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 combination showed efficacy in treating androgenetic alopecia (AGA), comparable to that of 3% minoxidil solution for mild to moderate AGA, observed in both male and female subjects. Efficacy was assessed through macroscopic expert panel evaluation (almost all subjects responded), microscopic evaluation (including hair count in the target area; terminal hair increase 8.3% vs. 8.68%, P=0.306; trichome index increase 13.8% vs. 31.5%, P=0.158), and patient self-report. Furthermore, the herbal extract combination demonstrated a good safety profile, with no eczema observed, compared to only one case in the minoxidil group. Currently, various molecules, products, and interventions, including herbal remedies and natural products, claim to promote hair growth in AGA patients. The hair regrowth efficacy of most such products stems from their mechanisms of inhibiting dihydrotestosterone (DHT) and anti-inflammation, as well as improving perifollicular vascularization and follicular nutrition; however, these mechanisms may not be clearly reported or may be caused by unknown mechanisms of action. Several randomized controlled trials have been conducted, but these trials are usually limited in sample size, short in duration (18 to 40 weeks), and have varied results. Most of these studies only use microscopic examination (such as hair counting) or only macroscopic assessment. In this study, the improvement in androgenetic alopecia (AGA) symptoms in patients receiving combination therapy with herbal extracts may be the result of multiple mechanisms working together, based on previously published research data, including inhibition of 5α-reductase, increasing hair anchoring and hair diameter by stimulating the production of extracellular matrix (ECM) in dermal papilla cells, and reducing micro-inflammation in the pathogenesis of male and female hair loss.

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Limitations[1]
Although this study was rigorously designed, covering all aspects of hair loss detection and employing a triple-blind randomized controlled trial with both macroscopic and microscopic assessments, there are still some limitations, including a small sample size and a short treatment period. Hair quality index is one of the methods for assessing hair growth, but it has not been widely accepted in hair research due to a lack of studies to validate its accuracy. Therefore, it is recommended to conduct a larger sample size and longer treatment duration randomized controlled trial, and to perform subgroup analysis based on gender and the combination of herbal extracts and minoxidil solution, to verify the efficacy and safety of this herbal extract combination as an alternative therapy for androgenetic alopecia.

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Conclusion[1]
Since there was no significant difference in clinical efficacy and safety between this herbal extract combination and 3% minoxidil solution, the herbal extract combination evaluated in this study may be a viable herbal medicine for the treatment of mild to moderate androgenetic alopecia in male and female patients. However, it is recommended to conduct a larger sample size and longer follow-up study 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.)