WO2022035849A1 - Methods reducing hair loss and/or increasing hair regrowth - Google Patents

Abstract

The present invention includes a method for treating at least one of hair thinning or hair loss in a patient comprising administering at a site in need of hair growth a fibroblast growth factor- 1 (FGF-1), mutants, and truncated versions thereof, in an amount effective to treat the hair thinning or hair loss.

Description

METHODS REDUCING HAIR LOSS AND/OR INCREASING HAIR REGROWTH

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application Serial No. 63/063,599, filed August 10, 2020, the entire contents of which are incorporated herein by reference.

TECHNICAL FIELD OF THE INVENTION

[0002] The present invention relates in general to the field of treating hair loss and promoting hair growth.

STATEMENT OF FEDERALLY FUNDED RESEARCH

[0003] None.

INCORPORATION-BY-REFERENCE OF MATERIALS FILED ON COMPACT DISC

[0004] The present application includes a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on , 2021, is named VENT2009WO_SeqList.txt and is , kilobytes in size.

BACKGROUND OF THE INVENTION

[0005] Without limiting the scope of the invention, its background is described in connection with hair loss.

[0006] Many different compositions and methods have been proposed to treat hair loss. One such patent is U.S. Patent No. 10,716,829, issued to Yang and entitled, “Method and composition for treatment of hair loss”. Briefly, this inventor is said to teach a method of treating a hair loss by administering a Notch signaling pathway activator to a subject in need thereof.

[0007] Another such patent is U.S. Patent No. 10,470,992, issued to Sekhavat and entitled, “Compositions for reducing hair loss and/or increasing hair regrowth”. This inventor teaches a composition comprising 2% to 5% minoxidil, 0.01% to 15% finasteride and 0.01% to 15% of a prostaglandin analog. In one embodiment, the prostaglandin analog is latanoprost, and the composition comprises 5% minoxidil, 0.1% finasteride and 0.03% latanoprost. The composition is said to reduce hair loss and/or increase regrowth of hair in a human subject. [0008] Another such patent is U.S. Patent No. 10,426,718, issued to Chen and entitled “Compositions and methods for treating hair loss and delaying aging of skin”. Briefly, this inventor is said to methods for inducing new hair growth, and methods of preventing hair loss, by applying an effective amount of a dihydromyricetin compound to the scalp.

[0009] Despite these improvements, what is needed is a long-term solution to hair thinning and/or loss.

SUMMARY OF THE INVENTION

[0010] In one embodiment, the present invention includes a method for treating at least one of hair thinning or hair loss in a patient comprising administering at a site in need of hair growth a fibroblast growth factor-1 (FGF-1) in an amount effective to treat the hair thinning or hair loss. In one aspect, the FGF-1 is an FGF-1 mutant or a truncated FGF-1. In another aspect, the truncated FGF-1 is an FGF-11 40, FGF-11 41, FGF-11454, or FGF-11455. In another aspect, the mutant FGF-1 comprises 1, 2, 3, 4, or 5 amino acid changes, or combinations thereof, selected from K9A, K12V, S17R, N18R, N18K, H21Y, R35E, L44F, A66C, Y94V, N95V, H102Y, F108Y, N114R, N114K, Cl 17V, L133A. In another aspect, the FGF-1 is administered subcutaneously, intradermally, or transcutaneous. In another aspect, the FGF-1 is formulated in a pharmaceutical composition formulated for immediate release, sustained release, microneedle patch, transdermal enhancers, or combinations thereof. In another aspect, the FGF-1 further comprises heparin, heparin like molecules, VEGF, TGF-a, TGF-J3, angiostatin, endostatin, and platelet-derived endothelial cell growth factor (PDGF), albumin, FGF-2, other growth factors that induce or enhance angiogenesis, or molecules that enhance the effects of FGF-1. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is administered to the subject every 8, 12, 16, 24, 36, 48, 60, 72 days, or at least one of: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 weeks. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, combinations thereof, is repeatedly administered to the subject at least once per week for 3, 4, 5, 6, 7 or more weeks. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is provided at 20, 50, 100, 200, 500 and 1,000 ug per injection site. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is via direct injection into the scalp or surrounding tissues. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, increases hair growth more than 25, 30, 33, 35, 40, 45, or 50% when compared to a non-treated scalp tissue. In another aspect, the FGF-1 is administered as a total of about 30 to about 60 subcutaneous injections, each in an amount about 0.05 to about 0.1 mL at intervals of about 1 cm to about 2 cm. In another aspect, at least one of the hair thinning and hair loss of the patient is caused by conditions such as male pattern hair loss, female pattern hair loss, alopecia areata, or traction alopecia.

[0011] In another embodiment, the present invention includes a method of treating at least one of hair thinning or hair loss in a patient comprising: identifying a patient with at least one of hair thinning or hair loss of the skin; administering at a site in need of hair growth a fibroblast growth factor- 1 (FGF-1) in an amount effective to treat the hair thinning or hair loss; and applying to the site in need of hair growth with the composition to treat a diminished microvascular blood flow of the scalp, wherein the FGF-1 at least one of hair thinning or hair loss, wherein the hair thinning or hair loss includes preventing additional hair loss, causing new hair growth, increasing the rate of growth, or increasing the thickness of hair. In one aspect, the FGF-1 is an FGF-1 mutant or a truncated FGF-1. In another aspect, the truncated FGF-1 is an FGF-I1 40, FGF-1 M4i, FGF-1 M54, or FGF-11.155. In another aspect, the mutant FGF-1 comprises 1, 2, 3, 4, or 5 amino acid changes, or combinations thereof, selected from K9A, K12V, S17R, N18R, N18K, H21Y, R35E, L44F, A66C, Y94V, N95V, H102Y, F108Y, N114R, N114K, C117V, L133A. In another aspect, the FGF-1 is administered subcutaneously, intradermally, or transcutaneous. In another aspect, the FGF-1 is formulated in a pharmaceutical composition formulated for immediate release, sustained release, microneedle patch, transdermal enhancers, or combinations thereof. In another aspect, the FGF-1 further comprises heparin, heparin like molecules, VEGF, TGF-a, TGF-J3, angiostatin, endostatin, and platelet-derived endothelial cell growth factor (PDGF), albumin, FGF-2, other growth factors that induce or enhance angiogenesis, or molecules that enhance the effects of FGF-1. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is administered to the subject every 8, 12, 16, 24, 36, 48, 60, 72 days, or at least one of: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 weeks. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, combinations thereof, is repeatedly administered to the subject at least once per week for 3, 4, 5, 6, 7 or more weeks. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is provided at 20, 50, 100, 200, 500 and 1,000 ug per injection site. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is via direct injection into the scalp or surrounding tissues. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, increases hair growth more than 25, 30, 33, 35, 40, 45, or 50% when compared to a non-treated scalp tissue. In another aspect, the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof are administered as a total of about 30 to about 60 subcutaneous injections, each in an amount about 0.05 to about 0.1 mL at intervals of about 1 cm to about 2 cm. In another aspect, at least one of the hair thinning and hair loss of the patient is caused by conditions such as male patern hair loss, female patern hair loss, alopecia areata, or traction alopecia.

BRIEF DESCRIPTION OF THE DRAWINGS

[0012] For a more complete understanding of the features and advantages of the present invention, reference is now made to the detailed description of the invention along with the accompanying figures and in which:

[0013] FIG. 1 is a photograph that shows the scalp of a subject on Day 0.

[0014] FIG. 2 is a photograph that shows the scalp of the subject 3 months later, which shows significant new hair growth.

[0015] FIG. 3 is a photograph that shows the scalp of the subject after over one year, showing that hair continued to remain and grow after at least one year.

DETAILED DESCRIPTION OF THE INVENTION

[0016] While the making and using of various embodiments of the present invention are discussed in detail below, it should be appreciated that the present invention provides many applicable inventive concepts that can be embodied in a wide variety of specific contexts. The specific embodiments discussed herein are merely illustrative of specific ways to make and use the invention and do not delimit the scope of the invention.

[0017] To facilitate the understanding of this invention, a number of terms are defined below. Terms defined herein have meanings as commonly understood by a person of ordinary skill in the areas relevant to the present invention. Terms such as “a”, “an” and “the” are not intended to refer to only a singular entity but include the general class of which a specific example may be used for illustration. The terminology herein is used to describe specific embodiments of the invention, but their usage does not limit the invention, except as outlined in the claims.

[0018] Arteriogenesis is technically considered remodeling of pre-existing vascular channels (collaterals) or de novo artery formation, it can be stimulated by local changes in perfusion (shear stress), as well as cellular influx and proliferation, and associated with a 20-30 fold increase in blood flow. Vasculogenesis is technically considered on the one hand to encompass embryonic vascular development, and on the other hand to include de novo formation or remodeling of pre-existing vascular channels initiated by circulating vascular precursor cells; furthermore; it is considered to be ischemia and injury initiated. The term “angiogenesis” is meant to encompass all three technical terms. [0019] Angiogenesis is known to occur physiologically during zygote implantation, embryogenesis, post-embryonic growth, and during tissue repair and remodeling. Pathologically, uncontrolled angiogenesis is associated with a variety of diseases such as macular degeneration, diabetic retinopathy, inflammation, including arthritis and psoriasis, and cancer. One common aspect of adult angiogenesis is tissue hypoxia. In situations of tissue expansion, cells are typically dependent on the microvasculature for nutrients and oxygen supply, as well as removal of metabolic waste products. Accordingly, during tissue growth, cells begin to “sense” a lack of oxygen. This triggers a cascade of events that culminates in angiogenesis. During pathological conditions, such as the conditions associated with hypoxic and/or ischemic intestinal disease, the lack of oxygen is induced through hypoperfusion. In some pathological conditions, the normal angiogenic response to hypoxia is absent or substantially diminished.

[0020] As used herein, the terms “treating,” “treatment,” “therapeutic,” or “therapy” do not necessarily mean total cure or abolition of the disease or condition, but rather, include any alleviation of any undesired signs or symptoms of a disease or condition, to any extent, can be considered treatment and/or therapy. It is entirely possible that “treatment” consists of a temporary improvement of the endplate vasculature that requires repeated treatment over time to continue the regenerative process. Furthermore, treatment may include acts that may worsen the patient's overall feeling of well-being or appearance.

[0021] As used herein, the phrase “therapeutically effective amount” refers to a compound as used herein to indicate an amount of an active compound, or pharmaceutical agent, that elicits the biological or medicinal response indicated, e.g., FGF-1. This response may occur in a tissue, system, animal or human and includes alleviation of the symptoms of the disease being treated. The exact formulation, route of administration and dosage for the composition and pharmaceutical compositions disclosed herein can be chosen by the individual physician in view of the patient's condition. (See e.g., Fingl et al. 1975, in “The Pharmacological Basis of Therapeutics”, Chapter 1, and updates thereof, or Remington's Pharmaceutical Sciences, Mace Publishing Company, Philadelphia, Pa., 17th ed. (1985) and updates thereof, relevant portions incorporated herein by reference). Therapeutic treatments can be achieved with small molecule organic drugs or biologies, such as proteins. Typically, the dose range of a small molecule therapeutic agent is administered from about 0.5 to 1000 pg/kg, or 1 to 500 pg/kg, or 10 to 500 pg/kg, or 50 to 100 pg/kg of the patient's body weight per dose. The dose of a therapeutic protein growth factor, such as truncated forms of FGF-1, can be administered to the patient intravenously or intraarterially as either a bolus dose or by infusion from about 0.1 to 100 pg/kg of the patient's body weight, or 0.3 to 30 pg/kg, or 1 to 3 pg/kg of the patient's body weight per dose. To achieve localized targeted dosing, FGF-1 can be injected either directly into or adjacent to the scalp. Localized dose ranges can be from 10 ng/cm³ to 1 mg/cm³, or 100 ug/cm³ to 100 ug/cm³ or 1 ug/cm³ to 10 ug/cm³ of target scalp tissue. Local doses can be administered at each area in need of treatment for hair loss, such as the scalp, but also facial tissue. The dosage may be a single one or a series of two or more given in the course of one or more weeks or months, as is needed by the patient.

[0022] As used herein, the term “FGF-1” refers to those native FGF-1, FGF-1 mutants or FGF-1 variants of FGF-1 that are still biologically active (that is, they activate FGF-1 receptors), but have a sequence that is altered from the natural FGF-1 that encodes a 154 or 155 amino acid protein of SEQ ID NO:2. The present invention includes the use of FGF-1¹⁵'¹⁵⁵ and mutants thereof. The skilled artisan will recognize that the mature form of FGF-1 has the first 14 amino acids cleaved after synthesis and prior to export. For examples, truncated FGF-1 proteins for use with the present invention include: synthetic genes that encode a 140 or 141 amino acid protein of SEQ ID NO: 1, which is the mature form of human (i.e., the first 14 amino acids have been cleaved), also referred to as: FGF-1¹'¹⁴⁰, FGF-1¹'¹⁴¹, FGF-1¹⁰'¹⁴⁰, FGF-1¹⁰'¹⁴¹, FGF-1¹'¹⁴⁰, FGF- 112-140, pGp_ | ^{l 2}'^l4 _anc| _mature FGF-1 with point mutations including, for example, one or more of the following: K9A, K12V, S17R, N18R, N18K, H21Y, R35E, L44F, A66C, Y94V, N95V, H102Y, F108Y, N114R, N114K, Cl 17V, L133A of the mature form of the human FGF-1 (FGF- l¹⁵'¹⁵⁵) which has the amino acid sequence: MFNLPP GNYKKPKLLY CSNGGHFLRI LPDGTVDGTRDRSDQHIQLQ LSAESVGEVY IKSTETGQYL AMDTDGLLYG SQTPNEECLF LERLEENHYN TYISKKHAEK NWFVGLKKNG SCKRGPRTHY GQKAILFLPL PVSSD (SEQ ID NO: 1). The full length human FGF-1 is UniProtKB - P05230 (FGF1 HUMAN), Entrez Gene: 2246) SEQ ID NO: 2 MAEGEITTFT ALTEKF NLPP GNYKKPKLLY CSNGGHFLRI LPDGTVDGTRDRSDQHIQLQ LSAESVGEVY IKSTETGQYL AMDTDGLLYG SQTPNEECLF LERLEENHYN TYISKKHAEK NWFVGLKKNG SCKRGPRTHY GQKAILFLPL PVSSD (SEQ ID NO:2). Mutant FGF-1 s with one or more amino acid insertions, deletions or substitutions are introduced by standard genetic engineering techniques, such as site-directed, deletion, and insertion mutagenesis. The wild type FGF-1 three-dimensional conformation is known to be marginally stable with denaturation occurring either at or near physiologic temperature. FGF-1 binding to heparin increases the thermal inactivation temperature by approximately 20°C, thus, in certain embodiments the mutant FGF-1 is combined with heparin. Further, the FGF-1 of the present invention can also be formulated with a therapeutically approved USP heparin, or a mutant heparin. The truncations, insertions, deletions or substitutions of the mutant FGF-1 tends to enhance half-life, which is further enhanced by the inclusion of heparin. Further, mutant heparins can also be used to further enhance the half-life or activity of the mutant FGF-1 used herein. However, heparin is an anti-coagulant that can promote bleeding as a function of increasing concentration. In addition, some individuals have been immunologically sensitized to heparin by previous therapeutic exposure, which can lead to heparin-induced thrombocytopenia and thrombotic events. Mutations that extend the storage stability in vitro and biologic activity in vivo would allow FGF-1 to be formulated and dosed in the absence of exogenous heparin. These include mutations that decrease the rate of oxidative inactivation, such as replacement of one or more of the three cysteine residues by either serine or other compatible residues. In particular, as has been described by others, substitution of cysteine 117 by serine is known to substantially increase the half-life of human FGF-1 by decreasing the rate of oxidative inaction. Other mutations have been described that increase conformational stability by making amino acid changes in internal buried and/or external exposed amino acid residues. In the case of repeat dosing regimens, FGF-ls exhibiting greater stability and life-time might effectively decrease the frequency and number of repeated doses needed to achieve sustained exposure and greater efficacy. These stabilized mutants would allow longer duration dosing from slow release polymeric matrices and delivery systems.

[0023] The pharmaceutical composition(s) of FGF-1, mutant FGF-1, truncated FGF-1 (including but not limited to, e.g.: FGF-1 H®, FGF-11-141, FGF-11.154, or FGF-11 55), or both mutant and truncated FGF-1, may be formulated for injection and administered by injection, e.g., intraperitoneal, intramuscular, or intravenous injection. Such compositions can have a pH of between 6.5 and 8.5 or between 6.8 and 7.8. Excipients/carriers/other ingredients can include a sterile aqueous buffer, an isotonizing agent, a microbicidal agent or preservative, a chelating agent, a solubility enhancing agent such as dimethylsulfoxide, and/or other ingredients. The isotonizing agent can be, e.g., sorbitol, glycerine, polyethylene glycol, propylene glycol, glucose and sodium chloride. The microbicidal agent/preservative can be, e.g., para-oxy benzoic acid esters, benzyl alcohol, para-chloro-meta-xylenol, chlorocresol, phenetyl alcohol, sorbic acid and salts thereof, thimerosal, chlorobutanol, etc. The chelating agent can be, for example, sodium edetate, sodium citrate or the sodium salt of condensed phosphoric acid. Other forms of FGF-1 may also be used, e.g., non-natural variants of FGF-1 that are still biologically active (activate FGF-1 receptors) but have a sequence that is not found in nature. For examples, truncated mutant FGF-1 proteins for use with the present invention include: synthetic genes that encode a 140 or 141 amino acid protein of SEQ ID NO:1, which is the mature form of human, also referred to as: FGF-11 40, FGF-I1 41, FGF-110-140, FGF-110-141, FGF-11.140, FGF-112-140, FGF-I12- 141, and mature FGF-1 with point mutants including, for example, one or more of the following: K9A, K12V, S17R, N18R, N18K, H21Y, R35E, L44F, A66C, Y94V, N95V, H102Y, F108Y, N114R, N114K, Cl 17V, L133A of the mature form of the human FGF-1 (FGF-116-155). The full length human FGF-1 is UniProtKB - P05230 and its gene sequence (FGF1 HUMAN), Entrez Gene: 2246, are incorporated herein by reference. Mutant FGF-ls with one or more amino acid insertions, deletions or substitutions are introduced by standard genetic engineering techniques, such as site-directed, deletion, and insertion mutagenesis. The wild type FGF-1 three- dimensional conformation is known to be marginally stable with denaturation occurring either at or near physiologic temperature. FGF-1 binding to heparin increases the thermal inactivation temperature by approximately 20°C, thus, in certain embodiments the mutant FGF-1 is combined with heparin. Further, the mutant FGF-1 of the present invention can also be formulated with a therapeutically approved USP heparin, or a mutant heparin. The truncations, insertions, deletions or substitutions of the mutant FGF-1 tends to enhance half-life, which is further enhanced by the inclusion of heparin. Further, mutant heparins can also be used to further enhance the half-life or activity of the mutant FGF-1 used herein. However, heparin is an anti-coagulant that can promote bleeding as a function of increasing concentration. In addition, some individuals have been immunologically sensitized to heparin by previous therapeutic exposure, which can lead to heparin-induced thrombocytopenia and thrombotic events. Mutations that extend the storage stability in vitro and biologic activity in vivo would allow FGF-1 to be formulated and dosed in the absence of exogenous heparin. These include mutations that decrease the rate of oxidative inactivation, such as replacement of one or more of the three cysteine residues by either serine or other compatible residues. In particular, as has been described by others, substitution of cysteine 117 by serine is known to substantially increase the half-life of human FGF-1 by decreasing the rate of oxidative inaction. Other mutations have been described that increase conformational stability by making amino acid changes in internal buried and/or external exposed amino acid residues. In the case of repeat dosing regimens, FGF-ls exhibiting greater stability and life-time might effectively decrease the frequency and number of repeated doses needed to achieve sustained exposure and greater efficacy. These stabilized mutants would allow longer duration dosing from slow release polymeric matrices and delivery systems.

[0024] In other embodiments it may be desirable to utilize an angiogenesis-stimulating protein for administration in a therapeutically effective amount. Said protein may be selected from proteins known to stimulate angiogenesis, including but not limited to TPO (thyroid peroxidase), SCF (stem cell factor), IL-1 (interleukin 1), IL-3, IL-6, IL-7, IL-11, flt-3L (fms-like tyrosine kinase 3 ligand), G-CSF (granulocyte-colony stimulating factor), GM-CSF (granulocyte monocyte-colony stimulating factor), EPO (erythropoietin), FGF-1, FGF-2, FGF-4, FGF-5, FGF-20, IGF (insulin-like growth factor), EGF (epidermal growth factor), NGF (nerve growth factor), LIF (leukemia inhibitory factor), PDGF (platelet-derived growth factor), BMPs (bone morphogenetic protein), activin-A, VEGF (vascular endothelial growth factor), VEGF-B, VEGF-C, VEGF-D, P1GF, and HGF (hepatocyte growth factor). In some embodiments, administration of the angiogenesis-stimulating protein is performed by injection directly into a vertebral body. In some embodiments, the angiogenic-stimulating protein is co-administered with stem or progenitor cells.

[0025] In some embodiments a carrier solution or containing/metering device may be desired. Appropriate carrier solutions may be selected based on properties such as viscosity, ease of administration, ability to bind solution over a period of time, and general affinity for the agent delivered. Said solutions may be modified or additives incorporated for modification of biological properties. Starting solutions may include certain delivery polymers known to one who is skilled in the art. These could be selected from, for example: polylactic acid (PLA), poly- L-lactic acid (PLLA), poly-D-lactic acid (PDLA), polyglycolide, polyglycolic acid (PGA), polylactide-co-glycolide (PLGA), polydioxanone, polygluconate, polylactic acid-polyethylene oxide copolymers, polyethylene oxide, modified cellulose, collagen, polyhydroxybutyrate, polyhydroxpriopionic acid, polyphosphoester, poly (alpha-hydroxy acid), polycaprolactone, polycarbonates, polyamides, polyanhydrides, polyamino acids, polyorthoesters, polyacetals, polycyanoacrylates, degradable urethanes, aliphatic polyester polyacrylates, polymethacrylate, acryl substituted cellulose acetates, non-degradable polyurethanes, polystyrenes, polyvinyl fluoride, polyvinyl imidazole, chlorosulphonated polyolefin, and polyvinyl alcohol.

[0026] As used herein, the term "transdermal enhancer" refers to compounds that increase permeability and/or accelerate the delivery of the FGF-1, mutants or variants taught herein through the skin at a location in need of treatment for hair thinning or hair loss. Non-limiting examples of transdermal enhancer include: alcohols including ethyl, isopropyl, butyl and benzyl alcohols; dihydric alcohols including ethylene glycol, diethylene glycol, or propylene glycol dipropylene glycol and trimethylene glycol; polyhydric alcohols including glycerin, sorbitol and polyethylene glycol; polyethylene glycol ethers of aliphatic alcohols (including cetyl, lauryl, oleyl and stearyl) polyoxyethylene-4-lauryl ether, polyoxyethylene-2-oleyl ether and polyoxyethylene-10-oleyl ether; vegetable, animal and fish fats and oils including cotton seed, com, safflower, olive and castor oils, squalene, and lanolin; fatty acid alcohols including oleyl alcohol and its derivatives; fatty acid amides including oleamide and its derivatives; urea and urea derivatives such as allantoin; polar solvents including dimethyldecylphosphoxide, methyloctylsulfoxide, dimethyllaurylamide, dodecylpyrrolidone, isosorbitol, dimethylacetonide, dimethylsulfoxide, decylmethylsulfoxide and dimethylformamide; salicylic acid; benzyl nicotinate; fatty acid esters including propyl oleate, decyl oleate, isopropyl palmitate, glycol palmitate, glycol laurate, dodecyl myristate, isopropyl myristate and glycol stearate; aliphatic surfactants including lauryl sulfate salts, esters of sorbitol and sorbitol anhydride such as polysorbate; or oleic and linoleic acids, triacetin, ascorbic acid, panthenol, butylated hydroxy toluene, tocopherol, tocopherol acetate, and/or tocopheryl linoleate.

[0027] In various embodiments, treatment of scalp tissue can include the use of a biocompatible or biodegradable implant. Biodegradable implants can contain a biodegradable delivery system, or carrier, as well as angiogenic factors; said angiogenic factors could be capable of stimulating sufficient neovascularization to overcome local hypoxia. One preferred angiogenic factor FGF- 1, mutant FGF-1, truncated FGF-1, or both mutant and truncated FGF-1. However, other recombinant naturally derived, in vitro derived, and in vivo derived angiogenic factors may also be used. In some embodiments, the biodegradable implant that contains the angiogenic factors contains a carrier. The carrier is preferably chosen so as to remain within the implanted site for a prolonged period and slowly release the angiogenic factors contained therein to the surrounding environment. This mode of delivery allows said angiogenic factors to remain in therapeutically effective amounts within the site for a prolonged period. By providing said angiogenic factors within a carrier, the advantage of releasing said angiogenic factors directly into the target area is realized. In some embodiments, the implant's carrier is provided in an injectable form. Injectability allows the carrier to be delivered in a minimally invasive and preferably percutaneous method. In some embodiments, the injectable carrier is a gel. In others, the injectable carrier comprises hyaluronic acid (HA).

[0028] In various embodiments, angiogenic treatments can be used in conjunction with other treatments, such as introduction and/or injection of stem cells, which may be embryonic stem cells or adult stem cells. Such angiogenic treatments could be used to prepare tissues for subsequent injection of stem cells, or angiogenic compounds could be injected concurrently with and/or after introduction of such cells. With regards to scalp tissue, growth factors, synthetic or treated allograft or xenograft tissue for scaffold (or extra-cellular matrix) and stem cells (each of which could be used separately or in varying levels of in combination with each other) could be utilized to “engineer” or otherwise modify the scalp tissue with the goal of regenerating living tissue within the scalp.

[0029] In addition, it may be determined that a combination of stem cells, engineered tissue, scaffold, growth factors, or combinations thereof, would be enhanced by combining angiogenic factors such as FGF-1 in its native state or through an FGF-1 mutant (through protein engineering technology) or any other appropriate angiogenic factor to decrease intestinal tissue damage.

[0030] Example 1. Treatment of Hair Loss with FGF-1.

[0031] Male subject had a family history of balding. Subject had used other treatments without success. These other treatments included: Finasteride, Minoxidil and other topical treatments including injections in scalp of platelet rich plasma (PRP). A concentration of 1.5M platelets/microliter is used. O.lml/cm² was injected. After three weeks there was no improvement from the PRP.

[0032] The subject was treated by subcutaneous administration as follows. FGF-1 via ~50 injections over the affected area using an insulin syringe. It was injected by a physician skilled in the field. No side effects reported other than standard pain associated with subcutaneous injections of the scalp, which are consistent with the procedure. There were 5 rows of 10 injections each of 0. Iml/inj ection for a total of 5 ml total. The concentration of FGF-1 used was Img/mL in phosphate buffered saline (PBS).

[0033] FIG. 1 is a photograph that shows the scalp of a subject on Day 0.

[0034] FIG. 2 is a photograph that shows the scalp of the same subject 3 months later, which shows significant new hair growth.

[0035] FIG. 3 is a photograph that shows the scalp of same subject after over one year, showing that hair continued to remain and grow after at least one year. While the single treatment did not fully restore the hairline, the treatment halted the receding hair line and led to new growth and regrowth, which was maintained over time.

[0036] It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method, kit, reagent, or composition of the invention, and vice versa. Furthermore, compositions of the invention can be used to achieve methods of the invention. [0037] It will be understood that particular embodiments described herein are shown by way of illustration and not as limitations of the invention. The principal features of this invention can be employed in various embodiments without departing from the scope of the invention. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures described herein. Such equivalents are considered to be within the scope of this invention and are covered by the claims.

[0038] All publications and patent applications mentioned in the specification are indicative of the level of skill of those skilled in the art to which this invention pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

[0039] The use of the word “a” or “an” when used in conjunction with the term “comprising” in the claims and/or the specification may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.” The use of the term “or” in the claims is used to mean “and/or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and “and/or.” Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the device, the method being employed to determine the value, or the variation that exists among the study subjects.

[0040] As used in this specification and claim(s), the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited features, elements, components, groups, integers, and/or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and/or steps. In embodiments of any of the compositions and methods provided herein, “comprising” may be replaced with “consisting essentially of’ or “consisting of’. As used herein, the term “consisting” is used to indicate the presence of the recited integer (e.g., a feature, an element, a characteristic, a property, a method/process step or a limitation) or group of integers (e.g., feature(s), element(s), characteristic(s), property(ies), method/process steps or limitation(s)) only. As used herein, the phrase “consisting essentially of’ requires the specified features, elements, components, groups, integers, and/or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and/or steps as well as those that do not materially affect the basic and novel characteristic(s) and/or function of the claimed invention.

[0041] The term “or combinations thereof’ as used herein refers to all permutations and combinations of the listed items preceding the term. For example, “A, B, C, or combinations thereof’ is intended to include at least one of: A, B, C, AB, AC, BC, or ABC, and if order is important in a particular context, also BA, CA, CB, CBA, BCA, ACB, BAC, or CAB. Continuing with this example, expressly included are combinations that contain repeats of one or more item or term, such as BB, AAA, AB, BBC, AAABCCCC, CBBAAA, CABABB, and so forth. The skilled artisan will understand that typically there is no limit on the number of items or terms in any combination, unless otherwise apparent from the context.

[0042] As used herein, words of approximation such as, without limitation, “about”, “substantial” or “substantially” refers to a condition that when so modified is understood to not necessarily be absolute or perfect but would be considered close enough to those of ordinary skill in the art to warrant designating the condition as being present. The extent to which the description may vary will depend on how great a change can be instituted and still have one of ordinary skill in the art recognize the modified feature as still having the required characteristics and capabilities of the unmodified feature. In general, but subject to the preceding discussion, a numerical value herein that is modified by a word of approximation such as “about” may vary from the stated value by at least ±0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 10, 12 or 15%, or as understood to be within a normal tolerance in the art, for example, within 2 standard deviations of the mean. Unless otherwise clear from the context, all numerical values provided herein are modified by the term about.

[0043] Additionally, the section headings herein are provided for consistency with the suggestions under 37 CFR 1.77 or otherwise to provide organizational cues. These headings shall not limit or characterize the invention(s) set out in any claims that may issue from this disclosure. Specifically and by way of example, although the headings refer to a “Field of Invention,” such claims should not be limited by the language under this heading to describe the so-called technical field. Further, a description of technology in the “Background of the Invention” section is not to be construed as an admission that technology is prior art to any invention(s) in this disclosure. Neither is the “Summary” to be considered a characterization of the invention(s) set forth in issued claims. Furthermore, any reference in this disclosure to “invention” in the singular should not be used to argue that there is only a single point of novelty in this disclosure. Multiple inventions may be set forth according to the limitations of the multiple claims issuing from this disclosure, and such claims accordingly define the invention(s), and their equivalents, that are protected thereby. In all instances, the scope of such claims shall be considered on their own merits in light of this disclosure, but should not be constrained by the headings set forth herein.

[0044] All of the compositions and/or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

[0045] To aid the Patent Office, and any readers of any patent issued on this application in interpreting the claims appended hereto, applicants wish to note that they do not intend any of the appended claims to invoke paragraph 6 of 35 U.S.C. § 112, U.S.C. § 112 paragraph (1), or equivalent, as it exists on the date of filing hereof unless the words “means for” or “step for” are explicitly used in the particular claim.

[0046] For each of the claims, each dependent claim can depend both from the independent claim and from each of the prior dependent claims for each and every claim so long as the prior claim provides a proper antecedent basis for a claim term or element.

Claims

What is claimed is:

1. A method for treating at least one of hair thinning or hair loss in a patient comprising administering at a site in need of hair growth a fibroblast growth factor- 1 (F GF- 1) in an amount effective to treat the hair thinning or hair loss.

2. The method of claim 1, wherein the FGF-1 is an FGF-1 mutant or a truncated FGF-1.

3. The method of claim 1, wherein the truncated FGF-1 is an FGF-11 40, FGF-11441, FGF- 11454, or FGF-11455.

4. The method of claim 1, wherein the mutant FGF-1 comprises 1, 2, 3, 4, or 5 amino acid changes, or combinations thereof, selected from K9A, K12V, S17R, N18R, N18K, H21Y, R35E, L44F, A66C, Y94V, N95V, H102Y, F108Y, N114R, N114K, Cl 17V, L133A.

5. The method of claim 1, wherein the FGF-1 is administered subcutaneously, intradermally, or transcutaneous.

6. The method of claim 1, wherein the FGF-1 is formulated in a pharmaceutical composition formulated for immediate release, sustained release, microneedle patch, transdermal enhancers, or combinations thereof.

7. The method of claim 1, wherein the FGF-1 further comprises heparin, heparin like molecules, VEGF, TGF-a, TGF-P, angiostatin, endostatin, and platelet-derived endothelial cell growth factor (PDGF), albumin, FGF-2, other growth factors that induce or enhance angiogenesis, or molecules that enhance the effects of FGF-1.

8. The method of claim 1, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is administered to the subject every 8, 12, 16, 24, 36, 48, 60, 72 days, or at least one of: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 weeks.

9. The method of claim 1, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, combinations thereof, is repeatedly administered to the subject at least once per week for 3, 4, 5, 6, 7 or more weeks.

10. The method of claim 1, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is provided at 20, 50, 100, 200, 500 and 1,000 ug per injection site.

11. The method of claim 1, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is via direct injection into the scalp or surrounding tissues.

12. The method of claim 1, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, increases hair growth more than 25, 30, 33, 35, 40, 45, or 50% when compared to a non-treated scalp tissue.

13. The method of claim 1, wherein the FGF-1 is administered as a total of about 30 to about 60 subcutaneous injections, each in an amount about 0.05 to about 0.1 mL at intervals of about 1 cm to about 2 cm.

14. The method of claim 1, wherein at least one of the hair thinning and hair loss of the patient is caused by conditions such as male pattern hair loss, female pattern hair loss, alopecia areata, or traction alopecia.

15. A method of treating at least one of hair thinning or hair loss in a patient comprising: identifying a patient with at least one of hair thinning or hair loss of the skin; administering at a site in need of hair growth a fibroblast growth factor- 1 (FGF-1) in an amount effective to treat the hair thinning or hair loss; and applying to the site in need of hair growth with the composition to treat a diminished microvascular blood flow of the scalp, wherein the FGF-1 at least one of hair thinning or hair loss, wherein the hair thinning or hair loss includes preventing additional hair loss, causing new hair growth, increasing the rate of growth, or increasing the thickness of hair.

16. The method of claim 15, wherein the FGF-1 is an FGF-1 mutant or a truncated FGF-1.

17. The method of claim 15, wherein the truncated FGF-1 is an FGF-11440, FGF-11441, FGF- 11454, or FGF-11455.

18. The method of claim 15, wherein the mutant FGF-1 comprises 1, 2, 3, 4, or 5 amino acid changes, or combinations thereof, selected from K9A, K12V, S17R, N18R, N18K, H21Y, R35E, L44F, A66C, Y94V, N95V, H102Y, F108Y, N114R, N114K, Cl 17V, L133A.

19. The method of claim 15, wherein the FGF-1 is administered subcutaneously, intradermally, or transcutaneous.

20. The method of claim 15, wherein the FGF-1 is formulated in a pharmaceutical composition formulated for immediate release, sustained release, microneedle patch, transdermal enhancers, or combinations thereof.

21. The method of claim 15, wherein the FGF-1 further comprises heparin, heparin like molecules, VEGF, TGF-a, TGF-P, angiostatin, endostatin, and platelet-derived endothelial cell growth factor (PDGF), albumin, FGF-2, other growth factors that induce or enhance angiogenesis, or molecules that enhance the effects of FGF-1.

22. The method of claim 15, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is administered to the subject every 8, 12, 16, 24, 36, 48, 60, 72 days, or at least one of: 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 weeks. 17

23. The method of claim 15, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, combinations thereof, is repeatedly administered to the subject at least once per week for 3, 4, 5, 6, 7 or more weeks.

24. The method of claim 15, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is provided at 20, 50, 100, 200, 500 and 1,000 ug per injection site.

25. The method of claim 15, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, is via direct injection into the scalp or surrounding tissues.

26. The method of claim 15, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof, increases hair growth more than 25, 30, 33, 35, 40, 45, or 50% when compared to a non-treated scalp tissue.

27. The method of claim 15, wherein the FGF-1, FGF-1 mutant, truncated FGF-1, or combinations thereof are administered as a total of about 30 to about 60 subcutaneous injections, each in an amount about 0.05 to about 0.1 mL at intervals of about 1 cm to about 2 cm.

28. The method of claim 15, wherein at least one of the hair thinning and hair loss of the patient is caused by conditions such as male pattern hair loss, female pattern hair loss, alopecia areata, or traction alopecia.