treat age-related hair loss. These treatments can be administered during the lithium treatment "holidays.” Alternatively, these treatments can be administered prior to or subsequent to a pulse lithium treatment.
  • Cytokines regulate the activity of Dermal Papillae which is believed to be the target of androgen regulation of hair growth.
  • Interleukin- 1 alpha decreases responses to androgen in cultured dermal papilla cells (Boivin et al., 2006, Exp Dermatol. 15:784-793).
  • TGF-betal may mediate androgen-induced hair growth suppression, since in culture, human dermal papilla cells (DPCs) from androgenetic alopecia (AGA) subjects that transiently expressing androgen receptor were co-cultured with keratinocytes (KCs), and secreted TGF- betal that inhibited KC growth (Inui et al, 2003, J Investig Dermatol Symp Proc. 8:69-71).
  • DPCs human dermal papilla cells
  • AGA androgenetic alopecia
  • adjuvants and/or other stimulators of local cytokines are used in conjunction with the intermittent lithium treatment or pulse lithium treatment.
  • one rationale for administering adjuvants and'Or other stimulators of local cytokines in conjunction with the intermittent lithium treatment or pulse lithium treatment is that the production of local cytokines may induce changes in the follicle cell cycle and recruit new FSCs to follicles.
  • Melatonin is a protein hormone secreted by the pineal gland modulates hair growth, pigmentation and/or molting in many species. Human scalp hair follicles in anagen are important sites of extra-pineal melatonin synthesis. Melatonin may also regulate hair Follicle Cycle control, since it inhibits estrogen receptor-alpha expression (Fischer et al., 2008, Pineal Res. 44: 1 -15). These treatments can be administered during the lithium treatment "holidays". Alternatively, these treatments can be administered prior to or subsequent to a pulse lithium treatment.
  • Minoxidil use is further complicated by the fact that it is messy, leaves a residue, and requires daily application.
  • side effects from persistent finasteride or minoxidil treatment - such as sexual dysfunction - are another reason why subjects may benefit from a reduced treatment duration or treatment at a lower dose as may be enabled by the combination treatments described herein.
  • both minoxidil and finasteride are effective only for as long as it is taken; the hair gained or maintained is lost within 6-12 months of ceasing therapy. See, e.g., Rossi, ed., 2004, Australian Medicines Handbook. Sydney: Australian Medicines Handbook. Even when effective, these drugs do no create hair follicles of the kind that were there before balding, and the resultant hair follicles are smaller and less dense.
  • These treatments can be administered during the lithium treatment "holidays.' * Alternatively, these treatments can be administered prior to or subsequent to a pulse lithium treatment.
  • Androgen receptor inhibitors are also useful for stimulating scalp hair growth and inhibiting beard and moustache hair (Hu LY, et al., 2007, Bioorg Med Chem Lett. 2007 17:5983-5988). These treatments can be administered during the lithium treatment
  • a candidate subject for intermittent lithium treatment i.e., alternating lithium treatment with "vacation/holiday” periods
  • a pulse lithium treatment for inhibiting hair growth or removing hair is any subject who has or has had a condition, disease or disorder associated with excess hair or unwanted hair, or who wishes to inhibit the growth of hair or remove hair.
  • permanent hair removal is desired.
  • the subject may be any subject, preferably a human subject, including male, female, and transsexual subjects.
  • the subject is a human adolescent.
  • the subject is undergoing puberty.
  • the subject is a middle-aged adult.
  • the subject is a premenopausal adult.
  • the subject is undergoing menopause.
  • the subject is elderly.
  • the subject is a human of 1 year old or less, 2 years old or less, 2 years old, 5 years old, 5 to 10 years old, 10 to 15 years old, e.g., 12 years old, 15 to 20 years old, 20 to 25 years old, 25 to 30 years old, 30 years old or older, 30 to 35 years old, 35 years old or older, 35 to 40 years old, 40 years old or older, 40 to 45 years old, 45 to 50 years old, 50 years old or older, 50 to 55 years old, 55 to 60 years old, 60 years old or older, 60 to 65 years old, e.g., 65 years old, 65 to 70 years old, 70 to 75 years old, 75 to 80 years old, 80 to 85 years old, 85 to 90 years old, 90 to 95 years old or 95 years old or older.
  • the subject is not a female subject.
  • the subject is not pregnant or expecting to become pregnant.
  • the subject is not breastfeeding.
  • the intermittent lithium treatment or a pulse lithium treatment is delivered to an area of unwanted hair, for example, the head (e.g., the eyebrow, nose and nares, upper lip, lower lip, chin, cheeks, ears, or forehead) or another part of the body, such as, e.g., the chest, breast (e.g., the nipples), abdomen, neck, back, arms, armpits (site of axillary hair), legs, hands, feet, buttocks, or genitals.
  • hair removal from a wounded or scarred area of the skin is desired.
  • hair removal from darkly pigmented skin or a darkly pigmented area of the skin such as a mole, freckle, or the genital area, is desired.
  • subjects who are candidates for intermittent lithium treatment or a pulse lithium treatment include those afflicted with hypertrichosis (excess hair not localized to the androgen-dependent areas of the skin), including generalized congenital hypertrichosis (congenital hypertrichosis lanuginosa), acquired generalized hypertrichosis (acquired hypertrichosis lanuginosa), patterned acquired hypertrichosis, localized congenital hypertrichosis, localized acquired hypertrichosis, paradoxical hypertrichosis, and "werewolf syndrome,” pili multigemini, excess hair in androgen-dependent areas of the skin, idiopathic hirsutism, post-menopausal facial hair, axillary hair, back hair, ear hair, or any other disease, disorder, or form of unwanted hair or excessive hair as discussed infra and/or known in the art. 1
  • the subject has excess hair caused by a genetic or hereditary disease or disorder.
  • the subject has excess hair growth caused by or associated with medication, such as chemotherapy (e.g., anti-cancer therapy, anti-viral therapy, psychological therapy), steroid therapy (e.g., systemic androgenic steroid therapy or corticosteroid therapy), antihypertensive drugs (e.g., minoxidil), or cyclosporine, radiation, trauma, endocrine dysfunction (such as, e.g., adrenal virilism, basophilic adenoma of the pituitary, masculinizing ovarian tumors, Stein-Leventhal syndrome), porphyria cutanea tarda, surgery, burning or other wound, stress, aging, an autoimmune disease or disorder, malnutrition (e.g., lanugo), an infection (such as, e.g.
  • a fungal, viral, or bacterial infection a fungal, viral, or bacterial infection
  • dermatitis a fungal, viral, or bacterial infection
  • psoriasis a fungal, viral, or bacterial infection
  • eczema a form of excessive hair growth known in the art.
  • Any other disease or disorder associated with unwanted hair or excessive hair known in the art is also contemplated.
  • a candidate subject is any subject who wishes to remove or prevent excess hair or unwanted hair, or who wishes to inhibit the growth of hair or remove hair, for example, to have less hair, slower-growing hair, shorter hair, and/or thinner hair.
  • the candidate is a subject who wishes to decrease hair pigmentation.
  • the subject is not affected by a condition or disease or disorder associated with excessive hair.
  • males In addition to MPHL and post-puberty male pattern body hair growth, males also manifest varying degrees of growth of new or increased terminal hair in middle age (over 35 years). Males begin to grow terminal hair on the ears, particularly around the auricular canal; and have increased hair growth (length and density) in the nares, and increased hair growth of eyebrows (hair length and curl.) In some cases, males develop terminal hairs on the skin covering the central prominence of the nose.
  • any combination treatment described in Section 5.3 may be used to reduce unwanted terminal hair in adult males.
  • the combination treatment comprises intermittent lithium treatment or a pulse lithium treatment in combination with a drug such as, e.g. , eflornithine (trade name: Vaniqa, which is formulated as eflornithine hydrochloride, 13.9%) or 5-fluorouracil (e.g.
  • the combination treatment for hair removal comprises intermittent lithium treatment or a pulse lithium treatment in combination with one or more antiandrogen drugs.
  • a combination treatment for hair removal may also comprise intermittent lithium treatment or a pulse lithium treatment with one or more of hair bleaching, shaving, waxing, sugaring, threading, plucking, use of an abrasive material, laser, electrolysis or electrology, use of an epilation device, use of friction, exfoliation, burning, intense pulsed light ("IPL"; e.g. , Flashlamp or EpiLight), use of a mechanical device (e.g. , epilators such as Epilady, Emjoi, etc.), or use of a chemical depilatory (e.g. , Nair ®).
  • These treatments for use in combination with lithium treatment can be administered during the lithium treatment "holidays.” Alternatively, these treatments can be administered prior to or subsequent to a pulse lithium treatment.
  • Vaniqa eflornithine hydrochloride 13.9%
  • other ornithine decarboxylase inhibitors polyamine derivatives
  • 5-fluorouracil 5-FU, Efudex 5% cream
  • certain antiandrogens inhibit hair growth.
  • These drugs (described in Section 5.3 supra) and the other drugs described in Section 5.3 can be used with, or as an adjunct to laser hair removal or to electrolysis, depilatory creams, plucking and waxing.
  • Other products that suppress terminal hair have been described: (a) Vicente RA, et al, 2009, J Eur Acad Dermatol Venereol.
  • Human skin and hair have features that are relatively unique among terrestrial mammals.
  • any method known in the art may be used to evaluate the safety and efficacy of an intermittent lithium protocol or pulse lithium protocol, or of the combination treatments described in Section 5.3.
  • a human skin xenograft model is used.
  • an intermittent lithium treatment or pulse lithium treatment may be administered with a full thickness excision, laser, inflammatory stimulus, or dermabrasion procedure for integumental perturbation described in the examples of Sections 7, 9, 1 1 , 13, and 24-36 below.
  • a synergistic effect of an intermittent lithium treatment or pulse lithium treatment on another treatment for restoring or enhancing hair growth may be measured as an improvement over a control subject receiving only one of the two treatments (i.e., the intermittent lithium treatment or pulse lithium treatment alone or the second treatment alone).
  • the intermittent lithium treatment or a pulse lithium treatment is used in combination treatments (e.g. , described in Section 5.3 supra) to reduce unwanted hair growth. Success of such treatments can be measured using an animal model, e.g., the human xenograft mouse model described herein, by:
  • Section 5.3 may be measured as an improvement over a control subject receiving only one of the two treatments (i.e., the intermittent lithium treatment or pulse lithium treatment alone or the second treatment alone).
  • Another animal model for use in evaluating treatment that may more closely mimic the biology of human skin and hair is a guinea pig model (see, Stenn & Paus, 2001 , Physiol. Revs. 81 : 449-494).
  • the methods for evaluating treatment in animals described elsewhere in this section and in the examples in Section 25 below may be applied to guinea pigs according to methods known in the art. See also, e.g., Kramer et al., 1990, Dermatol Monatsschr 176:417-20; and Simon et al., 1987, Ann Plast Surg 19:519-23.
  • Other animal models that may be of use in evaluating the treatments described herein include pig, cat, or stumptailed macaque models.
  • an intermittent lithium treatment or pulse lithium treatment may be administered in combination with minoxidil, finasteride, laser therapy, or a dermabrasion procedure for integumental perturbation to improve hair growth, as described in the Examples of Sections 6-13, 24, 25, 35, and 36 below.
  • Success of treatment aimed at improving hair growth can be measured by: • increased terminal hair formation (e.g., measuring new hair growth as an increased number of fibers in an affected area of the skin, or increased thickness (e.g. , diameter) or length of hair fibers)
  • telogen e.g., measured examination of a biopsy
  • the intermittent lithium treatment or pulse lithium treatment increases hair count by 5% or more, by 10% or more, by 15% or more, by 20% or more, by 25% or more, by 30% or more, by 40% or more, by 50% or more, by 75% or more, or by 100% or more.
  • the intermittent lithium treatment or pulse lithium treatment increases terminal hair by 5% or more, by 10% or more, by 15% or more, by 20% or more, by 25% or more, by 30% or more, by 40% or more, by 50% or more, by 75% or more, or by 100% or more.
  • the intermittent lithium treatment or pulse lithium treatment increases hair thickness by 5% or more, by 10% or more, by 15% or more, by 20% or more, by 25% or more, by 30% or more, by 40% or more, by 50% or more, by 75% or more, or by 100% or more.
  • Such an improvement in hair count, terminal hair, or hair thickness may be measured after 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or one year or longer after initiation of the intermittent lithium treatment or pulse lithium treatment.
  • a synergistic effect of an intermittent lithium treatment or pulse lithium treatment on another treatment for restoring or enhancing hair growth may be measured as an improvement over a control subject receiving only one of the two treatments (i.e.. the intermittent lithium treatment or pulse lithium treatment alone or the second treatment alone).
  • the intermittent lithium treatment or a pulse lithium treatment is used in combination treatments (e.g. , described in Section 5.3 supra) to reduce unwanted hair growth.
  • Any appropriate method for testing the safety and efficacy of such treatments may be used, for example, as described in the examples of Sections 14-23 below (see also sections on safety in the clinical protocol in Section 36). Success of such treatments can be measured by:
  • the intermittent lithium treatment or pulse lithium treatment decreases hair count by 5% or more, by 10% or more, by 15% or more, by 20% or more, by 25% or more, by 30% or more, by 40% or more, by 50% or more, by 75% or more, or by 100% or more.
  • the intermittent lithium treatment or pulse lithium treatment decreases terminal hair by 5% or more, by 10% or more, by 15% or more, by 20% or more, by 25% or more, by 30% or more, by 40% or more, by 50% or more, by 75%o or more, or by 100% or more.
  • the intermittent lithium treatment or pulse lithium treatment decreases hair thickness by 5% or more, by 10% or more, by 15% or more, by 20% or more, by 25% or more, by 30% or more, by 40% or more, by 50% or more, by 75% or more, or by 100% or more.
  • Such an improvement in hair count, terminal hair, or hair thickness may be measured after 1 day, 2 days, 3 days, 5 days, 1 week, 2 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or one year or longer after initiation of the intermittent lithium treatment or pulse lithium treatment.
  • a synergistic effect of an intermittent lithium treatment or pulse lithium treatment on another treatment for removing unwanted hair may be measured as an improvement over a control subject receiving only one of the two treatments (i.e., the intermittent lithium treatment or pulse lithium treatment alone or the second treatment alone).
  • the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical minoxidil and to apply the Lithium gluconate 8% gel to affected area of the scalp for one week (to synchronize the hair follicle cells in G2/M phase arrest). After one w r eek, treatment with lithium gluconate is discontinued and treatment with topical minoxidil foam is re-started and he is evaluated after three weeks.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers; and the patient's subjective evaluation of hair growth.
  • the treated area of affected scalp is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc.); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc.); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is treated with 10 cycles of the protocol: alternating topical lithium gluconate (1 week) with minoxidil foam treatment (3 weeks in which finasteride treatment is continued). Response to therapy is measured by the methods described above.
  • EXAMPLE 2 INTERMITTENT TREATMENT OF HAMILTON-NORWOOD TYPE VI MALE-PATTERN ALOPECIA USING LITHIUM GLUC NATE DEVICE ALTERNATING WITH TOPICAL MINOXIDIL
  • the bald and transitional areas of the subject ' s scalp are prepared by shaving and then treated with a fractional and non-ablative Erbium- YAG laser with an emission at 1540-1 550 nm (set to 50- 70 J/cm 2 , treatment level of 8- 10, and 8 passes) and the subject is provided with a topical preparation of Lithium gluconate 8% gel ( Lithioderm 8% gel) and instructed to discontinue topical minoxidil and to apply the Lithium gluconate 8% gel to treated area of the scalp for one week. After one week, treatment with lithium gluconate is discontinued and treatment with topical minoxidil foam is re-started and he is evaluated after three weeks.
  • Lithium gluconate 8% gel Lithioderm 8% gel
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers; and the patient's subjective evaluation of hair growth.
  • the treated area of affected scalp is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc. ); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is optionally treated with 10 more cycles, e.g., to increase hair density, for example: 1 week of topical lithium gluconate followed by 3 weeks of minoxidil foam treatment, with optional laser treatment.
  • Response to therapy is measured by the methods described above.
  • Pre-treatment In preparation for a hair transplant procedure, the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical minoxidil and to apply the Lithium gluconate 8% gel to the to the occipital and deep temporal areas area of the scalp from which follicles will be harvested for transplant for one week (to synchronize the hair follicle cells in G2/M phase arrest). After one week, treatment with lithium gluconate is discontinued and treatment with topical minoxidil foam is re-started and he is evaluated after three weeks.
  • Lithium gluconate 8% gel Lithioderm 8% gel
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers; and the patient ' s subjective evaluation of hair growth.
  • the treated area of affected scalp is biopsied and studied for new follicle growth, bifurcating follicles undergoing follicle division; follicles growing new hair fibers.
  • the subject is treated with 10 cycles of the protocol: alternating topical lithium gluconate ( 1 week) with minoxidil foam treatment (3 weeks). Response to therapy is measured by the methods described above.
  • Post-transplantation The area of scalp that was pre-treated with lithium and minoxidil is used as a source for the transplanted follicles. After hair follicle implantation, treatment with lithium gluconate is initiated for one week, and then discontinued and followed by treatment with topical minoxidil foam for three weeks.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers; and the patient's subjective evaluation of hair growth.
  • the treated area of affected scalp is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc. ); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc.); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is treated with 10 cycles of the protocol : alternating topical lithium gluconate ( 1 week) with minoxidil foam treatment (3 weeks).
  • the subject is provided a treatment in which the bald crown scalp receives in order: (i) Erbium- YAG laser; (ii) lithium; and (iii) estrogen during the phase where follicle stem cells are reorganizing and reforming hair follicles.
  • the estrogen treatment reprograms the follicle stem cells to alter their sensitivity/response to androgens. In effect, this could be described as rendering follicle stem cells either (a) '"female-type” with respect to crown scalp follicle cells or (b) "occipital scalp type" in terms of their lack of response to androgens by involution.
  • EXAMPLE 4 INTERMITTENT PRE-TREATMENT OF "DONOR” SCALP AREAS USING LITHIUM GLUCONATE DEVICE ALTERNATING WITH TOPICAL MINOXIDIL IN PREPARATION FOR A HAIR TRANSPLANT PROCEDURE IN A PATIENT WITH HAMILTON-NORWOOD TYPE VII MALE-PATTERN ALOPECIA
  • Pre-treatment In preparation for a hair transplant procedure, the subject is administered a fractional and non-ablative laser therapy using an Erbium- YAG laser with an emission at 1540-1550 nm (set to 50-70 J/cm 2 , treatment level of 8-10, and 8 passes) and the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical minoxidil and to apply the Lithium gluconate 8% gel to the to the occipital and deep temporal areas area of the scalp from which follicles will be harvest for transplant for one week. After one week, treatment with lithium gluconate is discontinued and treatment with topical minoxidil foam is re-started and he is evaluated after three weeks.
  • Lithium gluconate 8% gel Lithioderm 8% gel
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers.
  • the treated area of affected scalp is biopsied and studied for new follicle growth, bifurcating follicles undergoing follicle division; follicles growing new hair fibers.
  • the subject is optionally treated with 10 more cycles, e.g., to increase hair density, for example: topical lithium gluconate (1 week) with minoxidil foam treatment (3 weeks), with optional laser treatment.
  • Response to therapy is measured by the methods described above.
  • Post-transplantation The area of scalp that was treated with lithium and minoxidil is used as a source for the transplanted follicles. After hair follicle implantation, treatment with lithium gluconate is initiated for one week, and then discontinued and followed by treatment with topical minoxidil foam for three weeks.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers.
  • the treated area of affected scalp is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc.); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is treated with 1 0 cycles of the protocol: alternating Erbium- YAG laser treatment followed by topical lithium gluconate ( 1 week) with minoxidil foam treatment (3 weeks).
  • EXAMPLE 5 INTERMITTENT TREATMENT OF A POSTMENOPAUSAL PATIENT WITH DIFFUSE THINNING OF SCALP HAIR USING LITHIUM GLUCONATE GEL ALTERNATING WITH TOPICAL MINOXIDIL
  • a female human subject 65 years old, complains of diffuse thinning of scalp hair growth after menopause. She describes continued hair loss despite treatment with topical minoxidil foam 2% and oral finasteride 1 mg/day.
  • the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical minoxidil and to apply the Lithium gluconate 8% gel to affected area of the scalp for one week (to synchronize the hair follicle cells in G2/M phase arrest). After one week, treatment with lithium gluconate is discontinued and treatment with topical minoxidil foam is re-started and she is evaluated after three weeks.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers.
  • the treated area of affected scalp is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen. etc.); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is treated with 10 cycles of the protocol: alternating topical lithium gluconate ( I week) with minoxidil foam treatment (3 weeks). Response to therapy is measured by the methods described above.
  • EXAMPLE 6 INTERMITTENT TREATMENT OF A POSTMENOPAUSAL PATIENT WITH DIFFUSE THINNING OF SCALP HAIR USING A LITHIUM GLUCONATE DEVICE ALTERNATING WITH TOPICAL MINOXIDIL
  • the subject is administered a fractional and non-ablative laser therapy using an Erbium- YAG laser with an emission at 1540- 1550 nm (set to 50-70 J/cirf , treatment level of 8- 10, and 8 passes) and the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical minoxidil and to apply the Lithium gluconate 8% gel to the treated area of the scalp for one week. After one week, treatment with lithium gluconate is discontinued and treatment with topical minoxidil foam is re-started and she is evaluated after three weeks.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers.
  • the treated area of affected scalp is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc. ); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division: follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is optionally treated with 10 more cycles, e.g. , to increase hair density, for example: topical lithium gluconate (1 week) followed by minoxidil foam treatment (3 weeks), with optional laser treatment.
  • Response to therapy is measured by the methods described above.
  • a female human subject 65 years old, with Fitzgerald Type II skin complains of diffuse thinning of scalp hair growth after menopause. She describes continued hair loss despite treatment with topical minoxidil 2% foam and oral finasteride 1 mg/day.
  • Pre-treatment In preparation for a hair transplant procedure, the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical minoxidil and to apply the Lithium gluconate 8% gel to the occipital and deep temporal areas area of the scalp from which follicles will be harvested for transplant for one week (to synchronize the hair follicle cells in G2/M phase arrest). After one week, treatment with lithium gluconate is discontinued and treatment with topical minoxidil foam is re-started and she is evaluated after three weeks.
  • Lithium gluconate 8% gel Lithioderm 8% gel
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers.
  • the treated area of affected scalp is biopsied and studied for new follicle growth, bifurcating follicles undergoing follicle division; follicles growing new hair fibers.
  • the subject is treated with 10 cycles of the protocol: alternating topical lithium gluconate ( 1 week) with minoxidil foam treatment (3 weeks). Response to therapy is measured by the methods described above.
  • Post-transplantation The area of scalp that was pre-treated with lithium and minoxidil is used as a source for the transplanted follicles. After hair follicle implantation, treatment with lithium gluconate is initiated for one week, and then discontinued and followed by treatment with topical minoxidil foam for three weeks.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers.
  • the treated area of affected scalp is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc.); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • EXAMPLE 8 INTERMITTENT PRE-TREATMENT OF "DONOR" SCALP AREAS OF A POST-MENOPAUSAL PATIENT WITH DIFFUSE THINNING OF SCALP HAIR USING A LITHIUM GLUCONATE DEVICE ALTERNATING WITH TOPICAL MINOXIDIL IN PREPARATION FOR A HAIR TRANSPLANT PROCEDURE
  • a Caucasian female human subject (with Fitzgerald Type II skin, wherein Type I is the lightest and Type VI is the darkest), 65 years old, complains of diffuse thinning of scalp hair growth after menopause. She describes continued hair loss despite treatment with topical minoxidil 2% foam and oral finasteride 1 mg/day.
  • Pre-treatment In preparation for a hair transplant procedure, the subject is administered a fractional and non-ablative laser therapy using an Erbium- YAG laser with an emission at 1540-1550 nm (set to 50-70 J/cm% treatment level of 8-10, and 8 passes) and the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical minoxidil and to apply the Lithium gluconate 8% gel to the occipital and deep temporal areas area of the scalp from which follicles will be harvested for transplant for one week (to synchronize the hair follicle cells in G2/M phase arrest ). After one week, treatment with lithium gluconate is discontinued and treatment with topical minoxidil foam is re-started and she is evaluated after three weeks.
  • Lithium gluconate 8% gel Lithioderm 8% gel
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers.
  • the treated area of affected scalp is biopsied and studied for new follicle growth, bifurcating follicles undergoing follicle division; follicles growing new hair fibers.
  • the subject is optionally treated with 10 more cycles, e.g. , to increase hair density, for example: alternating Erbium- YAG laser treatment plus topical lithium gluconate ( 1 week) with minoxidil foam treatment (3 weeks). Response to therapy is measured by the methods described above.
  • Post-transplantation The area of scalp that was pre-treated with lithium and minoxidil is used as a source for the transplanted follicles. After hair follicle implantation, treatment with lithium gluconate is initiated for one week, and then discontinued and followed by treatment with topical minoxidil foam for three weeks, with or without laser treatment.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of scalp); thickness of fibers; length of hair fibers.
  • the treated area of affected scalp is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc.); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division: follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is optionally treated with 10 cycles of the protocol: alternating topical lithium gluconate ( 1 week) with minoxidil foam treatment (3 weeks), with or without laser treatment.
  • a female human subject 50 years old, complains of moustache hair growth after menopause. Vaniqa® (eflornithine hydrochloride 13.9%) is being applied without much relief.
  • the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical eflornithine and to apply the Lithium gluconate 8% gel to affected area of the face for one week (to synchronize the hair follicle cells in G2/M phase arrest). After one week, treatment with lithium gluconate is discontinued and treatment with topical eflomithine is re-started and she is evaluated after three weeks.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of facial skin); thickness of fibers; length of hair fibers; and the patient's subjective evaluation of hair growth inhibition.
  • the treated area of affected face is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc. ); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc.); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is optionally treated with 10 more cycles, e.g., to decrease hair density, for example: alternating topical lithium gluconate (1 week) with eflomithine foam treatment (3 weeks). Response to therapy is measured by the methods described above.
  • EXAMPLE 10 INTERMITTENT TREATMENT OF POST MENOPAUSAL FACIAL HAIR USING LITHIUM GLUCONATE DEVICE ALTERNATING WITH TOPICAL EFLORNITHINE
  • a Caucasian female human subject (with Fitzgerald Type II skin, wherein Type I is the lightest and Type VI is the darkest), 50 years old, complains of moustache hair growth after menopause. Vaniqa® (eflomithine hydrochloride 13.9%) is being applied without much relief.
  • the subject is administered a fractional and non-ablative laser therapy using an Erbium- YAG laser with an emission at 1540-1550 nm (set to 50-70 J/cm 2 , treatment level of 8-10, and 8 passes) and the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical eflomithine and to apply the Lithium gluconate 8% gel to the treated area of the face for one week (to synchronize the hair follicle cells in G2/M phase arrest). After one week, treatment with lithium gluconate is discontinued and treatment with topical eflomithine is re-started and she is evaluated after three weeks.
  • Lithium gluconate 8% gel Lithioderm 8% gel
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of facial skin); thickness of fibers; length of hair fibers.
  • the treated area of affected face is biopsied and studied for distribution of follicles in various stages of Follicle Cycle ( anagen, catagen, etc. ); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is optionally treated with 10 more cycles, e.g.. to decrease hair density, for example: topical lithium gluconate ( 1 week) followed by with etlomithine foam treatment ( 3 weeks), with or without laser treatment. Response to therapy is measured by the methods described above.
  • a female human subject 50 years old, complains of moustache hair growth after menopause. Vaniqa (eflornithine hydrochloride 13.9%) is being applied without much relief.
  • the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical eflornithine and to apply the Lithium gluconate 8% gel to affected area of the face for one week (to synchronize the hair follicle cells in G2/M phase arrest). After one week, treatment with lithium gluconate is discontinued and treatment with topical 5-fluorouracil (5-FU, Efudex 5% cream) is started and she is evaluated after three weeks.
  • 5-fluorouracil 5-FU, Efudex 5% cream
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of facial skin); thickness of fibers; length of hair fibers.
  • the treated area of affected face is biopsied and studied for new follicle growth, bifurcating follicles undergoing follicle division; follicles growing new hair fibers.
  • the subject is treated with 10 cycles of the protocol: alternating topical lithium gluconate ( 1 week) with 5-FU cream treatment (3 weeks). Response to therapy is measured by the methods described above.
  • EXAMPLE 12 INTERMITTENT TREATMENT OF POST MENOPAUSAL FACIAL HAIR USING LITHIUM GLUCONATE DEVICE ALTERNATING WITH TOPICAL 5-FLUOROURACIL
  • a Caucasian female human subject (with Fitzgerald Type II skin, wherein Type I is the lightest and Type VI is the darkest), 50 years old, complains of moustache hair growth after menopause. Vaniqa (eflornithine hydrochloride 13.9%) is being applied without much relief.
  • the subject is administered a fractional and non-ablative laser therapy using an Erbium-YAG laser with an emission at 1540-1550 nm (set to 50-70 J/cm 2 , treatment level of 8- 10.
  • Lithium gluconate 8% gel Lithium gluconate 8% gel
  • Lithioderm 8% gel topical preparation of Lithium gluconate 8% gel
  • eflornithine topical eflornithine
  • topical 5-fluorouracil 5-FU, Efudex 5% cream
  • [00461J Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of facial skin); thickness of fibers; length of hair fibers.
  • the treated area of affected face is biopsied and studied for new follicle growth, bifurcating follicles undergoing follicle division; follicles growing new hair fibers.
  • the subject is optionally treated with 10 more cycles, e.g. , to decrease hair density, for example: topical lithium gluconate ( 1 week) followed by 5-FU cream treatment (3 weeks), with or without laser treatment.
  • 10 more cycles e.g. , to decrease hair density
  • topical lithium gluconate 1 week
  • 5-FU cream treatment 3 weeks
  • Response to therapy is measured by the methods described above.
  • a female human subject 30 years old, complains of axillary hair growth and irritation from using razors to shave her armpits.
  • the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) ) and instructed to discontinue topical eflornithine and to apply the Lithium gluconate 8% gel to the affected area of the axilla for one week (to synchronize the hair follicle cells in G2/ phase arrest).
  • Treatment with lithium gluconate is discontinued and treatment with topical Vaniqa (eflornithine hydrochloride 13.9%) is started and she is evaluated after three weeks.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of axillary skin); thickness of fibers; length of hair fibers.
  • the treated area of affected axillary skin is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc. ); distribution of follicle cells in various stages of cell cycle (e.g. G2. M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • EXAMPLE 14 INTERMITTENT TREATMENT OF AXILLARY HAIR USING LITHIUM GLUCONATE DEVICE ALTERNATING WITH TOPICAL EFLORNITHINE
  • a Caucasian female human subject (with Fitzgerald Type 11 skin, wherein Type 1 is the lightest and Type VI is the darkest), 30 years old, complains of axillary hair growth and irritation from using razors to shave her armpits.
  • the subject is administered a fractional and non-ablative laser therapy using an Erbium- Y AG laser with an emission at 1540- 1550 nm (set to 50-70 J/cm 2 , treatment level of 8-10, and 8 passes) and the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical eflomithine and to apply the Lithium gluconate 8% gel to the treated area of the axilla for one week (to synchronize the hair follicle cells in G2/M phase arrest).
  • Treatment with lithium gluconate is discontinued and treatment with topical Vaniqa (eflomithine hydrochloride 13.9%) is started and she is evaluated
  • Response to therapy is detem ined by measuring new hair growth (increased number of fibers in an affected area of axillary skin); thickness of fibers; length of hair fibers.
  • the treated area of affected axillary skin is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc. ); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is optionally treated with 10 more cycles, e.g. , to decrease hair density, for example: topical lithium gluconate (T week) followed by eflomithine foam treatment (3 weeks), with or without laser treatment.
  • 10 more cycles e.g. , to decrease hair density
  • T week topical lithium gluconate
  • eflomithine foam treatment 3 weeks
  • Response to therapy is measured by the methods described above.
  • the treated area of affected back skin is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc. ); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • Follicle Cycle anagen, catagen, etc.
  • follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is treated with 10 cycles of the protocol: alternating topical lithium gluconate ( 1 week) with eflornithine foam treatment (3 weeks). Response to therapy is measured by the methods described above.
  • a Caucasian male human subject (with Fitzgerald Type II skin, wherein Type I is the lightest and Type VI is the darkest), 30 years old, complains of excess back hair growth. Repeated waxings have had short term effects and are painful.
  • the subject is administered a fractional and non-ablative laser therapy using an Erbium- YAG laser with an emission at 1540-1550 nm (set to 50-70 J/cm 2 , treatment level of 8-10, and 8 passes) and the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed apply it to the treated area of the skin for one week (to synchronize the hair follicle cells in G2/M phase arrest). After one week, treatment with lithium gluconate is discontinued and treatment with topical Vaniqa (eflornithine hydrochloride 13.9%) is started and he is evaluated after three weeks.
  • Lithium gluconate 8% gel Lithioderm 8% gel
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of back skin); thickness of fibers; length of hair libers.
  • the treated area of affected back skin is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc.); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair libers, follicles with no hair fibers.
  • the subject is optionally treated with 10 more cycles, e.g. , to decrease hair density, for example: topical lithium gluconate (1 week) followed by eflornithine foam treatment (3 weeks), with or without laser treatment. Response to therapy is measured by the methods described above. 22.
  • EXAMPLE 17 INTERMITTENT TREATMENT OF EAR HAIR
  • a male human subject 55 years old, complains of ear (auricle) hair growth.
  • the subject is provided with a topical preparation of Lithium gluconate 8% ge! (Lithioderm 8% gel) and instructed discontinue topical eilornithine and to apply the Lithium gluconate 8% gel to affected area of the ear for one week (to synchronize the hair follicle cells in G2/M phase arrest).
  • Treatment with lithium gluconate is discontinued and treatment with topical eilornithine is re-started and he is evaluated after three weeks.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of ear ); thickness of fibers; length of hair fibers.
  • the treated area of affected ear is biopsied and studied for distribution of follicles in various stages of Follicle Cycle (anagen, catagen, etc.); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ): new follicle growth, bifurcating follicles; follicles undergoing follicle division; follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is treated with 10 cycles of the protocol: alternating topical lithium gluconate (1 week) with eilornithine foam treatment (3 weeks). Response to therapy is measured by the methods described above.
  • a Caucasian male human subject (with Fitzgerald Type II skin, wherein Type I is the lightest and Type VI is the darkest), 55 years old, complains of ear (auricle) hair growth.
  • the subject is administered a fractional and non-ablative laser therapy using an Erbium- YAG laser with an emission at 1540-1550 nm (set to 50-70 J/cm , treatment level of 8-10, and 8 passes) and the subject is provided with a topical preparation of Lithium gluconate 8% gel (Lithioderm 8% gel) and instructed to discontinue topical eflomithine and to apply the Lithium gluconate 8% gel to the treated area of the ear for one week ( to synchronize the hair follicle cells in G2/M phase arrest). After one week, treatment with lithium gluconate is discontinued and treatment with topical eilornithine is re-started and he is evaluated after three weeks.
  • Response to therapy is determined by measuring new hair growth (increased number of fibers in an affected area of ear); thickness of fibers; length of hair fibers.
  • the treated area of affected ear is biopsied and studied for distribution of follicles in various stages of Follicle Cycle fanagen, catagen. etc.); distribution of follicle cells in various stages of cell cycle (e.g. G2, M, etc. ); new follicle growth, bifurcating follicles; follicles undergoing follicle division: follicles growing new hair fibers, follicles with no hair fibers.
  • the subject is optionally treated with 10 more cycles, e.g., to decrease hair density, for example: topical lithium gluconate ( 1 week) followed by eflornithine foam treatment (3 weeks), with or without laser treatment.
  • 10 more cycles e.g., to decrease hair density
  • eflornithine foam treatment 3 weeks
  • Response to therapy is measured by the methods described above.
  • the treatments presented in the examples in Sections 7, 9, 1 1, 13, 15, 17, 19, 21 , and 23 may alternatively be accomplished by applying an ablative laser treatment in place of the non-ablative laser treatment.
  • the application of Lithium gluconate is sterile and, optionally, the treatment area is covered by a bandage.
  • ablative laser treatment may accomplished using an Erbium-YAG laser at 2940 nm or a C0 2 laser at 10,600 nm.
  • Lidocaine HCL 2% with Epinephrine 1 : 100,000 are injected to anesthetize the surface of the area to be treated.
  • An Ultrapulse (fractional mode) CO? laser is used to disrupt the epidermis and dermis to approximately 100 to 500 ⁇ in depth.
  • the Ultrapulse laser produces an effect that is similar to that of dermabrasion yet the disruption produced delivers a greater amount of energy deeper into the skin in a non-scaring fractional ablation.
  • the treated area is a 1.5 cm x 1.5 cm square.
  • the Ultrapulse is set to deliver up to 350 mJ, up to 52.5 Watts, using pattern size #8, density #4, and fill the square treatment site with up to 5 passes.
  • Lidocaine HCL 2% with Epinephrine 1 .100,000 are injected to anesthetize the surface of the area to be treated.
  • An Ultrapulse C0 2 laser (ablative mode) is used to disrupt the epidermis and dermis to approximately 100 to 500 ⁇ in depth.
  • the Ultrapulse laser produces an effect that is similar to that of dermabrasion yet the disruption produced delivers a greater amount of energy deeper into the skin in a non-scaring ablation that resembles the dermabrasion.
  • the treated area is a 1.5 cm x 1 .5 cm square.
  • Ultrapulse is set to deliver up to 500 mJ in 1 msec, 1 Watts, using a spot size of 3 mm at 2 Hz to fill the square treatment site, which may require up to 15 passes.
  • Lidocaine HCL 2% with Epinephrine 1 : 100,000 are injected to anesthetize the surface of the area to be treated.
  • the ablative erbium laser is set to deliver up to 5 Joules 240 msec in of energy at level 3 so that in up to 15 passes it will produce a disruption up to 500 ⁇ deep.
  • the treated area is a 1.5 cm x 1.5 cm square.
  • Lidocaine HCL 2% with Epinephrine 1 : 100,000 is injected to anesthetize the surface of the area to be treated.
  • Standard dermabrasion using the Aseptico Econo-Dermabrader from Tiemann and Company, is performed to a depth of approximately 150 ⁇ , that includes removal the entire epidermis and disruption of the papillary dermis (detectable by a shiny, whitish appearance) inducing the formation of small pools of blood in the treated area.
  • Each dermabraded area is a 1.5 cm x 1.5 cm square.
  • a Bell Hand dermabrasion device may be used.
  • Sections 28-32 which present mouse studies using dermabrasion, and the protocols for use in humans in the examples of Sections 35 and 36.
  • dermabrasion was carried out using a microdermabrasion device. While dermabrasion in humans may also be carried out using a microdemiabrasion device, where sterile conditions are preferential, a dermabrasion device is preferably used.
  • This protocol is adapted from the IACUC VA protocol. Specifically, 4 week old male SCID mice are obtained from Charles River and allowed to acclimate for at least 1 week. In preparation for surgery, mice are anesthetized with ketamine (80 mg/kg)/xylazine (20 mg/kg) delivered i.p. in a volume ⁇ 100 ⁇ , and monitored by toe pinch to determine the surgical plane of anesthesia. Full thickness adult human skin (measuring approximately 1.5 cm x 2 cm; removed during surgical procedures from the CHTN, NDR1 or cadaver scalp skin from ABS) is sutured into a full thickness skin excision site on the dorsal surface of the mouse. The grafts are bandaged and allowed to heal for at least 5 weeks.
  • mice After healing and successful "'take, 1 ' prior to wounding, the human skin is analyzed using in vivo confocal microscopy, histology and/or photography to determine the ''contror or "pre-wounded" state of the skin graft. Prior to wounding, mice are anesthetized with ketamine (80
  • mice may be required to test the differences between full thickness and split thickness human scalp xenografts.
  • mice receive vehicle alone or the lithium composition, delivered systemically or topically, or neither vehicle nor lithium composition, for 5 consecutive days, (the lithium composition chosen is the one determined to be most efficacious in the C57BL/6J model, with efficacy determined to be increased number and/or size of neogenic hair follicles).
  • One dose of the lithium composition is delivered, using the most efficacious dose as described above, systemically and, in a separate experiment, a dose is delivered topically. Additionally, in vivo confocal microscopy, histology and/or photography is performed daily (until the end of the experiment) following scab detachment in order to monitor hair follicle neogenesis (confocal microscopy is noninvasive but does require anesthesia). An additional set of mice are treated with the lithium composition or vehicle or neither, with the exception that the xenografted mice are not wounded, in order to assess the effect of the lithium composition in the absence of wounding.
  • mice At approximately 2 weeks post-scab detachment, all mice are anesthetized with ketamine (80 mg/kg)/xylazine (20 mg/kg) delivered i.p. in a volume of ⁇ 100 ⁇ , and monitored by toe pinch to determine the surgical plane of anesthesia. Subsequently, they have a terminal blood draw (to detect drug in the plasma), and are euthanized. The wound is then removed, which is trisected with one-third taken for biochemistry, one third for determination of lithium levels in the skin using mass spectrometry, and one third for
  • mice are needed for the optimization of microdermabrasion settings and split thickness versus full thickness xenografts. Considering that the "take" rate of human skin xenografts is approximately 50%, the total number of mice to optimally receive human skin grafts is approximately 500.
  • This example provides a protocol for characterizing and comparing the percutaneous absorption pharmacokinetics of four formulations containing a lithium salt, in human cadaver skin, using the in vitro skin finite dose model.
  • This model is a well- established tool for the study of percutaneous absorption and the determination of the pharmacokinetics of topically applied drugs.
  • the model uses human cadaver skin mounted in specially designed diffusion chambers allowing the skin to be maintained at a temperature and humidity that match typical in vivo conditions.
  • a dose (e.g. , 0.1 gram) of formulation is applied to the top of the partial thickness skin or dermis and drug absorption is measured by monitoring its rate of appearance in the reservoir solution bathing the other surface of the skin.
  • compositions of the formulations are provided in Table 2 below.
  • the formulations were tested initially for stability in solution at 4 °C, 25 °C and 40 °C. All formulations were stable solutions or emulsions at the temperatures tested.
  • the excipients selected for the formulations were based on levels approved for topical drug formulations and each excipient was selected for its viscosity-enhancing properties or its ability to enhance permeation through tissues. Methylparaben was added to the formulations for its preservative activity.
  • Lithium Lithium Gluconate Gel, Lithium gluconate 8%
  • hydrogel Carbopol 980 1.5%
  • Dermal tissue was prepared by heating the full thickness skin at 40 °C for 20 minutes in de- ionized water and removing the epidermis using sterile forceps. All cells were mounted in a diffusion apparatus in which the dermal bathing solution was stirred magnetically at approximately 600 R.PM and its skin surface temperature maintained at 32.0° ⁇ 1.0 °C.
  • PBS Human Receptor Solution
  • the results for the percutaneous absorption of Lithium Chloride are shown in Figures 6-10.
  • the water/oil (w/o) emulsion 28A displayed the lowest percent permeated through the dermis, with 60% released over a 12 hour time period.
  • This emulsion was significantly different than the other prototypes, in that the drug was entrapped within lanolin alcohol lipid spheres and dispersed in a non-aqueous continuous medium (mineral oil).
  • the emulsion 35 A' demonstrated relatively rapid permeation through the dermis, with 100% released in approximately 5 hours at 32 °C.
  • the formulation BX is a neutral hydrogel, with its gel-like consistency produced by the presence of high molecular weight hydroxyethyl cellulose (HEC).
  • EXAMPLE 23 IN VIVO TIME-COURSE ASSESSMENT OF PERMEATION AND RESIDENCE TIME OF TOPICALLY ADMINISTERED LITHIUM IONS (AS A FUNCTION OF FORMULATION TYPE) THROUGH MOUSE SKIN TREATED WITH DERMABRASION AND FULL-THICKNESS EXCISION
  • This example provides an assessment of the rate of permeation and residence time of lithium ions provided in various formulations in an in vivo mouse model developed for follicle neogenesis. Based on the data, appropriate formulations are selected for an in vivo mouse experiment to assess neogenesis. Formulations that have an adequate rate of permeation through the dermis and longest residence time are selected as formulations to enroll in an in vivo model for neogenesis. It is postulated that lithium ions can induce differentiation of stem cells into neogenic hair follicles.
  • Formulations selected in this experiment were : 35A ⁇ 35BX and BV-001 -003A with their respective compositions as shown in Table 2 supra. 28.1 EXPERIMENTAL DESIGN
  • mice 24 C57/BL 6 mice were enrolled in each group. There were 6 groups in total, with 3 groups enrolled for dermabrasion (DA) and 3 for FTE treated skin. A different formulation was enrolled in each of the three groups for DA and FTE.
  • DA dermabrasion
  • FTE FTE
  • Dosing for the DA groups was started at day 0, immediately after debriding the mouse skin with dermabrasion, and continued to day 5. Scab formation on the wound occurs approximately at day 1 and thus the formulations are delivered on top of scabbed wounds.
  • Dosing for the FTE groups was started at approximately day 7, or when the scab detached from the wound.
  • the formulations were delivered to the re-epithelialized skin for five days.
  • Each wound was dosed with a formulation volume of 0.1 ml, or 0.1 g since the density of each formulation was determined to be approximately 1 g/ml. Dosing was accomplished with a 100 microliter Wiretrol device. Post-dosing, the wound was covered with a non-stick Tegaderm bandage.
  • Blood levels were an order of magnitude lower than in skin, possibly because the formulation used, in which the Li ion is complexed with CarboPol 980 to form a polymer, enhances its residence in the skin, in contrast to Li ion in, for example, saline, which is expected to be highly water soluble.
  • EXAMPLE 24 IN VIVO SKIN AND PLASMA DISTRIBUTION OF LI WITH SUBCUTANEOUSLY DELIVERED LITHIUM CHLORIDE IN A DOSE- ESCALATING FASHION IN C57/BLK MICE. WITH THEIR SKIN TREATED WITH DERMABRASION OR FULL THICKNESS SKIN EXCISION
  • the skin and the corresponding plasma concentrations of Li ions were determined following subcutaneous administration of lithium chloride at increasing dose concentrations.
  • T his protocol can also be adapted to determine follicular neogenesis as a function of increasing dose concentrations of lithium.
  • mice were treated with either DA or FTE or unwounded (see Table 3 below), and dosed subcutaneously with 0.1 ml of a formulation containing increasing concentrations of lithium chloride in isotonic saline.
  • mice that received FTE treatment as a mode of wounding were dosed on the day of scab detachment (e.g. , on day 10- 1 5 post-wounding) with a single dose of lithium chloride for 5 days.
  • the mice were sacrificed and the wound was biopsied and analyzed for Li concentration.
  • blood was drawn and centrifuged into RBC and plasma and assayed for lithium levels.
  • Lithium concentrations were measured by the validated bioanalytical ICP method provided below.
  • LOQ for Li in the assay 50 mM.
  • the calibration process involved using lithium standards that were dissolved in purified water.
  • the primary lithium standard was diluted to make a set of lithium working standards.
  • One set of these working standards was used to generate the calibration curve.
  • a separate set was used to prepare the QC samples described in the section entitled "Accuracy. " The method was validated for all three murine matrices.
  • the final calibration curve covered the concentration range from 0.05-50
  • Precision was measured as inter-injection variability, by analyzing six separate injections of the same QC sample, removed from the same vial. Analyte
  • Li concentrations in skin and blood increased in a dose-related fashion.
  • the concentrations of Li in RBC were negligible (data not shown).
  • Li concentrations in skin at trough (24 h post dosing on day 5) were 0.0001 -0.0009 mM Li.
  • Li concentrations in blood at peak ( 1 hr post dosing on day 5) were between 0.695 mM - 1.059 mM Li (see Figures 17 and 18), and trough concentrations in blood were 0.02-0.09 mM Li.
  • the data show that when lithium chloride is delivered subcutaneously, lithium ions extract to the skin and the plasma in a linear dose-related fashion, although plasma concentrations were many-fold higher than in skin.
  • the lithium concentration in skin at 5 days correlates in a linear dose-related fashion, with no difference observed between wounded skin and non-wounded skin (see Figure 15B).
  • the skin samples were obtained 1 hour post-dosing, in other words at "'peak" skin concentrations.
  • the data demonstrate that lithium does distribute to the skin, where it may play a role in stem cell modulation toward differentiation into de novo hair follicles.
  • EXAMPLE 25 IN VIVO DISTRIBUTION OF LI WITH ONCE DAILY TOPICALLY ADMINISTERED LITHIUM FORMULATIONS IN C57 BLK MICE, WITH THEIR SKIN TREATED WITH DERMABRASION OR FULL THICKNESS EXCISION
  • the purpose of the experiment in this example was to evaluate the absorption of Li ions into the skin and blood compartments in an in vivo mouse model developed for follicle neogenesis, with once/day topical administration of two lithium formulations, a Lithium Gluconate Hydrogel and a Lithium Chloride l lydrogel (see Table 10 below).
  • This example provides a protocol for and assessment of the rate of permeation and residence time of lithium ions provided in the formulations, which can be adapted for an in vivo experiment to assess HF neogenesis.
  • lithium gluconate referred to herein as “lithium chloride”
  • CarboPol 980 CarboPol 980

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Abstract

La présente invention concerne des traitements intermittents au lithium, ou un traitement ponctuel au lithium, pour moduler la pousse des poils et des cheveux chez un sujet humain. L'invention concerne également l'utilisation de compositions contenant des composés libérant des ions lithium, ainsi que des adjuvants et de dispositifs servant à leur administration. Le protocole de traitement intermittent implique de multiples cycles de traitement au lithium et interrompus par des « pauses » dans le traitement. Pour le protocole ponctuel, une dose de lithium est administrée sur un bref laps de temps. Le ou les traitements au lithium peuvent être utilisés en association avec d'autres traitements permettant de renforcer ou d'inhiber la pousse des poils ou des cheveux. Lesdits traitements combinés peuvent impliquer des traitements physiques ou mécaniques provoquant une perturbation tégumentaire (par exemple l'électrologie (électrolyse), le laser, la lumière intense pulsée, la dermabrasion, etc.); une stimulation immunitaire (par exemple par des adjuvants, des antigènes, des cytokines, des facteurs de croissance, etc.); et/ou des traitements chimiques (provoquant, par exemple, une perturbation tégumentaire ou favorisant ou inhibant la pousse des poils ou des cheveux); et/ou des traitements chirurgicaux (par exemple une greffe de cheveux), en vue de favoriser ou d'inhiber la pousse des poils ou des cheveux. Ledit ou lesdits traitements combinés peuvent être administrés pendant les cycles de traitement intermittent au lithium, ou durant les « pauses » entre ces cycles; ou simultanément, préalablement et/ou postérieurement à un traitement ponctuel au lithium.
PCT/US2010/048457 2009-09-11 2010-09-10 Traitements intermittents et ponctuels faisant appel au lithium pour moduler la pousse des poils et des cheveux Ceased WO2011031990A1 (fr)

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WO2011119748A1 (fr) * 2010-03-24 2011-09-29 Allergan, Inc. Compositions contenant un prosamide pour traiter la perte des cheveux, les cheveux clairsemés, et la décoloration des cheveux
WO2015049549A1 (fr) * 2013-10-01 2015-04-09 Beauty Thru Science Limited Procédé pour stimuler la repousse des cheveux du cuir chevelu
KR20150044029A (ko) * 2012-08-31 2015-04-23 바이오랩 세너스 팔마씨우티카 엘티디에이. 중합 피나스테리드 및 미녹시딜 나노입자 및 이의 제조 공정, 이를 포함한 수성 현탁액, 약학적 조성물 및 이 조성물의 용도
WO2015150738A1 (fr) * 2014-04-02 2015-10-08 Microarray Limited Pansement comprenant des électrodes
WO2015176161A1 (fr) * 2014-05-23 2015-11-26 Triple Hair Inc. Compositions permettant de réduire la chute des cheveux et/ou d'augmenter la repousse des cheveux
WO2016065268A1 (fr) * 2014-10-24 2016-04-28 Beautopia Llc Traitement combiné d'inhibition de la perte des cheveux
EP2648676A4 (fr) * 2010-12-06 2016-05-04 Follica Inc Procédés destinés à traiter la calvitie et à favoriser la croissance des cheveux
US10039703B2 (en) 2015-07-08 2018-08-07 Triple Hair Inc. Composition comprising resveratrol and melatonin for reducing hair loss and/or increasing hair regrowth
IT201700031017A1 (it) * 2017-03-21 2018-09-21 Bioenx S R L Composizioni utili per il trattamento dell'alopecia androgenica comprendenti acido etidronico
US11696883B2 (en) 2014-05-23 2023-07-11 Triple Hair Inc. Compositions for reducing hair loss and/or increasing hair regrowth

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WO2006105109A2 (fr) * 2005-03-29 2006-10-05 The Trustees Of The University Of Pennsylvania Procedes pour la generation de nouveaux follicules capillaires, de traitement de la calvitie, et l'elimination de poils
US20080269732A1 (en) * 2004-05-19 2008-10-30 Ostern Co., Ltd. Low Power Laser Irradiator for Treating Alopecia

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US20030083381A1 (en) * 2000-03-31 2003-05-01 Hiroki Kumagai Hair growth or hair formation controlling agents
US20080269732A1 (en) * 2004-05-19 2008-10-30 Ostern Co., Ltd. Low Power Laser Irradiator for Treating Alopecia
WO2006105109A2 (fr) * 2005-03-29 2006-10-05 The Trustees Of The University Of Pennsylvania Procedes pour la generation de nouveaux follicules capillaires, de traitement de la calvitie, et l'elimination de poils

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WO2011119748A1 (fr) * 2010-03-24 2011-09-29 Allergan, Inc. Compositions contenant un prosamide pour traiter la perte des cheveux, les cheveux clairsemés, et la décoloration des cheveux
US12263323B2 (en) 2010-12-06 2025-04-01 Follica, Inc. Methods for treating baldness and promoting hair growth
US11207511B2 (en) 2010-12-06 2021-12-28 Follica, Inc. Methods for treating baldness and promoting hair growth
AU2011338530B2 (en) * 2010-12-06 2017-06-15 Follica, Inc. Methods for treating baldness and promoting hair growth
EP3178465A1 (fr) 2010-12-06 2017-06-14 Follica, Inc. Procédé de traitement de calvitie et favorisant la pousse des cheveux
EP2648676A4 (fr) * 2010-12-06 2016-05-04 Follica Inc Procédés destinés à traiter la calvitie et à favoriser la croissance des cheveux
JP2015526484A (ja) * 2012-08-31 2015-09-10 バイオラブ・サヌス・ファーマセウティカ・エルティーディーエー． フィナステリドおよびミノキシジルポリマーナノ粒子、その調製方法、それを含有する水性懸濁液、医薬組成物、ならびにその使用
EP2891486A4 (fr) * 2012-08-31 2016-03-23 Biolab Sanus Farmacéutica Ltda Nanoparticule polymère de finastéride et de minoxidil, procédé pour sa préparation, suspension aqueuse la contenant, composition pharmaceutique, et son utilisation
KR102118582B1 (ko) 2012-08-31 2020-06-04 바이오랩 세너스 팔마씨우티카 엘티디에이. 중합 피나스테리드 및 미녹시딜 나노입자 및 이의 제조 공정, 이를 포함한 수성 현탁액, 약학적 조성물 및 이 조성물의 용도
KR20150044029A (ko) * 2012-08-31 2015-04-23 바이오랩 세너스 팔마씨우티카 엘티디에이. 중합 피나스테리드 및 미녹시딜 나노입자 및 이의 제조 공정, 이를 포함한 수성 현탁액, 약학적 조성물 및 이 조성물의 용도
AU2013308356B2 (en) * 2012-08-31 2017-09-28 Biolab Sanus Farmaceutica Ltda. Polymeric finasteride and minoxidil nanoparticles, method for preparing same, aqueous suspension containing same, pharmaceutical composition and use thereof
WO2015049549A1 (fr) * 2013-10-01 2015-04-09 Beauty Thru Science Limited Procédé pour stimuler la repousse des cheveux du cuir chevelu
WO2015150738A1 (fr) * 2014-04-02 2015-10-08 Microarray Limited Pansement comprenant des électrodes
WO2015176161A1 (fr) * 2014-05-23 2015-11-26 Triple Hair Inc. Compositions permettant de réduire la chute des cheveux et/ou d'augmenter la repousse des cheveux
US10470992B2 (en) 2014-05-23 2019-11-12 Triple Hair Inc. Compositions for reducing hair loss and/or increasing hair regrowth
US11696883B2 (en) 2014-05-23 2023-07-11 Triple Hair Inc. Compositions for reducing hair loss and/or increasing hair regrowth
US10195199B2 (en) 2014-10-24 2019-02-05 Beautopia Llc Combined hair loss inhibition treatment
US10744138B2 (en) 2014-10-24 2020-08-18 Garuhda Llc Combined hair loss inhibition treatment
WO2016065268A1 (fr) * 2014-10-24 2016-04-28 Beautopia Llc Traitement combiné d'inhibition de la perte des cheveux
US10039703B2 (en) 2015-07-08 2018-08-07 Triple Hair Inc. Composition comprising resveratrol and melatonin for reducing hair loss and/or increasing hair regrowth
WO2018172417A1 (fr) 2017-03-21 2018-09-27 Bioenx S.R.L. Compositions utiles pour le traitement de l'alopécie androgénique comprenant de l'acide éthidronique
IT201700031017A1 (it) * 2017-03-21 2018-09-21 Bioenx S R L Composizioni utili per il trattamento dell'alopecia androgenica comprendenti acido etidronico

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