MCH-1R (melanin-concentrating hormone receptor 1): binding IC50 = 0.3 ± 0.1 nM (n=10); functional EC50 (calcium mobilization, FLIPR) = 3.9 ± 1.2 nM (n=3); IP3 turnover EC50 = 88.7 ± 16 nM (n=4).
MCH-2R (melanin-concentrating hormone receptor 2): binding IC50 = 1.5 ± 0.9 nM (n=8); functional EC50 (calcium mobilization, FLIPR) = 0.1 ± 0.1 nM (n=3); functional EC50 (calcium mobilization, aequorin assay) = 31 nM; IP3 turnover EC50 = 2.7 ± 0.6 nM (n=3). [1]
MCH targets the melanin-concentrating hormone receptors, specifically MCH1R and MCH2R. It is a potent peptide agonist for these G-protein-coupled receptors. MCH exhibits high binding affinity for both receptor subtypes, with IC50 values of 0.3 nM and 1.5 nM for MCH1R and MCH2R, respectively. The compound is highly sensitive to MCH2R in CHO cell lines. Activation of these receptors by MCH triggers intracellular signaling cascades that mediate its various physiological effects, particularly the stimulation of appetite.

MCH (human, mouse, rat) (CAS#: 128315-56-0) activates MCH-2R stably expressed in CHO cells, inducing intracellular calcium mobilization as measured by FLIPR, with an EC50 of 0.1 ± 0.1 nM (n=3). The modified peptide Phe13Tyr19-MCH and salmon MCH also activate MCH-2R with reduced potency (EC50 = 0.2 ± 0.1 nM and 0.8 ± 0.1 nM, respectively). [1]
In HEK293-AEQ17 cells stably expressing MCH-2R, MCH elicits a dose-dependent calcium response with an EC50 of 31 nM, comparable to MCH-1R signaling under similar conditions. The response is specific; no activation is seen with other neuropeptides including NGE and NEI. [1]
MCH potently stimulates inositol phosphate (IP3) turnover through MCH-2R with an EC50 of 2.7 ± 0.6 nM (n=3), whereas MCH-1R shows an EC50 of 88.7 ± 16 nM (n=4) in stably expressing HEK293-AEQ17 cells. [1]
MCH-2R-mediated calcium mobilization is not sensitive to pertussis toxin (PTX) treatment (up to 5000 ng/ml), whereas MCH-1R signaling is reduced in a dose-dependent manner to about 50% of control. This indicates that MCH-2R couples exclusively to Gq/11 pathways, not Gi/o. [1]
MCH-2R does not inhibit forskolin-stimulated cAMP production, confirming lack of Gi/o coupling. [1]
Binding affinity: MCH displaces [125I]Phe13Tyr19-MCH from MCH-2R-containing membranes with an IC50 of 1.5 ± 0.9 nM (n=8); for MCH-1R, IC50 = 0.3 ± 0.1 nM (n=10). Salmon MCH shows much lower binding affinity for MCH-2R (IC50 = 436.7 ± 143.8 nM, n=3) compared to MCH-1R (IC50 = 0.2 ± 0.1 nM, n=5). [1]
In vitro, MCH demonstrates potent agonist activity at MCH receptors. It exhibits binding IC50 values of 0.3 nM for MCH1R and 1.5 nM for MCH2R. Functional activity is confirmed by EC50 values of 3.9 nM and 0.1 nM at MCH1 and MCH2 receptors, respectively. These in vitro studies typically use cell lines expressing the recombinant receptors to assess binding affinity and receptor activation. The compound's high potency in vitro reflects its strong interaction with its target receptors and its ability to activate downstream signaling pathways.

In vivo, MCH (human, mouse, rat) is a potent orexigenic peptide that increases food intake. Animal studies have demonstrated that central administration of MCH stimulates feeding behavior. Its effects on appetite regulation have been extensively studied, confirming its role as a key player in the central control of energy homeostasis. The peptide's in vivo activity is directly related to its agonist action at MCH receptors in the brain, making it a significant target for research into obesity and metabolic disorders.

Membrane binding assay (scintillation proximity assay, SPA): Membranes from COS-7 cells transiently transfected with MCH-2R or CHO cells stably expressing MCH-2R were prepared by hypotonic lysis and stored at -80°C. Wheat germ agglutinin-polyvinyltoluene SPA beads (0.25 mg per well) were mixed with 1-10 μg membrane protein and 200 μl binding buffer (50 mM Tris pH 7.4, 8 mM MgCl2, 12% glycerol, 0.1% BSA, protease inhibitors: 4 μg/ml leupeptin, 40 μg/ml bacitracin, 5 μg/ml aprotinin, 100 μM 4-(2-aminoethyl)benzenesulfonyl fluoride) in 96-well plates. After coating beads with membranes for 20 min, various concentrations of test compounds (MCH) in DMSO (final DMSO 1-2%) and 25 nCi of [125I]Phe13Tyr19-MCH were added. Plates were equilibrated at room temperature for 3 hours, then read on a TopCount. Specific binding was defined as total binding minus nonspecific binding in the presence of 500 nM unlabeled MCH. IC50 calculations were performed using Prism 3.0. [1]
In vitro enzyme/receptor binding assays for MCH are performed to determine its affinity for MCH1R and MCH2R. Standard protocols involve competitive binding experiments using membrane preparations from cells that overexpress the human MCH receptors. Radiolabeled MCH or a specific antagonist is used as the tracer ligand. Increasing concentrations of unlabeled MCH are incubated with the receptor membranes, and the displacement of the tracer is measured. The IC50 values are calculated from the resulting competition curves, with MCH showing values of 0.3 nM for MCH1R and 1.5 nM for MCH2R. These assays are performed in a suitable buffer at physiological pH.

Calcium mobilization assay using FLIPR (fluorometric imaging plate reader): MCH-2R-expressing CHO cells were seeded at 5×10^4 cells/well in black-wall clear-bottom 96-well plates one day before assay. Cells were incubated with 100 μl/well assay buffer (Hanks' balanced salt solution, 0.5% BSA, 20 mM Hepes, 2.5 mM probenecid, pH 7.4) containing 2 μM fluo-4 AM, 0.04% pluronic acid, and 1% FBS for 60 min in a CO2 incubator. After washing four times with assay buffer, the plate was placed in the FLIPR, and 50 μl/well of agonist solution (MCH in assay buffer with 1% DMSO final) was added. Fluorescence output was measured; basal fluorescence was 10,000-15,000, maximal response peak 40,000-50,000. [1]
Aequorin bioluminescence assay: HEK293-AEQ17 cells stably expressing MCH-2R were incubated with 10 μM coelenterazine cp and 300 μM reduced glutathione in ECB buffer (140 mM NaCl, 20 mM KCl, 20 mM Hepes-NaOH pH 7.4, 5 mM glucose, 1 mM MgCl2, 1 mM CaCl2, 0.1 mg/ml BSA) to charge apo-aequorin. Cells were harvested, washed, resuspended to 500,000 cells/ml. 100 μl (5×10^4 cells) was injected into a 96-well test plate, and integrated light emission recorded over 30 s (0.5-s units). Then 20 μl lysis buffer (0.1% Triton X-100 final) was injected, and light emission recorded over 10 s. Fractional response was calculated as the ratio of the initial integrated response to the total integrated luminescence including lysis response. [1]
Inositol phosphate turnover assay: HEK293-AEQ17 cells stably expressing MCH-1R or MCH-2R were plated in 12-well dishes (2.75×10^5 cells/well) and incubated with 4.8 μCi [3H]-myo-inositol overnight at 37°C. After stimulation with MCH for 1 h at 37°C, the reaction was stopped by replacing medium with 800 μl of 10 mM formic acid. Cell extracts were collected after 15 min at 4°C and neutralized with 100 μl of 50 mM ammonium hydroxide. The inositol phosphate fraction was isolated by anion exchange chromatography as described by Berridge et al. (1983). [1]
Pertussis toxin (PTX) sensitivity assay: Stable HEK293-AEQ17 cells expressing human MCH-1R or MCH-2R were treated for 4 hours with PTX at 0, 150, 500, or 5000 ng/ml in growth medium before the aequorin assay for calcium mobilization. [1]
In vitro cellular assays for MCH are conducted using cell lines, such as CHO cells, that are engineered to express MCH1R or MCH2R. Cells are seeded in multi-well plates and allowed to adhere. They are then incubated with various concentrations of MCH, and receptor activation is measured by quantifying downstream second messengers, such as intracellular calcium mobilization or cAMP levels. The functional potency (EC50) is determined from the dose-response curves. For MCH, EC50 values of 3.9 nM and 0.1 nM are observed at MCH1 and MCH2 receptors, respectively. These assays confirm the compound's agonist activity.

In vivo animal studies for MCH typically involve administration of the peptide to rodents, often via intracerebroventricular (ICV) injection to bypass the blood-brain barrier. Following administration, food intake is monitored over a defined period, usually several hours. The orexigenic effect of MCH is assessed by comparing the food consumption of treated animals to that of vehicle-treated controls. These studies have consistently shown that MCH increases food intake in a dose-dependent manner. The compound's effects on other parameters, such as body weight and energy expenditure, may also be evaluated in longer-term studies.

Specific pharmacokinetic data for the MCH peptide are not extensively detailed in standard reference sources, as it is primarily a research tool. As a neuropeptide, it is expected to have a short half-life in the systemic circulation due to rapid degradation by proteases. When administered centrally (e.g., ICV), its effects are localized to the central nervous system. For peripheral administration, the peptide faces challenges such as poor bioavailability and rapid clearance. These properties are typical for peptide-based compounds. Its in vivo effects on food intake are observed after direct central administration.

Comprehensive toxicology data for MCH as a research peptide are not typically reported in general references, as it is not a clinical drug. As an endogenous neuropeptide, its toxicological profile is expected to be related to its pharmacological effects, particularly the dysregulation of feeding behavior and energy balance. At high doses, central administration could lead to excessive food intake and associated metabolic disturbances. Standard safety assessments would include evaluation of off-target effects, as MCH primarily acts on its specific receptors, but at high concentrations, it could potentially interact with other receptors or pathways.

[1]. Identification and characterization of a second melanin-concentrating hormone receptor, MCH-2R. Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7564-9.

MCH (human, mouse, rat) (CAS#: 128315-56-0) is derived from a precursor that also produces neuropeptides NGE and NEI, but those peptides do not activate MCH-2R. [1]
In mammals, MCH is produced primarily in neurons of the lateral hypothalamus with far-reaching projections throughout the brain. MCH mRNA is upregulated in leptin-deficient ob/ob mice and upon fasting. Injection of MCH into rat lateral ventricles increases food consumption (reference to other studies). MCH-deficient mice are lean and hypophagic with increased metabolic rate, while MCH-overexpressing transgenic mice develop obesity and insulin resistance (referenced). [1]
Northern blot analysis of human tissues using a radiolabeled MCH-2R probe detected a 4.4 kb mRNA transcript specifically in brain. In situ hybridization of rhesus monkey brain showed MCH-2R mRNA expression in cerebral cortex, hippocampus, hypothalamus, caudate nucleus, putamen, and thalamus. In adjacent coronal sections of African green monkey hypothalamus, MCH-2R was abundant in anterior and lateral hypothalamic areas but barely detectable in dorsomedial hypothalamus, whereas MCH-1R showed strong expression in dorsomedial hypothalamus. Both receptors were abundant in ventromedial hypothalamic nuclei. [1]
The human MCH-2R gene was mapped to chromosome 6q16.2-16.3 by fluorescence in situ hybridization using PAC clone 60p05, and confirmed by radiation hybrid analysis. This region is associated with cytogenetic abnormalities in obese patients and neuropsychiatric disorders such as autism and schizophrenia. The SIM1 gene (human homolog of Drosophila single-minded) is located within 1 Mb of MCH-2R. [1]
MCH (human, mouse, rat) is an endogenous cyclic neuropeptide that plays a pivotal role in the central regulation of appetite and energy homeostasis. It is a potent agonist at MCH1R and MCH2R, with high binding affinities. The peptide increases food intake in vivo, making it a key orexigenic factor. MCH and its receptors are important targets for research into obesity, metabolic syndrome, and related disorders. The compound is widely used in preclinical studies to understand the neurobiology of feeding and to develop potential therapeutic interventions. It is not approved as a drug but is a standard research tool.

C89H139N27O24S4

2099.48246

2385.1

128315-56-0

MCH(human, mouse, rat) TFA

24868207

Typically exists as solid at room temperature

6.547

33

34

59

165

5130

18

CC(C[C@H]1C(NCC(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(N2CCC[C@H]2C(N[C@H](C(N[C@H](C(N[C@H](C(N[C@H](C(O)=O)C(C)C)=O)CCC(N)=O)=O)CC3=CNC4=CC=CC=C34)=O)CSSC[C@H](NC([C@@H](NC([C@@H](NC([C@@H](NC([C@@H](NC([C@@H](NC([C@@H](N)CC(O)=O)=O)CC5=CC=CC=C5)=O)CC(O)=O)=O)CCSC)=O)CC(C)C)=O)CCCNC(N)=N)=O)C(N[C@H](C(N1)=O)CCSC)=O)=O)=O)CCCNC(N)=N)=O)CC6=CC=C(O)C=C6)=O)C(C)C)=O)CCCNC(N)=N)=O)=O)C

MWLPXSMFEPHETN-QRXCLJFASA-N

InChI=1S/C105H160N30O26S4/c1-53(2)42-70-86(144)118-50-80(138)119-64(24-16-36-114-103(108)109)90(148)133-83(55(5)6)100(158)130-73(45-58-28-30-60(136)31-29-58)93(151)124-69(26-18-38-116-105(112)113)101(159)135-39-19-27-78(135)99(157)132-77(98(156)128-74(46-59-49-117-63-23-15-14-22-61(59)63)95(153)121-66(32-33-79(107)137)91(149)134-84(56(7)8)102(160)161)52-165-164-51-76(97(155)123-68(35-41-163-10)88(146)126-70)131-87(145)65(25-17-37-115-104(110)111)120-92(150)71(43-54(3)4)127-89(147)67(34-40-162-9)122-96(154)75(48-82(141)142)129-94(152)72(44-57-20-12-11-13-21-57)125-85(143)62(106)47-81(139)140/h11-15,20-23,28-31,49,53-56,62,64-78,83-84,117,136H,16-19,24-27,32-48,50-52,106H2,1-10H3,(H2,107,137)(H,118,144)(H,119,138)(H,120,150)(H,121,153)(H,122,154)(H,123,155)(H,124,151)(H,125,143)(H,126,146)(H,127,147)(H,128,156)(H,129,152)(H,130,158)(H,131,145)(H,132,157)(H,133,148)(H,134,149)(H,139,140)(H,141,142)(H,160,161)(H4,108,109,114)(H4,110,111,115)(H4,112,113,116)/t62-,64-,65-,66-,67-,68-,69-,70-,71-,72-,73-,74-,75-,76-,77-,78-,83-,84-/m0/s1

(2S)-2-[[(2S)-5-amino-2-[[(2S)-2-[[(3S,6S,9S,12S,18S,21S,24R,29R,32S)-24-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-3-carboxypropanoyl]amino]-3-phenylpropanoyl]amino]-3-carboxypropanoyl]amino]-4-methylsulfanylbutanoyl]amino]-4-methylpentanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3,12-bis(3-carbamimidamidopropyl)-6-[(4-hydroxyphenyl)methyl]-18-(2-methylpropyl)-21-(2-methylsulfanylethyl)-2,5,8,11,14,17,20,23,31-nonaoxo-9-propan-2-yl-26,27-dithia-1,4,7,10,13,16,19,22,30-nonazabicyclo[30.3.0]pentatriacontane-29-carbonyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-5-oxopentanoyl]amino]-3-methylbutanoic acid

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.)