Compositions and methods for treating hair loss or promoting hair growth

Abstract

Methods for preventing or treating hair loss or promoting hair growth or regrowth are provided. More specifically, the present invention relates to compositions for preventing or treating hair loss and / or promoting hair growth on the scalp and / or skin of a subject in need thereof. The compositions comprise an effective amount of a polypeptide or a nucleic acid molecule encoding the polypeptide, wherein the polypeptide comprises an amino acid sequence having an EGF-like domain of thrombomodulin or a conservative variant thereof.

Description

[Technical Field]

[0001] Sequence Listing In accordance with 37 CFR § 1.831-835, this application contains a computer readable Sequence Listing that has been submitted electronically in XML format and is incorporated by reference in its entirety. The XML file was created on February 13, 2023, has the file name 211140US-Sequence Listing-20230213, and is 4.75kb in size.

[0002] The present disclosure relates to maintaining and strengthening hair, particularly preventing and / or slowing hair loss and promoting hair growth and / or regrowth on the scalp and / or skin of mammals. [Background technology]

[0003] Hair loss, also known as alopecia, is caused by an interruption in the body's hair production cycle. In mammals, hair goes through three cycles:

[0004] (i) Anagen: The active stage in which the hair follicle penetrates deep into the dermis and the cells of the hair bulb rapidly divide and differentiate to form a hair.

[0005] (ii) Catagen phase: The catagen phase in which the hair follicle regresses upwards through the dermis and hair growth ceases.

[0006] (iii) Telogen: The resting stage in which the regressed hair follicle contains a small secondary hair germ beneath which is a dense bulb of dermal papilla cells.

[0007] The onset of a new anagen phase is manifested by rapid proliferation in the hair germ, enlargement of the dermal papilla, and synthesis of basement membrane components. The hair cycle is then repeated over and over again until, as a result of the onset of male pattern baldness, most of the hair follicles spend an increasing proportion of their time in the resting phase, and the hairs produced are thinner, shorter, and less noticeable (this is known as the terminal to vellus hair transition).

[0008] Hair loss can occur anywhere on the body, but it is most commonly on the scalp. On average, a human scalp has 100,000 hairs, which undergo cycles of growth, rest, shedding, and regrowth. Signs and symptoms of scalp hair loss can include gradual thinning of hair on the crown, circular or patchy baldness, sudden hair loss, and patchy scaling across the scalp. In general, excessive hair loss can be the result of genetics, hormonal changes, medical conditions (such as cancer chemotherapy), or part of normal aging.

[0009] As the population ages and modern society experiences increasingly stressful lives, more and more people are at risk from hair loss. It is estimated that 40% of 35-year-old men in the United States experience some noticeable hair loss. This condition worsens as men age. By the age of 60, it is estimated that 65% of adult men in the United States will suffer from noticeable hair loss. Women also suffer from hair loss. Hair loss in women differs from that in men and typically involves noticeable thinning of hair across the scalp and hairline. The American Academy of Dermatology reports that 40% of women experience noticeable hair loss by the age of 40. While excessive hair loss on the scalp is most often medically benign, it can have significant cosmetic consequences and cause significant emotional and psychosocial pressure on individuals, profoundly affecting their self-confidence and social life. For example, hair loss may not be considered normal or acceptable for women, in part due to social norms that may pressure women to look their best. Even slight hair thinning, if noticeable, can be undesirable for a woman's self-image and how others may judge her.

[0010] Current treatments for hair loss and / or baldness include drug therapy and hair transplantation. However, widespread dissatisfaction indicates that they are not sufficient to promote relatively involuted and / or dormant hair into an active stage, likely due to the diverse reasons for hair loss and the complexity of the mechanisms of hair growth.

[0011] Medications such as Minoxidil® (Rogaine), Finasteride® (Propecia), and Dutasteride® (Avodart) are approved treatments for hair loss. However, these medications have drawbacks because, to be effective, patients must take the medication on a daily basis. Furthermore, medications only work long-term if patients continue to take the medication. Any discontinuation is likely to result in the loss of acquired hair. Additionally, some of these medications are not approved for use by women due to safety concerns.

[0012] Available invasive hair loss treatments include surgical procedures such as follicular unit transplantation, bald spot reduction surgery, or scalp flaps. These surgical procedures carry the risk of potential skin necrosis or keloid scar tissue formation in the donor area. These surgical procedures also often result in unsatisfactory results, such as an unnatural appearance of hair growth depending on the direction of hair growth and a limited number of hair follicles that can be safely transplanted (fewer than 4,000 follicles can be transplanted per procedure, and procedures can be performed a maximum of three times over a patient's lifetime). Such surgical treatments are also inconvenient for patients, often requiring a 3-4 hour surgery, post-operative care, and approximately two weeks of disruption to normal daily activities immediately after surgery (even without complications).

[0013] Currently, there are also many different types of injections, such as injections of platelet-rich plasma (PRP) alone, used to treat hair loss. Extensive literature has already explored the use of PRP to improve tissue repair and healing after surgical procedures or injuries in sports medicine, orthopedics, oral surgery, and many other medical and surgical specialties. However, the results of these injections vary widely. While existing injection treatments can restore hair for some people, they lack the consistency to be effective for many other populations suffering from hair loss.

[0014] Despite various attempts as described above, current treatments for hair loss do not necessarily provide sufficient hair care effects, such as preventing hair loss and promoting hair growth in any area of ​​the scalp. Finasteride and minoxidil treatments have been touted as effective on both the vertex and frontal areas, but they only produce the most favorable results in the vertex area. Furthermore, most baldness treatments require a long period of time, such as 4 to 6 months, before the effects of treatment become apparent. Perhaps the only partially successful method for regrowing hair on the scalp of bald or balding individuals is through hair transplantation into the balding area. This is typically a painful procedure that is not always successful. Furthermore, even a casual observer can easily tell that a subject has undergone a hair transplant, and it can take months or even years for the hair regrowth to resemble the original, naturally growing hair.

[0015] It is therefore desirable to provide novel compositions that are effective in preventing hair loss and promoting hair growth and / or regrowth on the scalp and / or skin. Summary of the Invention

[0016] In view of the above, the present disclosure provides a composition that has hair loss prevention effects, hair growth promotion effects, and hair care effects in humans and other mammals, and is useful as a pharmaceutical, quasi-drug, or cosmetic composition for hair care. The composition comprises an effective amount of a polypeptide comprising an amino acid sequence having an EGF-like domain of thrombomodulin or a conservative variant thereof.

[0017] Thrombomodulin is an anticoagulant, endothelial cell membrane glycoprotein. Recombinant thrombomodulin proteins, TMD2 (containing an EGF-like structure), and TMD23 (containing TMD2 and a serine-threonine-rich domain), exhibit some activity in cellular behaviors.

[0018] In at least one embodiment of the present disclosure, the composition comprises an effective amount of a recombinant polypeptide comprising an amino acid sequence having an EGF-like domain of thrombomodulin or a conservative variant thereof. In other embodiments, the polypeptide is a recombinant polypeptide comprising an amino acid sequence of an EGF-like structure of thrombomodulin operably linked to a serine-threonine-rich domain of thrombomodulin or a conservative variant thereof. In at least one embodiment, the number of EGF-like structures is 1 to 6. In some embodiments, the number of EGF-like structures is 6.

[0019] The present invention provides a method for preventing or treating hair loss or promoting hair growth or regrowth, the method comprising administering to a subject in need thereof a therapeutically effective amount of a polypeptide or a nucleic acid molecule encoding the polypeptide, wherein the polypeptide comprises an amino acid sequence having an EGF-like domain of thrombomodulin or a conservative variant thereof.

[0020] In preferred embodiments, the polypeptide is a recombinant polypeptide comprising the amino acid sequence of an EGF-like structure of thrombomodulin operably linked to a serine-threonine-rich domain of thrombomodulin or a conservative variant thereof. In some embodiments, the number of EGF-like structures is six. In further embodiments, the recombinant polypeptide consists essentially of the amino acid sequence of six EGF-like structures of thrombomodulin operably linked to a serine-threonine-rich domain of thrombomodulin or a conservative variant thereof.

[0021] In at least one embodiment, the polypeptide is at least 35% identical to TMD23 having the amino acid sequence of SEQ ID NO:1 below. AWDCSVENGGCEHACNAIPGAPRCQCPAGAALQADGRSCTASATQSCNDLCEHFCVPNPDQPGSYSCMCETGYRLAADQHRCEDVDDCILEPSPCPQRCVNTQGGFECHCYPNYDLVDGECVEPVDPCFRANCEYQC QPLNQTSYLCVCAEGFAPIPHEPHRCQMFCNQTACPADCDPNTQASCECPEGYILDDGFICTDIDECENGGFCSGVCHNLPGTFECICGPDSALARHIGTDCDSGKVDGGDSGSGEPPPSPTPGSTLTPPAVGLVHS In some embodiments, the polypeptide consists essentially of TMD23. In some embodiments, the polypeptide is TMD23.

[0022] In at least one embodiment, the polypeptide consists essentially of BB101 having the amino acid sequence of SEQ ID NO:2 below. AWDCSVENGGCEHACNAIPGAPRCQCPAGAALQADGRSCTASATQSCNDLCEHFCVPNPDQPGSYSCMCETGYRLAADQHRCEDVDDCILEPSPCPQRCVNTQGGFECHCYPNYDLVDGECVEPVDPCFRANCEYQC QPLAQTSYLCVCAEGFAPIPHEPHRCQMFCAQTACPADCDPNTQASCECPEGYILDDGFICTDIDECENGGFCSGVCHNLPGTFECICGPDSALARHIGTDCDSGKVDGGDSGSGEPPPSPTPGSTLTPPAVGLVHS

[0023] In at least one embodiment, the polypeptide comprises a TME5 structure having an amino acid sequence that is at least 35% identical to SEQ ID NO:3 below. QMFCNQTACPADCDPNTQASCECPEGYILDDGFICTDIDE

[0024] In some embodiments, the polypeptide comprises a TME5 structure having an amino acid sequence consisting essentially of SEQ ID NO: 3. In some embodiments, the TME5 structure has the amino acid sequence of SEQ ID NO: 3.

[0025] In at least one embodiment, the polypeptide comprises a TME5C structure having an amino acid sequence at least 35% identical to SEQ ID NO:4 below. ECPEGYILDDGFICTDIDE

[0026] In some embodiments, the polypeptide comprises a TME5C structure having an amino acid sequence consisting essentially of SEQ ID NO: 4. In some embodiments, the TME5C structure has the amino acid sequence of SEQ ID NO: 4.

[0027] In at least one embodiment, the nucleic acid molecule is a polynucleic acid introduced from an exogenous source to increase expression of the polypeptide in a subject. In some embodiments, the nucleic acid molecule encoding the polypeptide is an mRNA.

[0028] In a further embodiment, the composition further comprises a pharmaceutically or cosmetically acceptable carrier.

[0029] In one aspect, the present disclosure provides a method for treating hair loss, the method comprising administering to a subject in need thereof a composition comprising: a) a recombinant polypeptide comprising the amino acid sequence of the EGF-like structure of thrombomodulin operably linked to a serine-threonine-rich domain of thrombomodulin or a conservative variant thereof; and b) a pharmaceutically or cosmetically acceptable carrier.

[0030] Common types of hair loss include male-pattern or female-pattern hair loss, alopecia areata, and thinning hair known as telogen effluvium. Male-pattern hair loss is caused by a combination of genetics and male hormones. The cause of female-pattern hair loss is unknown. Alopecia areata is caused by an autoimmune mechanism. Telogen effluvium is typically caused by a physically or emotionally stressful event. Telogen effluvium is also very common after pregnancy.

[0031] Less common causes of hair loss without inflammation and scarring include hair plucking, certain medications including chemotherapy, HIV / AIDS, hypothyroidism, and malnutrition including iron deficiency. Causes of hair loss that occur with scarring or inflammation include fungal infections, lupus erythematosus, radiation therapy, and sarcoidosis. Hair loss is diagnosed in part based on the area affected.

[0032] In at least one embodiment of the present disclosure, the hair loss is selected from the group consisting of regressive alopecia, androgenetic alopecia, male pattern baldness, female pattern baldness, alopecia areata, traction alopecia, anagen effluvium, telogen effluvium, cicatricial (scarring) alopecia, hair loss due to infection, hair loss due to chemotherapy, hair loss due to radiation, hair loss due to hormonal imbalance, hair loss due to medication, hair loss due to exposure to chemicals, hair loss due to stress, and any combination thereof.

[0033] In at least one embodiment of the present disclosure, the hair loss is regressive alopecia.

[0034] In at least one embodiment of the present disclosure, the hair loss is androgenetic alopecia (male- or female-pattern hair loss).

[0035] In some embodiments, the composition promotes hair growth and / or regrowth in a subject. In at least one embodiment of the present disclosure, administration of a composition of the present invention results in one or more of the following: cell proliferation, hair follicle regrowth, revitalized hair follicles, increased hair shaft size, and new hair growth.

[0036] The type of preparation that the composition of the present invention can be used in is not particularly limited as long as it is a preparation that can be applied to the skin, particularly the scalp, such as a gel, cream, paste, lotion, spray, suspension, solution, dispersion type ointment, hydrogel, liquid, emulsion, ointment, film, orabase, powder, liniment, etc. Furthermore, the composition of the present invention may be in any form, such as a hair tonic, hair clear lotion, hair care conditioner, hair care gel, hair care mousse, and shampoo.

[0037] The compositions of the present invention contain a pharmaceutically or cosmetically acceptable carrier to act as a diluent, dispersant, or excipient for the recombinant thrombomodulin of the present invention, thereby facilitating its dispersion when the composition is applied to the skin and / or scalp. Excipients other than or in addition to water can include liquid or solid emollients, solvents, moisturizers, thickeners, and powders.

[0038] In at least one embodiment, the pharmaceutically or cosmetically acceptable carrier is present in an amount of 0.001% to 99.999% by weight of the composition, for example, but not limited to, 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.0 8%, 0.09%, 0.1%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.9999%, and can form the remainder of the composition in the absence of other pharmaceutical or cosmetic adjuvants.

[0039] In at least one embodiment of the present application, the composition may be in the form of an aqueous, aqueous / alcoholic or oily solution, a lotion or serum type dispersion, an anhydrous or lipophilic gel, an emulsion of liquid or semi-liquid consistency obtained by dispersing a fatty phase in an aqueous phase (O / W) or vice versa (W / O), or a suspension or emulsion of smooth, semi-solid or solid consistency of cream or gel type. These compositions are formulated according to the usual techniques known in the art.

[0040] In at least one embodiment of the present application, the compositions of the present disclosure may also contain additives and adjuvants conventional in the cosmetic, pharmaceutical, or dermatological fields, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, fragrances, fillers, disinfectants, odor absorbers, and dyes or colorants. In at least one embodiment, the compositions of the present disclosure are delivered locally or systemically to a subject.

[0041] In at least one embodiment, the compositions of the present disclosure may be administered by routes including subcutaneous and intradermal injection. In some embodiments, the compositions are administered to the scalp of the subject. In at least one embodiment, the method further comprises applying heat to the skin of the subject by infrared light or a hot pack. In other embodiments, the compositions are administered to the scalp of the subject. In further embodiments, the compositions are administered to the subject topically, intradermally, or subcutaneously. Preferably, the compositions of the present disclosure are administered by intradermal injection.

[0042] In at least one embodiment, the method of the present disclosure further comprises applying physical intervention, chemical intervention, or a combination thereof. In some embodiments, the physical intervention may be applied by any device to increase skin penetration. In some embodiments, the physical intervention is selected from the group consisting of needles, microneedles, microneedle patches, microneedling rollers, microneedling stamps, mesoguns, microsurgery devices, superficial injection devices, needle-free intradermal injections, shaving, temperature elevation, electroporation, ultrasound intervention, ultrasound-mediated microbubble cavitation, and any combination thereof.

[0043] In at least one embodiment, the compositions of the present disclosure may be administered topically, intradermally, or subcutaneously to a target site(s), such as the scalp, using various injection tools. In some embodiments, the compositions of the present disclosure may be administered to a subject via a transdermal delivery system, including, but not limited to, needles, microneedles, solid microneedles, coated microneedles, dissolving microneedles, hollow microneedles, microneedle patches with attached microneedles, microneedling (derma) rollers, microneedling (derma) stamps, mesoguns, microsurgery devices, shaving, superficial injection devices, electroporation, ultrasound-mediated microbubble cavitation by applying nanoparticles or microparticles that enhance cavitation to the skin and then irradiating the area of ​​the skin with ultrasound energy, needleless intradermal injection, and any combination thereof.

[0044] In at least one embodiment, the compositions of the present invention may be administered topically, intradermally, or subcutaneously onto the target site(s), such as by applying or spraying directly onto the skin.

[0045] In at least one embodiment of the present disclosure, a polypeptide is at least 35% identical to a TMD23 having the amino acid sequence of SEQ ID NO: 1. In some embodiments, a polypeptide of the present disclosure is at least 35% to 99%, 40% to 90%, 55% to 80%, or 50% to 75% identical to the amino acid sequence of SEQ ID NO: 1. In some embodiments, a polypeptide is at least 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99.5% identical to the amino acid sequence of SEQ ID NO: 1.

[0046] In at least one embodiment, the polypeptide comprises a TME5 structure having an amino acid sequence at least 35% to 99%, 40% to 90%, 55% to 80%, or 50% to 75% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, the polypeptide is at least 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99.5% identical to the amino acid sequence of SEQ ID NO: 3.

[0047] In at least one embodiment, the polypeptide comprises a TME5C structure having an amino acid sequence at least 35% to 99%, 40% to 90%, 55% to 80%, or 50% to 75% identical to the amino acid sequence of SEQ ID NO: 4. In some embodiments, the polypeptide is at least 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 99.5% identical to the amino acid sequence of SEQ ID NO: 4.

[0048] The dosage of the composition of the present invention varies depending on age, individual differences, disease state, composition form, application method, etc., and cannot be clearly defined. The effective amount of TMD23, ​​BB101, or a variant thereof varies depending on age, individual differences, disease state of the subject, composition form, application method, etc., and is not particularly limited. Generally, when administered to humans, the effective amount is 1 cm on the skin surface, which is the target of hair growth treatment. 2 TMD23, ​​BB101, or variants thereof can be administered in amounts of 0.05 μg to 1 mg per cm of skin surface, for example. 2 The present disclosure is not limited to, but can be administered in an amount of about 0.05, 0.1, 0.5, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 μg per cm of the skin surface targeted for hair growth treatment. 2 In further embodiments, TMD23, ​​BB101, or variants thereof may be used at a concentration of 0.05 μg / mL to 200 μg / mL when administered to a subject in need thereof.

[0049] The frequency of administration of the composition of the present invention is not particularly limited. In some embodiments, the composition of the present invention is preferably administered to the affected area several times at regular intervals (e.g., every 2 to 6 weeks or every 3 to 5 weeks) to maintain excellent effects. In other words, the composition can be administered repeatedly. The frequency of administration may be adjusted depending on the condition of the subject's hair and the desired treatment results.

[0050] In some further embodiments, the methods of the present disclosure further comprise administering a second active agent, which is used sequentially, simultaneously, or separately with the composition of the present invention.

[0051] Other objects and features of the present invention will become apparent from the following detailed description considered in conjunction with the accompanying drawings. It is to be understood, however, that the drawings are for illustrative purposes only and are not designed as a definition of the limits of the invention, for which reference should be made to the appended claims. It should be further understood that the drawings are not necessarily drawn to scale, and that, unless otherwise indicated, they are merely intended to conceptually illustrate the structures and procedures described herein. [Brief explanation of the drawings]

[0052] The present disclosure can be more fully understood by reading the following description of the embodiments and with reference to the accompanying drawings, in which:

[0053] [Figure 1] FIG. 1 shows the effect of BB101 or TMD23 treatment on cell viability.

[0054] [Figure 2] FIG. 2 is a photograph showing the effect of BB101 or TMD23 compared to finasteride on hair growth in mice 1 to 11 days after administration.

[0055] [Figure 3] FIG. 3 is a photograph showing the effect of BB101 or TMD23 compared to finasteride on hair growth in mice 13 to 17 days after administration. DETAILED DESCRIPTION OF THE INVENTION

[0056] The technical solutions exemplified in the embodiments of the present disclosure will now be described more clearly and completely, and it will be apparent that the described embodiments are merely a part of the embodiments of the present disclosure and are not intended to be exhaustive. The present disclosure can also be implemented or applied as described in different embodiments. All other embodiments obtained by those skilled in the art without creative work are within the scope of the present disclosure.

[0057] It is further noted that, as used in this disclosure, the singular forms "a," "an," and "the" include plural referents unless expressly or unambiguously limited to one referent. The term "or" is used synonymously with the term "and / or" unless the context clearly dictates otherwise.

[0058] As used herein, the terms "comprising," "comprises," "include," "including," "have," "having," "contain," "containing," and any other variations thereof, are intended to cover a non-exclusive inclusion. For example, if the subject matter is described as "comprises" a certain limitation, unless otherwise specified, the subject matter may further include other ingredients, elements, components, structures, regions, parts, devices, systems, steps, or connections, etc., and should not exclude other limitations.

[0059] As used herein, the terms "prevent" or "prevention" refer to preventative or avoidant measures against a disease, symptom, or illness, and include, but are not limited to, the application or administration of one or more active agents to a subject, who has not yet been diagnosed as a patient suffering from the disease, symptom, or illness, but who may be susceptible to or prone to the disease, symptom, or illness. The preventative measures of the present disclosure are provided to avoid, prevent, or postpone the occurrence of a disease, symptom, or illness, such as hair loss.

[0060] As used herein, the terms "treat" or "treatment" refer to obtaining a desired pharmacological, physiological, and / or cosmetic effect, e.g., restoring hair growth. The effect may be preventative in that a disease, appearance, disorder, or symptom is completely or partially prevented, and / or therapeutic in that a partial or complete treatment is provided for the condition and / or adverse effects resulting from a disease or disorder.

[0061] As used herein, the term "administering" or "administration" refers to placing an active agent inside a subject by a method or route that results in at least partial localization of the active agent at a desired site to produce a desired effect. The active agents described herein may be administered by any suitable route known in the art. For example, the compositions of the present disclosure are administered to a subject by intradermal administration.

[0062] As used herein, the term "effective amount" refers to an amount sufficient to provide a hair growth promoting effect of the composition at a reasonable benefit-to-harm ratio. Within the scope of sound medical judgment, the amount and rate of active ingredient will depend on the condition of the affected area, the severity of the symptoms, the cause of the disease, the duration of treatment, the specific active ingredient(s) contained, the concentration, the method of administration used, and the overall health of the patient. The effective amount may vary depending on factors such as the disease, the subject's tolerance to the administration, and other drugs administered to the subject.

[0063] As used herein, the term "recombinant" may refer to the alteration of genetic material through human intervention. For example, recombination may refer to the manipulation of DNA or RNA within a cell or virus or expression vector by the techniques of molecular biology (recombinant DNA technology), including cloning and recombination. Recombination may also refer to the manipulation of DNA or RNA within a cell or virus by random or directed mutagenesis methods. A "recombinant" nucleic acid may be described by reference to its difference from its naturally occurring counterpart ("wild-type"). A recombinant protein may refer to a protein expressed by recombinant DNA technology. A recombinant protein has a similar amino acid sequence and maintains the same activity or function as its parent protein.

[0064] As used herein, the term "intradermal" means within the dermal tissue of the skin, and "subcutaneous" means administering the composition into the fatty tissue of the skin.

[0065] Numerical ranges used herein are inclusive and combinable, and subranges may be derived from any numerical value within the ranges herein as a minimum or maximum value. For example, a numerical range of "0.05 μg to 1 mg" is understood to include any subrange between the minimum value of 0.05 μg and the maximum value of 1 mg, such as subranges of 0.05 μg to 0.1 μg, 0.3 μg to 5 μg, 1 μg to 10 μg, 15 μg to 500 μg, 10 μg to 1000 μg, and 30 μg to 80 μg. In addition, multiple numerical values ​​used herein may be optionally selected as maximum and minimum values ​​to derive numerical ranges. For example, the numerical ranges of 0.1 μg to 50 μg, 0.1 μg to 100 μg, and 50 μg to 100 μg may be derived from the numerical values ​​of 0.1 μg, 50 μg, and 100 μg.

[0066] As used herein, the term "about" generally refers to a numerical value that is intended to encompass a variation of ±20%, ±10%, ±5%, ±1%, ±0.5%, or ±0.1% from a given value or range. Such variations in numerical values ​​may occur, for example, due to experimental error, typical errors in measuring or handling procedures to make a compound, composition, concentrate, or formulation, differences in the source, manufacture, or purity of starting materials or components used in the present disclosure, or similar considerations. Alternatively, the term "about" may mean within an acceptable standard error of the mean as considered by one of ordinary skill in the art. Unless explicitly stated otherwise, all numerical ranges, amounts, values, and percentages, such as those relating to amounts of materials, lengths of time, temperatures, operating conditions, ratios of amounts, etc., disclosed herein, are to be understood as being modified in all instances by the term "about."

[0067] As used herein, the term "hair" in this disclosure means any hair on the scalp, head, face, and body, including but not limited to hair growing on the scalp, but including eyelashes, eyebrows, beard, ear hair, nose hair, chest hair, pubic hair, etc.

[0068] As used herein, the term "hair growth / regeneration" refers to inducing new hair cycle growth, lengthening the anagen phase, increasing the hair growth rate, and thickening the hair.

[0069] As used herein, the terms "hair loss" and "thinning hair" refer to a decrease in hair density, a shortened hair growth phase, a decrease in hair shaft thickness, and a decrease in the number of hairs, which may be due to aging, genetic factors, or otherwise, and which may be caused by factors in both men and women of all ages and apply to both men and women of all ages.

[0070] As used herein, the terms "patient" and "subject" are used interchangeably. The term "subject" refers to a human or other animal. Examples of subjects include, but are not limited to, humans, monkeys, mice, rats, woodchucks, ferrets, rabbits, hamsters, cows, horses, pigs, deer, dogs, cats, foxes, wolves, chickens, emus, and ostriches. In at least one embodiment of the present disclosure, the subject is a mammal, e.g., a primate such as a human.

[0071] As used herein, the term "nucleic acid molecule" refers to a deoxyribonucleotide or ribonucleotide sequence in double- or single-stranded form, and generally includes naturally occurring nucleotides or artificial chemical mimetics, the source of which is not limited herein. As used herein, the term "nucleic acid" includes natural or non-natural genes, polynucleotides, oligonucleotides, RNA, mRNA, DNA, cDNA, or nucleic acid analogs, but the present disclosure is not limited thereto.

[0072] As used herein, the term "nucleic acid analog" refers to a compound or artificial nucleic acid that is structurally or functionally equivalent to natural RNA or DNA. In some embodiments, a nucleic acid analog may have at least one portion of the nucleotide (e.g., the nucleobase, the phosphate backbone, or the pentose sugar) modified to alter the structure of the nucleic acid.

[0073] A number of examples are used to illustrate the present disclosure, and the following examples should not be considered as limiting the scope of the present disclosure. [Example]

[0074] For a more detailed description of the present disclosure, the disclosed methods are provided and explained in detail with reference to the following examples. Materials used in this disclosure but not annotated herein are commercially available.

[0075] Example 1: Effect of TMD23 and BB101 on cell viability

[0076] 1x10 per well 4 Hair follicle dermal papilla cells (HFDPCs) were cultured in 96-well plates for 12 hours and exposed to 1 mM testosterone. The cells were then treated with different concentrations of BB101 and TMD23. 24 hours after treatment, cell viability was assessed by MTT and presented as a percentage of the DMSO control.

[0077] As shown in Figure 1, the results showed that treatment with either BB101 or TMD23 significantly increased cell viability compared to the control group.

[0078] Example 2: Hair growth in mice accelerated by TMD23

[0079] Six-week-old male C57BL / 6 mice were purchased from the Laboratory Animal Center. The space allocation for 10 animals was 29 x 18 x 13 cm. 3 All animals were maintained in a temperature-controlled (22-24°C) and humidity-controlled (50-60%) environment with a 12-hour light / dark cycle for at least 1 week. They were allowed free access to standard laboratory animal feed and RO water for mice. All aspects of this study, including animal housing, experimentation, and disposal, were conducted in accordance with the Guide for the Care and Use of Laboratory Animals (National Academy Press, Washington, DC, 1996).

[0080] After housing the animals for one week to allow them to acclimate to the laboratory environment, 7-week-old mice were divided into five groups of three mice each: 50 μg / mL BB101 treatment group, 200 μg / mL BB101 treatment group, 50 μg / mL TMD23 treatment group, and 200 μg / mL TMD23 treatment group.

[0081] To evaluate the effect of hair growth, the hair on the back of each mouse was shaved using electric animal clippers 1 day before the start of the experiment. The back was divided into two areas, and testosterone was applied topically to the shaved skin of each mouse in both areas once daily throughout the experiment. The test drugs TMD23, ​​BB101, and finasteride were applied topically to the shaved skin in the lower area using a transdermal drug delivery system, Dermastamp, once daily, at two stamps per dose (approximately 20 μL). The upper area received water via the drug delivery system as a solution control. Under ether anesthesia, mice were photographed with a digital camera on days 1, 9, 11, 13, 15, and 17 to assess skin darkening. The animal care and protocol for this study followed IACUC guidelines (Institutional Animal Care and Use Committee LAC-2022-0137).

[0082] Results showed that 17 days after administration, skin darkening was comparable between the experimental and positive (finasteride) treatment groups, indicating that TMD23 and BB101 are effective in accelerating hair growth and treating hair loss (Figures 2 and 3).

[0083] Although several embodiments of the present disclosure have been described in detail above, those skilled in the art may make various modifications and changes to the illustrated embodiments without substantially departing from the teachings and advantages of the present disclosure. Accordingly, such modifications and changes are intended to be encompassed within the scope of the present disclosure as set forth in the appended claims.

Claims

1. A method for preventing or treating hair loss or promoting hair growth or regrowth, comprising administering a composition to a subject in need thereof, wherein the composition comprises an effective amount of a polypeptide or a nucleic acid molecule encoding the polypeptide, and the polypeptide comprises an amino acid sequence having an EGF-like domain of thrombomodulin or a conservative variant thereof.

2. The method of claim 1, wherein the polypeptide is a recombinant polypeptide comprising the amino acid sequence of the EGF-like structure of thrombomodulin operably linked to a serine-threonine rich domain of thrombomodulin or its conservative variant.

3. The method of claim 2, wherein the number of EGF-like structures is 1 to 6.

4. The method of claim 3, wherein the recombinant polypeptide essentially consists of the amino acid sequence of the six EGF-like structures of thrombomodulin operably linked to the serine-threonine-rich domain of thrombomodulin or its conservative variant.

5. 2. The method of claim 1, wherein the polypeptide is at least 35% identical to TMD23 having the amino acid sequence of SEQ ID NO:

1.

6. 2. The method of claim 1, wherein the polypeptide consists essentially of BB101 having the amino acid sequence of SEQ ID NO:

2.

7. 2. The method of claim 1, wherein the polypeptide comprises a TME5 structure having an amino acid sequence that is at least 35% identical to SEQ ID NO:

3.

8. 2. The method of claim 1, wherein the polypeptide comprises a TME5C structure having an amino acid sequence that is at least 35% identical to SEQ ID NO:

4.

9. 2. The method of claim 1, wherein the nucleic acid molecule is a polynucleic acid introduced from an exogenous source to increase expression of the polypeptide in a subject.

10. 10. The method of claim 1, wherein the hair loss is selected from the group consisting of baldness, regressive alopecia, androgenetic alopecia, male pattern baldness, female pattern baldness, alopecia areata, traction alopecia, anagen effluvium, telogen effluvium, cicatricial (scarring) alopecia, hair loss due to infection, hair loss due to chemotherapy, hair loss due to radiation, hair loss due to hormonal imbalance, hair loss due to medication, hair loss due to exposure to chemicals, hair loss due to stress, and any combination thereof.

11. 10. The method of claim 1, wherein the composition is administered to the skin of the subject.

12. 10. The method of claim 1, wherein the composition is administered topically, intradermally, or subcutaneously to the subject.

13. 10. The method of claim 1, further comprising applying physical intervention, chemical intervention, or a combination thereof.

14. 14. The method of claim 13, wherein the physical intervention is selected from the group consisting of a needle, a microneedle, a microneedle patch, a microneedling roller, a microneedling stamp, a mesogun, a microsurgery device, shaving, a superficial injection device, a needle-free intradermal injection, temperature elevation, electroporation, ultrasound intervention, ultrasound-mediated microbubble cavitation, and any combination thereof.

15. 10. The method of claim 1, wherein the composition further comprises a pharmaceutically or cosmetically acceptable carrier.

16. 10. The method of claim 1, wherein the composition is formulated in a form selected from the group consisting of a gel, a cream, a paste, a lotion, a spray, a suspension, a solution, a dispersion ointment, a hydrogel, a liquid, an emulsion, an ointment, a film, an orabase, a powder, and a paint.

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