Physicochemical Properties
| Molecular Formula | C97H173N51O24 |
| Molecular Weight | 2437.74 |
| Exact Mass | 2436.388 |
| CAS # | 141237-50-5 |
| PubChem CID | 16141162 |
| Appearance | Typically exists as solid at room temperature |
| Density | 1.6±0.1 g/cm3 |
| Index of Refraction | 1.719 |
| LogP | -17.61 |
| Hydrogen Bond Donor Count | 54 |
| Hydrogen Bond Acceptor Count | 35 |
| Rotatable Bond Count | 97 |
| Heavy Atom Count | 172 |
| Complexity | 5420 |
| Defined Atom Stereocenter Count | 18 |
| SMILES | C[C@H]([C@@H](C(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCNC(=N)N)C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCCNC(=N)N)C(=O)O)N)O |
| InChi Key | SJDURFRPNNLLOO-LYAKTKFASA-N |
| InChi Code | InChI=1S/C97H173N51O24/c1-46(133-72(156)60(27-30-66(98)150)142-79(163)59(25-13-41-130-96(118)119)145-86(170)70(101)47(2)149)71(155)134-51(17-5-33-122-88(102)103)73(157)137-58(24-12-40-129-95(116)117)81(165)148-65(44-68(100)152)85(169)141-55(21-9-37-126-92(110)111)76(160)136-52(18-6-34-123-89(104)105)74(158)135-53(19-7-35-124-90(106)107)75(159)138-57(23-11-39-128-94(114)115)80(164)147-64(43-48-45-132-50-16-4-3-15-49(48)50)84(168)140-56(22-10-38-127-93(112)113)78(162)144-62(29-32-69(153)154)82(166)139-54(20-8-36-125-91(108)109)77(161)143-61(28-31-67(99)151)83(167)146-63(87(171)172)26-14-42-131-97(120)121/h3-4,15-16,45-47,51-65,70,132,149H,5-14,17-44,101H2,1-2H3,(H2,98,150)(H2,99,151)(H2,100,152)(H,133,156)(H,134,155)(H,135,158)(H,136,160)(H,137,157)(H,138,159)(H,139,166)(H,140,168)(H,141,169)(H,142,163)(H,143,161)(H,144,162)(H,145,170)(H,146,167)(H,147,164)(H,148,165)(H,153,154)(H,171,172)(H4,102,103,122)(H4,104,105,123)(H4,106,107,124)(H4,108,109,125)(H4,110,111,126)(H4,112,113,127)(H4,114,115,128)(H4,116,117,129)(H4,118,119,130)(H4,120,121,131)/t46-,47+,51-,52-,53-,54-,55-,56-,57-,58-,59-,60-,61-,62-,63-,64-,65-,70-/m0/s1 |
| Chemical Name | (4S)-4-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S,3R)-2-amino-3-hydroxybutanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-oxopentanoyl]amino]propanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-carbamimidamidopentanoyl]amino]-4-oxobutanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-[[(2S)-1-[[(2S)-5-amino-1-[[(1S)-4-carbamimidamido-1-carboxybutyl]amino]-1,5-dioxopentan-2-yl]amino]-5-carbamimidamido-1-oxopentan-2-yl]amino]-5-oxopentanoic acid |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets |
Antagonism of HIV-1 Rev function by binding to the Rev-responsive element (RRE) [1] Antagonism of the CXCR4 co-receptor (by blocking anti-CXCR4 monoclonal antibody 12G5 binding) [1] |
| ln Vitro |
HIV-1 Rev (34-50) is a 17-amino acid peptide with anti-HIV-1 activity that is derived from the Rev response element (RRE) binding domain of HIV-1 [1]. The peptide Rev34-50 (sequence: TRQARRNRRRRWRERQR) at 10 µM partially inhibited infection by the CXCR4-tropic HIV-1IIIB strain in a MAGI assay, but showed no activity against the CCR5-tropic HIV-1Ba-L strain [1] The modified peptide Rev34-50-A4C (Rev34-50 with AAAC added at the C-terminus to stabilize α-helical structure) showed enhanced anti-HIV-1 activity against HIV-1IIIB compared to the parental Rev34-50 in the MAGI assay [1] In a flow cytometric binding assay, both Rev34-50 and Rev34-50-A4C inhibited the binding of a phycoerythrin-conjugated anti-CXCR4 monoclonal antibody (12G5) to the surface of MT-2 cells in a dose-dependent manner (tested up to 100 µM) [1] In persistently HIV-1-infected PM1-CCR5 cells, Rev34-50 at 1 µM and 10 µM showed no inhibitory effect on viral p24 antigen production for either HIV-1IIIB (CXCR4-tropic) or HIV-1Ba-L (CCR5-tropic) strains [1] In contrast, Rev34-50-A4C at 10 µM significantly inhibited viral p24 production from both PM1-CCR5/IIIB and PM1-CCR5/Ba-L persistently infected cells [1] Addition of a protein transduction domain (PTD) from HIV-1 Tat to the C-terminus of Rev34-50-A4C (forming Rev34-50-A4C-PTD) hardly enhanced its inhibitory effect on p24 production compared to Rev34-50-A4C alone [1] The peptide Rev34-50-A4C (10 µM) reduced the infectivity-to-p24 ratio of HIV-1Ba-L virions released from treated, persistently infected cells, suggesting a potential effect on virion integrity or post-entry events [1] Intracellular expression of Rev34-50 in MT-2 cells via a transduction vector conferred moderate resistance against HIV-1IIIB infection, while expression of Rev34-50-A4C conferred higher resistance [1] Fluorescence microscopy showed that EGFP-fused Rev34-50 and Rev34-50-A4C peptides accumulated dominantly in the nucleus when expressed in 293T cells [1] |
| Cell Assay |
For anti-HIV-1 activity in the early phase (MAGI assay), HeLa CD4/CCR5/LTR-β-galactosidase (MAGI/CCR5) cells were used. The peptides were added to cells, which were then infected with HIV-1IIIB or HIV-1Ba-L. After 48 hours, β-galactosidase activity was measured to quantify infection [1] For evaluation of CXCR4 antagonism (flow cytometry), MT-2 cells were resuspended in PBS containing 1% fetal calf serum and preincubated with peptides at 4°C for 30 minutes. A phycoerythrin-conjugated mouse anti-human CXCR4 monoclonal antibody (12G5) was then added and incubated at 4°C for a further 30 minutes. Cells were washed, fixed with formaldehyde, and analyzed using a flow cytometer [1] For inhibitory effects on viral production (late phase), PM1-CCR5 cells persistently infected with HIV-1IIIB or HIV-1Ba-L (PM1-CCR5/IIIB or PM1-CCR5/Ba-L) were washed and resuspended in medium containing peptides. After 48 hours, the amount of viral p24 antigen in the culture supernatants was measured using a commercial ELISA kit [1] For infectivity assessment of secreted virus, supernatants from peptide-treated (10 µM, 24 h) PM1-CCR5/Ba-L cells were harvested. The infectivity was titrated using MAGI cells, and the p24 antigen amount was measured. The infectivity per p24 ratio was calculated [1] For intracellular expression studies, MT-2 cells were transfected with vectors encoding various Rev mutants (including Rev34-50 and Rev34-50-A4C) linked via an internal ribosomal entry site (IRES) to a neomycin resistance gene. Transfected cells were selected with G418. These transduced cells were then infected with HIV-1IIIB, and cell viability was assessed 5 days post-infection using an MTT assay [1] For subcellular localization, 293T or NP-2 cells were transfected with plasmids expressing C-terminally EGFP-fused peptides. After 48 hours, cells were fixed with formaldehyde and examined using a fluorescence microscope [1] Cytotoxicity of peptides was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay on relevant cell lines over several days [1] |
| Toxicity/Toxicokinetics |
The peptides Rev34-50 and Rev34-50-A4C showed no cytotoxicity over 5 days in the tested cell lines as evaluated by MTT assay [1] |
| References | [1]. Shimane K, et al. Rev-derived peptides inhibit HIV-1 replication by antagonism of Rev and a co-receptor, CXCR4. Int J Biochem Cell Biol. 2010 Sep;42(9):1482-8. |
| Additional Infomation |
HIV-1 Rev 34-50 is a 17-amino acid peptide derived from the arginine-rich RNA-binding/nuclear localization signal domain (amino acids 34-50) of the HIV-1 Rev protein [1] Rev34-50-A4C is a derivative of Rev34-50 with the sequence AAAC added to its C-terminus. This modification is intended to stabilize the α-helical structure of the peptide, which is known to be important for efficient binding to the RRE [1] These arginine-rich peptides exhibit cell permeability and nuclear localization properties [1] The proposed mechanism of action involves dual antagonism: 1) Inhibition of HIV-1 entry by blocking the CXCR4 co-receptor. 2) Inhibition of viral production by acting as a trans-dominant repressor of Rev function, likely through competitive binding to the RRE within the nucleus, thereby disrupting the nuclear export of unspliced/singly spliced viral mRNA [1] The study suggests that targeting Rev with inhibitors like these peptides may present a high genetic barrier for the development of HIV-1 resistance, as mutations in the Rev coding region could simultaneously affect the sequences and functions of the overlapping Tat and Env proteins [1] |
Solubility Data
| Solubility (In Vitro) | 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 |
| 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 (e.g. IP/IV/IM/SC) 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). 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) 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). 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 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.4102 mL | 2.0511 mL | 4.1022 mL | |
| 5 mM | 0.0820 mL | 0.4102 mL | 0.8204 mL | |
| 10 mM | 0.0410 mL | 0.2051 mL | 0.4102 mL |