RO27-3225 is a selective melanocortin-4 receptor (MC4R) agonist with EC₅₀ values of 0.6 nM at MC4R, 4.8 nM at MC3R, and >10,000 nM at MC1R and MC5R. [1]
Functions as a potent and selective MC4R agonist. [2][3]

RO27-3225 is a cyclic heptapeptide (Ac-Cys¹⁰-[D-Nal⁷,Arg⁸]-Cys⁹-NH₂) with 1,000-fold selectivity for MC4R over MC1R/MC5R. [1]
Mechanism: Activates MC4R to modulate central appetite regulation and peripheral anti-inflammatory pathways. [1][2]
Confers neuroprotection by promoting angiogenesis and suppressing neuroinflammation post-stroke. [3]

In freely feeding rats, intraperitoneal injection of RO27-3225 (0.01-1 mg/kg) dose-dependently reduced food intake by up to 90% at 1 mg/kg within 4 hours without inducing conditioned taste aversion. This effect was absent in MC4R knockout mice. [1]
In a rat hemorrhagic shock model, intravenous administration of RO27-3225 (90 μg/kg) restored mean arterial pressure (MAP) from 30±5 mmHg to 85±10 mmHg within 15 minutes, improved organ perfusion, and reduced plasma TNF-α and IL-6 levels by >50%. [2]
In mice with middle cerebral artery occlusion (MCAO), daily intraperitoneal injection of RO27-3225 (0.5 mg/kg for 14 days) increased neurogenesis (BrdU⁺/DCX⁺ cells by 2.5-fold), enhanced PDGFRβ⁺ pericytes recruitment, and reduced microglial activation (Iba1⁺ cells decreased by 40%). [3]

Food intake study: Rats/mice received RO27-3225 (0.01, 0.1, 1 mg/kg) or vehicle via intraperitoneal (i.p.) injection. Food consumption was measured 1-4h post-dosing. Taste aversion assessed by sucrose preference after drug pairing. [1]
Hemorrhagic shock model: Rats subjected to 45% blood loss received RO27-3225 (90 μg/kg) or vehicle intravenously at 60 min post-shock. Hemodynamics monitored for 6h; organs harvested for histology/cytokine analysis. [2]
Stroke recovery model: MCAO mice treated with RO27-3225 (0.5 mg/kg i.p. daily for 14 days) starting 24h post-surgery. Brains analyzed by immunohistochemistry and ELISA at day 15. [3]

RO27-3225 (1 mg/kg intraperitoneal injection) did not induce conditioned taste aversion, nor did it impair motor coordination in the rotarod test. [1]
In rats with hemorrhagic shock, no acute toxicity was observed at 90 μg/kg intravenous injection. [2]
In MCAO mice, no behavioral abnormalities or deaths were reported after 14 days of intraperitoneal treatment at 0.5 mg/kg. [3]

[1]. A novel selective melanocortin-4 receptor agonist reduces food intake in rats and mice without producing aversive consequences. J Neurosci. 2000 May 1;20(9):3442-8.

[2]. Selective melanocortin MC4 receptor agonists reverse haemorrhagic shock and prevent multiple organ damage. Br J Pharmacol. 2007 Mar;150(5):595-603.

[3]. Effects of RO27-3225 on neurogenesis, PDGFRβ+ cells and neuroinflammation after cerebral infarction. Int Immunopharmacol. 2020 Feb 11;81:106281.

Studies using non-selective agonists and antagonists of melanocortin-3 receptor (MC3R) and MC4R have demonstrated the important role of the melanocortin system in controlling food intake in the central nervous system. This paper describes a novel compound exhibiting highly selective agonist activity towards the MC4 receptor, while showing minimal activity towards the MC3 receptor. Central injection of this selective agonist into rats increased Fos-like immunoreactivity in the paraventricular nucleus, central amygdala, nucleus of the solitary tract, and posterior pole, a neuronal activation pattern similar to that induced by non-selective MC3/4R agonists. Furthermore, central injection of this compound into rats or peripheral injection into db/db mice lacking functional leptin receptors suppressed food intake through a mechanism without disease or other non-specific effects. Conversely, a related selective MC4R antagonist significantly increased food intake in rats upon central administration. These results support the hypothesis that brain MC4R is closely associated with food intake and weight control, and provide evidence that selective activation of MC4R leading to anorexia is not secondary to an aversion effect. [1]

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Background and Objectives: Melanocortin plays a life-saving role in circulatory shock, which may be mediated by the MC4 receptor. To directly understand the role of the melanocortin MC4 receptor in hemorrhagic shock, we investigated the effects of two novel selective MC4 receptor agonists. Methods: A severe hemorrhagic shock model was established in rats under general anesthesia. Rats were then treated with the non-selective agonist [Nle4, D-Phe7]-melanocyte-stimulating hormone (NDP-MSH) or the selective MC4 agonists RO27-3225 and PG-931, respectively. Cardiovascular and respiratory function were continuously monitored for 2 hours; survival rate was recorded within 24 hours. Free radicals in the blood were measured by electron spin resonance spectroscopy; tissue damage was histologically assessed at 25 minutes or 24 hours after treatment. Main Results: All rats receiving saline treatment died within 30–35 minutes. NDP-MSH, RO27-3225 and PG-931 treatments all dose-dependently (13-108 nmol kg-1 intravenous injection) restored cardiovascular and respiratory function and improved survival. These three melanocortin agonists also significantly reduced circulating free radical levels compared with saline-treated shock rats. All of these effects could be prevented by pretreatment with the selective MC4 receptor antagonist HS024 via intraperitoneal injection. In addition, RO27-3225 treatment prevented morphological and immunocytochemical changes in the heart, lungs, liver and kidneys in the early (25 min) and late (24 h) stages. Conclusion and significance: Stimulation of MC4 receptors can reverse hemorrhagic shock, reduce multi-organ damage and improve survival. Our results suggest that selective MC4 receptor agonists may have a protective effect against multi-organ failure following circulatory shock. [2] Cerebral infarction imposes a severe social and economic burden on patients due to its high incidence and mortality, and existing treatments are limited. RO27-3225 is a highly selective melanocortin receptor 4 agonist that can alleviate damage caused by various neurological diseases, such as cerebral hemorrhage, traumatic brain injury, and chronic neurodegenerative diseases. However, the effect of RO27-3225 on cerebral infarction remains unclear. This study used a mouse model of transient middle cerebral artery occlusion (tMCAO) and administered RO27-3225 or saline via intraperitoneal injection. Results showed that on day 7 after tMCAO, RO27-3225 increased the number of Nestin+/BrdU+ cells and dicortin (DCX)+/BrdU+ cells in the subventricular zone (SVZ), as well as the number of DCX+/BrdU+ cells in the peri-infarct region. In addition, on day 3 after tMCAO, RO27-3225 reduced the number of activated microglia (Iba1+ cells with a specific morphology) in the peri-infarct area and decreased the expression levels of Iba1, TNFα, IL6 and iNOS proteins, while increasing the number of PDGFRβ+ cells. Finally, mice treated with RO27-3225 showed a significant reduction in infarct volume, brain water content and neurological deficits after cerebral infarction. Therefore, RO27-3225 can improve the prognosis after cerebral infarction, partly through regulating neurogenesis in the SVZ, survival of PDGFRβ+ cells and neuroinflammation in the peri-infarct area. Our study suggests that RO27-3225 is a potential new therapy for cerebral infarction. [3]

C39H52N12O6

784.906987190247

898.406

1373926-49-8

274682-89-2;1057258-86-2 (free base isomer);1373926-49-8 (TFA);1051970-60-5 (3 TFA);

146026285

Typically exists as solid at room temperature

10

13

22

64

1470

4

CCCC(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC3=CNC4=CC=CC=C43)C(=O)N(C)CC(=O)N.C(=O)(C(F)(F)F)O

XBNXUPIBUGBMCO-WYDLTDSDSA-N

InChI=1S/C39H52N12O6.C2HF3O2/c1-3-10-34(53)47-31(19-26-21-43-23-46-26)37(56)49-30(17-24-11-5-4-6-12-24)36(55)48-29(15-9-16-44-39(41)42)35(54)50-32(38(57)51(2)22-33(40)52)18-25-20-45-28-14-8-7-13-27(25)28;3-2(4,5)1(6)7/h4-8,11-14,20-21,23,29-32,45H,3,9-10,15-19,22H2,1-2H3,(H2,40,52)(H,43,46)(H,47,53)(H,48,55)(H,49,56)(H,50,54)(H4,41,42,44);(H,6,7)/t29-,30-,31-,32-;/m0./s1

(2S)-N-[(2S)-1-[(2-amino-2-oxoethyl)-methylamino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]-2-[[(2S)-2-[[(2S)-2-(butanoylamino)-3-(1H-imidazol-5-yl)propanoyl]amino]-3-phenylpropanoyl]amino]-5-(diaminomethylideneamino)pentanamide;2,2,2-trifluoroacetic acid

RO27-3225; 1373926-49-8; RO273225; RO27-3225; (2S)-N-[(2S)-1-[(2-amino-2-oxoethyl)-methylamino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]-2-[[(2S)-2-[[(2S)-2-(butanoylamino)-3-(1H-imidazol-5-yl)propanoyl]amino]-3-phenylpropanoyl]amino]-5-(diaminomethylideneamino)pentanamide;2,2,2-trifluoroacetic 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.)