ABSTRACT
Pharmaceutically attractive methylene lactone- and methylene lactam-based spiro compounds have been synthesized by employing amido-functionalized γ-hydroxylactam as a common intermediate. Development of a new route for bifurcated synthesis of two types of N,O-spiro compounds was accomplished by treatment of the intermediate under acidic conditions, leading to potent cytotoxic methylene lactone-based spiro compounds. New methylene lactam-based N,N-spiro compounds could be delivered via N-tert-butyloxycarbonyl protection of the terminal amide moiety of the intermediate followed by lactam cyclization under basic conditions.
Subject(s)
Antineoplastic Agents/pharmacology , Lactams/pharmacology , Lactones/pharmacology , Spiro Compounds/pharmacology , Animals , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Cell Line, Tumor , Cell Proliferation/drug effects , Crystallography, X-Ray , Drug Screening Assays, Antitumor , Lactams/chemistry , Lactones/chemistry , Mice , Models, Molecular , Molecular Structure , Spiro Compounds/chemical synthesis , Spiro Compounds/chemistryABSTRACT
Thermospermine, a structural isomer of spermine, is widely spread in the plant kingdom and has recently been shown to play a key role in the repression of xylem differentiation in vascular plants. However, a standard high-performance liquid chromatography (HPLC) protocol for detecting polyamines as their dansyl derivative cannot distinguish themospermine from spermine. These isomers become separated from each other after benzoylation. In this chapter, we describe a simple protocol for extraction, benzoylation, and HPLC detection of thermospermine and spermine with other polyamines from plant material.
Subject(s)
Chromatography, High Pressure Liquid , Plants/chemistry , Spermine/analogs & derivatives , Spermine/analysis , Arabidopsis/chemistry , Plant Extracts/analysisABSTRACT
Thermospermine is a structural isomer of spermine, which is one of the polyamines studied extensively in the past, and is produced from spermidine by the action of thermospermine synthase encoded by a gene named ACAULIS5 (ACL5) in plants. According to recent genome sequencing analyses, ACL5-like genes are widely distributed throughout the plant kingdom. In Arabidopsis, ACL5 is expressed specifically during xylem formation from procambial cells to differentiating xylem vessels. Loss-of-function mutants of ACL5 display overproliferation of xylem vessels along with severe dwarfism, suggesting that thermospermine plays a role in the repression of xylem differentiation. Studies of suppressor mutants of acl5 that recover the wild-type phenotype in the absence of thermospermine suggest that thermospermine acts on the translation of specific mRNAs containing upstream open reading frames (uORFs). Thermospermine is a novel type of plant growth regulator and may also serve in the control of wood biomass production.
