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In vitro inhibition of hyaluronidase by sodium copper chlorophyllin complex and chlorophyllin analogs.

McCook JP, Dorogi PL, Vasily DB, Cefalo DR - Clin Cosmet Investig Dermatol (2015)

Bottom Line: The oxidized form of copper isochlorin e4 disodium salt had substantial hyaluronidase inhibitory activity at 100 µg/mL but not at 10 µg/mL.Ascorbate derivatives did not enhance the hyaluronidase inhibitory activity of sodium copper chlorophyllin.Copper isochlorin e4 analogs were always the dominant components of the small molecule content of the commercial lots tested; oxidized copper isochlorin e4 was found in increased concentrations in older compared to newer lots tested.

View Article: PubMed Central - PubMed

Affiliation: Discovery Partners, LLC, Frisco, TX, USA.

ABSTRACT

Background: Inhibitors of hyaluronidase are potent agents that maintain hyaluronic acid homeostasis and may serve as anti-aging, anti-inflammatory, and anti-microbial agents. Sodium copper chlorophyllin complex is being used therapeutically as a component in anti-aging cosmeceuticals, and has been shown to have anti-hyaluronidase activity. In this study we evaluated various commercial lots of sodium copper chlorophyllin complex to identify the primary small molecule constituents, and to test various sodium copper chlorophyllin complexes and their small molecule analog compounds for hyaluronidase inhibitory activity in vitro. Ascorbate analogs were tested in combination with copper chlorophyllin complexes for potential additive or synergistic activity.

Materials and methods: For hyaluronidase activity assays, dilutions of test materials were evaluated for hydrolytic activity of hyaluronidase by precipitation of non-digested hyaluronate by measuring related turbidity at 595 nm. High-performance liquid chromatography and mass spectroscopy was used to analyze and identify the primary small molecule constituents in various old and new commercial lots of sodium copper chlorophyllin complex.

Results: The most active small molecule component of sodium copper chlorophyllin complex was disodium copper isochlorin e4, followed by oxidized disodium copper isochlorin e4. Sodium copper chlorophyllin complex and copper isochlorin e4 disodium salt had hyaluronidase inhibitory activity down to 10 µg/mL. The oxidized form of copper isochlorin e4 disodium salt had substantial hyaluronidase inhibitory activity at 100 µg/mL but not at 10 µg/mL. Ascorbate derivatives did not enhance the hyaluronidase inhibitory activity of sodium copper chlorophyllin. Copper isochlorin e4 analogs were always the dominant components of the small molecule content of the commercial lots tested; oxidized copper isochlorin e4 was found in increased concentrations in older compared to newer lots tested.

Conclusion: These results support the concept of using the hyaluronidase inhibitory activity of sodium copper chlorophyllin complex to increase the hyaluronic acid level of the dermal extracellular matrix for the improvement of the appearance of aging facial skin.

No MeSH data available.


Related in: MedlinePlus

HPLC chromatogram of sodium copper chlorophyllin, item number P5521 from Food Ingredient Solutions, lot number 200/98 (“old lot”).Notes: HPLC analysis was performed using YMC C30 column, (250×4.6 mm), 405 nm. The gradient program used was: 0–50% B over 30 minutes, 50–100% B over 10 minutes, 100% B for 5 minutes and 100–0% B over 5 minutes. Solvent A: methanol:water: acetic acid (90:10:0.5) and Solvent B: tertbutyl methyl ether:methanol:acetic acid (100:10:0.5) at a flow rate of 1.1 mL/min.Abbreviation: HPLC, high-performance liquid chromatography.
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f3-ccid-8-443: HPLC chromatogram of sodium copper chlorophyllin, item number P5521 from Food Ingredient Solutions, lot number 200/98 (“old lot”).Notes: HPLC analysis was performed using YMC C30 column, (250×4.6 mm), 405 nm. The gradient program used was: 0–50% B over 30 minutes, 50–100% B over 10 minutes, 100% B for 5 minutes and 100–0% B over 5 minutes. Solvent A: methanol:water: acetic acid (90:10:0.5) and Solvent B: tertbutyl methyl ether:methanol:acetic acid (100:10:0.5) at a flow rate of 1.1 mL/min.Abbreviation: HPLC, high-performance liquid chromatography.

Mentions: Analysis of old and new lots from two commercial suppliers, Food Ingredient Solutions and Spectrum Chemical, showed varying levels of copper isochlorin e4 and oxidized copper isochlorin e4, as shown in Table 4. “New” lots were analyzed within 1 year of production while “old” lots were 4–5 years post-production and had been stored at room temperature (59–86°F). In addition to the disodium salts of copper isochlorin e4 and oxidized copper isochlorin e4 analogs, smaller percentages of sodium salts of copper chlorin e6, copper chlorin p6, oxidized chlorin p6, oxidized chlorin e6, chlorin e4, and isochlorin e4 were identified in some of the lots. However, the disodium salt of copper isochlorin e4 was always the dominant component of the small molecule content. The disodium salt of oxidized copper isochlorin e4 appears to increase in concentration in the older lots as expected. A representative HPLC chromatogram of an “old” lot from Food Ingredient Solutions is presented in Figure 3.


In vitro inhibition of hyaluronidase by sodium copper chlorophyllin complex and chlorophyllin analogs.

McCook JP, Dorogi PL, Vasily DB, Cefalo DR - Clin Cosmet Investig Dermatol (2015)

HPLC chromatogram of sodium copper chlorophyllin, item number P5521 from Food Ingredient Solutions, lot number 200/98 (“old lot”).Notes: HPLC analysis was performed using YMC C30 column, (250×4.6 mm), 405 nm. The gradient program used was: 0–50% B over 30 minutes, 50–100% B over 10 minutes, 100% B for 5 minutes and 100–0% B over 5 minutes. Solvent A: methanol:water: acetic acid (90:10:0.5) and Solvent B: tertbutyl methyl ether:methanol:acetic acid (100:10:0.5) at a flow rate of 1.1 mL/min.Abbreviation: HPLC, high-performance liquid chromatography.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4536759&req=5

f3-ccid-8-443: HPLC chromatogram of sodium copper chlorophyllin, item number P5521 from Food Ingredient Solutions, lot number 200/98 (“old lot”).Notes: HPLC analysis was performed using YMC C30 column, (250×4.6 mm), 405 nm. The gradient program used was: 0–50% B over 30 minutes, 50–100% B over 10 minutes, 100% B for 5 minutes and 100–0% B over 5 minutes. Solvent A: methanol:water: acetic acid (90:10:0.5) and Solvent B: tertbutyl methyl ether:methanol:acetic acid (100:10:0.5) at a flow rate of 1.1 mL/min.Abbreviation: HPLC, high-performance liquid chromatography.
Mentions: Analysis of old and new lots from two commercial suppliers, Food Ingredient Solutions and Spectrum Chemical, showed varying levels of copper isochlorin e4 and oxidized copper isochlorin e4, as shown in Table 4. “New” lots were analyzed within 1 year of production while “old” lots were 4–5 years post-production and had been stored at room temperature (59–86°F). In addition to the disodium salts of copper isochlorin e4 and oxidized copper isochlorin e4 analogs, smaller percentages of sodium salts of copper chlorin e6, copper chlorin p6, oxidized chlorin p6, oxidized chlorin e6, chlorin e4, and isochlorin e4 were identified in some of the lots. However, the disodium salt of copper isochlorin e4 was always the dominant component of the small molecule content. The disodium salt of oxidized copper isochlorin e4 appears to increase in concentration in the older lots as expected. A representative HPLC chromatogram of an “old” lot from Food Ingredient Solutions is presented in Figure 3.

Bottom Line: The oxidized form of copper isochlorin e4 disodium salt had substantial hyaluronidase inhibitory activity at 100 µg/mL but not at 10 µg/mL.Ascorbate derivatives did not enhance the hyaluronidase inhibitory activity of sodium copper chlorophyllin.Copper isochlorin e4 analogs were always the dominant components of the small molecule content of the commercial lots tested; oxidized copper isochlorin e4 was found in increased concentrations in older compared to newer lots tested.

View Article: PubMed Central - PubMed

Affiliation: Discovery Partners, LLC, Frisco, TX, USA.

ABSTRACT

Background: Inhibitors of hyaluronidase are potent agents that maintain hyaluronic acid homeostasis and may serve as anti-aging, anti-inflammatory, and anti-microbial agents. Sodium copper chlorophyllin complex is being used therapeutically as a component in anti-aging cosmeceuticals, and has been shown to have anti-hyaluronidase activity. In this study we evaluated various commercial lots of sodium copper chlorophyllin complex to identify the primary small molecule constituents, and to test various sodium copper chlorophyllin complexes and their small molecule analog compounds for hyaluronidase inhibitory activity in vitro. Ascorbate analogs were tested in combination with copper chlorophyllin complexes for potential additive or synergistic activity.

Materials and methods: For hyaluronidase activity assays, dilutions of test materials were evaluated for hydrolytic activity of hyaluronidase by precipitation of non-digested hyaluronate by measuring related turbidity at 595 nm. High-performance liquid chromatography and mass spectroscopy was used to analyze and identify the primary small molecule constituents in various old and new commercial lots of sodium copper chlorophyllin complex.

Results: The most active small molecule component of sodium copper chlorophyllin complex was disodium copper isochlorin e4, followed by oxidized disodium copper isochlorin e4. Sodium copper chlorophyllin complex and copper isochlorin e4 disodium salt had hyaluronidase inhibitory activity down to 10 µg/mL. The oxidized form of copper isochlorin e4 disodium salt had substantial hyaluronidase inhibitory activity at 100 µg/mL but not at 10 µg/mL. Ascorbate derivatives did not enhance the hyaluronidase inhibitory activity of sodium copper chlorophyllin. Copper isochlorin e4 analogs were always the dominant components of the small molecule content of the commercial lots tested; oxidized copper isochlorin e4 was found in increased concentrations in older compared to newer lots tested.

Conclusion: These results support the concept of using the hyaluronidase inhibitory activity of sodium copper chlorophyllin complex to increase the hyaluronic acid level of the dermal extracellular matrix for the improvement of the appearance of aging facial skin.

No MeSH data available.


Related in: MedlinePlus