KR101433419B1 - Hydroxycinnamoyl-amino acidyl-hydroxamic acid derivatives - Google Patents

Abstract

The present invention relates to a hydroxy cinnamic acid-amino acid-hydroxyamine acid derivative. More particularly, the present invention relates to a hydroxy-cinnamic acid-amino acid-hydroxyamine acid derivative having an antioxidative effect and a whitening effect, and its use for skin whitening and anti-aging cosmetics.

Description

HYDROXYCINNAMOYL-AMINO ACIDYL-HYDROXAMIC ACID DERIVATIVES < / RTI >

The present invention relates to a hydroxy-cinnamic acid-amino acid-hydroxyamine acid derivative having an antioxidative effect. More particularly, the present invention relates to compounds of formula (I) having the following antioxidative and melanin synthesis inhibitory effects and their use in cosmetics.

(I)

In the above formula (I), R ¹ = R ² = OH and R ³ = H; R ¹ = OCH ₃ , R ² = OH and R ³ = H; Or R ¹ = R ³ = OCH ₃ and R ² = OH; And R ⁴ is phenylalanine proline.

Amino acids are basic constituent units of proteins, and have biological activity with various side chains besides amine groups and carboxy groups. The side chains of amino acids can be largely divided into hydrophobic side chains and hydrophilic side chains. Amino acids having a hydrocarbon chain or a ring are hydrophobic, and alanine (alanine), glycine, isoleucine, leucine, Methionine, phenylalanine, tryptophan, proline, valine, and the like.

Peptides are polymeric molecules of amino acids, and generally polymers of more than 100 amino acids are classified as proteins. The peptide bond is an amide (-CONH-) bond generated by a condensation reaction between a carboxy group (-COOH) and an amine group (-NH ₂ ). That is, a peptide refers to a compound in which amino acid, which is a basic unit constituting a protein, is linked by two or more amide bonds.

The peptide is usually a polymer having 2 to 50 amino acids bound thereto, and has a molecular weight of 100,000 or less. The number of amino acids 2, 3, 4, etc. constituting the peptide is referred to as a dipeptide, a tripeptide, or a tetrapeptide depending on the number of amino acids. About 10 or fewer amino acid polymers are referred to as oligopeptides, and more than one amino acid is linked by an amide bond is referred to as a polypeptide.

Hydroxycinnamic acid is known to have excellent antioxidative properties by virtue of its radical scavenging ability. It is abundant in greenish-yellow vegetables such as spinach, corn, cabbage, and tomatoes, coffee beans and olive oil.

Cinnamic acid is a metabolite of phenylalanine or tyrosine that is present in most plants. Among them, the hydroxamicamic acid breaks the OH bond and emits a hydrogen radical (H) to form a phenoxy radical. At this time, since the whole molecule has a resonance structure and can maintain a relatively stable form of radical, It plays a role of eliminating free radicals with high reactivity around them.

The mechanisms by which free radicals are produced vary widely. Most of the creatures living on Earth, whether plants or plants, require oxygen to sustain life. However, since more oxygen than necessary becomes active oxygen, it oxidizes and destroys the lipid layer constituting the cell membrane, oxidizes the coenzyme contained in the protein and loses its activity, or mutates by oxidizing DNA or RNA, Lt; / RTI > In addition, it promotes the melanin generation mechanism and may cause local skin pigmentation such as freckles and stains.

In other words, reactive radicals, including active oxygen, are the major cause of adult diseases such as cancer and Alzheimer's disease by destroying normal cellular metabolism, and are the main factors promoting aging and skin aging.

Recently, it has become an important technical task to develop materials that are biocompatible and have high antioxidant ability in accordance with expectation of an extension of human life and increase in socioeconomic demand for beauty. In the food industry, various antioxidants are used as additives to prevent food corruption. In the case of synthetic antioxidants, their use has been limited due to stability problems.

Therefore, the development of biocompatible antioxidant peptides is actively under way. It is obtained by hydrolyzing proteins present in nature using enzymes, or screening peptides having high antioxidative activity through peptide libraries.

The most widely used antioxidant peptides are carnosine and glutathione, each having the ability to trap free radicals and trap lipid radicals because they have histidine imidazole and cysteine sulfur atoms.

However, these antioxidant peptides have weak antioxidative ability at low concentration and antioxidative ability only at very high concentration.

What is important in the development of antioxidants is to develop substances that are highly biocompatible with high antioxidant capacity and stable even in long-term storage. That is, it effectively eliminates free radicals, inhibits the oxidation of fats, and is biocompatible so that there is no side effect to the skin.

In order to synthesize an antioxidant satisfying these criteria, the art has tried to chemically modify a substance known to have antioxidant ability to secure activity, biocompatibility and storage stability.

Hydroxycinnamic acid is a type of phenolic acid that acts as a carcinostatic agent and is known as a potent antioxidant in vivo and ex vivo.

Accordingly, various hydroxycinnamic acid derivatives have been developed and used. Representative examples thereof include a caffeic acid-mercaptotriazole derivative (Korean Patent No. 10-0849021) developed as an oxidation stabilizer for vitamin C, (Korean Patent No. 10-0668171), a cinnamic acid-guanidide complex (Korean Patent Application Publication No. 1997-0006281) which is an effective cardiovascular treatment agent, a cosmetic and dermatological composition having light stability Cinnamic acid-polyalkylene polyamine derivatives (Korean Patent No. 10-0295299), and the like.

However, when these derivatives are used alone, their use is limited due to the insufficient antioxidant capacity or the possibility of a safety risk.

The present inventors have found that by binding an amino acid to anhydroxycinnamic acid having an antioxidative ability and thereby enhancing the antioxidant ability and introducing a hydroxyamine acid capable of forming a metal complex at the C-terminal, an enzyme such as tyrosinase Thereby confirming the effect of inhibiting melanin synthesis. Thus, the present invention has been accomplished.

A basic object of the present invention is to provide a compound of the formula (I)

(I)

In the above formula (I), R ¹ = R ² = OH and R ³ = H; R ¹ = OCH ₃ , R ² = OH and R ³ = H; Or R ¹ = R ³ = OCH ₃ and R ² = OH; And R ⁴ is phenylalanine or proline.

A still further object of the present invention is to provide a cosmetic composition for skin whitening and anti-aging comprising the compound of formula (I).

The basic object of the present invention can be achieved by providing a compound of the formula (I)

(I)

In the above formula (I), R ¹ = R ² = OH and R ³ = H; R ¹ = OCH ₃ , R ² = OH and R ³ = H; Or R ¹ = R ³ = OCH ₃ and R ² = OH; And R ⁴ is phenylalanine or proline.
In the compound of the above formula (I), when R ⁴ is phenylalanine, it is a compound of the following formula (I-1), and when R ⁴ is proline, the compound has the following formula (I-2).

The compounds of formulas (I-1),

(I-2).

The compound of formula (I) of the present invention can be obtained by introducing a biocompatible and antioxidative moiety into a hydroxycinnamic acid known as a substance having an antioxidative effect, The inhibitory ability, biocompatibility and stability are improved.

The compound of formula (I) is a compound wherein an amino acid derivative is bonded to a compound of formula (II).

(II)

In the formula (II), when R ¹ = R ² = OH and R ³ = H, caffeic acid is obtained. When R ¹ = OCH ₃ , R ² = OH and R ³ = Ferulic acid, and when R ¹ = R ³ = OCH ₃ and R ² = OH, it is a sinapic acid.

Compounds of formula (I) of the present invention show improved free radical scavenging ability as compared to conventional radical scavengers as a result of DPPH (diphenylpicrylhydrazyl) radical scavenging test.

Also, as a result of lipid autoxidation test, the compound of the above formula (II) and the peptide are synergized to exhibit greatly improved fat oxidation inhibition ability.

Another object of the present invention described above can be attained by providing a cosmetic composition for skin whitening comprising a compound of the following formula (I) and a physiologically acceptable cosmetic base.

(I)

In the above formula (I), R ¹ = R ² = OH and R ³ = H; R ¹ = OCH ₃ , R ² = OH and R ³ = H; Or R ¹ = R ³ = OCH ₃ and R ² = OH; And R ⁴ is phenylalanine or proline.
In the compound of formula (I), when R ⁴ is phenylalanine, the compound of formula (I-1) and the compound of formula (I-2) when R ⁴ is proline.

Since the compound of formula (I) scavenges free radicals, it inhibits the action of active oxygen in the body and also reduces melanin production as the C-terminal hydroxyamine acid inhibits tyrosinase activity, have.

The skin whitening cosmetic composition of the present invention can be manufactured into a formulation such as a skin lotion, an emulsion, a cream, a pack, an essence and the like, and further, it can be used as an oil, a water, a surfactant, a humectant, a lower alcohol, a thickener, , Preservatives, fragrances, and the like.

In particular, considering that ultraviolet rays are a major factor in the formation of melanin, the skin whitening cosmetic composition of the present invention may include an ultraviolet screening agent, a light scattering agent, and the like.

Another object of the present invention described above can be attained by providing a cosmetic composition for preventing skin aging comprising a compound of the following formula (I) and a physiologically acceptable cosmetic base.

(I)

In the above formula (I), R ¹ = R ² = OH and R ³ = H; R ¹ = OCH ₃ , R ² = OH and R ³ = H; Or R ¹ = R ³ = OCH ₃ and R ² = OH; And R ⁴ is phenylalanine or proline.
In the compound of formula (I), when R ⁴ is phenylalanine, the compound of formula (I-1) and the compound of formula (I-2) when R ⁴ is proline.

Since the compound of formula (I) inhibits autoxidation of fat, it has an effect of preventing cell membrane destruction and preventing skin aging.

The cosmetic composition for preventing skin aging according to the present invention may be formulated into a form such as a skin lotion, an emulsion, a cream, a pack, a serum or the like, and may further contain at least one selected from the group consisting of oil, water, a surfactant, a humectant, a lower alcohol, , Preservatives, fragrances, and the like.

In particular, considering that ultraviolet rays are a major factor in melanin formation and wrinkle formation, the skin anti-aging cosmetic composition of the present invention may include a sunscreen agent, a light scattering agent, and the like.

According to the present invention, by introducing a biocompatible amino acid into a conventional antioxidant, hydroxycinnamic acid, the free radical scavenging ability is improved, the autoxidation inhibiting ability of the fat is enhanced through a synergistic action, and the hydroxy- A novel composition having antioxidative and whitening effects can be produced by effectively inhibiting the activity of cinacalcetin.

FIG. 1 shows the results of measuring the free radical scavenging activity of hydrocinamic acid and the hydrocinamic acid-amino acid-hydroxyamine acid derivative into which the amino acid-hydroxyamine acid of the present invention is introduced.
FIG. 2 shows the results of measuring the inhibition of autolipid oxidation of hydrocinamic acid and the hydrocinamic acid-amino acid -hydroxyamine acid derivative into which the amino acid-hydroxyamine acid of the present invention is introduced.
FIG. 3 shows the results of measuring the ability of hydrocinamic acid and the hydrocinamic acid-amino acid-hydroxyamine acid derivative to which the amino acid-hydroxyamine acid of the present invention is introduced to inhibit melanin formation.
FIG. 4 shows the results of measuring the inhibitory activity of tyrosinase enzyme activity of the hydrocinamic acid-amino acid-hydroxyamine acid derivative into which the amino acid-hydroxyamine acid of the present invention is introduced.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, it should be understood that the following examples are intended to illustrate specific embodiments of the present invention and are not intended to limit or limit the scope of the present invention.

Example  1. Solid-phase amino acid- Hydroxyamine acid  synthesis

Two equivalents of Fmoc-hydroxamic acid were coupled for 48 hours on the polystyrene resin to which the linker had been introduced by solid phase synthesis, and Fmoc-amino acid was then coupled It was reacted for 2 hours under methylpyrrolidone (NMP, N -methylpyrrolidone) - reagent BOP (benzotriazole-1-yloxy- tris (dimethylamino) -phosphoniumhexafluoro-phosphate), HOBt (1-Hydroxybenzotriazole) , and DIEA (Diisopropylethylamine) and N Peptide was synthesized. Fmoc was removed by treatment with 20% piperidine / NMP (piperidine / NMP) for 2 min each for 3 min and 7 min.

Example  2. Amino acid- Hydroxyamine acid  Introduced new Hydroxy Cinnamic acid -amino acid- Hydroxyamine acid  Preparation of derivative antioxidants

The C-terminal of Example 1 was prepared by introducing hydroxycinnamic acid in the same manner as Fmoc-amino acid on a polystyrene resin synthesized with an amino acid of hydroxamic acid, the reaction time being 5 hours, Hydroxy-amino acid derivatives were prepared. The desired material was then separated from the resin by treatment with 5% trifluoroacetic acid (TFA) / dichloromethane (DCM) and then precipitated with ether to give the hydroxycinnamic acid-amino acid-hydroxy Amino acid-hydroxamic acid (HCA-AA-NHOH) derivatives.

Example  3. Hydroxy Cinnamic acid -amino acid- Hydroxy  Identification of Amine Acid Derivatives

After confirming the purity of the product obtained in Example 2 using HPLC, it was confirmed to be a hydroxycinnamic acid-amino acid-hydroxyamine acid derivative using ESI-MS. The chemical formulas of the synthesized compounds are shown in Table 1. As a result of HPLC analysis, it was confirmed that this substance was a pure substance. Mass spectrometry of ESI-MS confirmed that it was a hydroxy cinnamic acid-amino acid-hydroxyamine acid derivative. The results in Table 1 below are higher than the original molecular weight because they are cationized.

Determination of the yield, purity and mass of the hydroxycinamic acid-peptide derivative Yield
(%) water
(%) ESI-MS Calculated value Actual value Caffeic acid-proline-hydroxamic acid 48 98 293 ([M + H] < ⁺ >) [ 293.0 Caffeic acid-phenylalanine-hydroxyamine acid 31 95 343 ([M + H] < ⁺ >) [ 343.1 Ferulic acid-proline-hydroxamic acid 43 99 307 ([M + H] < ⁺ >) [ 307.1 Ferulic acid-phenylalanine-hydroxyamine acid 37 90 357 ([M + H] < ⁺ >) [ 357.0 Synapic acid-proline-hydroxamic acid 45 91 337 ([M + H] < ⁺ >) [ 337.1 Cinnamic acid-phenylalanine-hydroxyamine acid 35 95 387 ([M + H] < ⁺ >) [ 387.1

Example  4. The Hydroxy Cinnamic acid -amino acid- Hydroxy  Free of amine acid derivatives Radical Scatters  Test

The substance obtained in Example 2 was added to a DPPH radical solution (0.1 M) dissolved in methanol, reacted at 25 ° C for 10 minutes, and the absorbance at 516 nm was measured. The free radical scavenging activity was measured by the following equation.

% DPPH radical scavenging ability = (A-B) / A x 100

Where A is the absorbance of the sample to which the material of Example 2 is not added and B is the absorbance of the sample to which the material of Example 2 is added. The free radical scavenging activity of each of the antioxidant peptides and hydroxycinnamic acid introduced by this equation was measured and compared. The results are shown in FIG. As can be seen from Fig. 1, the DPPH radical scavenging activity of caffeic acid-proline-hydroxyamine acid was greatly increased compared with caffeic acid, and other derivatives also maintained or slightly increased the radical scavenging activity of hydroxycinnamic acid.

Example  5. The Hydroxy Cinnamic acid -amino acid- Hydroxy  Lipoic oxidation of an amine acid derivative Inhibition  Test

Linolenic acid and Tween 20 were mixed in a phosphate buffer solution (0.1 M, pH 7.0) to prepare a 50 mM linoleic acid emulsion. The distilled water, the phosphate buffer solution and the emulsion were mixed, and the material obtained in Example 2 was added and reacted at 50 ° C for 10 hours. Then, the reaction solution was collected and washed with 75% ethanol containing FeCl ₂ and NH ₄ ⁺ SCN ^- Solution. Iron ions, which were present as Fe ^{2 +} , were oxidized to Fe ^{3 +} by the lipid peroxy radical (ROO) produced by the autoxidation of fat and formed a complex of SCN ^- and red color. The absorbance at 500 nm was measured, and the ability to inhibit lipid oxidation was measured by the following equation.

% Local autoxidation inhibition = (A-B) / A x 100

Where A is the absorbance of the sample to which the material of Example 2 is not added and B is the absorbance of the sample to which the material of Example 2 is added.

The results are shown in Fig. As can be seen in FIG. 2, both hydroxycinamic acid-amino acid-hydroxyamine acid derivatives showed improved lipid oxidation inhibition over hydroxycinnamic acid.

Example  6. The Hydroxy Cinnamic acid -amino acid- Hydroxyamine acid  Cytotoxicity Testing of Derivatives

HaCaT cells were cultured on a 96-well plate, and 1-100 uM of the hydroxycinamic acid-peptide derivative of the present invention was added thereto. After 12 hours, MTT treatment was performed to measure cell viability. MTT is formed by the intracellular mitochondria forming a formazan substance. It is known that MTT, originally yellow, is converted into a substance called formazan, which is an insoluble matter of violet through metabolism by mitochondria, , And the survival rate of adipocyte was measured by the following equation.

% Cell survival rate = (A-B) / A x 100

Where A is the absorbance of the sample to which the material of Example 2 is not added and B is the absorbance of the sample to which the material of Example 2 is added. Cell viability was measured and compared. No cytotoxicity was seen when treated up to 100 μM in both hydroxy-cinnamic acid-amino acid-hydroxyamine acid derivatives.

Example  7. The Hydroxy Cinnamic acid -amino acid- Hydroxyamine acid  Derivatives of tyrosinase Low performance  Measure

Hydroxycinamic acid-amino acid-hydroxyamine acid derivative dissolved in DMSO or distilled water was added to mushroom tyrosinase in 70 uL of phosphate buffer solution (0.1 M, pH 6.8), and 10 mM L-DOPA After incubation at 37 ° C for 10 minutes, the absorbance was measured at 475 nm, and tyrosinase enzyme activity was measured by the following equation.

% Tyrosinase activity = (A-B) / A x 100

Where A is the absorbance of the sample to which the material of Example 2 is not added and B is the absorbance of the sample to which the material of Example 2 is added.

As can be seen from FIG. 3, it can be seen that the hydroxycinamic acid-amino acid -hydroxyamine acid derivative effectively inhibits tyrosinase enzyme activity.

Example  8. The Hydroxy Cinnamic acid -amino acid- Hydroxyamine acid  Melanogenesis of derivatives Inhibition  Measure

Melanin inhibition assay was performed using rat melanoma cells (B16F1). As can be seen from FIG. 4, it can be confirmed that the hydroxycinamic acid-amino acid -hydroxyamine acid derivative effectively inhibits melanin production in melanocytes. In addition, it was confirmed that it is much more effective than arbutin, which is a commercially widely used whitening agent.

Claims (7)

A compound of the formula (I-1) or a compound of the formula (I-2)

The compounds of formulas (I-1),

The compound of formula (I-2)
In the above formula (I), R ¹ = R ² = OH and R ³ = H; R ¹ = OCH ₃ , R ² = OH and R ³ = H; Or R ¹ = R ³ = OCH ₃ and R ² = OH. WHAT IS CLAIMED IS: 1. A cosmetic composition for skin whitening comprising a compound of the formula (I-1) or a compound of the formula (I-2) shown below and a physiologically acceptable cosmetic base:

The compounds of formulas (I-1),

The compound of formula (I-2)
In the above formula (I), R ¹ = R ² = OH and R ³ = H; R ¹ = OCH ₃ , R ² = OH and R ³ = H; Or R ¹ = R ³ = OCH ₃ and R ² = OH. The skin whitening cosmetic composition according to claim 2, wherein the composition is a formulation selected from the group consisting of lotion, milk, cream, pack, and serum. The skin whitening cosmetic composition according to claim 2, further comprising an ultraviolet screening agent or a light scattering agent. A cosmetic composition for preventing skin aging comprising a compound of the following formula (I-1) or a compound of the following formula (I-2) and a physiologically acceptable cosmetic base:

The compounds of formulas (I-1),

The compound of formula (I-2)
In the above formula (I), R ¹ = R ² = OH and R ³ = H; R ¹ = OCH ₃ , R ² = OH and R ³ = H; Or R ¹ = R ³ = OCH ₃ and R ² = OH. 6. The cosmetic composition for preventing skin aging according to claim 5, wherein the composition is a formulation selected from the group consisting of lotion, milky lotion, cream, pack, and serum. 6. The cosmetic composition for preventing skin aging according to claim 5, further comprising an ultraviolet screening agent or a light scattering agent.

Images