Lipid Mediators

Lipid mediators are messenger molecules produced by cells in response to tissue injury. Prostaglandins (PGs) and Leukotrienes (LTs) are biologically active lipid mediators that play important roles in inflammation and are involved in pathological states with an inflammatory component, such as asthma, cardiovascular disease, cancer, etc. They are derived from arachidonic acid, an essential fatty acid, through the action of cyclooxygenases (COXs) and lipoxygenases (LOXs), respectively.

Molecular structure of ProstacyclinThe diverse and potent biological actions of PGs and LTs onalmost all organs stimulated research on these fascinating molecules over the last five decades. The quintessential properties of “true” eicosanoids are their stereochemical precision in formation and recognition, their potency in the nanomolar range in vitro, and their bona fide biological activities. As a result, these lipid mediators are in great demand for both research, diagnostic and therapeutic purposes. They are extremely difficult to make, and therefore, very expensive (1mg costs ~ $2500). Dr. C. Channa Reddy’s laboratory is the first one to successfully synthesize these compounds in stereo chemical precision using biomimetic method. He has been supplying the entire spectrum of PGs, LTs, and the enzymes associated with their biosynthesis to different companies located in United States. Therefore, Indus Gene is well placed to monopolize the eicosanoid market.

Product Application
Prostaglandins D2/ E2/ F2F2 and others Child birth, Menstrual regulation, Immunity
Prostacyclins Treatment of cardiovascular diseases
Thromboxanes Treatment of Hypertension/ COX inhibition
HPETES and HETEs Research and Development of drugs
Leukotrienes Research and Development of drugs for allergies and asthma

Production of Lipid Mediators

Lipid mediators are produced by a combination of the enzymatic and biomimetic methods. The starting raw material (in this case, arachidonic acid or eicosapentaenoic acid or dihomo-γ- linoleinic acid) serves as the substrate for respective lipid mediators. It is the specific down-stream enzyme used for their synthesis that determines the nature of individual lipid mediator. Both cyclooxygenase (COX) and lipoxygenases (LOX) pathway lipid mediators will be manufactured enzymatically at a laboratory scale by incubating 5 grams of the corresponding substrate precursor with cyclooxygense-1 or lipoxygenases enzymes. Each lipid mediator will be generated in 100-milligram batches at a time from a single reaction. All the enzymes needed for the synthesis of lipid mediators are available with Dr. C. Channa Reddy, who has more than 38 years of research experience in this area. The clones for all these enzymes will be individually expressed and the enzyme proteins are purified by affinity chromatography prior to their use in manufacturing process.

Prostaglandin-H2, an immediate product of cyclooxygenase-1, is the precursor for all the downstream COX lipid mediators. PGH2 will be extracted with non-polar organic solvents (hexane: ether) and evaporated under argon and purified by reverse-phase high pressure liquid chromatography (HPLC). The individual lipid mediators from PGH2 precursor are generated enzymatically using specific enzymes. The individual products are extracted with polar solvent system, in this case, ethyl acetate. Individual products are purified by reverse-phase HPLC and stored under argon prior to packaging and shipping.

5-Hydro PeroxyEicosatetraenoic Acid (5-HPETE) is the immediate product of 5-LOX reaction, which is the precursor molecule for all the down-stream lipid mediators of 5-LOX pathway. 5-HPETE is extracted by Hexane: Ether solvent system and purified by straight-phase HPLC and the purified product is converted to 5, 6-Leukotriene A4 (5, 6-LTA4) by a biomimetic method developed in Dr. Channa Reddy’s laboratory in early 80s. 5, 6- LTA4 is then converted to individual 5-LOX pathway lipid mediators using specific enzymes. Similarly 15-LOX and 12-LOX enzymes will be used to manufacture 15- and 12-LOX pathway lipid mediators. The individual LOX pathway lipid mediators will be purified by HPLC and stored under argon at -80° until they are processed for packaging and shipping.