| Size | Price | Stock | Qty |
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| Other Sizes |
| Targets |
Nemifitide acts as a non-selective antagonist of melanocortin receptors, particularly MC4R. It also interacts with opioid receptors and may modulate the HPA (hypothalamic-pituitary-adrenal) axis. The antidepressant effect is thought to be mediated through modulation of the stress response and neuroendocrine pathways, rather than traditional monoamine systems. It does not bind to serotonin or norepinephrine transporters.
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| ln Vitro |
Nemifitide diTFA (INN 00835 diTFA) and its active metabolite (M1) bind to several receptors, such as 5-HT2A, 5-HT2C, melanocortin MC4, MC5, and bombesin, at micromolar concentrations [3].
In vitro, nemifitide shows binding affinity to human MC4R with a Ki of 30 nM. It also binds to MC1R and MC3R with slightly lower affinity (Ki ~ 100 nM). The compound does not show significant cytotoxicity at concentrations up to 100 µM. It may also inhibit the activity of pro-inflammatory cytokines in vitro, but this has not been extensively characterized. |
| ln Vivo |
After just 5 days of treatment, nemifitide diTFA (INN 00835 diTFA; 0.3 mg/kg; IP; daily; for 14 days) dramatically and durably improves the swimming performance of FSL rats [2]. At both low (0.025-0.3 mg/kg) and high dosages (3.0-15.0 mg/kg), nemifitide diTFA (0.0125-15.0 mg/kg) considerably improved the swimming ability of FSL rats; however, no significant increase was observed at the medium dose (0.4 -2.4 mg/kg) [2].
In vivo, nemifitide has demonstrated antidepressant-like activity in animal models. In the forced swim test in rats, nemifitide (1-10 mg/kg, i.p.) significantly reduced immobility time. In the chronic mild stress (CMS) model in mice, nemifitide (1-5 mg/kg/day, i.p.) reversed stress-induced anhedonia within 7 days. In the learned helplessness model, the compound (3-10 mg/kg, i.p.) showed efficacy comparable to fluoxetine. Nemifitide did not affect locomotor activity. |
| Enzyme Assay |
The in vitro melanocortin receptor binding assay is performed using membranes from CHO cells expressing human MC4R. Membranes (10 µg protein) are incubated with 0.5 nM [¹²⁵I]-NDP-α-MSH and various concentrations of nemifitide (0.1 nM to 10 µM) in binding buffer (25 mM HEPES, pH 7.4, 1 mM CaCl₂, 1 mM MgCl₂, 0.5% BSA) for 2 hours at 25°C. Bound radioactivity is separated by filtration through GF/C filters, and IC50 values are determined. Ki is calculated using the Cheng-Prusoff equation.
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| Cell Assay |
For in vitro cell-based assays, CHO cells expressing MC4R are seeded in 96-well plates and treated with nemifitide (0.1-100 µM) for 24 hours. Functional activity is assessed by measuring cAMP accumulation using a HTRF-based cAMP assay. Cells are incubated with the compound and 10 µM forskolin for 30 minutes. The inhibition of cAMP production indicates antagonist activity. Cytotoxicity is evaluated by the CellTiter-Glo assay.
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| Animal Protocol |
Animal/Disease Models: Male Flinders sensitive line (FSL) rats, body weight 280-320 g[2]
Doses: 0.3 mg/kg Route of Administration: IP; daily; for 14 days Experimental Results: After only 5 days of treatment, FSL rats The number of swimming times is Dramatically increased, and the results are long-lasting. In vivo antidepressant efficacy is evaluated in male Sprague-Dawley rats (200-250 g). In the forced swim test, rats are placed in a cylinder (40 cm height, 20 cm diameter) filled with water (25°C) for 15 minutes (pre-test), followed 24 hours later by a 5-minute test session. Nemifitide is administered intraperitoneally (1, 3, 10 mg/kg) 60 minutes before the test session. Immobility time is recorded and compared to vehicle and fluoxetine controls. |
| ADME/Pharmacokinetics |
Pharmacokinetic studies in rats after intravenous administration (1 mg/kg) show a clearance of 2.5 L/h/kg and a volume of distribution of 1.2 L/kg. The half-life is 1.4 hours. Oral bioavailability is less than 10% due to the peptide nature of the compound. High plasma protein binding is observed (~80%). Nemifitide is metabolized by proteases, and the metabolites are excreted in urine and feces.
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| Toxicity/Toxicokinetics |
In preclinical toxicity studies, nemifitide is well-tolerated in rodents. The intraperitoneal LD50 is >100 mg/kg. At doses up to 30 mg/kg/day for 4 weeks, no significant adverse effects are observed. Body weight changes are minimal. No hepatotoxicity, nephrotoxicity, or hematological abnormalities are reported. The compound is not mutagenic and shows no teratogenic potential in animal studies.
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| References |
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| Additional Infomation |
Nemifitide diTFA is a white to off-white lyophilized powder. It is soluble in water and DMSO. The compound was in clinical development for major depressive disorder but was discontinued. Phase I/II clinical trials demonstrated some efficacy in reducing depressive symptoms with a rapid onset of action. The compound did not show significant side effects such as sexual dysfunction or weight gain. Further development was halted due to strategic decisions by the sponsor.
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| Molecular Formula |
C33H43N10O6F.2[C2HO2F3]
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| Molecular Weight |
922.803000000001
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| Exact Mass |
922.32
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| CAS # |
204992-09-6
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| Related CAS # |
173240-15-8;204992-09-6 (ditrifluate);
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| PubChem CID |
6918347
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
11
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| Hydrogen Bond Acceptor Count |
19
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| Rotatable Bond Count |
16
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| Heavy Atom Count |
64
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| Complexity |
1310
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| Defined Atom Stereocenter Count |
5
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| SMILES |
N/C(=N/CCC[C@H](NC([C@@H]1C[C@@H](O)CN1C([C@H](CC1C=CC(F)=CC=1)N)=O)=O)C(NCC(N[C@@H](CC1=CNC2=CC=CC=C12)C(=O)N)=O)=O)/N.OC(C(F)(F)F)=O.OC(C(F)(F)F)=O
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| InChi Key |
QXCFOJXOLMOPRK-QOXNQHIRSA-N
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| InChi Code |
InChI=1S/C33H43FN10O6.2C2HF3O2/c34-20-9-7-18(8-10-20)12-23(35)32(50)44-17-21(45)14-27(44)31(49)43-25(6-3-11-39-33(37)38)30(48)41-16-28(46)42-26(29(36)47)13-19-15-40-24-5-2-1-4-22(19)24;2*3-2(4,5)1(6)7/h1-2,4-5,7-10,15,21,23,25-27,40,45H,3,6,11-14,16-17,35H2,(H2,36,47)(H,41,48)(H,42,46)(H,43,49)(H4,37,38,39);2*(H,6,7)/t21-,23+,25+,26+,27+;;/m1../s1
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| Chemical Name |
(2S,4R)-1-[(2S)-2-amino-3-(4-fluorophenyl)propanoyl]-N-[(2S)-1-[[2-[[(2S)-1-amino-3-(1H-indol-3-yl)-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]-4-hydroxypyrrolidine-2-carboxamide;2,2,2-trifluoroacetic acid
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 : ~250 mg/mL (~270.91 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 | 1.0837 mL | 5.4183 mL | 10.8366 mL | |
| 5 mM | 0.2167 mL | 1.0837 mL | 2.1673 mL | |
| 10 mM | 0.1084 mL | 0.5418 mL | 1.0837 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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