MEDICAL DISCLAIMER: Educational research guidelines only. Lyophilized peptides are investigational chemical compounds and are NOT approved for human consumption, diagnosis, or therapy. Consult a licensed physician before any research application.
Humanin Dosage Chart, Schedule & Reconstitution Protocol
Quickstart Highlights
Humanin (HN) is a 24-amino-acid mitochondrial-derived peptide encoded in the MT-RNR2 (16S rRNA) region of mitochondrial DNA, discovered in 2001 as a neuronal survival factor against Alzheimer's toxicity [1]. It is broadly cytoprotective and anti-apoptotic: it binds the pro-apoptotic protein BAX, interacts with IGFBP-3 [2], and signals through a gp130/CNTFR/WSX-1 receptor complex to activate STAT3 [5]. Circulating humanin falls with age and tracks inversely with the GH/IGF-1 axis [6], which underlies its study as a longevity and metabolic peptide. The most-used research tool is the analog HNG (S14G-humanin), about 1,000-fold more potent than the native peptide [1]. Critically, there is no validated human dose, no human pharmacokinetics, and no completed clinical trial; reported dosing is preclinical (roughly 2-4 mg/kg HNG by injection in rodents) [4][8]. The subcutaneous reconstitution figures here are an educational reference only, not medical advice.
Reconstitute: Add 2 mL bacteriostatic water → 5 mg/mL concentration.
Typical dose: No validated human dose; ~250-1000 mcg per injection educational reference (research only)
Easy measuring: At 5 mg/mL, 1 unit = 0.01 mL = 0.0500 mg (50 mcg) on a U-100 insulin syringe.
Storage: Lyophilized powder stored frozen at -20 °C, protected from light and moisture. After reconstitution, refrigerate at 2-8 °C and use within roughly 3-4 weeks; avoid repeated freeze-thaw cycles, which degrade the peptide.
Half-life: Native humanin is short-lived in circulation (minutes range, on the order of 30 minutes); HNG and HNGF6A analogs were engineered for greater stability and longer half-life [4].
Route: No approved human route. Preclinical research used intraperitoneal, intravenous, subcutaneous, intracerebroventricular, and intranasal delivery; the subcutaneous model here is an educational measurement convention only.
Status: Not approved by the FDA or EMA for any use. Sold for laboratory research only; not a supplement and not intended for human consumption.
About Humanin
Humanin is a 24-amino-acid mitochondrial-derived peptide (MDP) encoded within the MT-RNR2 (16S rRNA) region of mitochondrial DNA, first identified as a neuronal survival factor that protects cells from familial Alzheimer's disease mutations and amyloid-beta toxicity [1]. It works largely as an anti-apoptotic, cytoprotective signal, binding the pro-apoptotic protein BAX and interacting with IGFBP-3 [2]. This page targets the question of Humanin dosage and reconstitution for educational reference only.\n\nImportant route note: there is no approved human Humanin product, no established human pharmacokinetics, and no completed clinical trial. All published dosing is preclinical and parenteral — rodent studies used intraperitoneal, intravenous, subcutaneous, intracerebroventricular, and intranasal delivery, most often with the potent analog HNG (S14G-humanin) at roughly 2-4 mg/kg [4][8]. The subcutaneous reconstitution figures below are an educational measurement convention used across this site, not a clinically validated human protocol.\n\nThis guide models a 10 mg vial reconstituted with 2.0 mL of bacteriostatic water (5 mg/mL, or 50 mcg per U-100 insulin-syringe unit) so that an illustrative research reference of 250-1000 mcg maps cleanly onto a syringe: 250 mcg ≈ 5 units, 500 mcg ≈ 10 units, and 1000 mcg ≈ 20 units.\n\nFrequency: Once daily subcutaneously in this educational model. Humanin is not FDA- or EMA-approved and is presented here for educational purposes only, not as medical advice.
Quick Protocol Navigation
Reconstitution Instruction & Mixing Step-by-Step
Lyophilized powder must be reconstituted carefully. Agitating peptide chains can shear disulfide bonds and render the peptide biologically inert.
Draw 2.0 mL of bacteriostatic water into a sterile syringe.
Inject the water slowly down the inner glass wall of the 10 mg Humanin vial; do not spray the stream directly onto the lyophilized powder, and never shake the vial.
Gently swirl or roll the vial until the solution is completely clear; the result is a 5 mg/mL concentration (50 mcg per U-100 insulin-syringe unit).
Store the reconstituted vial refrigerated at 2-8 °C and draw the chosen number of units per dose (250 mcg ≈ 5 units, 500 mcg ≈ 10 units, 1000 mcg ≈ 20 units).
Educational note: Humanin has no approved human route — preclinical research used intraperitoneal, intravenous, intracerebroventricular, and intranasal delivery in animals, so these subcutaneous figures are a measurement reference only; for the educational model, swab the site, inject slowly into subcutaneous tissue, and wait a few seconds before withdrawing the needle.
Interactive Humanin Syringe Calculator
Currently visualizing the 10 mg vial reconstituted with 2 mL bacteriostatic water. Adjust the target dose to dynamically render syringe units.
Reconstitution Calculation: 10mg dry powder in 2mL water yields 5.00 mg/mL. To evaluate a 250mcg dose, pull to 5.0 units (5 syringe ticks).
U-100 Syringe Representation
Educational reference visual. Assumes standard U-100 insulin syringe where 1.0 mL volume = 100 units.
Titration & Dose Escalation Schedules
| Phase | Dose per injection | Units (per injection) |
|---|---|---|
| Educational reference — low (research only) | 250 mcg | 5 units (0.05 mL) |
| Educational reference — mid | 500 mcg | 10 units (0.10 mL) |
| Educational reference — upper | 1000 mcg (1 mg) | 20 units (0.20 mL) |
Administration guidelines: Refer to guidelines | 2 mL Reconstitution
Research Supplies Quantity Planner
Scientific mathematical planning of syringes, bacteriostatic water and dry vials needed for extended research blocks using the 10 mg vial.
Peptide Vials (Humanin, 10 mg each):
- check8-week educational course at ~500 mcg once daily (56 doses ≈ 28 mg): about 3 vials.
- check12-week course at ~500 mcg daily (84 doses ≈ 42 mg): about 5 vials.
- check16-week course at ~500 mcg daily (112 doses ≈ 56 mg): about 6 vials.
- checkAdd one spare vial if titrating toward the 1000 mcg upper reference, which doubles peptide use.
Insulin Syringes (U-100):
- checkOne sterile syringe per injection; daily dosing uses about 56 syringes over 8 weeks.
- checkAbout 84 syringes over 12 weeks.
- checkAbout 112 syringes over 16 weeks.
- checkUse 0.3 mL/31 G syringes; at 50 mcg per unit, 500 mcg = 10 units and 1000 mcg = 20 units.
Bacteriostatic Water (30 mL bottles): Use 2 mL per vial for reconstitution.
- checkA single 30 mL multi-dose bottle covers an entire 8-16 week course (3-6 vials x 2 mL = 6-12 mL).
- checkEach 10 mg vial reconstitutes to 5 mg/mL (50 mcg per U-100 unit).
- checkDiscard each reconstituted vial after about 3-4 weeks refrigerated, even if peptide remains.
Alcohol Swabs: clean the vial septum and injection site before every use.
- checkTwo swabs per injection (vial top + skin): about 112 over 8 weeks.
- checkAbout 168 over 12 weeks.
- checkAbout 224 over 16 weeks.
- checkKeep a surplus 100-200 count box on hand for repeated vial-top cleaning.
Mechanism of Action (MOA)
Humanin is a mitochondrial-derived peptide (MDP): a short polypeptide translated from an open reading frame located within the MT-RNR2 (16S ribosomal RNA) region of mitochondrial DNA. Depending on whether it is read by cytoplasmic or mitochondrial ribosomes, it exists as a 24-amino-acid (MAPRGFSCLLLLTSEIDLPVKRRA) or 21-amino-acid peptide, both biologically active [1][5]. It was discovered in 2001 from surviving neurons in the occipital cortex of an Alzheimer's patient, where it abolished cell death triggered by mutant APP, presenilin-1, presenilin-2, and amyloid-beta [1]. Substituting the serine at position 14 with glycine yields HNG (S14G-humanin), an analog roughly 1,000-fold more potent and more stable that is used in most in vivo studies [1][4].\n\nHumanin acts through several converging pathways. Intracellularly, it binds the pro-apoptotic proteins BAX, tBID, and BimEL, preventing BAX translocation and oligomerization at the outer mitochondrial membrane and thereby blocking cytochrome c release and the intrinsic apoptosis cascade [5]. Extracellularly, it binds IGFBP-3 [2] and engages a trimeric cell-surface receptor composed of gp130, CNTF receptor alpha, and WSX-1, driving JAK2/STAT3 signaling that upregulates pro-survival genes; it can also signal via the formyl-peptide receptors FPRL1/FPRL2 [5]. The net effect is protection against oxidative stress, serum starvation, hypoxia, and endoplasmic-reticulum stress in cell and animal models [5].\n\nSystemically, circulating humanin declines with age in rodents and humans and is inversely related to the growth hormone/IGF-1 axis: long-lived, GH-deficient Ames mice show elevated humanin, while GH or IGF-1 administration lowers it [6]. Centrally, intracerebroventricular humanin infusion improves hepatic insulin sensitivity through a hypothalamic STAT3-dependent mechanism [3], and humanin and related MDPs are described as age-dependent regulators of apoptosis, insulin sensitivity, and inflammatory markers [7].\n\nPharmacokinetics: native humanin has a short circulating half-life in the minutes range (on the order of 30 minutes) because of rapid proteolysis, which is why more stable, potent analogs were engineered. Pharmacokinetic work in male rodents showed that HNG and the non-IGFBP-3-binding analog HNGF6A display longer half-lives than native peptide, and that IGFBP-3 binding influences clearance [4]. Documented research routes include intraperitoneal, intravenous, subcutaneous, intracerebroventricular, and intranasal delivery (the latter reaching the brain via olfactory/trigeminal pathways); there is no oral bioavailability data, and the peptide would be degraded in the gut.\n\nHonest status: there is no approved human Humanin product, no established human pharmacokinetics, and no completed clinical trial. The subcutaneous reconstitution scheme on this page is an educational measurement convention, not a clinically validated delivery method, and nothing here is medical advice.
Clinical Trial Efficacy Highlights
- starHashimoto and colleagues (2001, PNAS) identified Humanin from neurons that survived in an Alzheimer's patient's cortex and showed that the cDNA encoded a short polypeptide which abolished neuronal death caused by a wide spectrum of familial Alzheimer's disease genes (mutant APP, PS1, PS2) and by amyloid-beta; the same work reported that the S14G (HNG) substitution increased neuroprotective potency roughly 1,000-fold [1].
- starIkonen and colleagues (2003, PNAS) used a yeast two-hybrid screen to identify IGFBP-3 as a high-affinity humanin binding partner, confirmed the interaction by pull-down, ligand-blot and in vivo co-immunoprecipitation, and demonstrated that humanin and IGFBP-3 regulate cell survival and apoptosis (humanin blocks IGFBP-3-induced apoptosis in glial lines while acting synergistically to protect neurons from amyloid-beta) [2].
- starMuzumdar and colleagues (2009, PLoS ONE) showed in Sprague-Dawley and Zucker diabetic fatty rats that continuous intracerebroventricular humanin infusion (about 0.16 µg/kg/min) and intravenous HNGF6A improved peripheral and hepatic insulin sensitivity during hyperinsulinemic-euglycemic clamps; hypothalamic STAT3 inhibition abolished the effect, localizing the action to a central nervous system pathway, and a single HNGF6A dose lowered blood glucose in diabetic rats [3].
- starTajima and colleagues (2005) reported that the analog S14G-HN, given by intracerebroventricular injection at doses as low as ~50 pmol, prevented amyloid-beta-induced impairment of spatial working memory (Y-maze) and latent learning (water-finding task) in mice, supporting a direct neuroprotective/pro-cognitive effect [9].
- starNiikura and colleagues (2011, PLoS ONE) found that intranasal S14G-HN administered for 3 months reduced amyloid-beta accumulation and ameliorated memory deficits in triple-transgenic (3xTg-AD) mice at the early plaque-bearing stage, demonstrating efficacy by a noninvasive brain-delivery route in a chronic model [8].
- starLee and colleagues (2014, Aging Cell) demonstrated that humanin levels decline with age in rodents and humans and are regulated by the somatotropic axis: long-lived GH/IGF-1-deficient models show high humanin, whereas GH or IGF-1 treatment lowers it, linking the peptide to longevity-associated endocrine signaling [6].
- starCobb and colleagues (2016, Aging) characterized humanin and related small humanin-like peptides (SHLPs) as naturally occurring, age-dependent regulators of apoptosis, insulin sensitivity, reactive oxygen species, and inflammatory markers, reinforcing humanin's role as a systemic metaboloprotective MDP [7].
- starYen, Lee, Mehta and Cohen (2013, Journal of Molecular Endocrinology) reviewed the preclinical evidence that humanin protects cells from oxidative stress, serum starvation, and hypoxia in vitro and improves outcomes in cardiovascular and Alzheimer's disease models in vivo, while emphasizing that human interventional data are lacking [5].
Side Effects & Tolerability Profile
Clinical subjects transiently report mild side effects. Slowly escalating the titration dose represents the single most effective intervention to limit side effects.
- warningThere is no human safety data for exogenous Humanin: the entire profile derives from cell-culture and animal studies, so tolerability, long-term effects, and a maximum tolerated dose in humans are unknown [5].
- warningBecause humanin is a potent, broadly acting anti-apoptotic and cytoprotective factor, there is a theoretical concern that it could promote the survival of damaged or malignant cells; humanin signaling (BAX inhibition, STAT3 activation) overlaps pathways implicated in tumor cell survival [5].
- warningMetabolic effects are a key consideration: humanin and its analogs lower blood glucose and improve insulin sensitivity in animals, creating a theoretical risk of hypoglycemia and additive effects if combined with insulin or other glucose-lowering agents [3].
- warningHumanin binds IGFBP-3 and intersects the GH/IGF-1 axis, so interactions with growth-factor signaling are plausible and unstudied in humans [2][6].
- warningFor the educational subcutaneous model, injection-site reactions (redness, irritation, bruising, or infection) are possible; sterile technique, fresh syringes, and proper storage are essential.
- warningSTAT3 and cytokine-receptor (gp130/CNTFR/WSX-1) signaling can influence inflammation and immune responses, an additional theoretical concern with chronic exposure [5].
- warningResearch-grade peptide is not produced to pharmaceutical (GMP) standards, so purity, correct sequence, sterility, and endotoxin content are not guaranteed and can pose their own risks.
- warningRegulatory/research status: Humanin is NOT approved by the FDA, EMA, or any major regulator for any indication, is not a dietary supplement, and is sold strictly for laboratory research; it is not intended for human use, and nothing here is medical advice.
Subcutaneous Injection Technique
Most research peptides require subcutaneous injection into fatty tissue. Never inject directly into a blood vessel or deep muscle tissue unless clinically detailed.
1. Site Selection
Common locations include the abdomen (2 inches from navel), outer upper arms, or thighs.
2. Sanitization
Thoroughly clean the selected site, stopper and vial top using 70% isopropyl alcohol prep swabs.
3. Angle & Push
Pinch the skin and insert the needle at a 45 to 90-degree angle. Depress plunger smoothly.
4. Site Rotation
Rotate injection sites continuously to avoid lipodystrophy or tissue scarring.
Frequently Asked Questions
What is the typical Humanin dosage?expand_more
There is no validated human Humanin dosage. No clinical trial has established a safe or effective dose, and no human pharmacokinetics exist. All published dosing is preclinical and parenteral: rodent studies most often used the potent analog HNG (S14G-humanin) at roughly 2-4 mg/kg by intraperitoneal or subcutaneous injection, with central infusion studies using about 0.16 µg/kg/min intracerebroventricularly. The 250-1000 mcg per-injection figures on this page are an educational reconstitution reference only, not a recommendation.
Is Humanin FDA approved?expand_more
No. Humanin is not approved by the FDA, the EMA, or any other major regulator for any indication. It is not a dietary supplement and is sold strictly for laboratory research. It has never completed a human interventional trial, so it should be regarded as an experimental research compound and not used as medicine.
How do you reconstitute Humanin?expand_more
In this educational model, draw 2.0 mL of bacteriostatic water and inject it slowly down the inner wall of a 10 mg vial, then swirl gently (do not shake) until clear. That yields a 5 mg/mL solution, or 50 mcg per U-100 insulin-syringe unit, so 250 mcg is about 5 units, 500 mcg about 10 units, and 1000 mcg about 20 units. Store the reconstituted vial refrigerated and use it within about 3-4 weeks.
What is Humanin's half life?expand_more
Native humanin has a short circulating half-life in the minutes range (on the order of 30 minutes) because it is rapidly broken down by proteases. The engineered analogs HNG (S14G-humanin) and the non-IGFBP-3-binding HNGF6A were shown in rodent pharmacokinetic studies to be more stable and to have longer half-lives than the native peptide, which is why analogs dominate preclinical research [4].
Can Humanin be stacked with other peptides?expand_more
In research settings humanin is often grouped conceptually with other mitochondrial-derived peptides such as MOTS-c and the small humanin-like peptides (SHLPs), but there are no human studies of any humanin-containing combination, no data on interactions, and no established safety. Because it lowers glucose and improves insulin sensitivity in animals, combining it with insulin or glucose-lowering agents is a particular theoretical concern. This is educational information, not advice to combine compounds.
Related Guides & Tools
Step-by-step references for reconstituting, measuring, and storing Humanin, plus the universal dosing calculator.
Academic References & Study Citations
Hashimoto Y, Niikura T, Tajima H, et al. A rescue factor abolishing neuronal cell death by a wide spectrum of familial Alzheimer's disease genes and Abeta. Proc Natl Acad Sci U S A. 2001;98(11):6336-6341. View Scientific Paper →
Ikonen M, Liu B, Hashimoto Y, et al. Interaction between the Alzheimer's survival peptide humanin and insulin-like growth factor-binding protein 3 regulates cell survival and apoptosis. Proc Natl Acad Sci U S A. 2003;100(22):13042-13047. View Scientific Paper →
Muzumdar RH, Huffman DM, Atzmon G, et al. Humanin: a novel central regulator of peripheral insulin action. PLoS One. 2009;4(7):e6334. View Scientific Paper →
Chin YP, Keni J, Wan J, et al. Pharmacokinetics and tissue distribution of humanin and its analogues in male rodents. Endocrinology. 2013;154(10):3739-3744. View Scientific Paper →
Yen K, Lee C, Mehta H, Cohen P. The emerging role of the mitochondrial-derived peptide humanin in stress resistance. J Mol Endocrinol. 2013;50(1):R11-R19. View Scientific Paper →
Lee C, Wan J, Miyazaki B, et al. IGF-I regulates the age-dependent signaling peptide humanin. Aging Cell. 2014;13(5):958-961. View Scientific Paper →
Cobb LJ, Lee C, Xiao J, et al. Naturally occurring mitochondrial-derived peptides are age-dependent regulators of apoptosis, insulin sensitivity, and inflammatory markers. Aging (Albany NY). 2016;8(4):796-809. View Scientific Paper →
Niikura T, Sidahmed E, Hirata-Fukae C, Aisen PS, Matsuoka Y. A humanin derivative reduces amyloid beta accumulation and ameliorates memory deficit in triple transgenic mice. PLoS One. 2011;6(1):e16259. View Scientific Paper →
Tajima H, Kawasumi M, Chiba T, et al. A humanin derivative, S14G-HN, prevents amyloid-beta-induced memory impairment in mice. J Neurosci Res. 2005;79(5):714-723. View Scientific Paper →