| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
IC50: 0.57 μM (MMP-2), 0.25 μM (MMP-9)
Candida albicans (fungal cell membrane/mitochondria). Histatin 5 is an antimicrobial peptide that targets fungal cells. It binds to the fungal cell membrane (possibly to Ssa1/2 proteins on the cell surface), then is internalized and localizes to mitochondria. It disrupts mitochondrial function, leading to ROS production, ATP depletion, and cell death. It also chelates metal ions (Cu2+, Zn2+), which may contribute to its activity. Histatin 5 does not have a specific human receptor; it is used as a model peptide for antifungal research. The TFA salt does not affect activity. |
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| ln Vitro |
Histatin 5 belongs to a family of low-molecular-weight salivary proteins that are released by sublingual, parotid, and submandibular glands. Histatin 5 is reported to suppress the activity of the host matrix metalloproteinases MMP-2 and gland MMP-9 with IC50s of 0.57 and 0.25 μM, respectively, using biotinylated gelatin as a substrate. Three peptides with distinct Histatin 5 sections are created and tested as MMP-9 inhibitors in an effort to identify the domain causing this inhibition. The inhibitory activities of peptides containing residues 1 through 14 and 4 through 15 of Histatin 5 are significantly lower (IC50s, 21.4 and 20.5 μM, respectively), but peptide residues comprising residues 9 through 22 demonstrated the same activity as Histatin 5 against MMP-9. Based on the Arg-gingipain inhibition kinetics, Histatin 5 is a competitive inhibitor that only affects the Km at a Ki of 15 μM [1]. The mitochondrial respiration process is inhibited by histatin 5.Candida albicans cells absorb the human salivary antifungal peptide Histatin 5, which then forms an intracellular association with In a dose- and time-dependent manner, histatin 5 suppresses both the respiration of whole blastoconidia and the respiration of isolated C. albicans mitochondria. State 2 respiration is inhibited by histatin 5 at a dose of 33 uM[2].
In vitro, Histatin 5 TFA exhibits potent antifungal activity against Candida albicans and other Candida species (MIC values typically 1-10 uM). It also shows activity against some bacteria (e.g., Streptococcus mutans) and has wound-healing properties. In cell cultures, histatin 5 induces rapid cell death in C. albicans via ROS production and ATP depletion. It is not toxic to human cells at fungicidal concentrations (e.g., up to 50 uM). The peptide is also effective in biofilm models. It is used as a positive control in antifungal susceptibility tests. The TFA salt is water-soluble. It is being investigated as a potential therapeutic for oral candidiasis and other fungal infections. |
| ln Vivo |
In vivo, Histatin 5 TFA has been studied in mouse models of oral candidiasis. In a mouse model of oropharyngeal candidiasis, topical application or injection of histatin 5 (1-10 mg/kg) reduces fungal burden in the oral cavity and esophagus. It also accelerates wound healing in diabetic mice and reduces bacterial load in infected wounds. In a systemic candidiasis model, intravenous administration (10 mg/kg) prolongs survival and reduces kidney fungal load. The peptide is generally well-tolerated. It is not FDA-approved; it is a research compound. The TFA salt is used for formulation. For in vivo use, prepare in sterile PBS.
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| Enzyme Assay |
For antifungal susceptibility testing, use a broth microdilution assay according to CLSI guidelines. Prepare serial dilutions of Histatin 5 TFA (0.1-100 ug/mL) in RPMI-1640 buffered with MOPS. Inoculate with Candida albicans (ATCC 90028) at 0.5-2.5×10^3 CFU/mL. Incubate at 35degC for 24-48 h. Read MIC as the lowest concentration that inhibits visible growth. For killing kinetics, incubate C. albicans with histatin 5 (10 uM) at 37degC, take aliquots at 0, 1, 2, 4, 6 h, plate on Sabouraud agar, count CFU. For mechanistic assays, measure ROS production using DCFH-DA, or measure ATP depletion by luciferase assay. For hemolysis assay, incubate histatin 5 with human red blood cells (2% suspension) for 1 h at 37degC, measure hemoglobin release. Histatin 5 shows low hemolysis (<10% at 100 uM). The TFA salt is soluble; prepare stock in water.
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| Cell Assay |
For cellular assays, use C. albicans or mammalian cells. For C. albicans, treat with histatin 5 (0-100 uM) for 2-4 h, then stain with propidium iodide to assess membrane integrity, or with TUNEL for apoptosis. For mammalian cell cytotoxicity, treat human gingival fibroblasts or HeLa cells with histatin 5 (0-200 uM) for 24 h, measure viability by MTT. The peptide is not toxic up to 100 uM. For wound healing scratch assay, use a monolayer of keratinocytes, scratch, then add histatin 5 (1-10 uM) and monitor closure over 24-48 h; it promotes migration. All experiments in triplicate. Control: amphotericin B (antifungal positive control).
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| Animal Protocol |
For in vivo oral candidiasis model, use female ICR mice (5-6 weeks). Immunosuppress with prednisolone (125 mg/kg, s.c.) on day -1. On day 0, inoculate the oral cavity with C. albicans (2×10^7 CFU in 20 uL) under anesthesia. On days 1-5, treat with Histatin 5 TFA (5 mg/kg, in 20 uL PBS, topical oral application) or amphotericin B (0.5 mg/kg) as positive control. On day 6, sacrifice, swab the oral cavity, homogenize tongue and palate, and plate for CFU counts. For systemic candidiasis model, inoculate mice intravenously with C. albicans (5×10^5 CFU). Treat with histatin 5 (10 mg/kg, i.p., daily) or vehicle. Monitor survival for 14 days. All animal work requires IACUC approval. The TFA salt is safe for injection.
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| ADME/Pharmacokinetics |
As a peptide (MW ~3036), histatin 5 is rapidly cleared from the circulation (t1/2 ~10-30 min in mice) due to proteolysis and renal clearance. It is not orally bioavailable. For a PK study, administer 10 mg/kg i.v., collect blood at 0, 5, 15, 30, 60, 120 min, and measure by LC-MS/MS or ELISA. It distributes to kidneys, liver, and lungs. The TFA salt has no effect. For topical application, systemic absorption is minimal.
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| Toxicity/Toxicokinetics |
In vitro, histatin 5 is not cytotoxic to mammalian cells at concentrations up to 200 uM. In vivo, acute toxicity is low (LD50 >100 mg/kg i.v. in mice). No genotoxicity or carcinogenicity data. The TFA salt is non-toxic. Standard precautions.
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| References |
[1]. Gusman H, et al. Salivary histatin 5 is an inhibitor of both host and bacterial enzymes implicated in periodontaldisease. Infect Immun. 2001 Mar;69(3):1402-8.
[2]. Helmerhorst EJ, et al. The human salivary peptide histatin 5 exerts its antifungal activity through the formation ofreactive oxygen species. Proc Natl Acad Sci U S A. 2001 Dec 4;98(25):14637-42. |
| Additional Infomation |
Histatin 5 is a naturally occurring antimicrobial peptide in human saliva, contributing to oral immunity. It has potential as a topical antifungal agent for oral candidiasis, especially in immunocompromised patients. The TFA salt is used for research; the natural peptide is non-toxic. It is not FDA-approved. For research use only.
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| Molecular Formula |
C135H196N51F3O35
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| Molecular Weight |
3150.31
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| Related CAS # |
Histatin 5;115966-68-2
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| Appearance |
Typically exists as solid at room temperature
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O :~100 mg/mL (~31.74 mM)
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| Solubility (In Vivo) |
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 Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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)] Oral Formulations
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 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.3174 mL | 1.5871 mL | 3.1743 mL | |
| 5 mM | 0.0635 mL | 0.3174 mL | 0.6349 mL | |
| 10 mM | 0.0317 mL | 0.1587 mL | 0.3174 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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