Biochanin A; Acetyl tetrapeptide-3

The combination of biochanin A, acetyl tetrapeptide-3, and ginseng extracts has been shown to stimulate dermal papilla ECM proteins by increasing hydroxyproline, Collagen Type 3, and laminin, yielding a significant improvement in hair follicle size and hair anchoring. Increased hair growth has been observed after the seven-day culture by biochanin A and acetyl tetrapeptide-3compared to minoxidil in part of the activation of hair growth. Observed in in-vitro studies, biochanin A and acetyl tetrapeptide-3 appear to exert an anti-inflammatory effect by decreasing pro-inflammatory cytokines [1].

OBJECTIVE: We sought to evaluate the efficacy and safety profile of an herbal extract combination comprising biochanin A, acetyl tetrapeptide-3, and ginseng extracts, and compare this to 3% minoxidil solution for the treatment of andogenetic alopecia (AGA). METHODS: A 24-week, triple-blinded, randomized controlled study was conducted in male and female subjects (N=32) with mild to moderate AGA. All were randomized to receive twice-daily, 1mL applications of the herbal extract combination or 3% minoxidil solution. Clinical efficacy from photographic assessment and adverse reactions were evaluated. RESULTS: There were thirty-two subjects (16 male, mean age 41.3±13.8 years), with AGA onset and duration of 35.5±13.6 and 6.5±5.1 years, respectively. The herbal extract combination demonstrated a comparable efficacy to 3% minoxidil solution. Expert panel photographic assessment observed a response to both treatments in most patients at 24 weeks, with no statistically significant difference in an increase of terminal hair counts (8.3% [P=0.009] and 8.7% [P=0.002] at 24 weeks in the herbal extract combinations and the 3% minoxidil solution groups, respectively). No local adverse reactions from the herbal extract combination were observed, but one subject developed scalp eczema after using the 3% minoxidil solution. CONCLUSION: The non-significant difference in clinical efficacy and safety to 3% minoxidil solution suggests that the herbal extract combination evaluated here could potentially be an alternative treatment with for AGA. Further studies with larger groups and longer follow-up periods are recommended to verify our results [1].

Study design. This study aimed to compare the efficacy of an herbal extract combination to 3% minoxidil solution without a placebo group. The number of subjects was calculated using the t-test analysis with alpha error = 0.05, power of the test = 80 percent, and intercept = 2.5, which yielded a sample size of 32 subjects. They were then randomized using predetermined computer-generated blocked randomization lists to apply the assigned 1mL of herbal extracts (n=17) or 3% minoxidil solution (n=15) twice daily at approximately 12-hour intervals (total daily dose of 2mL) to the vertex area of the scalp for 24 weeks. The two treatment medications were prepared in identically-appearing, pre-packaged, and pre-labeled bottles. Efficacy, safety, and patient adherence were evaluated at 12 weeks and the end of 24 weeks. Adherence was assessed from the daily use of the study drugs. All subjects were also asked to return the bottles to measure the residual volumes of the two medications. The herbal extract combinatio evaluated here comprises acetyl tetrapeptide-3, biochanin A (red clover extracts), Panax Ginseng extract, and Salvia officinalis oil. Other ingredients include water, alcohol, panthenol, PEG-7 glyceryl cocoate, inositol, fragrance, PEG-40, hydrogenated castor oil, menthol and DMDM hydantoin. [1]

[1]. An Herbal Extract Combination (Biochanin A, Acetyl tetrapeptide-3, and Ginseng Extracts) versus 3% Minoxidil Solution for the Treatment of Androgenetic Alopecia: A 24-week, Prospective, Randomized, Triple-blind, Controlled Trial. J Clin Aesthet Dermatol. 2020 Oct;13(10):32-37.

The herbal extract combinations showed the efficacy in AGA treatment with comparable efficacy to the 3% minoxidil solution for treatment of mild-to-moderate AGA in both male and female participants. This was evaluated by a macrographic expert panel of global assessment (almost all response), microscopic evaluation including target area hair count (increase of terminal hair 8.3% vs. 8.68%, P=0.306), HMI (increase of 13.8% vs. 31.5%, P=0.158), and patient self assessment. Also, the satisfactory safety profile of the herbal extract combination was observed with no eczematous reaction and only one participant in the minoxidil group. A wide range of molecules, products, and interventions claim to promote hair growth in patients with AGA, including as phytopharmaceutical agents and natural products. The hair growth properties in most of these products are from the mechanism of DHT-inhibitory and anti-inflammatory activities, with improved perifollicular vascularization and hair follicle nutrition, or not be precisely reported or likely caused by unknown mechanism of action. There have been several randomized controlled trials that are usually studied in the limited sample sizes and durations (ranged from 18 to 40 weeks) with diverse results. The studies have mostly been evaluated using only microscopic evaluation, such as hair count or only macroscopic assessment. The improvements in AGA in the patients who received the herbal extract combination in our study could be the result of multiple mechanisms based on previously published study data, including the inhibition of 5α-reductase, increase of hair anchoring, and hair diameter via the stimulation of extracellular matrix (ECM) production of dermal papilla, and decrease of micro-inflammation in the pathogenesis of both male and female pattern hair loss.

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LIMITATIONS [1]
Despite thoroughly designed to cover all aspects of hair loss testing conducted in this triple-blinded, randomized trial, evaluated by both global and microscopic assessments, our study had limitations, including a small sample size and a short period of treatment. Hair Mass Index is one of the hair-growth evaluation methods, but it is not a well-accepted measurement in hair research due to a lack of studies that validate its accuracy. Hence, a larger sample size and a longer duration of randomized trial, with subgroup analysis according to gender and combination treatments of herbal extracts and minoxidil solution is recommended to verify the efficacy and safety profile of the herbal extract combination as an alternative treatment for AGA.

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CONCLUSION [1]
Due to the non-significant difference in clinical efficacy and safety to the 3% minoxidil solution, the herbal extract combination evaluated here could be a viable phytopharmaceutical for the treatment of mild to moderate AGA in both male and female patients. However, a study with a larger sample size and a longer follow-up is recommended to verify our results.

C22H39N9O5

509.61

509.307

C, 51.85; H, 7.71; N, 24.74; O, 15.70

827306-88-7

11752568

Typically exists as solid at room temperature

1.3±0.1 g/cm3

1075.4±65.0 °C at 760 mmHg

604.2±34.3 °C

0.0±0.3 mmHg at 25°C

1.562

-4.15

8

8

18

36

737

3

[C@@H](NC(=O)CNC(=O)[C@@H](NC(=O)C)CCCCN)(C(=O)N[C@H](C(=O)N)CCCCN)CC1NC=NC=1

RRJOMESUBQAYOA-BZSNNMDCSA-N

InChI=1S/C22H39N9O5/c1-14(32)29-17(7-3-5-9-24)21(35)27-12-19(33)30-18(10-15-11-26-13-28-15)22(36)31-16(20(25)34)6-2-4-8-23/h11,13,16-18H,2-10,12,23-24H2,1H3,(H2,25,34)(H,26,28)(H,27,35)(H,29,32)(H,30,33)(H,31,36)/t16-,17-,18-/m0/s1

(2S)-2-[[(2S)-2-[[2-[[(2S)-2-acetamido-6-aminohexanoyl]amino]acetyl]amino]-3-(1H-imidazol-5-yl)propanoyl]amino]-6-aminohexanamide

Acetyl tetrapeptide-3; Acetyl Tetrapeptide-3; 827306-88-7; Folixyl; Kollaren 6; Acetyl tetrapeptide-1; Uxi hair growth peptide; Ac-lys-gly-his-lys-NH2; ; Uxi hair growth peptide; D1HW9N9QBX; Kollaren 6; Folixyl

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)