对于拟肽与他们抗利什曼原虫活性混合赖氨酸型/精氨酸型单体及评价合成的有效方法
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Summary
Abstract
Introduction
Protocol
Representative Results
Discussion
Disclosures
The authors have nothing to disclose.
Acknowledgements
Materials
| Polypropylene solid phase extraction cartridges with two frits | Crawford Scientific | 12131017 and 12131015 | 20 ml and 6 ml cartridges used in this preparation |
| Trifluoroacetic acid | Tokyo Chemical Industry (Europe) | T0431-100g | >98%; CAUTION: can cause severe burns and respiratory irritation |
| <em>N,N'</em>-diisopropylcarbodiimide | Sigma Aldrich | 38370-100ML | >98%; CAUTION: hazardous to eyes, skin and via respiratory inhalation, may also cause sensitisation |
| Dimethylformamide | Fischer Scientific | 10346180 | HPLC grade; CAUTION: suspected teratogen |
| Acetonitrile | Fischer Scientific | 10407440 | HPLC grade |
| Dichloromethane | Fischer Scientific | 10354263 | 99.8%; CAUTION: suspected carcinogen |
| Bromacetic acid | Sigma Aldrich | 17000-100G | >99%; CAUTION: causes burns and hazardous to skin, eyes and respiratory tract |
| (<em>S</em>)-(-)-alpha methylbenzylamine | Sigma Aldrich | 115568-100G | 98%; CAUTION: harmful if swallowed, toxic in contact with skin, causes severe skin burns and eye damage, used to synthesise the Nspe monomer |
| <em>N</em>-Boc 1,4-diaminobutane | Tokyo Chemical Industry (Europe) | A1373-25g | >98%; CAUTION: causes severe skin burns and eye damage, used to synthesise the NLys monomer |
| <em>N</em>-Boc 1,2-diaminoethane | Tokyo Chemical Industry (Europe) | A1371-25g | >97%; CAUTION: causes severe skin burns and eye damage, used to synthesise the Nae monomer |
| 1,2-diaminobutane | Sigma Aldrich | D13208-100G | 99%; CAUTION: flammable liquid and vapour, harmful if swallowed or inhaled, toxic in contact with skin, causes severe skin burns and eye damage, used in the synthesis of the NhArg monomer |
| 1,4-diaminoethane | Sigma Aldrich | 03550-250ML | >99.5%; CAUTION: flammable liquid and vapour, harmful if swallowed or inhaled, toxic in contact with skin, causes severe skin burns and eye damage, used in the synthesis of the NnArg monomer<br /> <br /> |
| 1H-pyrazole-1-carboxamidine HCl | Sigma Aldrich | 402516-10G | as HCl salt; CAUTION: harmful if swallowed and may cause an allergic skin reaction, causes serious eye damage |
| Hydrazine monohydrate | Sigma Aldrich | 207942-5G | reagent grade; CAUTION: suspected carcinogen, fatal if inhaled and causes severe burns to skin and eyes |
| 2-acetyl dimedone | Novabiochem | 8510150005 | Dde-OH |
| Rink Amide resin | Novabiochem | 8551190005 | 100-200 mesh, high loading |
| Piperidine | Sigma Aldrich | 411027-1L | >99.5%, a controlled substance, so adequate permission must be obtained before purchase; CAUTION: highly flammable liquid and vapour, harmful if swallowed and toxic in contact with skin or if inhaled, causes severe skin burns and eye damage, harmful to aquatic life with long lasting effects |
| Triisopropylsilane | Sigma Aldrich | 233781-50G | 98%; CAUTION: flammable liquid and vapour, causes skin irritation and serious eye irritation |
| alamarBlue | ThermoFischer | DAL1025 | Not classified as hazardous |
| Schneider's Insect Medium | Sigma Aldrich | S9895-1L | Powdered, medium must be made prior to use following manafacturers instructions; allow to warm to room temperature before use in biological assays |
| 96 well plates | VWR | 734-1793 | Flat bottom (to allow fluorescence measurement from the bottom), tissue-culture treated |
| Solvent reservoirs | VWR | 613-1182 | Used with multi channel pipette |
| Multi channel pipette | Eppendorf | 3122000043 | |
| Pipette tips | Starlab Group | S1111-3810, S1113-1810, S1111-6810 | Volume of tip dependent on pipette used. 10 µl, 10 – 200 µl and 1,000 µl recommended for assays |
| 25 cm<sup>3</sup> cell culture flasks | VWR | 734-2312 | |
| 50 ml centrifuge tubes | VWR | 525-0791 | |
| dimethylsulphoxide (molecular biology grade) | Sigma Aldrich | D8418-50ML | Not classified as hazardous |
| Heat Inactivated Fetal Bovine Serum | ThermoFischer | 10082139-100mL | Gibco |
| Penicillin/Streptomycin | ThermoFischer | 15140148-20mL | Abbreviation: P/S |
| Amphotericin B | Sigma Aldrich | 46006-100mg | Amphotericin B trihydrate, VetranalTM analytical standard |
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Cite This Article
Bolt, H. L., Denny, P. W., Cobb, S. L. An Efficient Method for the Synthesis of Peptoids with Mixed Lysine-type/Arginine-type Monomers and Evaluation of Their Anti-leishmanial Activity. J. Vis. Exp. (117), e54750, doi:10.3791/54750 (2016).