Integrins (by binding to MEPE's RGD-like sequence) and cell adhesion receptors involved in osteoprogenitor attachment.

MEPE belongs to the family of secreted glycophosphoproteins known as SIBLING (Small Integrin-Binding Ligand, N-linked Glycoprotein). MEPE controls osteoblast activity and bone mass [1]. In just two to four hours, dentonin (3–30 μg/ml) dramatically boosts cell proliferation and exhibits improved cell adhesion promotion. On the other hand, there are no appreciable variations in the quantity of attached cells[2]. In comparison to all other treatments, dentonin (3-30 μg/ml; 2-24 hours) considerably increases cell areas and promotes osteoblast spreading[2].

---

Dentonin (3-30 microg/mL; 2-24 h) significantly increases cell numbers and enhances osteoblast spreading, exhibiting increased cell areas in adhesion assays.

In DPSCs (dental pulp stem cells), Dentonin promotes proliferation suggesting potential for pulp repair; it plays a role in phosphate homeostasis and bone metabolism regulation.

A typical protocol involves coating plates with Dentonin solution (3-30 microg/mL in PBS) overnight at 4degC, then blocking with BSA. Cells are seeded and incubated for 2-24 hours at 37degC. Adherent cells are fixed, stained with crystal violet, and counted under microscope. Non-adherent cells are removed by gentle washing.

Osteoprogenitor cells or DPSCs are seeded in Dentonin-coated plates at 2-5×10⁴ cells/well in alpha-MEM medium. After 2-24 hours, adherent cells are quantified via MTT assay or fluorescence staining. Alkaline phosphatase activity and matrix mineralization are assessed after 7-14 days of culture.

Animal studies typically involve local injection (0.5-2 mg/kg) or implantation of Dentonin-soaked scaffolds in bone defect models (e.g., femoral condyle or calvarial defect in rats). Bone healing is evaluated by micro-CT, histology, and biomechanical testing after 4-8 weeks. Dosing frequency is once daily or every other day for 2-4 weeks. For pulp repair, AC-100 is applied directly to exposed pulp in animal models.

Pharmacokinetic data specific to Dentonin are limited; as a peptide, it is expected to have a short plasma half-life (minutes to hours) and low oral bioavailability. It primarily undergoes proteolytic degradation.

In preclinical studies, Dentonin shows low toxicity; no significant adverse effects on mature osteoblasts or systemic toxicity at effective doses (3-30 microg/mL in vitro and mg/kg in vivo) have been reported. Standard toxicity studies for this research chemical are not publicly available.

[1]. Andrew P Sprowson, et al. ASARM-truncated MEPE and AC-100 enhance osteogenesis by promoting osteoprogenitor adhesion. J Orthop Res. 2008 Sep;26(9):1256-62.
[2]. N Six, et al. Dentonin, a MEPE fragment, initiates pulp-healing response to injury.J Dent Res. 2007 Aug;86(8):780-5.

AC-100 is a novel synthetic peptide derived from an endogenous human protein (a fragment of extracellular phosphoglycoprotein produced by bone and tooth cells). It was developed by Acologix.

---

Pharmacological Indications
Its investigation is underway for the treatment of periodontal disease.Mechanism of Action
In several preclinical models, AC-100 has demonstrated potent and selective activity against tooth tissue and bone formation, but its exact mechanism remains unclear. One study (PMID: 15153459) showed that AC-100 enhances the proliferation of dental pulp stem cells (DPSCs) by downregulating P16 and upregulating the ubiquitin ligase E3A and human ubiquitin-associated protein SUMO-1. Enhanced cell proliferation requires the intact RGD and SGDG motifs in the peptide. This study suggests that AC-100 promotes the proliferation of dental pulp stem cells (DPSCs) and has a potential role in pulp repair.

---

Pharmacodynamics
In animal models, AC-100 promotes cartilage regeneration. This study used a goat model of femoral condyle (knee articular cartilage) osteochondral defects to evaluate the effect of AC-100 on cartilage regeneration. Cylindrical defects were created in the knee articular cartilage and filled with collagen sponges containing two different doses of AC-100 or collagen sponges soaked in saline (control group). Postoperative treatment included intra-articular injection of the test drug at weeks 1, 2, and 3 postoperatively. Cartilage regeneration was assessed in one group at 84 and 168 days postoperatively. Results showed that AC-100 application promoted cartilage defect repair in a dose-dependent manner. Compared with the saline control group, the group receiving higher doses of AC-100 had higher healing scores. AC-100 demonstrated good safety in this study as no inflammatory response was observed.

---

Dentonin (also known as AC-100/TPX-100) is a 23-amino acid peptide (residues 242-264 of MEPE). Mechanism involves promoting cell adhesion and survival of osteoprogenitors, not affecting mature osteoblasts. It is in research stage for bone regeneration, osteoarthritis, and endodontic restoration. No FDA approval; primarily used in preclinical research. Molecular formula: C107H160N30O42; molecular weight: ~2538.59.

C107H160N30O42

2538.59112548828

2538.133

400090-20-2

Dentonin TFA

145722058

Typically exists as solid at room temperature

-18.1

38

45

84

179

5900

21

CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(=O)O)C(=O)NCC(=O)N[C@@H](CCC(=O)N)C(=O)N3CCC[C@H]3C(=O)N[C@@H](CC4=CC=CC=C4)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)O)NC(=O)CNC(=O)[C@H](CCCNC(=N)N)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C(C)O)N

PRFOXOFIVVOCCT-GRDAUCDVSA-N

InChI=1S/C107H160N30O42/c1-6-51(4)86(135-99(171)67(43-83(155)156)128-96(168)63(39-75(111)143)127-97(169)65(41-81(151)152)121-78(146)46-116-87(159)55(23-15-33-115-107(113)114)122-92(164)58(28-31-79(147)148)124-91(163)57(26-29-73(109)141)125-93(165)60(36-50(2)3)126-98(170)66(42-82(153)154)131-102(174)85(112)52(5)140)103(175)134-70(49-139)105(177)137-35-17-25-72(137)101(173)130-62(38-54-20-11-8-12-21-54)95(167)133-69(48-138)89(161)118-47-77(145)120-64(40-80(149)150)88(160)117-45-76(144)119-59(27-30-74(110)142)104(176)136-34-16-24-71(136)100(172)129-61(37-53-18-9-7-10-19-53)94(166)123-56(22-13-14-32-108)90(162)132-68(106(178)179)44-84(157)158/h7-12,18-21,50-52,55-72,85-86,138-140H,6,13-17,22-49,108,112H2,1-5H3,(H2,109,141)(H2,110,142)(H2,111,143)(H,116,159)(H,117,160)(H,118,161)(H,119,144)(H,120,145)(H,121,146)(H,122,164)(H,123,166)(H,124,163)(H,125,165)(H,126,170)(H,127,169)(H,128,168)(H,129,172)(H,130,173)(H,131,174)(H,132,162)(H,133,167)(H,134,175)(H,135,171)(H,147,148)(H,149,150)(H,151,152)(H,153,154)(H,155,156)(H,157,158)(H,178,179)(H4,113,114,115)/t51-,52?,55-,56-,57-,58-,59-,60-,61-,62-,63-,64-,65-,66-,67-,68-,69-,70-,71-,72-,85-,86-/m0/s1

(2S)-2-[[(2S)-6-amino-2-[[(2S)-2-[[(2S)-1-[(2S)-5-amino-2-[[2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2S)-1-[(2S)-2-[[(2S,3S)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-3-hydroxybutanoyl]amino]-3-carboxypropanoyl]amino]-4-methylpentanoyl]amino]-5-oxopentanoyl]amino]-4-carboxybutanoyl]amino]-5-carbamimidamidopentanoyl]amino]acetyl]amino]-3-carboxypropanoyl]amino]-4-oxobutanoyl]amino]-3-carboxypropanoyl]amino]-3-methylpentanoyl]amino]-3-hydroxypropanoyl]pyrrolidine-2-carbonyl]amino]-3-phenylpropanoyl]amino]-3-hydroxypropanoyl]amino]acetyl]amino]-3-carboxypropanoyl]amino]acetyl]amino]-5-oxopentanoyl]pyrrolidine-2-carbonyl]amino]-3-phenylpropanoyl]amino]hexanoyl]amino]butanedioic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.

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.

---

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).

View More

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)

---

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).

View More

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

---

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.

---

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