KR20220043455A - A composition for prevention or treatment of hair loss comprising quinazolinone deravitives - Google Patents

Abstract

The present invention relates to a composition for preventing, treating or alleviating hair loss comprising a quinazolinone derivative or a pharmaceutically acceptable salt thereof as an active ingredient. A compound of the present invention promotes hair growth by proliferating dermal papilla cells and converting the resting state of the hair growth cycle into an anagen phase.

Description

A composition for preventing or treating hair loss comprising a quinazolinone derivative {A COMPOSITION FOR PREVENTION OR TREATMENT OF HAIR LOSS COMPRISING QUINAZOLINONE DERAVITIVES}

The present invention relates to a composition for preventing, treating or improving hair loss comprising a quinazolinone derivative or a pharmaceutically acceptable salt thereof as an active ingredient. The compound of the present invention promotes hair growth by proliferating dermal papilla cells and converting the resting state of the hair growth cycle to the anagen phase.

Human hair is an aggregate of about 100,000 individual hairs, and hair is produced by hair follicles. The hair follicle serves as a repository of stem cells capable of generating all the cell lines needed to reconstruct the hair follicle itself, the epithelium and the sebaceous glands. Hair follicles are composed of dermal papilla cells (DPC), hair germinal matrix cells, outer root sheath cells (ORS), inner root sheath cells (IRS), and the like.

The dermal papilla cells are the core cells responsible for the development and growth of hair, and are located at the bottom of the hair root, which is the subcutaneous root of the hair. In fact, when dermal papilla cells are collected from the scalp and transplanted after culturing, new hair grows. However, there are several difficulties in using dermal papilla cells as a treatment for hair loss. First, it is difficult to separate from the scalp, the culture conditions are difficult, and it is difficult to culture a sufficient amount of cells. In addition, there is a limit in that the hair regeneration ability is significantly reduced when a lot of culture.

The outer root sheath cells are cells that help the growth and development of hair and protect the hair, and are in contact with the basal layer, the innermost layer of the epidermis. Together with the inner root sheath, it protects the hair generated in the follicle, the lower part of the hair follicle, until keratinization is completed, and serves to transport it to the epidermis. Outer root sheath cells are created by cell division near the hair bulb, transport hair to the epidermis and fall off from the scalp after fulfilling their role.

Hair goes through a three-stage cycle of anagen, catagen, and telogen, where it grows, maintains, and falls out. It is known that in the case of adults, hair grows on average about 0.3 mm per day during the growth period, which is the period of hair growth, grows by about 1 cm per month, and usually lasts for 3 to 7 years. In general, for 10 to 14 days after the growth phase, overall hair follicle cell apoptosis (apoptosis) occurs and hair follicles shrink during the catagen phase, which is a phase in which the hair follicles are reduced, and the telogen phase, which is a phase to prepare for the next growth phase with an average of 3 months. The hair falls out after passing through, and the reason the length of the hair is different depending on the part is that the duration of the growth phase, which is a unique feature of the hair follicle, is different for each part.

As such, human hair always maintains a constant number of hairs because it has a constant hair growth cycle. However, as hair loss progresses, the dermal papilla present at the root becomes smaller, and when the dermal papilla becomes smaller, the thickness of the hair becomes thinner and the hair growth cycle becomes shorter at the same time. Therefore, as hair loss progresses, the hair becomes very thin, and the hair growth cycle becomes shorter and falls out after a little growth.

It is known that factors such as genetic causes and the action of male hormones, as well as severe physical and mental stress such as endocrine disease, nutritional deficiency, drug use, childbirth, fever, and surgery, are known to act as the cause of hair loss. Recently, as well as male pattern hair loss, the female hair loss population is increasing due to the increase in stress caused by changes in diet, social environment, etc., and the age is also decreasing.

Currently, the most frequently used hair loss treatments in Korea include finasteride (trade name: Propecia®), dutasteride (trade name: Avodart®), and minoxidil (trade name: Minoxidil® or Rogaine®). Finasteride and dutasteride are 5α-reductase inhibitors that inhibit the conversion of testosterone to 5α-dihydrotestosterone (DHT). However, this product exhibits side effects such as decreased libido, erectile dysfunction, and loss of driving and performance. Meanwhile, in the case of minoxidil, the mechanism has not yet been fully elucidated, but it is a potassium channel opener that hyperpolarizes cell membranes, and through vasodilation and potassium channel opening, provides oxygen, blood, nutrients, etc. to the hair follicles. It is believed to keep hair follicles healthy by increasing their supply. However, this product also has side effects such as itching, erythema, skin irritation, eye irritation, etc. at the application site, and unwanted hair growth in other parts of the body other than the head is also observed. In addition, although hair transplantation has been attempted recently for severe hair loss patients, high cost and side effects after the procedure are pointed out as limitations.

An object of the present invention is to provide a composition capable of preventing, treating or improving hair loss by proliferating dermal papilla cells and inducing the growth phase of the hair growth cycle to promote hair thickness and length growth.

The present invention provides a composition for preventing, treating or improving hair loss comprising a compound of Formula I or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula I]

In the above formula,

R ₁ is hydrogen, oxygen, hydroxy, halo, alkyl, alkenyl, alkynyl, nitro, cyano, amino, or alkoxy;

R ₂ , R ₃ , R ₄ , and R ₅ are each independently hydrogen, hydroxy, halo, alkyl, alkenyl, alkynyl, nitro, cyano, amino, or alkoxy;

X is alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or cycloalkylalkyl;

Y is -C(=O)NH- or -C(=O)NH-(R _a )-C(=O)NH-,

Z is alkyl, alkenyl, alkynyl, or -CH(COOH)(R _b )-;

R _a and R _b are each independently alkyl, alkenyl, or alkynyl;

Ring A is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, optionally substituted with halo, alkyl or alkoxy, and

는 단일결합 또는 이중결합이다.

is a single bond or a double bond.

The composition may be a pharmaceutical composition, a cosmetic composition or a food composition.

In one embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof proliferates dermal papilla cells and converts the resting state of the hair growth cycle to the anagen phase so that the hair can grow thicker and longer to prevent, treat or can be used for improvement.

Since the composition of the present invention promotes the growth of thickness and length of hair by inducing proliferation of dermal papilla cells and entering the growth phase of the hair growth cycle, it can be usefully used for preventing, treating or improving hair loss due to various causes.

1 and 2 are graphs showing the results of iDPC proliferation according to the treatment with the compound of Formula II.
3 and 4 are graphs showing cytotoxicity in primary DPC and ORS cells according to the treatment concentration of the compound of Formula II.
5 to 10 are graphs showing the expression levels of Sox2, Versican (VCAN), CD133, Corin, β-catenin, and Wnt5a in iDPC according to the treatment with the compound of Formula II.
11 is a photograph of the mustache hair length of mice treated with the compound of Formula II.
12 is a graph showing the length of the mustache hair in mice after culturing according to the treatment with the compound of Formula II.
13 is a photograph of the length of pig hair follicles treated with the compound of Formula II.
14 is a graph showing the hair length of pig hair follicles after culturing according to treatment with the compound of Formula II.
15 is a photograph of the degree of hair regeneration according to the application concentration of the compound of Formula II.
16 is a graph showing the degree of hair regeneration according to the application concentration of the compound of Formula II.

Hereinafter, embodiments and examples of the present invention will be described in detail with reference to the accompanying drawings so that those of ordinary skill in the art to which the present invention pertains can easily carry out. However, the present application may be embodied in various forms and is not limited to the embodiments and examples described herein.

Throughout this specification, when a part "includes" a component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.

The present invention is characterized in that it was discovered for the first time that the compound of formula (I) prevents or treats hair loss or promotes hair growth. In one embodiment, the present invention provides a pharmaceutical composition for preventing or treating hair loss comprising a compound of Formula I or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula I]

In the above formula,

R ₁ is hydrogen, oxygen, hydroxy, halo, alkyl, alkenyl, alkynyl, nitro, cyano, amino, or alkoxy;

R ₂ , R ₃ , R ₄ , and R ₅ are each independently hydrogen, hydroxy, halo, alkyl, alkenyl, alkynyl, nitro, cyano, amino, or alkoxy;

X is alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or cycloalkylalkyl;

Y is -C(=O)NH- or -C(=O)NH-(R _a )-C(=O)NH-,

Z is alkyl, alkenyl, alkynyl, or -CH(COOH)(R _b )-;

R _a and R _b are each independently alkyl, alkenyl, or alkynyl;

Ring A is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, optionally substituted with halo, alkyl or alkoxy, and

는 단일결합 또는 이중결합이다.

is a single bond or a double bond.

In one embodiment,

R ₁ is hydrogen, oxygen, or alkyl;

R ₂ , R ₃ , R ₄ , and R ₅ are each independently hydrogen, halo, or alkoxy;

X is alkyl or cycloalkylalkyl,

Y is -C(=O)NH- or -C(=O)NH-(R _a )-C(=O)NH-,

Z is alkyl or -CH(COOH)(R _b )-,

R _a and R _b are alkyl,

Ring A is aryl or heteroaryl, optionally substituted with halo or alkoxy, and

는 단일결합 또는 이중결합일 수 있다.

may be a single bond or a double bond.

In another embodiment,

R ₁ is hydrogen, oxygen, or C ₁ -C ₆ alkyl,

R ₂ , R ₃ , R ₄ , and R ₅ are each independently hydrogen, halo, or C ₁ -C ₆ alkoxy;

X is C ₁ -C ₆ alkyl or C ₄ -C ₁₀ cycloalkylC ₁ -C ₆ alkyl,

Y is -C(=O)NH- or -C(=O)NH-(R _a )-C(=O)NH-,

Z is C ₁ -C ₆ alkyl or —CH(COOH)(R _b )—,

R _a and R _b are C ₁ -C ₆ alkyl,

Ring A is phenyl, pyridinyl, or indole, optionally substituted with halo or C ₁ -C ₆ alkoxy, and

는 단일결합 또는 이중결합일 수 있다.

may be a single bond or a double bond.

In another embodiment,

R ₁ is hydrogen, oxygen, or C ₁ -C ₆ alkyl,

R ₃ is hydrogen or C ₁ -C ₆ alkoxy,

R ₄ is hydrogen, halo, or C ₁ -C ₆ alkoxy;

R ₂ and R ₅ are hydrogen,

X is C ₁ -C ₆ alkyl or C ₄ -C ₁₀ cycloalkylC ₁ -C ₆ alkyl,

Y is -C(=O)NH- or -C(=O)NH-(R _a )-C(=O)NH-,

Z is C ₁ -C ₆ alkyl or —CH(COOH)(R _b )—,

R _a and R _b are C ₁ -C ₆ alkyl,

Ring A is phenyl, pyridinyl, or indole, optionally substituted with halo or C ₁ -C ₆ alkoxy, and

는 단일결합 또는 이중결합일 수 있다.

may be a single bond or a double bond.

In one embodiment, the compound may be a compound having a structure of Formula II below.

[Formula II]

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating hair loss comprising the compound of Formula II or a pharmaceutically acceptable salt thereof as an active ingredient. The chemical name of the compound of formula II is "N-[2-(5-chloro-1H-indol-3-yl)ethyl]-6-(4-oxo-3,4-dihydroquinazolin-3-yl)hexanamide".

In another embodiment, the compound of the present invention may be selected from the group consisting of the following compounds.

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As used herein, "pharmaceutically acceptable salt" refers to a salt that is pharmaceutically acceptable and retains the desired pharmacological activity of the parent compound. The salt is not particularly limited as long as it is pharmaceutically acceptable, and for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, maleic acid, succinic acid, methanesulfonic acid , benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, and the like can be used.

"Hair loss" of the present invention, regardless of the cause, refers to a state in which there is no hair in the site where hair should normally exist, alopecia areata, hereditary androgen hair loss, telogen hair loss, traumatic hair loss, hair growth wall, pressure hair loss, It may be anagen phase alopecia, alopecia pityis, syphilitic hair loss, seborrheic hair loss, symptomatic hair loss, scarring hair loss, or congenital hair loss, but is not limited thereto.

The compound of the present invention promotes the proliferation of dermal papilla cells and induces the growth phase of hair, thereby preventing or treating hair loss.

"Treatment" of the present invention refers to all methods of beneficially changing hair loss symptoms such as delaying or stopping hair loss by administration of the composition of the present invention, improving the symptoms of hair loss, or promoting hair growth and hair growth, such as lengthening or increasing the number of hair. means action.

The pharmaceutical composition of the present invention may be administered parenterally or orally according to a desired method, and the dosage may vary depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of disease, etc. Its scope is diverse. In addition, the therapeutically effective amount of the composition may vary depending on the method of administration, the target site, and the condition of the patient.

The present invention also provides a cosmetic composition for preventing or improving hair loss comprising the compound of Formula I or a pharmaceutically acceptable salt thereof as an active ingredient. Specifically, the cosmetic composition may be, for example, a cosmetic for hair, and the formulation is not particularly limited, and may be appropriately selected according to the purpose.

For example, the cosmetic composition is formulated as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray. may be, but is not limited thereto. More specifically, it may be a cleaning agent such as shampoo, conditioner, body cleanser, etc., hair dressing agent such as hair tonic, gel or mousse, and a cosmetic composition for hair such as a hair conditioner or hair dye.

In one embodiment of the present invention, the cosmetic composition may contain various suitable bases and additives as necessary, and the types and amounts of these components can be easily selected by the inventor. If necessary, it may contain acceptable additives, for example, may further include components such as preservatives, pigments, and additives conventional in the art.

In another embodiment, the present invention also provides a food composition for preventing or improving hair loss comprising a compound of Formula I or a pharmaceutically acceptable salt thereof as an active ingredient, for example, a health functional food. The "health functional food" means a food manufactured and processed using raw materials or ingredients useful for the human body, and "functionality" means a health purpose such as regulating nutrients or physiological action with respect to the structure and function of the human body. It means to consume for the purpose of obtaining useful effects.

Justice

Unless defined otherwise, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Moreover, numerical values recited herein are intended to include the meaning of “about” unless expressly stated otherwise. Definitions of residues and substituents used herein are provided below. Unless otherwise specified, each residue has the following definitions and is used in the sense commonly understood by one of ordinary skill in the art.

The term “alkyl,” as used herein, is a hydrocarbon having primary, secondary, tertiary and/or quaternary carbon atoms, substituted or unsubstituted, and saturated, which may be straight-chain, branched, cyclic, or combinations thereof. contains aliphatic groups. For example, an alkyl group has 1 to 20 carbon atoms (ie, C1-C20 alkyl), 1 to 10 carbon atoms (ie, C1-C10 alkyl), or 1 to 6 carbon atoms (ie, C1-C6 alkyl). ) can have Unless otherwise defined, in a preferred embodiment, alkyl refers to C1-C6 alkyl. Examples of suitable alkyl groups include methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, - CH(CH3)2), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, -CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH3)3), 1-pentyl (n-pentyl, - CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (- CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2 -pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3) )2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3- dimethyl-2-butyl (-CH(CH3)C(CH3)3), and octyl (-(CH2)7CH3). Moreover, the term “alkyl” as used throughout the specification, examples and claims is intended to include both unsubstituted and substituted alkyl groups, the latter of which include haloalkyl groups such as trifluoromethyl, and the like, hydrocarbons, refers to an alkyl moiety having a substituent replacing a hydrogen on one or more carbons of the backbone.

As used herein, the term "Cx-y" or "Cx-Cy", when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, haloalkyl, cycloalkyl, alkenyl, alkynyl or alkoxy, is within a chain is meant to include groups containing from x to y carbons. C0 alkyl means hydrogen when the substituent is at the terminal position, and a bond when it is internal. Also, for example, a C1-C6 alkyl group contains 1 to 6 carbon atoms in the chain.

The term "alkenyl," as used herein, has primary, secondary, tertiary and/or quaternary carbon atoms, and includes straight-chain, branched and cyclic, or combinations thereof, one or more regions of unsaturation; That is, it is a hydrocarbon having a carbon-carbon sp2 double bond. For example, an alkenyl group has 2 to 20 carbon atoms (i.e., C2-C20 alkenyl), 2 to 12 carbon atoms (i.e., C2-C12 alkenyl), 2 to 10 carbon atoms (i.e., C2-C20 alkenyl) C10 alkenyl), or 2 to 6 carbon atoms (ie, C2-C6 alkenyl). Examples of suitable alkenyl groups include, but are not limited to, vinyl (-CH=CH2), allyl (-CH2CH=CH2), cyclopentenyl (-C5H7), and 5-hexenyl (-CH2CH2CH2CH2CH=CH2). it is not

The term "alkynyl," as used herein, has primary, secondary, tertiary and/or quaternary carbon atoms, and includes straight-chain, branched and cyclic groups, or combinations thereof, and includes one or more carbon- It is a hydrocarbon with a carbon sp triple bond. For example, an alkynyl group has 2 to 20 carbon atoms (i.e., C2-C20 alkynyl), 2 to 12 carbon atoms (i.e., C2-C12 alkynyl), 2 to 10 carbon atoms (i.e., C2-C2- C10 alkynyl), or 2 to 6 carbon atoms (ie, C2-C6 alkynyl). Examples of suitable alkynyl groups include, but are not limited to, acetylenic (-C≡CH) and propargyl (-CH2C≡CH).

As used herein, the term "alkoxy" refers to an alkyl group attached to the parent compound through an oxygen atom, which may be represented by -O-alkyl, wherein the alkyl group is as defined herein and may be substituted or unsubstituted. . The alkyl group of the alkoxy group has, for example, 1 to 20 carbon atoms (ie C1-C20 alkoxy), 1 to 12 carbon atoms (ie C1-C12 alkoxy), 1 to 10 carbon atoms (ie C1-C20 alkoxy). C10 alkoxy), or 1 to 6 carbon atoms (ie, C1-C6 alkoxy). Examples of suitable alkoxy groups may be methoxy (-O-CH3 or -OMe), ethoxy (-OCH2CH3 or -OEt), and t-butoxy (-OC(CH3)3 or -O-tBu), etc. However, the present invention is not limited thereto.

The term “cycloalkyl,” as used herein, refers to a non-aromatic, saturated or unsaturated ring, wherein each atom of the ring is a carbon, substituted or unsubstituted monocyclic, bicyclic or polycyclic. Cycloalkyl may be a polycyclic cycloalkyl consisting of two or more rings in which one or more carbons are common to adjacent rings. A polycyclic cycloalkyl may be a fused ring system, a spirocyclic ring system or a bridged ring system, wherein at least one of the rings is cycloalkyl and the other ring is, for example, cycloalkyl, aryl, heteroaryl, as defined herein; and/or heterocycloalkyl. Examples of suitable cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

The term “cycloalkylalkyl,” as used herein, refers to an alkyl group in which at least one of the hydrogen atoms of the alkyl group as defined above has been replaced by a cycloalkyl.

As used herein, the term “heterocycloalkyl” refers to a substituted or unsubstituted monocyclic, bicyclic or polycyclic, non-aromatic, saturated or partially saturated ring containing one or more heteroatoms in the ring. Heterocycloalkyl may be a polycyclic heterocycloalkyl consisting of two or more rings in which one or more atoms are common to adjacent rings. A polycyclic heterocycloalkyl may be a fused ring system, a spirocyclic ring system or a bridged ring system, wherein at least one ring is heterocycloalkyl and the other ring is, for example, cycloalkyl, aryl, heterocycle as defined herein. aryl, and/or heterocycloalkyl. Examples of suitable heterocycloalkyls include piperidinyl, piperazinyl, pyrrolidinyl, morpholinyl, lactonyl, lactamyl, azetidinyl, dihydropyridinyl, dihydroindolyl, tetrahydropyridinyl (piperidinyl nyl), tetrahydrothiophenyl, sulfur-oxidized tetrahydrothiophenyl, indolenyl, 4-piperidinyl, 2-pyrrolidonyl, tetrahydrofuranyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl , decahydroquinolinyl, octahydroisoquinolinyl, 6H-1,2,5-thiadiazinyl, 2H,6H-1,5,2-dithiazinyl, pyranyl, chromenyl, xanthenyl, Phenoxatinyl, 2H-pyrrolyl, 3H-indolyl, 4H-quinolizinyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, 4aH-carbazolyl, Carbazolyl, β-Carbolinyl, Phenanthridinyl, Acridinyl, Phenanthrolinyl, Phenazinyl, Phenothiazinyl, Furazanyl, Phenoxazinyl, Isochromanyl, Cromanyl, Imidazolidinyl, Pyrazolyl dinyl, pyrazolinyl, quinuclidinyl, and oxazolidinyl, and the like.

As used herein, the term “aryl” refers to an aromatic hydrocarbon group in which each atom of the ring is a carbon, substituted or unsubstituted monocyclic, bicyclic or polycyclic. The aryl ring may preferably be a 6- to 14-membered ring or a 6- to 10-membered ring, more preferably a 6-membered ring. Aryl can be polycyclic aryl having two or more rings in which two or more carbons are common to two adjacent rings. Examples of suitable aryl groups include, but are not limited to, phenyl, naphtharenyl, phenanthryl, anthryl, and anilinyl.

As used herein, the term “heteroaryl” refers to an aromatic group that is monocyclic, bicyclic or polycyclic, substituted or unsubstituted, containing one or more heteroatoms in the ring. Non-limiting examples of suitable heteroatoms that may be contained in the aromatic ring include N, O and S. Heteroaryl may be a polycyclic heteroaryl consisting of two or more rings in which one or more atoms are common to adjacent rings. Examples of suitable heteroaryl groups include benzofuranyl, benzothiophenyl, pyrrolo, furanyl, thiophenyl, imidazolyl, indolyl, isoindolyl, isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, qui nolinyl, isoquinolinyl, pyrazolyl, pyridinyl, pyrazinyl, pyridazinyl, and pyrimidinyl, and the like.

As used herein, the term “amino” refers to —NH2 in which a hydrogen atom is substituted or unsubstituted with a substituent such as alkyl, aryl, etc., wherein the substituent such as alkyl, aryl, etc. substituted with a hydrogen atom is as defined herein. and may be substituted or unsubstituted. Examples of suitable amino groups include, but are not limited to, -NH2, -N(CH3)2, -N(CH3)-CH2CH2-N(CH3)2, and the like.

As used herein, the terms “halo” and “halogen” both mean halogen and include chloro, fluoro, bromo, and iodo.

As used herein, the term “substituted,” eg, “substituted alkyl,” means that one or more hydrogen atoms of the alkyl are each independently replaced by a non-hydrogen substituent. A substituent may be any of the substituents described herein, for example, halogen, hydroxyl, alkyl, hydroxyalkyl, haloalkyl, cyanoalkyl, alkoxyalkyl, carbonyl (eg, carboxyl, alkoxycarbonyl, formyl, or acyl). ), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro , azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, aryl, or heteroaryl. . A substituted moiety on the hydrocarbon chain may optionally itself be substituted.

The present invention will be described in more detail through the following examples, but the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[Example 1] Measurement of proliferation of dermal papilla cells and superficial root sheath cells

cell culture

Immortalized dermal papilla cells (iDPC) in DMEM (Hyclone, Cat No. SH30243.01) containing 10% FBS (Hyclone, Cat No. SH30084.03) and 1% PS (Hyclone, Cat No. SV30010) at 37 ° C. , and cultured in humidified conditions containing 5% CO ₂ . And Follicle Dermal Papilla cell Growth media (Promocell, Heidelberg, Germany) cells were prepared by culturing in medium conditions.

In addition, with prior consent, human hair follicles were extracted and outer root sheath cells (ORS cells) were isolated (Yonsei University College of Medicine, 2018-0141). ORS cells were cultured in a medium condition of EpiLife medium (Gibco) containing 1% antibiotic (penicillin/streptomycin) and EpiLife™ Defined Growth Supplement (EDGS; Gibco). All cells were each cultured in a 5% carbon dioxide incubator at 37°C.

To check the degree of cell proliferation, the immortalized hDPC and primary hDPC passaged the day before the test were dispensed in a 96-well plate at a concentration of 2Х10 ³ cells/well, and cultured in DMEM medium and Follicle Dermal Papilla cell growth media for 24 hours, respectively. In addition, 3D iDPC spheroids were passaged with 2D monolayer cells and dispensed in an ultra low attachment 96-well round bottom plate at a concentration of 2x10 ³ cells/well to form 3D spheroids after 24 hours. ORS cells were aliquoted in a 12-well plate at a concentration of 1x10 ⁵ cells/well and cultured for 24 hours.

Preparation and application of test and control substances

Compound of formula II (N-[2-(5-chloro-1H-indol-3-yl)ethyl]-6-(4-oxo-3,4-dihydroquinazolin-3-yl)hexanamide, 10 mM in DMSO, The final concentration of OTAVA Chemicals) was used by diluting a DMSO stock solution with a culture medium, and a negative (solvent) control used in the test method to measure the change in cell proliferation according to the concentration of the compound of Formula II was prepared with DMSO and the test substance The same dilution process as the concentration was carried out.

iDPC was treated with the compound of Formula II at concentrations of 1 and 10 uM and cultured for 48 hours (FIG. 1), and iDPC 3D spheroids and 2D monolayer cells were treated with a single concentration of 10 uM and cultured for 24 hours (FIG. 2). In addition, primary DPC was treated according to the concentration (0.001 ~ 100 uM) and then cultured for 48 hours (Fig. 3). In the case of ORS cells, cell proliferation occurs slowly unlike DPC, so 1x10 cells in a collagen-coated 12-well plate After aliquoting at a concentration of ⁵ cells/well, after 24 hours, the cells were treated with concentrations of 1 and 10 uM and cultured for 48 hours (FIG. 4).

CCK-8 absorbance measurement and cell count confirmation

After each culture condition, for CCK-8 absorbance measurement, 20 μL of CCK8 reagent was added to each well of a 96-well plate, incubated at 37°C for 1-2 hours, and then 450 using an ELISA reader. The degree of cell proliferation was confirmed by measuring the absorbance at nm. In addition, in the case of ORS cells, the number of cells in each well was directly counted instead of CCK-8 absorbance measurement. The results were compared with the negative control group (100%), and the values of the experimental group treated with the compound of Formula II were expressed relatively. In each cell group, the value of the negative (solvent) control group was set to “100%”, and the values of the experimental group were calculated relative to each (Cell viability, percentage %), and the results were summarized in a graph.

As a result, as shown in FIGS. 1 and 2, in iDPC, when the compound of Formula II was treated at a concentration of 10 uM, proliferation was promoted in both dermal papilla cells 2D and 3D spheroid, and as shown in FIGS. 3 and 4, primary DPC and In ORS cells, it was confirmed that the compound of Formula II was not toxic to cells despite treatment with different concentrations.

[Example 2] Changes in gene expression in dermal papilla cells

The iDPC was aliquoted into 3x10 ⁵ cells in a 60 mm dish, and after 24 hours, DMSO (the same dose as the compound of Formula II) and the compound of Formula II were treated at a concentration of 10 uM for 48 hours with TRIzol reagent (Invitrogen, Grand Island, NY, USA) was used to isolate total RNA. 1st cDNA was synthesized using isolated RNA, oligo(dT), HelixCriptTM Thermo Reverse Transcriptase system (NanoHelix, Madison, WI, USA), and quantitative RT-PCR was performed with SYBRGreen qPCR master mix (Takara, Shiga, Japan) and DP Changes in Sox2, Versican (VCAN), CD133, Corin, β-catenin, and Wnt5a mRNA known as makers were measured.

As a result, as shown in FIGS. 5 to 10, the expression of representative genes Sox2, Versican (VCAN), CD133, Corin, β-catenin, and Wnt5a of dermal papilla cells by the compound of Formula II in iDPC was significantly higher than in the negative control group. increase was confirmed.

[Example 3] Measurement of the growth effect of mouse mustache and pig hair follicles

Isolation and Culture of Hair Follicles from Mouse Mustache and Pig Hair

In order to measure the hair growth efficacy of the compound of Formula II, a 5-week-old mouse C3H/HeJ male was purchased from Orient Bio and euthanized using CO ₂ , and then the left and right upper labia (mystacial pads) of the mouse were separated. 1% penicillin / streptomycin (Gibco Inc, NY, USA) containing William E medium (William E medium, Gibco Inc, NY, USA) was put. Pig hair follicles aged 90-125 days old were observed under a dissecting microscope from the skin of pigs purchased from Apure Co., Ltd. and carefully separated together with vibrissa follicles of mice. The separated hair follicles were stored in a petri dish containing William E medium until now. In each well of a 48-well plate, 2 mM L-glutamine (L-glutamine, Gibco Inc, NY, USA), 10 μg/ml insulin (insulin, Sigma MO, USA), 50 nM hydrocortisone (hydrocortisone, Sigma MO, USA) and 250 μl of William E medium (William E medium, Gibco Inc, NY, USA) containing 1% penicillin/streptomycin, put one hair follicle into one well, 37 ° C, 5% CO ₂ incubator cultured in Eight hair follicles were used in one experimental group, and mouse mustache hair follicles were cultured for 2 days, and pig hair follicles were exchanged on the 2nd day during 5 days of culture. The compound of Formula II was treated at a concentration of 10 uM, and the negative (solvent) control (DMSO) was treated with the same amount as the volume added to the concentration of 10 μM of the drug in William E medium.

Measurement of growth of mouse mustache and pig hair follicles

The morphology of hair follicles in culture was photographed using a microscope. Using the Image J program, mouse mustache and pig hair lengths were measured on days 0, 2, and 5, respectively, and the change in hair follicle length was obtained for each treatment period of the compound of Formula II, and the growth rate was compared with the average length of the control group. was measured.

As a result, the hair growth efficacy of the compound of Formula II was confirmed by measuring the length of the hair follicles while culturing for 2 to 5 days after separating the mustaches of 5-week-old mice and 90-120-day-old pig hair follicles.

11 and 12, when the compound of Formula II was treated at a concentration of 10 μM in order to compare the length difference (%) of hair follicles between the group treated with the compound of Formula II and the control group, a negative control (“100%”) ), it was confirmed that the mouse mustache increased on the 2nd day. Also, as shown in FIGS. 13 and 14 , the growth effect of the hair shaft was significantly increased on both the 2nd and 5th days of pig hair follicles.

[Example 4] Confirmation of hair growth promoting effect in experimental animals

Since it was confirmed that the compound of Formula II promotes the proliferation of dermal papilla cells and the expression of representative genes, whether the hair cycle can be induced from the resting phase to the anagen phase when applied topically to the back of mice, thereby exhibiting an actual hair growth promoting effect Confirmed.

Specifically, in order to confirm the hair growth promoting effect of topical treatment of the compound of Formula II, 7-week-old male C3H/HeNCr1jOri mice were purchased and treated with a negative control group (DMSO) and a compound of Formula II (0.01 - 1% concentration) divided and used in the experiment. When the mouse reached the age of 7 weeks, in order to check the effect of hair growth in the mouse, the hair of the mouse was first removed using a mouse clipper. The compound of Formula II (0.01 - 1% concentration) was applied daily to the back of the mice from which the hair was removed for a total of 10 days. After 14 days, the hairs on the back were removed with a razor, and the weight of the collected hairs was measured and compared with the hair weight of the negative control group.

As a result, as shown in FIGS. 15 and 16, when the compound of Formula II was applied, the hair cycle was induced to the anagen phase by the compound of Formula II, and when visually confirmed, the hair grew significantly compared to the negative control, and the When the weight was measured, it was confirmed that the weight of the hair increased by about 1-3 times or more according to the concentration of the treated compound of Formula II compared to the negative control.

Claims (10)

A pharmaceutical composition for preventing or treating hair loss comprising a compound of Formula I or a pharmaceutically acceptable salt thereof:
[Formula I]

In the above formula,
R ₁ is hydrogen, oxygen, hydroxy, halo, alkyl, alkenyl, alkynyl, nitro, cyano, amino, or alkoxy;
R ₂ , R ₃ , R ₄ , and R ₅ are each independently hydrogen, hydroxy, halo, alkyl, alkenyl, alkynyl, nitro, cyano, amino, or alkoxy;
X is alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or cycloalkylalkyl;
Y is -C(=O)NH- or -C(=O)NH-(R _a )-C(=O)NH-,
Z is alkyl, alkenyl, alkynyl, or -CH(COOH)(R _b )-;
R _a and R _b are each independently alkyl, alkenyl, or alkynyl;
Ring A is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, optionally substituted with halo, alkyl or alkoxy, and

is a single bond or a double bond.
According to claim 1,
R ₁ is hydrogen, oxygen, or alkyl;
R ₂ , R ₃ , R ₄ , and R ₅ are each independently hydrogen, halo, or alkoxy;
X is alkyl or cycloalkylalkyl,
Y is -C(=O)NH- or -C(=O)NH-(R _a )-C(=O)NH-,
Z is alkyl or -CH(COOH)(R _b )-,
R _a and R _b are alkyl,
Ring A is aryl or heteroaryl, optionally substituted with halo or alkoxy, and

A pharmaceutical composition for preventing or treating hair loss, characterized in that it is a single bond or a double bond.
According to claim 1,
R ₁ is hydrogen, oxygen, or C ₁ -C ₆ alkyl,
R ₂ , R ₃ , R ₄ , and R ₅ are each independently hydrogen, halo, or C ₁ -C ₆ alkoxy;
X is C ₁ -C ₆ alkyl or C ₄ -C ₁₀ cycloalkylC ₁ -C ₆ alkyl,
Y is -C(=O)NH- or -C(=O)NH-(R _a )-C(=O)NH-,
Z is C ₁ -C ₆ alkyl or —CH(COOH)(R _b )—,
R _a and R _b are C ₁ -C ₆ alkyl,
Ring A is phenyl, pyridinyl, or indole, optionally substituted with halo or C ₁ -C ₆ alkoxy, and

A pharmaceutical composition for preventing or treating hair loss, characterized in that it is a single bond or a double bond.
According to claim 1,
R ₁ is hydrogen, oxygen, or C ₁ -C ₆ alkyl,
R ₃ is hydrogen or C ₁ -C ₆ alkoxy,
R ₄ is hydrogen, halo, or C ₁ -C ₆ alkoxy;
R ₂ and R ₅ are hydrogen,
X is C ₁ -C ₆ alkyl or C ₄ -C ₁₀ cycloalkylC ₁ -C ₆ alkyl,
Y is -C(=O)NH- or -C(=O)NH-(R _a )-C(=O)NH-,
Z is C ₁ -C ₆ alkyl or —CH(COOH)(R _b )—,
R _a and R _b are C ₁ -C ₆ alkyl,
Ring A is phenyl, pyridinyl, or indole, optionally substituted with halo or C ₁ -C ₆ alkoxy, and

A pharmaceutical composition for preventing or treating hair loss, characterized in that it is a single bond or a double bond.
A pharmaceutical composition for preventing or treating hair loss comprising a compound selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof:

;

;

;

;

;

;

;

;

;

;

;

;

;

;

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;

;

;

;

;

;

;

;

;

;

;

;

;

;

;

; and

.
A pharmaceutical composition for preventing or treating hair loss comprising a compound of Formula II or a pharmaceutically acceptable salt thereof:
[Formula II]

The hair loss according to any one of claims 1 to 6, wherein the hair loss is alopecia areata, hereditary androgenic alopecia, telogen hair loss, traumatic hair loss, hair growth wall, pressure hair loss, anagen phase hair loss, alopecia areata, syphilitic hair loss, seborrheic hair loss , A pharmaceutical composition for preventing or treating hair loss, characterized in that it is selected from the group consisting of symptomatic hair loss, scarring hair loss and congenital hair loss.
The pharmaceutical composition for preventing or treating hair loss according to any one of claims 1 to 6, wherein the compound promotes proliferation of dermal papilla cells to induce hair growth phase.
A cosmetic composition for preventing or improving hair loss comprising a compound of Formula I or a pharmaceutically acceptable salt thereof:
[Formula I]

In the above formula,
R ₁ is hydrogen, oxygen, hydroxy, halo, alkyl, alkenyl, alkynyl, nitro, cyano, amino, or alkoxy;
R ₂ , R ₃ , R ₄ , and R ₅ are each independently hydrogen, hydroxy, halo, alkyl, alkenyl, alkynyl, nitro, cyano, amino, or alkoxy;
X is alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, or cycloalkylalkyl;
Y is -C(=O)NH- or -C(=O)NH-(R _a )-C(=O)NH-,
Z is alkyl, alkenyl, alkynyl, or -CH(COOH)(R _b )-;
R _a and R _b are each independently alkyl, alkenyl, or alkynyl;
Ring A is cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, optionally substituted with halo, alkyl or alkoxy, and

is a single bond or a double bond.
The method according to any one of claims 10 to 11, wherein the hair loss is alopecia areata, hereditary androgen hair loss, telogen hair loss, traumatic hair loss, hair growth wall, pressure hair loss, anagen hair loss, alopecia areata, syphilitic hair loss, seborrheic hair loss, symptomatic hair loss , A cosmetic composition for preventing or improving hair loss, characterized in that it is selected from the group consisting of scarring hair loss and congenital hair loss.

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