Compositions and methods for treatment of androgenetic alopecia

Abstract

Methods and compositions for stimulating hair growth are disclosed. A composition for stimulating hair growth comprises hyaluronic acid. Also provided herein are methods and compositions for treating androgenetic alopecia (AGA). Methods for stimulating hair growth and / or treating AGA comprise administering such compositions into the skin of a patient.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480024053.9 (22) Application Date 2024.04.02 (30) Priority Data 63 / 457928 2023.04.07 US (85) PCT International Application Entering National Phase Date 2025.09.30 (86) PCT International Application Application Data PCT / US2024 / 022632 2024.04.02 (87) PCT International Application Publication Data WO2024 / 211282 EN 2024.10.10 (71) Applicant Amplifica Holding Group Co., Ltd. Address California, USA (72) Inventors F. Fazio W. Ahmed L. Massi (74) Patent Agency China Patent Agency (Hong Kong) Limited 72001 Patent Attorney Duan Jiarong Peng Chang (51) Int.Cl. A61K 31 / 728 (2006.01) A61P 17 / 00 (2006.01) (54) Invention Title Composition and Method for Treating Androgenetic Alopecia (57) Abstract Discloses methods and compositions for stimulating hair growth. The composition for stimulating hair growth contains hyaluronic acid. This document also provides methods and compositions for treating androgenetic alopecia (AGA). The method for stimulating hair growth and / or treating AGA involves applying such compositions to the skin of a patient. Claims (2 pages), Description (22 pages), Drawings (1 page), CN 121057583 A, 2025.12.02, CN 1 21 05 75 83 A 1. A method for treating androgenetic alopecia (AGA) in a patient in need, the method comprising: applying to the patient's skin at least twice a week a composition comprising up to about 1% by weight of hyaluronic acid; wherein the hyaluronic acid is non-crosslinked; and wherein the hyaluronic acid has an average molecular weight of up to about 100 kDa; thereby treating AGA. 2. The method of claim 1, wherein the composition is applied to the patient on several consecutive days of the week. 3. The method of claim 1, wherein the composition is administered via intradermal injection. 4. The method of claim 3, wherein the composition is injected into the skin to a depth of about 0.1 mm or more. 5. The method of claim 3, wherein the composition is applied at or near the stem cell region of the hair follicle. 6. The method of claim 1, wherein the method comprises an injection area of ​​about 4 cm² or larger. 7. The method of claim 1, wherein the composition is applied to the skin comprising one or more of the frontotemporal region of the patient's scalp and the vertex region of the patient's scalp.8. The method of claim 1, wherein treatment with the composition improves or increases one or more of: a) target area hair count (TAHC); b) target area hair width (TAHW); and c) hair darkness. 9. The method of claim 1, wherein the patient's hair loss onset occurred approximately three to five years prior to application of the composition. 10. The method of claim 1, wherein the patient's hair loss onset occurred approximately ten years prior to application of the composition. 11. The method of claim 1, wherein the patient has previously undergone hair transplantation. 12. The method of claim 11, wherein the patient underwent hair transplantation approximately one to two months prior to application of the composition. 13. The method of claim 1, wherein the AGA is mild or moderate AGA. 14. The method of claim 1, wherein the patient has received a modified Norwood-Hamilton Scale score III or a modified Norwood-Hamilton Scale score IV. 15. The method of claim 8, wherein the improvement or increase is measured approximately 10 to 160 days after application of the composition. 16. The method of claim 1, wherein the patient has previously experienced hair loss. 17. The method of claim 7, wherein the hyaluronic acid stimulates hair growth in the frontotemporal region and / or the vertex region of the patient's scalp. 18. A method of stimulating hair growth in the skin of a patient in need, the method comprising: applying to the skin of the patient at least twice a week a composition comprising up to about 1% by weight of hyaluronic acid; wherein the hyaluronic acid is non-crosslinked; and wherein the hyaluronic acid has an average molecular weight of up to about 100 kDa, thereby stimulating hair growth in the skin of the patient. 19. A method of applying a composition for hair growth to a patient in need: applying to the skin of the patient at least twice a week a composition comprising up to about 1% by weight of hyaluronic acid; wherein the hyaluronic acid is non-crosslinked; and wherein the hyaluronic acid has an average molecular weight of up to about 100 kDa. 20. The method of claim 18 or 19, wherein the composition is administered to the patient on several consecutive days of the week. 21. The method of claim 18 or 19, wherein the composition is administered via intradermal injection. 22. The method of claim 21, wherein the composition is injected into the skin to a depth of about 0.1 mm or more. 23. The method of claim 21, wherein the composition is administered at or near the stem cell region of the hair follicle.24. The method of claim 18 or claim 19, wherein the method comprises an injection area of ​​about 4 cm² or larger. 25. The method of claim 18 or claim 19, wherein the composition is applied to the skin comprising one or more of the frontotemporal region and the apical region of the patient's scalp. 26. The method of claim 18 or claim 19, wherein the composition improves or increases one or more of: a) target area hair count (TAHC); b) target area hair width (TAHW); and c) hair darkness. 27. The method of claim 18 or claim 19, wherein the patient's hair loss onset occurred about three to five years prior to the application of the composition. 28. The method of claim 18 or claim 19, wherein the patient's hair loss onset occurred about ten years prior to the application of the composition. 29. The method of claim 18 or claim 19, wherein the patient has previously undergone hair transplantation. 30. The method of claim 29, wherein the patient has undergone hair transplantation about one to two months prior to the application of the composition. 31. The method of claim 18 or claim 19, wherein the patient suffers from androgenetic alopecia (AGA). 32. The method of claim 31, wherein the AGA is mild to moderate AGA. 33. The method of claim 18 or claim 19, wherein the patient has received a modified Norwood-Hamilton Scale score III or a modified Norwood-Hamilton Scale score IV. 34. The method of claim 26, wherein the improvement or increase is measured from about 10 days to about 160 days after application of the composition. Claims 2 / 2 Page 3 CN 121057583 A Composition and method for treating androgenetic alopecia

[0001] Reference to related applications

[0002] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 457,928, filed April 7, 2023, the contents of which are incorporated herein by reference. Background Art

[0003] Hair loss often has negative social and psychological effects on individuals suffering from hair loss. Many factors are believed to contribute to hair loss, including genetics, hormones, environmental exposure, medication, psychological stress, and nutrition. One known treatment is hair transplantation, which requires anesthesia, is expensive, time-consuming, and sometimes painful. Other methods include massage and acupuncture, but these have not been shown to be effective. Hormones and other medications have been used to treat hair loss; however, these treatments often cause undesirable side effects, such as hair growth in unwanted areas.

[0004] Therefore, there is a need for an effective therapy to stimulate hair growth. Summary of the Invention

[0005] This document provides a method for treating androgenetic alopecia (AGA) in patients of need. In some embodiments, the method may include applying the composition described herein to the patient's skin. In some embodiments, the method may include applying the disclosed composition to the patient's skin via intradermal injection. In some embodiments, the composition may contain up to about 1% by weight of hyaluronic acid. For example, the composition may contain about 0.2% by weight to about 0.4% by weight of hyaluronic acid. In some embodiments, the composition may be applied to the subject at least twice a week. In some embodiments, the composition may be applied about three times a week. In some embodiments, the composition may contain uncrosslinked hyaluronic acid. In some embodiments, the average molecular weight of the hyaluronic acid may be up to about 100 kDa.

[0006] In some embodiments, the method of treating AGA may include applying the composition to the patient for several consecutive days of the week. In some embodiments, the composition may be applied for three consecutive days of the week. In some embodiments, the composition may be injected into the skin to a depth of about 0.1 mm or greater. In some embodiments, the composition may be injected into the skin to a depth of about 0.6 mm to about 0.8 mm. In some embodiments, the injection area may be about 4 cm² or larger. In some embodiments, the composition may be applied at or near the stem cell region of hair follicles in the scalp of the subject.

[0007] In some embodiments, the composition may be applied to the skin in one or more of the frontotemporal region and the vertex region of the patient's scalp.

[0008] In some embodiments, treatment with the composition may improve or increase the target area hair count (TAHC); target area hair width (TAHW); and / or hair darkness.

[0009] In some embodiments, the composition is applied to a patient who has experienced hair loss episodes for about three to five years prior to application of the composition. In some embodiments, the composition is applied to a patient who has experienced hair loss episodes for about ten years prior to application of the composition. In some embodiments, the patient has previously undergone hair transplantation. In some embodiments, the patient has undergone hair transplantation for about one to two months prior to application of the composition. In some embodiments, the patient is a male patient.

[0010] In some embodiments, the compositions disclosed herein may be used to treat mild or moderate AGA. In some embodiments, the compositions disclosed herein can be used to treat patients who have received a modified Norwood-Hamilton Scale score III or a modified Norwood-Hamilton Scale score IV. In some embodiments, improvements or increases in TAHC, TAHW, and / or hair tint are measured approximately 10 days to approximately 160 days after application of the composition.

[0011] In some embodiments, the patient has previously experienced hair loss.

[0012] In some embodiments, the compositions of this disclosure can stimulate hair growth in the frontotemporal region and / or the vertex region of the patient's scalp.

[0013] Methods for stimulating hair growth in the skin of a patient are also provided herein. This method may include applying the compositions of this disclosure to the patient's skin. In some embodiments, the compositions may be applied to the patient via intradermal injection. In some embodiments, the compositions of this disclosure may contain up to 1% by weight of hyaluronic acid, such as about 0.2% by weight to about 0.4% by weight of hyaluronic acid. In some embodiments, the compositions of this disclosure may be applied at least twice a week. For example, the compositions may be applied about three times a week. In some embodiments, the compositions may contain uncrosslinked hyaluronic acid. In some embodiments, the average molecular weight of the hyaluronic acid may be up to about 100 kDa.

[0014] This disclosure provides a method for administering a composition for hair growth to a patient in need. This method may include applying the compositions of this disclosure to the patient's skin. In some embodiments, the compositions may be applied via intradermal injection. In some embodiments, the composition may contain up to about 1% by weight of hyaluronic acid, such as about 0.2% by weight to about 0.4% by weight of hyaluronic acid. The composition may be applied at least twice a week, for example, about three times a week. In some embodiments, the composition may contain uncrosslinked hyaluronic acid. In some embodiments, the average molecular weight of the hyaluronic acid may be up to about 100 kDa.

[0015] In some embodiments, the method for hair growth may include applying the composition to a patient for several consecutive days. In one embodiment, the composition may be applied for three consecutive days within a week. In some embodiments, the composition may be injected into the skin to a depth of about 0.1 mm or more, for example, about 0.6 mm to about 0.8 mm. In some embodiments, the injection area may be about 4 cm² or more. In some embodiments, the composition of this disclosure may be applied at or near the stem cell region of the hair follicle.

[0016] In some embodiments, the composition for hair growth may be applied to the skin at one or more of the frontotemporal region and the vertex region of the patient's scalp.

[0017] In some embodiments, treatment with the composition can improve or increase the target area hair count (TAHC); target area hair width (TAHW); and / or hair darkness.

[0018] In some embodiments, the composition is administered to a patient who has experienced approximately three to five years of hair loss episodes prior to its administration. In some embodiments, the composition is administered to a patient who has experienced approximately ten years of hair loss episodes prior to its administration. In some embodiments, the patient has previously undergone hair transplantation. In some embodiments, the patient...The patient had undergone hair transplantation approximately one to two months prior to application of the composition. In some embodiments, the patient is male.

[0019] In some embodiments, the composition can be used for hair growth in patients with AGA. AGA can be mild or moderate. In some embodiments, the compositions disclosed herein can be used to treat hair loss in patients who have received a modified Norwood-Hamilton Scale score III or a modified Norwood-Hamilton Scale score IV. In some embodiments, improvement or increase in TAHC, TAHW, and / or hair darkness is measured approximately 10 to 160 days after application of the composition. Brief Description of the Drawings

[0020] The following detailed description of embodiments of compositions and methods for stimulating hair growth, page 2 / 22, CN 121057583 A, will be better understood when read in conjunction with the accompanying drawings. However, it should be understood that the invention is not limited to the precise arrangement shown.

[0021] In the accompanying drawings:

[0022] The figure is a diagram showing the characteristics associated with each modified Norwood-Hamilton scale score. Detailed Description

[0023] I. Compositions

[0024] In some embodiments, the hair growth stimulating composition comprises hyaluronic acid. In some embodiments, the composition described in U.S. Patent 11,337,993 may be used in this disclosure (the entire contents of which are incorporated herein by reference).

[0025] 1. Hyaluronic Acid (HA)

[0026] HA is an endogenous, naturally occurring glycosaminoglycan. HA is present in the extracellular matrix and is widely distributed throughout the body, with the highest abundance in the skin (approximately 50%). HA is primarily produced by mesenchymal cells, and its functions in the body are to retain moisture; to be responsible for skin hydration and viscoelasticity; to facilitate the transport of ionized solubilities and nutrients; and to protect the skin from free radical damage (Erickson and Stern, 2012).

[0027] In some embodiments, the composition that may be used to stimulate hair growth comprises hyaluronic acid. Hyaluronic acid is a natural ligand of CD44 and has now been found to be pro-inflammatory and stimulate hair growth. Hyaluronic acid is a natural linear polymer containing disaccharide repeating units β-1,4-D-glucuronic acid and β-1,3-N-acetyl-D-glucosamine, as shown in formula (I): (I).

[0028] These units are linked together to form long linear molecules, which form random coils as they become longer. At concentrations above 0.1%, HA chains form a continuous network. HA can be chemically enhanced to create crosslinks between HA molecules with the aim of modulating the physical properties of the product.

[0029] HA has been used in a variety of medical applications, including aesthetic (i.e., dermal fillers), ophthalmic (viscoelastic) and osteoarthritis applications.Studies have been conducted in the United States. A variety of commercial HA products are available, including dermal fillers such as Juvéderm and Restylane; viscoelastic products such as Healon and NuVisc; and osteoarthritis products such as Hyalgan and Orthovisc. HA-based mesotherapy (intradermal injection of small aliquots) has been well-documented in the literature as a safe treatment for skin regeneration. Reported adverse events associated with HA are mild and transient and include injection site pain, itching, stinging, erythema, edema, ecchymosis, purpura, bruising, swelling, hematoma, dark circles, petechiae, dermatographia and pinpoint bleeding, allergic reactions, urticaria, and papule formation. Risks associated with HA dermal fillers are generally mild and transient, but rare and serious complications, including infection, embolism, and tissue necrosis, have occurred due to vascular occlusion. These events are associated with deep injection of cross-linked HA products. This invention provides a non-cross-linked form of hyaluronic acid in phosphate-buffered saline (PBS) for use in the methods described herein.

[0030] In some embodiments, hyaluronic acid has a low average molecular weight. In some embodiments, "low average molecular weight" as used herein refers to the range of about 15,000 Da to about 40,000 Da. In some embodiments, hyaluronic acid has a medium average molecular weight. In some embodiments, "medium average molecular weight" as used herein refers to the range of about 75,000 Da to about 350,000 Da. In some embodiments, hyaluronic acid has a high average molecular weight. In some embodiments, "high average molecular weight" as used herein refers to about 950,000 Da and greater.

[0031] In some embodiments, the average molecular weight of hyaluronic acid is about 4,000 Da or less to about 10,000 Da. In some embodiments, the average molecular weight of hyaluronic acid is about 10,000 Da to about 100,000 Da. In some embodiments, the average molecular weight of hyaluronic acid is about 100,000 Da to about 1,500,000 Da or greater. In some embodiments, the average molecular weight of hyaluronic acid may be about 40 kDa to about 100 kDa. In one embodiment, the average molecular weight of hyaluronic acid is about 60 kDa.

[0032] In some embodiments, the average molecular weight of hyaluronic acid is between about 1 kDa and about 10 kDa, between about 10 kDa and about 50 kDa, between about 50 kDa and about 100 kDa, between about 100 kDa and about 150 kDa, between about 200 kDa and about 250 kDa, between about 300 kDa and about 350 kDa, between about 400 kDa and about 450 kDa.Between approximately 500 kDa and approximately 550 kDa, between approximately 600 kDa and approximately 650 kDa, between approximately 700 kDa and approximately 750 kDa, between approximately 800 kDa and approximately 850 kDa, between approximately 900 kDa and approximately 1000 kDa, between approximately 1000 kDa and approximately 1100 kDa, between approximately 1100 kDa and approximately 1200 kDa, between approximately 1200 kDa and approximately 1300 kDa, between approximately 1300 kDa and approximately 1400 kDa, between approximately 1400 kDa and approximately 1500 kDa, between approximately 1 kDa and approximately 100 kDa, between approximately 100 kDa and approximately 250 kDa, between approximately 250 kDa and approximately 500 kDa, between approximately 500 kDa and approximately 750 kDa Between, between about 750 kDa and about 1000 kDa, between about 1000 kDa and about 1250 kDa, between about 1250 kDa and about 1500 kDa, between about 1 kDa and about 250 kDa, between about 1 kDa and about 500 kDa, between about 100 kDa and about 500 kDa, between about 250 kDa and about 750 kDa, between about 500 kDa and about 1000 kDa, between about 750 kDa and about 1250 kDa, or between about 1000 kDa and about 1500 kDa.

[0033] In some embodiments, the average molecular weight of hyaluronic acid can be as high as about 100 kDa. For example, the average molecular weight of hyaluronic acid can be between approximately 1 kDa and approximately 10 kDa, between approximately 5 kDa and approximately 15 kDa, between approximately 10 kDa and approximately 20 kDa, between approximately 15 kDa and approximately 25 kDa, between approximately 20 kDa and approximately 30 kDa, between approximately 25 kDa and approximately 35 kDa, between approximately 30 kDa and approximately 40 kDa, between approximately 35 kDa and approximately 45 kDa, between approximately 40 kDa and approximately 50 kDa, between approximately 45 kDa and approximately 55 kDa, between approximately 50 kDa and approximately 60 kDa, and between approximately 55 kDa. Between approximately 65 kDa, between approximately 60 kDa and approximately 70 kDa, between approximately 65 kDa and approximately 75 kDa, between approximately 70 kDa and approximately 80 kDa, between approximately 75 kDa and approximately 85 kDa, between approximately 80 kDa and approximately 90 kDa, between approximately 85 kDa and approximately 95 kDa, between approximately 90 kDa and approximately 100 kDa, between approximately 95 kDa and approximately 105 kDa, between approximately 100 kDa and approximately 200 kDa, between approximately 150 kDa and approximately 250 kDa, between approximately 200 kDa and approximately 300 kDa, between approximately...Between 250kDa and approximately 350kDa, between approximately 300kDa and approximately 400kDa, between approximately 350kDa and approximately 450kDa, between approximately 400kDa and approximately 500kDa, between approximately 450kDa and approximately 550kDa, between approximately 500kDa and approximately 600kDa, between approximately 550kDa and approximately 650kDa, between approximately 600kDa and approximately 700kDa, between approximately 650kDa and approximately 750kDa, between approximately 700kDa and approximately 800kDa, between approximately 750kDa and approximately 850kDa, between approximately 800kDa and approximately 900kDa, between approximately 850kDa and approximately 950kDa, between approximately 900kDa and approximately 1000kDa, or greater.

[0034] In some embodiments, the hyaluronic acid is not cross-linked. In some embodiments, the hyaluronic acid is substantially free of cross-links. In some embodiments, the composition comprises free hyaluronic acid, i.e., uncross-linked hyaluronic acid. Typically, free hyaluronic acid is not exposed to cross-linking conditions. In some embodiments, the composition may have free hyaluronic acid, and the free hyaluronic acid may be present at a concentration between 5% by weight and 95% by weight of the total hyaluronic acid. Specification 4 / 22 pages 7 CN 121057583 A

[0035] In some embodiments, the hyaluronic acid is cross-linked. Cross-linking can improve the lifespan of hyaluronic acid, and in some embodiments, a certain degree of cross-linking may be desired. In some embodiments, the hyaluronic acid has sufficient cross-linking to last for about a week. However, regardless of the mechanism of action, hyaluronic acid is believed to effectively stimulate hair growth by interacting with CD44 receptors. Therefore, in some embodiments, it is desirable that the hyaluronic acid is not so extensively cross-linked that cross-linking interferes with the ability of hyaluronic acid to interact with CD44 receptors.

[0036] The hydroxyl (-OH), carboxyl (-COOH), and / or amide (-NHCOCH3) functional groups of hyaluronic acid can be crosslinked via ether bonds (R-O-R), ester bonds (R-COO-R), or carbodiimide, respectively. In some embodiments, hyaluronic acid is crosslinked with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), glutaraldehyde (GTA), poly(ethylene glycol) diglycidyl ether (PEGDE), ethylene glycol diglycidyl ether (EGDE), divinyl sulfonate (DVS), or pentaerythritol tetraacrylate (PT).

[0037] The crosslinking density of hyaluronic acid can be about 1 × 10⁻⁷ mol / cm³ or greater, about 2 × 10⁻⁷ mol / cm³ or greater, about3×10⁻⁷ mol / cm³ or greater, about 4×10⁻⁷ mol / cm³ or greater, about 5×10⁻⁷ mol / cm³ or greater, about 6×10⁻⁷ mol / cm³ or greater, about 7×10⁻⁷ mol / cm³ or greater, about 8×10⁻⁷ mol / cm³ or greater, about 9×10⁻⁷ mol / cm³ or greater, about 1×10⁻⁶ mol / cm³ or greater, about 2×10⁻⁶ mol / cm³ or greater, about 3×10⁻⁶ mol / cm³ or greater, about 4×10⁻⁶ mol / cm³ or greater, about 5×10⁻⁶ mol / cm³ or greater, about 6×10⁻⁶ mol / cm³ or greater, about 7×10⁻⁶ mol / cm³ or greater, about 8×10⁻⁶ mol / cm³ or greater, about 9×10⁻⁶ mol / cm³ or greater, or about 1×10⁻⁵ mol / cm³ or greater. In some embodiments, the crosslinking density of hyaluronic acid can be between about 1×10⁻⁷ mol / cm³ and about 1×10⁻⁵ mol / cm³, between about 1×10⁻⁷ mol / cm³ and about 1×10⁻⁶ mol / cm³, between about 1×10⁻⁷ mol / cm³ and about 5×10⁻⁷ mol / cm³, between about 5×10⁻⁷ mol / cm³ and about 1×10⁻⁶ mol / cm³, between about 1×10⁻⁷ mol / cm³ and about 2×10⁻⁷ mol / cm³, between about 2×10⁻⁷ mol / cm³ and about 4×10⁻⁷ mol / cm³, between about 4×10⁻⁷ mol / cm³ and about 6×10⁻⁷ mol / cm³. Between -7 mol / cm³, between approximately 6 × 10⁻⁷ mol / cm³ and approximately 8 × 10⁻⁷ mol / cm³, or between approximately 8 × 10⁻⁷ mol / cm³ and approximately 1 × 10⁻⁶ mol / cm³, between approximately 1 × 10⁻⁶ mol / cm³ and approximately 1 × 10⁻⁵ mol / cm³, between approximately 1 × 10⁻⁶ mol / cm³ and approximately 5 × 10⁻⁶ mol / cm³, between approximately 5 × 10⁻⁶ mol / cm³ and approximately 1 × 10⁻⁵ mol / cm³, between approximately 1 × 10⁻⁶ mol / cm³ and approximately 2 × 10⁻⁶ mol / cm³, between approximately 2 × 10⁻⁷ mol / cm³ and approximately 4 × 10⁻⁷ mol / cm³, between approximately 4 × 10⁻⁷ mol / cm³ and approximately 6 × 10⁻⁷ mol / cm³, between approximately 6 × 10⁻⁷ mol / cm³ and approximately 8 × 10⁻⁷ mol / cm³ Between about 8 × 10⁻⁷ mol / cm³ and about 1 × 10⁻⁵ mol / cm³, or between about 5 × 10⁻⁷ mol / cm³ and about 5 × 10⁻⁶ mol / cm³.

[0038] The compositions of the present invention may contain sufficient components to provide a therapeutic effect, such as stimulating hair growth in a patient in need.And / or an amount of hyaluronic acid used to treat androgenetic alopecia (AGA). In some embodiments, higher concentrations of hyaluronic acid can lead to undesirable inflammation. In some embodiments, the compositions of the present invention contain an amount of hyaluronic acid sufficient to provide a therapeutic effect, such as stimulating hair growth in a patient in need or treating AGA, and insufficient to provide an unacceptable or undesirable inflammatory response.

[0039] In some embodiments, the composition comprises about 0.001% by weight or more of hyaluronic acid, about 0.0025% by weight or more of hyaluronic acid, about 0.0050% by weight or more of hyaluronic acid, about 0.0075% by weight or more of hyaluronic acid, about 0.01% by weight or more of hyaluronic acid, about 0.025% by weight or more of hyaluronic acid, about 0.05% by weight or more of hyaluronic acid, about 0.075% by weight or more of hyaluronic acid, about 0.1% by weight or more of hyaluronic acid, about 0.25% by weight or more of hyaluronic acid, about 0.5% by weight or more of hyaluronic acid, about 0.75% by weight or more of hyaluronic acid, or about 1% by weight or more of hyaluronic acid. In some embodiments, the composition comprises between about 0.001% by weight and about 0.0025% by weight of hyaluronic acid, between about 0.0025% by weight and about 0.005% by weight of hyaluronic acid, and between about 0.0025% by weight of hyaluronic acid. (See specification 5 / 22 page 8 CN 121057583 A) Hyaluronic acid in the following weight percentages: approximately 0.0075 wt%, approximately 0.0025 wt% to approximately 0.01 wt%, approximately 0.001 wt% to approximately 0.01 wt%, approximately 0.005 wt% to approximately 0.0075 wt%, approximately 0.005 wt% to approximately 0.01 wt%, approximately 0.0075 wt% to approximately 0.01 wt%, approximately 0.0075 wt% to approximately 0.01 wt%, approximately 0.01 wt% to approximately 0.02 wt%, approximately 0.01 wt% to approximately 0.05 wt%, approximately 0.02 wt% to approximately 0.04 wt%, approximately 0.05 wt% to approximately 0.1 wt%, and approximately 0.04 wt% to approximately 0.06 wt%. Hyaluronic acid in the following proportions: approximately 0.06% to approximately 0.08% by weight; approximately 0.08% to approximately 0.1% by weight; approximately 0.1% to approximately 0.2% by weight; approximately 0.2% to approximately 0.3% by weight; approximately 0.1% to approximately 0.5% by weight; approximately 0.2% to approximately 0.4% by weight; approximately 0.5% to approximately 1% by weight; and so on.Hyaluronic acid in the range of about 0.3% to about 0.4% by weight, about 0.4% to about 0.5% by weight, about 0.5% to about 0.6% by weight, about 0.4% to about 0.6% by weight, about 0.6% to about 0.6% by weight, about 0.6% to about 0.7% by weight, about 0.7% to about 0.8% by weight, about 0.6% to about 0.8% by weight, about 0.8% to about 0.9% by weight, or about 0.8% to about 1% by weight. In some embodiments, the composition comprises about 0.1 wt%, about 0.11 wt%, about 0.12 wt%, about 0.13 wt%, about 0.14 wt%, about 0.15 wt%, about 0.16 wt%, about 0.17 wt%, about 0.18 wt%, about 0.19 wt%, about 0.2 wt%, about 0.21 wt%, about 0.22 wt%, about 0.23 wt%, about 0.24 wt%, about 0.25 wt%, about 0.26 wt%, about 0.27 wt%, about 0.28 wt%, about 0.29 wt%, about 0.3 wt%, about 0.31 wt%, about 0.32 wt%, about 0.33 wt%, about 0.34 wt%, about 0.35 wt%, about 0.36 wt%, about 0.37 wt%, about 0.38 wt%, about 0.39 wt%, about 0.4 wt%, about 0.41 wt%, about 0.42 wt%, about 0. 0.43 wt%, approximately 0.44 wt%, approximately 0.45 wt%, approximately 0.46 wt%, approximately 0.47 wt%, approximately 0.48 wt%, approximately 0.49 wt%, approximately 0.5 wt%, approximately 0.51 wt%, approximately 0.52 wt%, approximately 0.53 wt%, approximately 0.54 wt%, approximately 0.55 wt%, approximately 0.56 wt%, approximately 0.57 wt%, approximately 0.58 wt%, approximately 0.59 wt%, approximately 0.6 wt%, approximately 0.61 wt%, approximately 0.62 wt%, approximately 0.63 wt%, approximately 0.64 wt%, approximately 0.65 wt%, approximately 0.66 wt%, approximately 0.67 wt%, approximately 0.68 wt%, approximately 0.69 wt%, approximately 0.7 wt%, approximately 0.71 wt%, approximately 0.72 wt%, approximately 0.73 wt%, approximately 0.74 wt%, approximately 0.75 wt%, approximately 0.76 wt%, approximately 0 0.77 wt%, approximately 0.78 wt%, approximately 0.79 wt%, approximately 0.8 wt%, approximately 0.81 wt%, approximately 0.82 wt%, approximately 0.83 wt%, approximately 0.84 wt%, approximately 0.85 wt%, approximately 0.86 wt%, approximately 0.87 wt%, approximately 0.88 wt%, approximately 0.89 wt%, approximately 0.9 wt%, approximately 0.91 wt%, approximately 0.92 wt%Hyaluronic acid in amounts of approximately 0.93% by weight, approximately 0.94% by weight, approximately 0.95% by weight, approximately 0.96% by weight, approximately 0.97% by weight, approximately 0.98% by weight, approximately 0.99% by weight, or approximately 1% by weight or more.

[0040] As used throughout this specification, mcg / mL and ug / mL refer to micrograms per milliliter. In some embodiments, hyaluronic acid is used in compositions of about 10 mcg / mL or greater, about 15 mcg / mL or greater, about 20 mcg / mL or greater, about 25 mcg / mL or greater, about 30 mcg / mL or greater, about 35 mcg / mL or greater, about 40 mcg / mL or greater, about 45 mcg / mL or greater, about 50 mcg / mL or greater, about 55 mcg / mL or greater, about 60 mcg / mL or greater, about 65 mcg / mL or greater, about 70 mcg / mL or greater, about 75 mcg / mL or greater, about 80 mcg / mL or greater, about 85 mcg / mL or greater, about 90 mcg / mL, or more. The following concentrations of the composition are listed: mL or greater, about 95 mcg / mL or greater, about 100 mcg / mL or greater, about 110 mcg / mL or greater, about 120 mcg / mL or greater, about 130 mcg / mL or greater, about 140 mcg / mL or greater, about 150 mcg / mL or greater, about 160 mcg / mL or greater, about 170 mcg / mL or greater. (Instructions for use, page 6 / 22, 9, CN 121057583 A) Compositions with a concentration of approximately 180 mcg / mL or greater, compositions with a concentration of approximately 190 mcg / mL or greater, compositions with a concentration of approximately 200 mcg / mL or greater, compositions with a concentration of approximately 210 mcg / mL or greater, compositions with a concentration of approximately 220 mcg / mL or greater, compositions with a concentration of approximately 230 mcg / mL or greater, compositions with a concentration of approximately 240 mcg / mL or greater, compositions with a concentration of approximately 250 mcg / mL or greater, compositions with a concentration of approximately 260 mcg / mL or greater, compositions with a concentration of approximately 270 mcg / mL or greater, compositions with a concentration of approximately 280 mcg / mL or greater. L composition or larger, about 290 mcg / mL composition or larger, about 300 mcg / mL composition or larger, about 310 mcg / mL composition or larger, about 320 mcg / mL composition or larger, about 330 mcg / mL composition or larger, about 340 mcg / mL composition or larger, about 350 mcg / mL composition or larger, about 360 mcg / mL composition or larger, about 370 mcg / mL composition or larger, about 380 mcg / mL composition or larger, aboutCompositions with a concentration of 390 mcg / mL or greater, compositions with a concentration of about 400 mcg / mL or greater, compositions with a concentration of about 410 mcg / mL or greater, compositions with a concentration of about 420 mcg / mL or greater, compositions with a concentration of about 430 mcg / mL or greater, compositions with a concentration of about 440 mcg / mL or greater, compositions with a concentration of about 450 mcg / mL or greater, compositions with a concentration of about 460 mcg / mL or greater, compositions with a concentration of about 470 mcg / mL or greater, compositions with a concentration of about 480 mcg / mL or greater, compositions with a concentration of about 490 mcg / mL or greater, compositions with a concentration of about 500 mcg / mL or greater, compositions with a concentration of about 510 mcg / mL or greater, compositions with a concentration of about 520 mcg / mL or greater, compositions with a concentration of about 530 mcg / mL or greater. Compositions with concentrations of approximately 540 mcg / mL or greater, approximately 550 mcg / mL or greater, approximately 560 mcg / mL or greater, approximately 570 mcg / mL or greater, approximately 580 mcg / mL or greater, approximately 590 mcg / mL or greater, approximately 600 mcg / mL or greater, approximately 610 mcg / mL or greater, approximately 620 mcg / mL or greater, approximately 630 mcg / mL or greater, approximately 640 mcg / mL or greater, approximately 650 mcg / mL or greater, approximately 660 mcg / mL or greater, approximately 670 mcg / mL Composition L or larger, composition of about 680 mcg / mL or larger, composition of about 700 mcg / mL or larger, composition of about 710 mcg / mL or larger, composition of about 720 mcg / mL or larger, composition of about 730 mcg / mL or larger, composition of about 740 mcg / mL or larger, composition of about 750 mcg / mL or larger, composition of about 760 mcg / mL or larger, composition of about 770 mcg / mL or larger, composition of about 780 mcg / mL or larger, composition of about 790 mcg / mL or larger, composition of about 800 mcg / mL or larger, composition of about 810 mcg / mL or larger, composition of about 820 mcg / mL Or greater, about 830 mcg / mL composition or greater, about 840 mcg / mL composition or greater, about 850 mcg / mL composition or greater, about 860 mcg / mL composition or greater, about 870 mcg / mL composition or greater, about 880 mcg / mL composition or greater, about 890 mcg / mL composition or greater, about 900 mcg / mL composition or greater, about 910 mcg / mL composition or greater, about 920 mcg / mL composition or greater, about 930 mcg / mL composition or greater, about 940 mcg / mL composition or greater, about 950 mcg / mL composition or greater, about 960 mcg / mL composition or greater, aboutThe composition is present in amounts of 970 mcg / mL or greater, or about 980 mcg / mL or greater.

[0041] In some embodiments, hyaluronic acid is present in amounts of about 1 mcg / mL composition to about 250 mcg / mL composition, about 10 mcg / mL composition to about 250 mcg / mL composition, about 10 mcg / mL composition to about 200 mcg / mL composition, or 10 mcg / mL composition. Compositions of about 10 mcg / mL to about 150 mcg / mL, 10 mcg / mL to about 100 mcg / mL, about 25 mcg / mL to about 250 mcg / mL, about 25 mcg / mL to about 200 mcg / mL, about 25 mcg / mL to about 150 mcg / mL, about 25 mcg / mL to about 100 mcg / mL, about 50 mcg / mL to about 250 mcg / mL, about 50 mcg / mL to about 200 mcg / mL, about 50 mcg / mL to about 150 mcg / mL, about 50 mcg / mL to about 100 mcg / mL, about 75 mcg / mL to about 250 mcg / mL 0 mcg / mL composition, about 75 mcg / mL composition to about 200 mcg / mL composition, about 75 mcg / mL composition to about 150 mcg / mL composition, about 75 mcg / mL composition to about 100 mcg / mL composition, about 100 mcg / mL composition to about 250 mcg / mL composition, about 100 mcg / mL composition to about 200 mcg / mL composition, about 100 mcg / mL composition to about 150 mcg / mL composition, about 150 mcg / mL composition to about 250 mcg / mL composition, about 200 mcg / mL composition to about 250 mcg / mL composition, about 60 mcg / mL composition to about 80 mcg / mL composition, about 50 mcg / mL composition to about 75 mcg / mL composition The specification, page 7 / 22, 10 CN 121057583 A, contains a composition of about 1 mL, a composition of about 25 mcg / mL, a composition of about 75 mcg / mL, a composition of about 10 mcg / mL, a composition of about 50 mcg / mL, or a composition of about 10 mcg / mL, a composition of about 25 mcg / mL.

[0042] In some embodiments, hyaluronic acid is present in amounts ranging from about 1 mcg / mL to about 500 mcg / mL, a composition of about 10 mcg / mL to about 500 mcg / mL, a composition of about 10 mcg / mL to about 450 mcg / mL, a composition of about 10 mcg / mL to about 400 mcg / mL, a composition of about 10 mcg / mL to about 350 mcg / mL, and about...10 mcg / mL composition to about 300 mcg / mL composition, about 25 mcg / mL composition to about 500 mcg / mL composition, about 25 mcg / mL composition to about 450 mcg / mL composition, about 25 mcg / mL composition to about 400 mcg / mL composition, about 25 mcg / mL composition to about 350 mcg / mL composition, about 25 mcg / mL composition to about 300 mcg / mL composition, about 50 mcg / mL composition to about 500 mcg / mL composition, about 50 mcg / mL composition to about 450 mcg / mL composition, about 50 mcg / mL composition Compositions with a concentration of approximately 100 mcg / mL: 100 mcg / mL to 400 mcg / mL; 50 mcg / mL to 350 mcg / mL; 50 mcg / mL to 300 mcg / mL; 75 mcg / mL to 500 mcg / mL; 75 mcg / mL to 450 mcg / mL; 75 mcg / mL to 400 mcg / mL; 75 mcg / mL to 350 mcg / mL; 75 mcg / mL to 300 mcg / mL; 100 mcg / mL. Compositions of about 500 mcg / mL, compositions of about 100 mcg / mL to about 450 mcg / mL, compositions of about 100 mcg / mL to about 400 mcg / mL, compositions of about 100 mcg / mL to about 350 mcg / mL, compositions of about 100 mcg / mL to about 300 mcg / mL, compositions of about 150 mcg / mL to about 500 mcg / mL, compositions of about 150 mcg / mL to about 450 mcg / mL, compositions of about 150 mcg / mL to about 400 mcg / mL, compositions of about 150 mcg / mL The composition is present in amounts ranging from about 350 mcg / mL to about 300 mcg / mL, from about 200 mcg / mL to about 500 mcg / mL, from about 200 mcg / mL to about 450 mcg / mL, from about 200 mcg / mL to about 400 mcg / mL, from about 200 mcg / mL to about 350 mcg / mL, from about 200 mcg / mL to about 300 mcg / mL, or from about 200 mcg / mL to about 250 mcg / mL.

[0043] In some embodiments, hyaluronic acid is present in amounts ranging from about 1 mcg / mL to about 750 mcg / mL, from about...10 mcg / mL composition to about 750 mcg / mL composition, about 10 mcg / mL composition to about 700 mcg / mL composition, about 10 mcg / mL composition to about 650 mcg / mL composition, about 10 mcg / mL composition to about 600 mcg / mL composition, about 10 mcg / mL composition to about 550 mcg / mL composition, about 25 mcg / mL composition to about 750 mcg / mL composition, about 25 mcg / mL composition to about 700 mcg / mL composition, about 25 mcg / mL composition to about 650 mcg / mL composition, about 25 ... Compositions with a concentration of g / mL to about 600 mcg / mL, compositions with a concentration of about 25 mcg / mL to about 550 mcg / mL, compositions with a concentration of about 50 mcg / mL to about 750 mcg / mL, compositions with a concentration of about 50 mcg / mL to about 700 mcg / mL, compositions with a concentration of about 50 mcg / mL to about 650 mcg / mL, compositions with a concentration of about 50 mcg / mL to about 600 mcg / mL, compositions with a concentration of about 50 mcg / mL to about 550 mcg / mL, compositions with a concentration of about 75 mcg / mL to about 750 mcg / mL, compositions with a concentration of about 75 mcg / mL Compositions with a concentration of about 700 mcg / mL, compositions with a concentration of about 75 mcg / mL, compositions with a concentration of about 650 mcg / mL, compositions with a concentration of about 75 mcg / mL, compositions with a concentration of about 75 mcg / mL, compositions with a concentration of about 550 mcg / mL, compositions with a concentration of about 100 mcg / mL, compositions with a concentration of about 100 mcg / mL, compositions with a concentration of about 700 mcg / mL, compositions with a concentration of about 1 ... Compositions of about 550 mcg / mL, compositions of about 150 mcg / mL to about 750 mcg / mL, compositions of about 150 mcg / mL to about 700 mcg / mL, compositions of about 150 mcg / mL to about 650 mcg / mL, compositions of about 150 mcg / mL to about 600 mcg / mL, compositions of about 150 mcg / mL to about 550 mcg / mL, compositions of about 200 mcg / mL to about 750 mcg / mL, compositions of about 200 mcg / mL to about 700 mcg / mL, (See page 8 / 22 of the specification, CN 121057583 A) Compositions of about 200 mcg / mL to about 650 mcg / mL, compositions of about 200 mcg / mL to about 600 mcg / mL, (See page 11 of the specification, CN 121057583 A)200 mcg / mL composition to about 550 mcg / mL composition, about 300 mcg / mL composition to about 750 mcg / mL composition, about 300 mcg / mL composition to about 700 mcg / mL composition, about 300 mcg / mL composition to about 650 mcg / mL composition, about 300 mcg / mL composition to about 600 mcg / mL composition, about 300 mcg / mL composition to about 550 mcg / mL composition, about 400 mcg / mL composition to about 750 mcg / mL composition, about 400 mcg / mL composition to about 700 mcg / mL composition, about 400 mcg / mL composition to about 650 mcg / mL composition, about 400 mcg / mL composition to about 600 mcg / mL composition, about 400 mcg / mL composition to about 600 mcg / mL composition, about 400 mcg / mL composition to about 550 mcg / mL composition, about 400 mcg / mL composition to about 750 mcg / mL composition, about 400 mcg / mL composition to about 6 ...750 mcg / mL composition, about 400 mcg / mL composition to about 750 mcg / mL composition, about 400 mcg / mL composition to about 750 mcg / mL composition, about 400 mcg The composition is present in amounts ranging from approximately 500 mcg / mL to approximately 750 mcg / mL, from approximately 500 mcg / mL to approximately 700 mcg / mL, from approximately 500 mcg / mL to approximately 650 mcg / mL, from approximately 500 mcg / mL to approximately 600 mcg / mL, from approximately 500 mcg / mL to approximately 550 mcg / mL, from approximately 600 mcg / mL to approximately 750 mcg / mL, from approximately 600 mcg / mL to approximately 700 mcg / mL, from approximately 600 mcg / mL to approximately 650 mcg / mL, or from approximately 700 mcg / mL to approximately 750 mcg / mL.

[0044] In some embodiments, hyaluronic acid is present in amounts of about 100 mcg / ml, about 200 mcg / ml, about 300 mcg / ml, about 400 mcg / ml, about 500 mcg / ml, about 600 mcg / ml, about 700 mcg / ml, about 800 mcg / ml, about 900 mcg / ml, about 1000 mcg / ml, or greater.

[0045] In some embodiments, hyaluronic acid is expressed in quantities of about 1 mcg / mL to about 950 mcg / mL, about 10 mcg / mL to about 950 mcg / mL, about 10 mcg / mL to about 900 mcg / mL, about 10 mcg / mL to about 850 mcg / mL, about 10 mcg / mL to about 800 mcg / mL, about 25 mcg / mL to about 950 mcg / mL, about 25 mcg / mL to about 900 mcg / mL, about 25 mcg / mL to about 850 mcg / mL, about 25 mcg / mL to about 800 mcg / mL, about 50 mcg / mL to about950 mcg / mL composition, about 50 mcg / mL composition to about 900 mcg / mL composition, about 50 mcg / mL composition to about 850 mcg / mL composition, about 50 mcg / mL composition to about 800 mcg / mL composition, about 75 mcg / mL composition to about 950 mcg / mL composition, about 75 mcg / mL composition to about 900 mcg / mL composition, about 75 mcg / mL composition to about 850 mcg / mL composition, about 75 mcg / mL composition to about 800 mcg / mL composition, about 100 mcg / mL composition to about 95 ... The composition has various values: approximately 100 mcg / mL to about 850 mcg / mL; approximately 100 mcg / mL to about 800 mcg / mL; approximately 150 mcg / mL to about 950 mcg / mL; approximately 150 mcg / mL to about 900 mcg / mL; approximately 150 mcg / mL to about 850 mcg / mL; approximately 150 mcg / mL to about 800 mcg / mL; approximately 200 mcg / mL to about 950 mcg / mL; approximately 200 mcg / mL to about 900 mcg / mL. Compositions of about 200 mcg / mL to about 850 mcg / mL, compositions of about 200 mcg / mL to about 800 mcg / mL, compositions of about 300 mcg / mL to about 950 mcg / mL, compositions of about 300 mcg / mL to about 900 mcg / mL, compositions of about 300 mcg / mL to about 850 mcg / mL, compositions of about 300 mcg / mL to about 800 mcg / mL, compositions of about 400 mcg / mL to about 950 mcg / mL, compositions of about 400 mcg / mL to about 900 mcg / mL, compositions of about 4 00 mcg / mL composition to about 950 mcg / mL composition, about 400 mcg / mL composition to about 900 mcg / mL composition, about 400 mcg / mL composition to about 850 mcg / mL composition, about 400 mcg / mL composition to about 800 mcg / mL composition, about 500 mcg / mL composition to about 950 mcg / mL composition, about 500 mcg / mL composition to about 900 mcg / mL composition, about 500 mcg / mL composition to about 850 mcg / mL composition, about 500 mcg / mL composition to about 800 mcg / mL composition, about 600 mcg / mL composition to about950 mcg / mL composition, about 600 mcg / mL composition to about 900 mcg / mL composition, about 600 mcg / mL composition to about 850 mcg / mL composition, about 600 mcg / mL composition to about 600 mcg / mL composition, about 700 mcg / mL composition to about the specification page 9 / 22 12 CN 121057583 A The composition is present in amounts ranging from 950 mcg / mL, about 700 mcg / mL to about 900 mcg / mL, about 700 mcg / mL to about 850 mcg / mL, about 700 mcg / mL to about 800 mcg / mL, about 800 mcg / mL to about 950 mcg / mL, about 800 mcg / mL to about 900 mcg / mL, about 800 mcg / mL to about 850 mcg / mL, about 900 mcg / mL to about 950 mcg / mL, and about 1000 mcg / mL.

[0046] In some embodiments, hyaluronic acid is present in amounts ranging from about 1 mg / mL to about 10 mg / mL. For example, hyaluronic acid is available in concentrations of approximately 1.0 mg / ml, 1.1 mg / ml, 1.2 mg / ml, 1.3 mg / ml, 1.4 mg / ml, 1.5 mg / ml, 1.6 mg / ml, 1.7 mg / ml, 1.8 mg / ml, 1.9 mg / ml, 2 mg / ml, 2.1 mg / ml, 2.2 mg / ml, 2.3 mg / ml, 2.4 mg / ml, 2.5 mg / ml, 2.6 mg / ml, 2.7 mg / ml, 2.8 mg / ml, 2.9 mg / ml, 3 mg / ml, 3.1 mg / ml, 3.2 mg / ml, 3.3 mg / ml, 3.4 mg / ml, and 3.5 mg / ml. Approximately 3.6 mg / ml, approximately 3.7 mg / ml, approximately 3.8 mg / ml, approximately 3.9 mg / ml, approximately 4 mg / ml, approximately 4.1 mg / ml, approximately 4.2 mg / ml, approximately 4.3 mg / ml, approximately 4.4 mg / ml, approximately 4.5 mg / ml, approximately 4.6 mg / ml, approximately 4.7 mg / ml, approximately 4.8 mg / ml, approximately 4.9 mg / ml, approximately 5 mg / ml, approximately 5.5 mg / ml, approximately 6 mg / ml, approximately 6.5 mg / ml, approximately 7 mg / ml, approximately 7.5 mg / ml, approximately 8 mg / ml, approximately 8.5 mg / ml, approximately 9 mg / ml, approximately 9.5 mg / ml, approximately 10 mg / ml, or greater amounts are present.

[0047] In some embodiments, the composition according to embodiments of the present invention comprises commercially available hyaluronic acid.Compounds. For example, suitable commercially available hyaluronic acid compositions include, but are not limited to, hyaluronic acid sold under the trademarks JUVEDERM™, RESTYLANE-L™, CAPTIQUE™, BELOTERO BALANCE™, PREVELLE SILK™, ELEVESS™, HYLAFORM™, EUFLEXXA™, GEL-ONE™, HYALGAN™, ORTHOVISC™, MONOVISC™, SUPARTZ™, SYNVISC™, and SYNVISC-ONE™.

[0048] a. Polydispersity Index (PDI) of Hyaluronic Acid

[0049] The term “polydispersity index (PD)” as used herein characterizes in the art the width of a molecular weight distribution curve relating to the ratio of weight-average molecular weight Mw (which is the total weight of all polymer (hyaluronic acid) molecules in a sample divided by the total number of molecules) to number-average molecular weight Mn (which is the weight-average molecular weight of each molecule multiplied by its corresponding molecular weight, i.e., the weight statistical average molecular weight). Generally, a smaller PDI value indicates a narrower range of molecular weight distribution.

[0050] In some embodiments, the analysis of hyaluronic acid can be performed by size exclusion chromatography (SEC). In some embodiments, SEC may include separating the sample, monitoring the output with a concentration detector (such as a refractometer or UV absorbance detector), and comparing the elution time with the molar mass. In some embodiments, the analysis of hyaluronic acid may include using a multi-angle light scattering (MALS) device in series with a concentration detector. In some embodiments, MALS instruments, differential refractometers, and differential viscometers with SEC may be used to study the material properties of hyaluronic acid. A combination of these methods may be used to determine the absolute molecular weight, molecular weight distribution, and radius information of hyaluronic acid.

[0051] 2. Carrier and other additives

[0052] In some embodiments, the composition comprises a carrier medium. Such a carrier medium may be a biocompatible fluid suitable for injection into mammalian skin. In some embodiments, the carrier medium comprises an aqueous saline solution. In some embodiments, hyaluronic acid serves as both the carrier medium and the active ingredient.

[0053] In some embodiments, the composition comprises one or more additives. Such additives may include preservatives or biocides.

[0054] In some embodiments, the composition comprises a microemulsifier, a nanoemulsifier, solid lipid nanoparticles, a nanostructured lipid carrier, liposomes, or vesicles.

[0055] In some embodiments, the composition may comprise fatty acids (e.g., oleic acid), esters of fatty acids and alcohols (e.g., isopropyl myristate, isopropyl palmitate, ethyl oleate), medium-chain triglycerides, triacetin, or terpenes (e.g., limonene, thiocyanate ...Methol, cinoele). In some embodiments, the composition may contain surfactants. For example, suitable surfactants include, but are not limited to, TWEEN™ (polysorbate), CREMOPHOR™ (a mixture of polyethylene glycol glycerol hydroxystearate, PEG-40 castor oil, and polyoxyethylene 40 hydrogenated castor oil), TRANSCUTOL™ P (diethylene glycol monoethyl ether), PLUROL OLEIQUE™ (polyglycerol-3-oleate), PLUROL ISOSTEARIQUE™ (isostearate of polyglycerol and higher oligomers), and LABRASOL™ (a mixture of monoglycerides, diglycerides, and triglycerides of C8 and C10 fatty acids, and monoesters and diesters of PEG), as well as lecithin. In some embodiments, the composition may contain co-surfactants. For example, suitable co-surfactants include, but are not limited to, short-chain and medium-chain alcohols and polyglycerol derivatives, including ethanol, isopropanol, isopropyl myristate, and propylene glycol.

[0056] In some embodiments, the composition comprises one or more of soybean oil, jojoba oil, aloe vera oil, soybean phosphatidylcholine, water, polysorbate 80, ethanol, benzyl alcohol, isopropanol, glycerin, glyceryl monostearate, and propylene glycol.

[0057] II. Methods

[0058] This disclosure provides methods of treatment and administration, as well as methods of preparing the composition. In some embodiments, the methods described in U.S. Patent 11,337,993 can be used in this disclosure (the entire contents of which are incorporated herein by reference).

[0059] 1. Treatment Methods

[0060] This disclosure provides methods of treatment using the compositions described herein. As used herein, the term “treat / treatment / treating” means a therapeutic treatment in which the aim is to reverse, reduce, improve, inhibit, slow, or stop the progression or severity of a disease or condition (e.g., androgenetic alopecia or other forms of hair loss). The term "treatment" includes reducing or alleviating at least one side effect or symptom of a disease or condition (e.g., androgenetic alopecia or other forms of hair loss). Treatment is considered effective if one or more symptoms are alleviated, such as a decrease in the rate of hair loss. Alternatively, treatment is considered effective if the progression of the disease is reduced or stopped. That is, the term "treatment" includes not only improvement of symptoms but also cessation or at least slowing of the progression or worsening of symptoms compared to what would be expected without treatment. Beneficial or desired clinical outcomes include, but are not limited to, reduction of one or more symptoms, attenuation of the severity of the disease, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or attenuation of the disease state, and / or remission (whether partial or complete). For example, if the degree or amount of hair loss is reduced, the progression of hair loss is slowed or stopped, or hair regrowth is induced.Regeneration is considered an effective treatment.

[0061] In some embodiments, the compositions of this disclosure can be used to treat androgenetic alopecia (AGA). In some embodiments, the compositions of this disclosure can improve one or more metrics associated with AGA. In some embodiments, the compositions of this disclosure can be used to treat hair loss in patients with AGA. In some embodiments, the composition stimulates hair growth in patients with AGA.

[0062] Androgenetic alopecia (AGA) is a genetically predetermined disorder caused by an overreaction to androgens, characterized by progressive loss of terminal hair on the scalp at any time after puberty and affecting up to 50% of men and women. Elevated levels of androgens, including dihydrotestosterone (DHT), lead to increased activation of androgen receptors on hair follicles, which in turn leads to a shortening of the anagen (growth) phase in the normal hair growth cycle. Overactivation leads to miniaturization of hair follicles through the progressively shortened anagen phase, resulting in thinner and shorter hair follicles. In men, hair loss is significant in the vertex and frontotemporal regions; in women, hair loss typically occurs in the vertex region.

[0063] Methods for stimulating hair growth in a patient in need include applying a composition according to an embodiment of the invention to the surface of or into the patient's skin. A method for applying the composition for hair growth is also provided herein (pages 11 / 22, CN 121057583 A).

[0064] In some embodiments, the disclosed composition may be applied to the patient's skin. In some embodiments, the composition is applied topically by applying it to the surface of the patient's skin. In some embodiments, the composition is applied to the dermis or subepithelium of the patient's skin, for example, by injection as described herein. In one embodiment, the composition may be applied to the patient's skin by intradermal injection.

[0065] Assessment of a patient's hair growth may be based on patient self-assessment, physician assessment using standardized scales, global radiographic assessment, target area hair count (TAHC), target area hair width (TAHW; also referred to herein as hair diameter measurement), mean hair length measurement, mean hair diameter measurement, and / or hair weight measurement. In one embodiment, the composition can improve or increase the target area hair count (TAHC), target area hair width (TAHW), average hair length, average hair diameter, hair darkness, and / or hair weight. As used herein, the target area hair count refers to the number of hairs per cm² on a patient's scalp. In some embodiments, a higher TAHC value may indicate a better outcome. In some embodiments, the target area hair width (TAHW) may refer to the cumulative width of hairs in a patient's scalp region (target area) being treated with the composition of this disclosure. A higher TAHW value may indicate a better outcome.

[0066] In some aspects, the composition can improve or increase the target area hair count (TAHC), TAHW, average hair length, average hair diameter, hair darkness and / or hair weight by about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, about 1% to 10%, 15% to 25%, 20% to 30%, 25% to 35%, 30% to 40%, 35% to 45%, 40% to 50%, 45% to 55%, 50% to 60%, 55% to 65%, 60% to 70%, 65% to 75%, 70% to 80%, 75% to 85%, 80% to 90%, 85% to 95% or more. In some aspects, the composition can improve or increase the target area hair count (TAHC), TAHW, average hair length, average hair diameter, hair dullness, and / or hair weight by about 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or more.

[0067] The improvement or increase in the target area hair count (TAHC), TAHW, average hair length, average hair diameter, hair dullness, and / or hair weight can be measured at any time after treatment with the composition of this disclosure. For example, the improvement can be measured from about 10 days to about 160 days after application of the composition. In some embodiments, improvement may be measured after approximately 10 days, approximately 20 days, approximately 30 days, approximately 40 days, approximately 50 days, approximately 60 days, approximately 70 days, approximately 80 days, approximately 90 days, approximately 100 days, approximately 110 days, approximately 120 days, approximately 130 days, approximately 140 days, approximately 150 days, approximately 160 days, approximately 170 days, approximately 180 days, or longer.

[0068] In some embodiments, prior to treatment with the compositions of this disclosure, the patient has experienced hair loss for approximately 1 month, approximately 2 months, approximately 3 months, approximately 4 months, approximately 5 months, approximately 6 months, approximately 7 months, approximately 8 months, approximately 9 months, approximately 10 months, approximately 1 year, approximately 2 years, approximately 3 years, approximately 1 year, approximately 5 years, approximately 6 years, approximately 7 years, approximately 8 years, approximately 9 years, approximately 10 years, or longer. In some embodiments, the patient has experienced hair loss for approximately three to five years prior to application of the compositions disclosed herein.

[0069] In some embodiments, the patient has undergone a previous hair transplant procedure. In some embodiments, the patient has undergone hair transplantation for approximately 1 to 2 months, approximately 2 to 3 months, approximately 3 to 4 months, approximately 4 to 5 months, approximately 5 to 6 months, approximately 6 to 7 months, approximately 7 to 8 months, approximately 8 to 9 months, approximately 9 to 10 months, approximately 10 to 11 months, approximately 11 to 12 months, or longer.

[0070] In some embodiments, the patient is a male or female patient.

[0071] In some embodiments, the patient has mild or moderate AGA.

[0072] In some embodiments, the patient has received a modified Norwood-Hamilton Scale score of I, II, IIa, III, III vertex, IIIa, IV, V, Iva, VI, VII, or Va. The characteristics associated with each modified Norwood-Hamilton Scale score are presented in the figure. In some aspects, the patient has received a modified Norwood-Hamilton Scale score of III or a modified Norwood-Hamilton Scale score of IV.

[0073] In some embodiments, a method of stimulating hair growth in a patient in need comprises applying a composition according to an embodiment of the invention to the surface of or into the patient's skin. In some embodiments, the composition is applied topically by applying it to the surface of the patient's skin. In some embodiments, the composition is applied to the dermis or subepithelium of the patient's skin, for example by injection as described herein.

[0074] In some embodiments, the composition according to the embodiments of the invention is applied to the patient's skin once a day for one day, once a day for one week, once a day for one month, once a day for one year, twice a day for one day, twice a day for one week, twice a day for one month, twice a day for one year, once a week for one week, once a week for one month, once a week for one year, twice a week for one week, twice a week for one month, twice a week for one year, once a month for one month, once a month for two months, once a month for six months, once a month for one year, twice a month for one month, twice a month for two months, twice a month for two months, twice a month for two months, twice a month for four months, twice a month for six months, twice a month for one year, once a month for three months, once a month for six months, once a month for nine months, once a month for one year, once a month for four months, once a month for eight months, once a month for one year, once a month for six months, once a month for six months, once a month for one year, or applied as needed.

[0075] In some embodiments, the composition is applied at least once a week, at least twice a week, at least three times a week, at least four times a week, at least five times a week, at least six times a week, at least seven times a week, or more. In some embodiments, the composition is applied over several consecutive days of the week. In some embodiments, the composition is applied over two consecutive days, three consecutive days, four consecutive days, five consecutive days, six consecutive days, seven consecutive days, or longer. In some embodiments, the composition is applied every other day of the week. As a non-limiting example, the composition may be applied three times a week for three consecutive days.

[0076] As a non-limiting example, the method of this disclosure includes applying up to 1% hyaluronic acid three times a week. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least twice a week. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least three times a week. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least four times a week. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least five times a week. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least six times a week. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least seven times a week.

[0077] As a non-limiting example, the method of this disclosure includes applying up to 1% hyaluronic acid three times a week for three consecutive days. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least twice a week for two consecutive days. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least four times a week for four consecutive days. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least five times a week for five consecutive days. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least six times a week for six consecutive days. As a non-limiting example, the method of this disclosure may include applying up to 1% hyaluronic acid to the skin of a subject at least seven times a week for seven consecutive days.

[0078] As a non-limiting example, the method of this disclosure may involve intradermal injection of 0.3% hyaluronic acid three times a week. As a non-limiting example, the method of this disclosure may involve intradermal injection of 0.4% hyaluronic acid three times a week. As a non-limiting example, the method of this disclosure may involve intradermal injection of 0.2% hyaluronic acid three times a week. As a non-limiting example, the method of this disclosure can be performed by intradermal injection of 0.3% hyaluronic acid three times a week for three consecutive days. As an example of non-limiting specification 13 / 22 pages 16 CN 121057583 A, the method of this disclosure can be performed by intradermal injection of 0.4% hyaluronic acid three times a week for three consecutive days. As a non-limiting example, the method of this disclosure can be performed by intradermal injection of 0.2% hyaluronic acid three times a week for three consecutive days.

[0079] 2. Application

[0080] A method of applying a composition according to an embodiment of the invention comprises delivering the composition according to an embodiment of the invention to a hair follicle. In some embodiments, delivery is by topical application, i.e., applying the composition to the surface of the skin.This allows the composition to penetrate the skin. In some embodiments, the method of administration includes a step to enhance penetration before topical application of the composition. In some embodiments, delivery is performed by injection into the skin.

[0081] A method of administering the composition according to an embodiment of the invention includes injecting a therapeutic amount of the composition into the skin of a patient requiring treatment. In some embodiments, the composition according to an embodiment of the invention is administered as a bolus injection, which, as used herein, refers to a dose delivered over a time period of less than ten minutes. In some embodiments, the composition according to an embodiment of the invention is administered as an infusion, which, as used herein, refers to a dose delivered over a time period of about ten minutes or longer.

[0082] Such injections may be performed via a single needle, microneedle, or similar device, or an array of needles, microneedles, or similar devices. In some embodiments, the composition according to an embodiment of the invention is delivered via a conventional syringe. In some embodiments, subcutaneous delivery is performed via a hollow microneedle syringe. In some embodiments, subcutaneous delivery is performed via a microneedle patch coated with the composition according to an embodiment of the invention, for example, coated with the composition according to an embodiment of the invention by 3D printing. In some embodiments, the composition is delivered via a jet syringe. As used herein, the term "needle" refers to any such device used to pierce the skin and inject the composition according to an embodiment of the invention.

[0083] Preferably, the composition is applied near the patient's hair follicles. Thus, in some embodiments, the composition is applied by injecting a therapeutic amount of the composition into the patient's dermis (intradermal injection).

[0084] In some embodiments, the composition is applied by injecting a therapeutic amount of the composition into the patient's subcutaneous tissue. In some embodiments, the composition is applied to the patient's skin at a depth of about 0.4 mm to about 2 mm (i.e., about 0.4 mm to about 3 mm from the surface of the skin). In some embodiments, the composition is applied to the patient's skin in a depth of about 0.1 mm, 0.2 mm, 0.3 mm, 0.4 mm, about 0.5 mm, about 0.6 mm, about 0.7 mm, about 0.8 mm, about 0.9 mm, about 1 mm, about 1.1 mm, about 1.2 mm, about 1.3 mm, about 1.4 mm, about 1.5 mm, about 1.6 mm, about 1.7 mm, about 1.8 mm, about 1.9 mm, about 2 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm, about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, or about 3 mm. In some embodiments, the composition is applied to the patient's skin in a depth of about 0.1 mm to about 0.5 mm, about 0.1 mm to about 1 mm, about 0.5 mm to about 1 mm, about 1 mm to about 1.5 mm, or about 1 mm.Between 0.5mm and 2mm, between 2mm and 2.5mm, between 2.5mm and 3mm, between 1mm and 3mm, between 1.5mm and 3mm, between 0.4mm and 0.6mm, between 0.4mm and 0.8mm, between 0.4mm and 1mm, between 0.4mm and 1.2mm, between 0.4mm and 1.4mm, between 0.4mm and 1.6mm, between 0.4mm and 1.8mm, between 0.4mm and 2mm, between 0.4mm and 2.2mm. Between about 0.4mm and about 2.4mm, Between about 0.4mm and about 2.6mm, Between about 0.4mm and about 2.8mm, Between about 0.4mm and about 3mm, Between about 0.6mm and about 0.8mm, Between about 0.6mm and about 1mm, Between about 0.6mm and about 1.2mm, Between about 0.6mm and about 1.4mm, Between about 0.6mm and about 1.6mm, Between about 0.6mm and about 1.8mm, Between about 0.6mm and about 2mm, Between about 0.6mm and about 2.2mm, Between about 0.6mm and about 2.4mm, Between about 0.6mm and about 2.6mm, Between about 0.6mm and about 2.8mm, Between about 0.6mm and about 3mm, Between about 0.8mm and about 1mm, Between about 0.8mm and about 1.2mm, Between about 0.8mm... (Instruction manual 14 / 22) Page 17 CN 121057583 A Between approximately 1.4 mm, Between approximately 0.8 mm and approximately 1.6 mm, Between approximately 0.8 mm and approximately 1.8 mm, Between approximately 0.8 mm and approximately 2 mm, Between approximately 0.8 mm and approximately 2.2 mm, Between approximately 0.8 mm and approximately 2.4 mm, Between approximately 0.8 mm and approximately 2.6 mm, Between approximately 0.8 mm and approximately 2.8 mm, Between approximately 0.8 mm and approximately 3 mm, Between approximately 1 mm and approximately 1.2 mm, Between approximately 1 mm and approximately 1.4 mm, Between approximately 1 mm and approximately 1.6 mm, Between approximately 1 mm and approximately 1.8 mm, Between approximately 1 mm and approximately 2 mm, Between approximately 1 mm and approximately 2.2 mm, Between approximately 1 mm and approximately 2.4 mm, Between approximately 1 mm and approximately 2.6 mm, Between approximately 1 mm and approximately 2.8 mm, Between approximately 1 mm and approximately 3 mm, Between approximately 1.2 mm and approximately 1.4 mm Between, between about 1.2mm and about 1.6mm, between about 1.2mm and about 1.8mm, between about 1.2mm and about 2mm, between about 1Between 0.2mm and 2.2mm, between 1.2mm and 2.4mm, between 1.2mm and 2.6mm, between 1.2mm and 2.8mm, between 1.2mm and 3mm, between 1.4mm and 1.6mm, between 1.4mm and 1.8mm, between 1.4mm and 2mm, between 1.4mm and 2.2mm, between 1.4mm and 2.4mm, between 1.4mm and 2.6mm, between 1.4mm and 2.8mm, between 1.4mm and 3mm, between 1.6mm and 1.8mm, between 1.6mm and 2mm, between 1.6mm and 2.2mm, between 1.6mm and 2.4mm, between 1.6mm and 2.6mm, between 1.6mm and 2.8mm, between 1.6mm and 2.8mm, between 1.2mm and 2.2mm, between 1.6mm and 2.4mm, between 1.6mm and 2.6 ... Between 0.6mm and 3mm, between 1.8mm and 2mm, between 1.8mm and 2.2mm, between 1.8mm and 2.4mm, between 1.8mm and 2.6mm, between 1.8mm and 2.8mm, between 1.8mm and 3mm, between 2.0mm and 2.2mm, between 2.0mm and 2.4mm, between 2.0mm and 2.6mm, between 2.0mm and 2.8mm, between 2.0mm and 3mm, between 2.2mm and 2.4mm, between 2.2mm and 2.6mm, between 2.2mm and 2.8mm, between 2.2mm and 3mm, between 2.4mm and 2.6mm, between 2.0mm and 2.2mm, between 2.2mm and 2.8mm, between 2.2mm and 3mm, between 2.4mm and 2.6mm, between 2.0mm and 2.2mm, between 2.2mm and 2.8mm, between 2.2mm and 2.8mm, between 2.2mm and 3mm, between 2.4mm and 2.6mm, between 2.0mm and 2.2mm, between 2.2 ... The depth of injection is between 0.4 mm and 2.8 mm, between 2.4 mm and 3 mm, between 2.6 mm and 2.8 mm, between 2.6 mm and 3 mm, or between 2.8 mm and 3 mm. In some embodiments, the composition of this disclosure is injected above the skull.

[0085] In one aspect, the injection depth may vary depending on the thickness of the scalp in different regions of the scalp. Different regions of the scalp may have different thicknesses. For example, the average dermal thickness of the posterior scalp may be about 1443.86 μm, the thickness of the temporal scalp may be about 1349.52 μm, and / or the thickness of the anterior scalp may be about 1146.13 μm. In some embodiments, the injection depth may be from about 0.7 mm to about 2 mm.

[0086] In one embodiment, the composition is applied at the stem cell region of the hair follicle, adjacent to the stem cell region of the hair follicle, or near the stem cell region of the hair follicle.

[0087] In some embodiments, the injection area can be about 1 cm², about 2 cm², about 3 cm², about 4 cm², about 5 cm², about 6 cm², about 7 cm², about 8 cm², about 9 cm², about 10 cm², about 15 cm², about 20 cm², about 25 cm², about 30 cm², about 35 cm², about 40 cm², about 45 cm², about 50 cm², about 55 cm², about 60 cm², about cm², about 70 cm², about 75 cm², about 80 cm², about 85 cm², about 90 cm², about 95 cm², about 100 cm², 200 cm², 300 cm², 400 cm², 500 cm², 600 cm², 700 cm², or larger. In one embodiment, the injection area can reach the entire area of ​​the subject's scalp.

[0088] The composition can be injected in a row at intervals of about 0.2 cm to 0.3 cm. In some embodiments, the composition may be injected in multiple rows at approximately 1 cm intervals.

[0089] In some embodiments, the composition according to an embodiment of the invention may be administered via multiple injections. In some embodiments, the composition according to an embodiment of the invention may be administered via approximately 1 injection / cm² skin to approximately 1000 injections / cm² skin, approximately 200 injections / cm² skin to approximately 800 injections / cm² skin, or approximately 400 injections / cm² skin to approximately 650 injections / cm² skin. In some embodiments, the composition is applied via approximately 200 injections / cm² of skin, approximately 250 injections / cm² of skin, approximately 300 injections / cm² of skin, approximately 350 injections / cm² of skin, approximately 400 injections / cm² of skin, approximately 450 injections / cm² of skin, approximately 500 injections / cm² of skin, approximately 550 injections / cm² of skin, approximately 600 injections / cm² of skin, or approximately 650 injections / cm² of skin. The contents of U.S. Patent No. 11,337,993 are hereby incorporated by reference.

[0090] The compositions of this disclosure may be applied to one or more areas of the scalp. For example, the compositions may be applied to the frontal region (or central frontal hairline), the middle scalp (or posterior middle scalp), the temporal scalp (lateral), the vertex transition zone (or vertex transition point), and the crown (or vertex). In some embodiments, the frontal and temporal regions of the scalp may be collectively referred to as the frontotemporal region. In one embodiment, the composition is applied to the frontotemporal region of the scalp.

[0091] In some embodiments, local delivery is performed after or in conjunction with the application of iontophoresis. For example, iontophoresis may include applying a mild current (e.g., 0.1 mA / cm² to 1.0 mA / cm²) to increase the skin penetration of the composition. Without being bound by theory, it is believed that iontophoresis can modify [the skin] through electromigration, electroosmosis, and / or enhanced passive diffusion.

[0092] In some embodiments, local delivery is performed after or in conjunction with the application of electroporation. For example, electroporation may include applying short-duration (10 microseconds to 10 milliseconds) high-intensity, high-voltage (e.g., 50V to 1500V) electrical pulses to form aqueous pores in the lipid bilayer of the stratum corneum of the skin.

[0093] In some embodiments, local delivery is performed after or in conjunction with the application of ultrasound-enhanced permeation. For example, ultrasound-enhanced permeation may include applying sound waves at a high frequency (e.g., about 500kHz to 1250kHz) or a low frequency (e.g., about 20kHz to about 100kHz) or (starting at one of the high frequency or low frequency and progressing to the other of the high frequency or low frequency).

[0094] In some embodiments, local delivery is performed after or in conjunction with the application of laser ablation. Laser ablation may include generating photomechanical waves by ablating a target material (e.g., a polymer) placed on the surface of the skin.

[0095] In some embodiments, local delivery is performed after or in combination with magnetophoresis. Magnetophoresis may include applying a magnetic field, such as a pulsed electromagnetic field, to the skin.

[0096] In some embodiments, local delivery is performed after radiofrequency thermal ablation. Thermal ablation may include applying extreme heat (e.g., about 300 degrees Celsius for several microseconds) to the skin surface. Unbound by theory, thermal ablation is believed to cause partial evaporation of the stratum corneum to create micron-scale channels. Thermal ablation can be performed using commercially available devices, including VIADOR™ (Syneron Medical Ltd, Israel) and PASSPORT™ (Nitto-Denko, Japan). In some embodiments, thermal ablation may be performed using an erbium:yttrium-gallium-garnet (Er:YAG) laser emitted at 2,790 nm or a yttrium scandium-gallium-garnet (YSGG) laser emitted at 2,940 nm. In some embodiments, partial laser ablation may be applied to a sub-mm region to generate spots (e.g., 40 μm to 300 μm, density between 50 cm⁻¹ and 600 cm⁻¹) simulating a microneedle array pattern. In some embodiments, local delivery is performed after the application of the microneedle device.

[0097] 3. Production Method

[0098] A method for producing a composition according to an embodiment of the invention comprises preparing an effective amount of hyaluronic acid. In some embodiments, the effective amount of hyaluronic acid may be further mixed with an effective amount of osteopontin. In some embodiments, the average molecular weight of the hyaluronic acid is in the range of about 4,000 Da to 10,000 Da, in the range of about 10,000 Da to about 100,000 Da, in the range of about 15 kDa to about 50 kDa, in the range of about 75 kDa to about 350 kDa, or in the range of aboutThe range is from 20 kDa to 1350 kDa. In some embodiments, the average molecular weight of hyaluronic acid is greater than about 950 kDa. In some embodiments, an effective amount of hyaluronic acid is mixed with an effective amount of CD44 binding ligand. The method may also include the step of preparing a physiologically acceptable carrier medium by adding an active agent to the physiologically acceptable carrier medium. In some embodiments, the physiologically acceptable carrier medium is injectable.

[0099] In some embodiments, the production method includes the step of forming a microemulsion or nanoemulsion. The microemulsion or nanoemulsion may contain oil, water, surfactant and co-surfactant to form a colloidal dispersion with droplet sizes in the range of about 10 nm to about 100 nm. In some embodiments, the microemulsion or nanoemulsion may contain fatty acids (e.g., oleic acid), esters of fatty acids and alcohols (e.g., isopropyl myristate, isopropyl palmitate, ethyl oleate), medium-chain triglycerides, triacetin, or terpenes (e.g., limonene, methol, cinoele). In some embodiments, the microemulsion or nanoemulsion may contain surfactants. Suitable surfactants include, but are not limited to, TWEEN™ (polysorbate), CREMOPHOR™ (a mixture of polyethylene glycol glycerol hydroxystearate, PEG-40 castor oil, and polyoxyethylene 40 hydrogenated castor oil), TRANSCUTOL™ P (diethylene glycol monoethyl ether), PLUROL OLEIQUE™ (polyglycerol-3-oleate), PLUROL ISOSTEARIQUE™ (isostearate of polyglycerol and higher oligomers), and LABRASOL™ (a mixture of monoglycerides, diglycerides, and triglycerides of C8 and C10 fatty acids, and monoesters and diesters of PEG), as well as lecithin. In some embodiments, the microemulsion or nanoemulsion may contain a co-surfactant. Suitable co-surfactants include, for example, short-chain and medium-chain alcohols and polyglycerol derivatives, including ethanol, isopropanol, isopropyl myristate, and propylene glycol. In some embodiments, the formation of the microemulsion or nanoemulsion includes using a high-pressure homogenizer, a microfluidizer, and / or an ultrasonic generator.

[0100] In some embodiments, the production method includes the step of mixing a composition according to an embodiment of the invention with solid nanoparticles. The solid nanoparticles may contain inorganic materials, such as metal oxides (e.g., zinc oxide, titanium dioxide) or polymers that are solid at room temperature.

[0101] In some embodiments, the production method includes the step of mixing a composition according to an embodiment of the invention with solid lipid nanoparticles. The solid lipid nanoparticles may contain lipids that are solid at room temperature, the lipids having a surface...The surfactant's surface coating stabilizes the lipid into droplets having a size of less than about 100 nm when dispersed in water.

[0102] In some embodiments, the production method includes the step of mixing a composition according to an embodiment of the invention with a nanostructured lipid carrier. The nanostructured lipid carrier may comprise a fluid lipid phase embedded in a solid lipid matrix or positioned at the surface of solid flakes and a surfactant layer.

[0103] In some embodiments, the production method includes the step of mixing a composition according to an embodiment of the invention with liposomes. The liposomes may comprise spherical vesicles composed of amphiphilic phospholipids and cholesterol, which self-associate into multilayer, large monolayer, and small monolayer vesicles.

[0104] In some embodiments, the production method includes the step of mixing a composition according to an embodiment of the invention with flexible vesicles. The flexible vesicles may comprise materials that will associate into a bilayer structure and components that impart flexibility. In some embodiments, the flexible vesicles comprise ethosomes (i.e., phospholipids with a high proportion of ethanol), niosomes (i.e., nonionic surfactants), invasomes (i.e., mixtures of phospholipids, ethanol, and terpene permeability enhancers), SECosomes (i.e., surfactants, ethanol, and cholesterol), or PEVs (i.e., permeability enhancer vesicles). In some embodiments, the PEV may comprise oleic acid, limonene, or propylene glycol.

[0105] In some embodiments, the production method includes the step of mixing the composition according to embodiments of the invention with polymeric micelles or polymeric dendritic polymers. The polymeric micelles may be colloidal carriers having a hydrophilic shell and a hydrophobic core. The polymeric micelles may be nanoscale. The polymeric dendritic polymers may comprise branched polymer structures.

[0106] Examples

[0107] Example 1. Evaluation of hyaluronic acid (AMP-303) compared to saline control in male subjects with androgenetic alopecia

[0108] Study Design Explanation of Specification 17 / 22 pages 20 CN 121057583 A

[0109] Quantitative hair growth measurement has become the standard for confirming the efficacy of AGA hair growth products. To date, dosage forms commonly used to treat AGA are topically or orally administered. Intradermal injection may alter skin surface topology / contour, hair orientation, or other characteristics that may interfere with hair imaging or accurate serial imaging over time. To measure the effect of this route of administration on the utility of standard hair count, relevant measurements were evaluated. This study focuses on assessing what differences might occur between before and after injection due to AMP-303 compared to saline control during the study period. Local safety effects of intradermal injection of AMP-303 compared to saline control will be collected. In addition, this study will include three (3) differentThe study population of patients with androgenetic alopecia was refined for future research on androgenetic alopecia.

[0110] Study Product and Formulation

[0111] The study compound is hyaluronic acid (also referred to herein as AMP-303) for intradermal injection at a concentration of 0.3% ± 0.1%.

[0112] AMP-303 for intradermal injection consists of non-crosslinked hyaluronic acid and phosphate-buffered saline.

[0113] The control (injectable sterile saline) will be phosphate-buffered saline.

[0114] Both the AMP-303 mediator and the saline control will be supplied as injectable solutions. The same 1 mL syringe will be used for both AMP-303 and the control, with a 0.1 mL to 0.5 mL cutoff. AMP-303 and the saline control will be supplied as sterile products.

[0115] Study Objectives

[0116] The aim of this study is to evaluate the safety and tolerability of AMP-303 injected into the dermis of the scalp in male subjects with androgenetic alopecia, compared to a saline control. The study will also evaluate the impact of AMP-303 on hair counting assessment methods in male subjects with androgenetic alopecia, if any, compared to a saline control. Additionally, this study will be used to refine the intended user population.

[0117] Study Design

[0118] This study is a prospective, multicenter, randomized, saline-controlled, non-significant risk (NSR) early feasibility study.

[0119] The dosing regimen will consist of up to 2 mL of AMP-303 to be injected intradermally and a saline control; injections will be administered over 3 consecutive days.

[0120] The study will be randomized at each study site on day 0. Subjects will be randomly assigned to receive AMP-303 on the right or left side and a saline control on the contralateral side. All participants will receive intradermal injections of AMP-303 as a carrier and a saline control into the frontotemporal region of the scalp on days 0, 1, and 2. Participants will receive AMP-303 on one side of the frontotemporal region and a saline control on the other side. The injection area will be approximately 4 cm², and the injection volume will be up to 2 mL for each area. Injections will be limited to the frontotemporal region of the scalp. Each participant is expected to remain in the study for approximately 6 months (screening will be conducted upon withdrawal).

[0121] The visit schedule is as follows:

[0122] Visit 1: Screening (Day -30 to Day -1)

[0123] Visit 2: Day 0 (Injection / Baseline)

[0124] Visit 3: Day 1 (Injection)

[0125] Visit 4: Day 2 (Injection)

[0126] Visit 5: Day 14 (±3 days)

[0127] Visit 6: Day 30 (±7 days)

[0128] Visit 7: Day 60 (±7 days)

[0129] Visit 8: Day 90 (±7 days) Instruction Manual 18 / 22 Pages 21 CN 121057583 A

[0130] Visit 9: Day 120 (±7 days)

[0131] Visit 10: Day 150 (±7 days)

[0132] Approximately 36 subjects (12 subjects per group) were recruited to this study at up to 3 study sites.

[0133] Three (3) different groups were recruited to the study, each with approximately 12 male subjects: 1) recently started hair loss (defined as a hair loss history of about 3 to 5 years); 2) established hair loss (defined as a hair loss history of >10 years); and 3) post-transplantation. Group 3 recruited subjects who had completed a hair transplantation procedure involving bilateral frontotemporal regions. Eligible subjects had to (1) be 1 to 2 months post-transplantation, (2) have a non-significant post-transplantation healing process, and (3) have no active adverse events or medical problems in the designated post-transplantation injection area.

[0134] On day 0, eligible subjects were randomized to receive AMP-303 in the right or left frontotemporal region and a saline control on the contralateral side (“injection site”). AMP-303 and the saline control were administered via intradermal injection by a physician investigator at each randomization visit on days 0, 1, and 2. The physician, investigator, and subjects were unaware of the study product injected on each side. Subjects who discontinued the study were not replaced.

[0135] Potential Benefits and Risks

[0136] The benefits of intradermal injection of the AMP-303 mediator and this study include collecting data on the safety and tolerability of the mediator for future study products, determining the impact on hair count assessments (if any) due to possible surface topological effects associated with the intradermal injection of the AMP-303 mediator and the saline control, and refining the expected cohort for future studies. Potential risks associated with needle injection into the skin include injection site pain, erythema, bruising, and edema. These events are expected to be the result of intradermal injection of AMP-303 or the saline control; however, if the severity or incidence of any particular reaction or effect is greater than expected, it may be considered an adverse event (AE).

[0137] Minimization of Expected Risk

[0138] Risk is minimized under this protocol by: 1) a limited injection area on the scalp, consisting of a total area not exceeding 4 cm²; 2) low molecular weight, non-crosslinked hyaluronic acid; 3) limited to intradermal injection at a depth of approximately 0.6 mm to 0.8 mm; careful consideration of scalp anatomy; 4) AMP-303 and the saline control will be provided as sterile products in syringes intended for injection; 5) the injector will be a physician with experience in intradermal injection; 6) standard precautions for injectable materials will be followed; and / or 7) only male subjects will be recruited.

[0139] Inclusion Criteria

[0140] The following are the criteria for inclusion in the study:

[0141] 1. Adult males aged 18 to 45 years at the time of consent.

[0142] 2. Clinical diagnosis of mild to moderate androgenetic alopecia (AGA) in the temples and vertex areas, with a score of III or IV on the modified Norwood-Hamilton Scale, in subjects who have had hair loss for approximately 3 to 5 years since the onset of hair loss, or for at least 10 years since the onset of hair loss, or who have recently undergone bilateral frontotemporal hair transplantation.

[0143] 3. Willing to maintain the same hairstyle, hair length, and hair color throughout the study.

[0144] 4. Agree to continue general hair care products and protocols for at least 2 weeks before Day 0 and throughout the study.

[0145] 5. Willing to have hair shaved at the injection site at screening and at each follow-up visit.

[0146] 6. Willing to have a photograph taken to identify the subject.

[0147] 7. Written informed consent prior to any study-related procedures.

[0148] 8. Written authorization for the use and publication of health and research information.

[0149] 9. Able to follow the instructions for use of the study and complete the study assessment tools without assistance, and likely to complete all required visits. Instructions for Use 19 / 22 pages 22 CN 121057583 A

[0150] Exclusion Criteria

[0151] The following are the criteria for exclusion from the study:

[0152] 1. Any uncontrolled systemic disease.

[0153] 2. History of a positive result for human immunodeficiency virus (HIV) or active hepatitis B or hepatitis C.

[0154] 3. History or current evidence of drug or alcohol abuse within 12 months prior to the screening visit.

[0155] 4. Visible inflammatory skin disease, lesion, or condition of the scalp in the injection area.

[0156] 5. History of known or suspected malignancy (other than melanoma or skin cancer); treatment with chemotherapy or cytotoxic agents; or radiation to the scalp.

[0157] 6. Any dermatological condition of the scalp on the area of ​​baldness and thinning that may interfere with the study injection or study evaluation, such as fungal or bacterial infection, seborrheic dermatitis, psoriasis, eczema, folliculitis, scarring, skin cancer, tattooing, or scalp atrophy.

[0158] 7. A history of or active hair loss due to any condition other than AGA, such as, but not limited to, diffuse telogen effluvium, alopecia areata, cicatricial alopecia, trichotillomania, or other genetic disorders.

[0159] 8. A history of severe dietary changes, weight changes, or eating disorders that the investigator considers to have caused hair loss within 6 months prior to the screening visit.

[0160] 9. A history of scalp reduction, significant trauma with associated scarring, previous hair transplantation (Groups 1 and 2), hair weaving, and / or platelet-rich plasma (PRP) procedures on the scalp.

[0161] 10. The subject has received any other systemic therapy that the investigator believes could substantially affect the subject's hair growth, including but not limited to vitamins or homeopathic supplements for hair growth or hair health products or others.Steroid hormones (in any form), including anabolic steroids.

[0162] 11. Any of the following topical preparations or procedures on the scalp occurring during a specified period prior to day 0 (baseline)

[0163] 4 weeks: Topical over-the-counter (OTC) or cosmetic treatments for the scalp that may affect hair growth (e.g., brands such as Aminexil, Maxilene, Nioxin, Foltene); hair growth products containing saw palmetto, copper, etc., including hair growth devices such as Cappilus, Revian, Theradome, etc.; and / or topical scalp treatments that may have an adjunctive effect on hair growth, including but not limited to corticosteroids, pimecrolimus, tacrolimus, and retinoids.

[0164] 12 weeks: Topical scalp treatments for hair growth including minoxidil, hormone therapy, anti-androgens, or other agents known to affect hair growth.

[0165] 6 months: Laser, light source, or energy therapy, or microneedling of the scalp; oral retinoids, isotretinoin.

[0166] 12. Any of the following systemic pharmacological treatments or procedures occurred during a specified period prior to Day 0 (baseline).

[0167] 12 weeks: β-blockers, cimetidine, diazoxide, or corticosteroids (including intramuscular and intralesional injections) during 12 weeks prior to Visit 2 / baseline. Inhaled, intranasal, or ocular corticosteroids are permitted if stable. Stable is defined as a consistent dose and frequency for at least 4 weeks prior to Visit 2 / baseline.

[0168] 6 months: 5α-reductase inhibitors, such as finasteride (e.g., Propecia®) or dutasteride; retinoids, isotretinoin, vitamin A intake of approximately 10,000 IU / day, or cyclosporine therapy; and / or minoxidil.

[0169] 13. Known hypersensitivity to any component of the investigational product (AMP-303 mediator or sterile saline).

[0170] 14. Subjects are known to be non-compliant or are deemed unlikely by the investigator to comply with the study protocol requirements.

[0171] 15. Currently recruited for an investigational drug or device study, or participating in such a study within 30 days of entering this study.

[0172] 16. Patients with or in any of the following situations: which the investigator considers likely to place the patient at significant risk, confound the study results, or significantly interfere with the patient's participation in the study.

[0173] Injection Administration

[0174] Injection will be limited to the frontotemporal region of the scalp. On day 0, eligible subjects will be randomized to receive an injection (AMP-303 or saline control) into the right or left treatment area. Standard injection site preparation will be performed (e.g., cleaning and wiping with alcohol). The same injection technique should be used for each subject daily (days 0, 1, and 2).

[0175] Response Measurement:

[0176] Response measurements to be evaluated include adverse events (including injection adverse events), local skin reactivity: the severity (none, minimal, mild, moderate, severe) of erythema, edema, and bruising will be assessed by a physician investigator, and pain, itching, and burning / tingling will be assessed by the subject.

[0177] Vital signs such as blood pressure, pulse rate, and respiratory rate will also be measured.

[0178] Other response measurements include digital image analysis of non-vellus hair photographs; target area hair count (TAHC); target area hair width (TAHW); and / or hair darkness.

[0179] Standardized Photography and Digital Image Analysis (DIA)

[0180] A qualified supplier will provide instructions for use in taking photographs and processing digital photographs. Macroscopic hair imaging (1 cm² target area) of a representative area of ​​thinning will be obtained to record and measure scalp hair: length, width, growth rate (anagen / telogen ratio), follicular unit ratio, and color. Digital image analysis of the photographs will be used to calculate the target area hair count (TAHC), the target area hair width (TAHW), and hair darkness.

[0181] A modified Norwood-Hamilton classification will be used to classify the stages of male androgenetic alopecia (Figure) and will be used to determine eligibility.

[0182] Statistical Procedures

[0183] Descriptive statistics will be reported for all endpoints. The descriptive statistics will vary depending on the type of variable. Generally, they will include continuous variables: descriptive statistics will include the number of observations, mean, median, standard deviation, minimum, maximum, and 95% confidence interval. Categorical variables will include descriptive statistics, which will include the number in the category, total number of observations, percentage of subjects in each category, and 95% confidence interval.

[0184] Visits and Related Procedures

[0185] Assessments will be performed by the same assessors whenever possible throughout the study.

[0186] Visit 1: Screening (Day -30 to Day -1)

[0187] The screening visit may be conducted at any time from 30 days to 1 day prior to the baseline visit. After informed consent has been obtained, the procedures performed include inclusion / exclusion criteria; demographics; Fitzpatrick dermatography type; medical / surgical history; history of androgenetic alopecia; modified Norwood-Hamilton classification; concomitant medications and comorbidities; vital signs; standardized radiography; and review of the subject's instructions for use.

[0188] Visit 2: Baseline and Injection (Day 0)

[0189] If removal from prohibited medications is not required, screening and Day 0 may be conducted on the same date. At any time during the visit, information regarding changes in the patient's health status, concomitant medications, and comorbidities will be collected. The sequence of events will include inclusion / exclusion criteria (confirmation of eligibility); vital signs; standardized radiography (after the above assessments).Completed); randomization; intradermal injection; local skin reaction (approximately 15 minutes before and after injection); adverse events.

[0190] Visit 3: Day 1

[0191] At any time during the visit, information about adverse events, concomitant drug treatment, and complications will be collected from the patient. See page 21 / 22 of the instruction manual, 24 CN 121057583 A. Vital signs will be measured before intradermal injection; and local skin reaction (approximately 15 minutes before and after injection) will be measured.

[0192] Visit 4: Day 2

[0193] At any time during the visit, information about adverse events, concomitant drug treatment, and complications will be collected from the patient. Vital signs will be measured before intradermal injection; and local skin reactivity (approximately 15 minutes before and after injection) will be measured.

[0194] Visit 5: Day 14 (±3 days)

[0195] At any time during the visit, information about adverse events, concomitant drug treatment, and complications will be collected from the patient. Vital signs were measured, followed by local skin reaction and standardized radiography.

[0196] Visit 6: Day 30 (±7 days)

[0197] At any time during the visit, information about adverse events, accompanying drug treatments, and concurrent procedures was collected from the patient. Vital signs were measured, followed by local skin reaction and standardized radiography.

[0198] Visit 7: Day 60 (±7 days)

[0199] At any time during the visit, information about adverse events, accompanying drug treatments, and concurrent procedures was collected from the patient.

[0200] Vital signs were measured, followed by local skin reaction and standardized radiography.

[0201] Visit 8: Day 90 (±7 days)

[0202] At any time during the visit, information about adverse events, accompanying drug treatments, and concurrent procedures was collected from the patient. Vital signs were measured, followed by local skin reaction and standardized radiography.

[0203] Visit 9: Day 120 (±7 days)

[0204] Information about adverse events, accompanying drug treatments, and concurrent procedures will be collected from the patient at any time during the visit.

[0205] Vital signs will be measured, followed by local skin reaction and standardized photography.

[0206] Visit 10: Day 150 (±7 days)

[0207] Information about adverse events, accompanying drug treatments, and concurrent procedures will be collected from the patient at any time during the visit.

[0208] Vital signs will be measured, followed by local skin reaction and standardized photography. The subject then withdraws from the study.

[0209] Those skilled in the art will understand that changes can be made to the exemplary embodiments shown and described above without departing from their broad inventive concept. Therefore, it should be understood that the invention is not limited to the exemplary embodiments shown and described.The invention is not intended to cover modifications within the spirit and scope of the invention as defined by the claims. For example, specific features of exemplary embodiments may or may not be part of the claimed invention, and various features of the disclosed embodiments may be combined. Unless specifically set forth herein, the terms “a,” “an,” and “the” are not limited to a single element, but are instead to be understood as meaning “at least one.”

[0210] Furthermore, the specific order of steps should not be construed as a limitation on the claims to the extent that the method of the invention does not depend on the specific order of steps set forth herein. Any claim relating to the method of the invention should not be limited to performing its steps in the written order, and it will be readily understood by those skilled in the art that these steps may vary and still remain within the spirit and scope of the invention. Specification 22 / 22 pages 25 CN 121057583 A Figure 1 Specification Drawing 1 / 1 page 26 CN 121057583 A

Claims

1. A method for treating androgenetic alopecia (AGA) in patients in need, the method comprising: A composition containing up to about 1% by weight of hyaluronic acid shall be applied to the patient’s skin at least twice a week. The hyaluronic acid mentioned therein is non-crosslinked; and The hyaluronic acid described therein has an average molecular weight of up to about 100 kDa; This allows for the treatment of AGA.

2. The method of claim 1, wherein the composition is administered to the patient on several consecutive days of the week.

3. The method of claim 1, wherein the composition is administered via intradermal injection.

4. The method of claim 3, wherein the composition is injected into the skin at a depth of about 0.1 mm or more.

5. The method of claim 3, wherein the composition is applied at or near the stem cell region of the hair follicle.

6. The method according to claim 1, wherein the method comprises approximately 4 cm 2 Or a larger injection area.

7. The method of claim 1, wherein the composition is applied to the skin comprising one or more of the frontotemporal region of the patient's scalp and the vertex region of the patient's scalp.

8. The method of claim 1, wherein the composition is used to treat and improve or increase one or more of the following: a) Target area hair count (TAHC); b) Target area hair width (TAHW); and c) Hair darkness.

9. The method of claim 1, wherein the patient's hair loss onset occurred approximately three to five years prior to the application of the composition.

10. The method of claim 1, wherein the patient's hair loss onset occurred approximately ten years prior to the application of the composition.

11. The method of claim 1, wherein the patient has previously undergone hair transplantation.

12. The method of claim 11, wherein the patient has undergone the hair transplant about one to two months prior to the application of the composition.

13. The method of claim 1, wherein the AGA is mild or moderate AGA.

14. The method of claim 1, wherein the patient has received a modified Norwood-Hamilton Scale score III or a modified Norwood-Hamilton Scale score IV.

15. The method of claim 8, wherein the improvement or increase is measured from about 10 days to about 160 days after application of the composition.

16. The method of claim 1, wherein the patient has previously experienced hair loss.

17. The method of claim 7, wherein the hyaluronic acid stimulates hair growth in the frontotemporal region and / or the vertex region of the patient's scalp.

18. A method for stimulating hair growth in the skin of a patient in need, the method comprising: A composition containing up to about 1% by weight of hyaluronic acid is applied to the patient's skin at least twice a week. The hyaluronic acid described therein is non-crosslinked; and The hyaluronic acid described therein has an average molecular weight of up to about 100 kDa. This stimulates hair growth in the patient's skin.

19. A method of administering a composition for hair growth to a patient in need: A composition containing up to about 1% by weight of hyaluronic acid shall be applied to the patient’s skin at least twice a week. The hyaluronic acid described therein is non-crosslinked; and The hyaluronic acid therein has an average molecular weight of up to about 100 kDa.

20. The method of claim 18 or claim 19, wherein the composition is administered to the patient on several consecutive days of the week.

21. The method of claim 18 or claim 19, wherein the composition is administered via intradermal injection.

22. The method of claim 21, wherein the composition is injected into the skin at a depth of about 0.1 mm or more.

23. The method of claim 21, wherein the composition is applied at or near the stem cell region of the hair follicle.

24. The method according to claim 18 or claim 19, wherein the method comprises approximately 4 cm 2 Or a larger injection area.

25. The method of claim 18 or claim 19, wherein the composition is applied to the skin comprising one or more of the frontotemporal region of the patient's scalp and the apical region of the patient's scalp.

26. The method according to claim 18 or claim 19, wherein the composition improves or increases one or more of the following: a) Target area hair count (TAHC); b) Target area hair width (TAHW); and c) Hair darkness.

27. The method of claim 18 or claim 19, wherein the patient's hair loss onset occurred approximately three to five years prior to the application of the composition.

28. The method of claim 18 or claim 19, wherein the patient's hair loss onset occurred approximately ten years prior to the application of the composition.

29. The method of claim 18 or claim 19, wherein the patient has previously undergone hair transplantation.

30. The method of claim 29, wherein the patient has undergone the hair transplant approximately one to two months prior to the application of the composition.

31. The method of claim 18 or claim 19, wherein the patient suffers from androgenetic alopecia (AGA).

32. The method of claim 31, wherein the AGA is mild to moderate AGA.

33. The method of claim 18 or claim 19, wherein the patient has received a modified Norwood-Hamilton Scale score III or a modified Norwood-Hamilton Scale score IV.

34. The method of claim 26, wherein the improvement or increase is measured from about 10 days to about 160 days after application of the composition.

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