Pharmacology Pharmacology of Recombinant or Genetically Engineered Drugs Kamal Kishore, Pawan Krishan1 Departments of Pharmacy, M.J.P. Rohilkhand University, Bareilly-243 006, Uttar Pradesh, 1Pharmaceutical Sciences and Drug Research, Punjabi University, Patiala-147 002, Punjab, India Address for correspondence: Dr. Kamal Kishore; E-mail: [email protected] ABSTRACT Recombinant technology or genetic engineering is a modern method used for the synthesis of therapeutic agents. The central theme of recombinant technology is the process of “gene cloning” which consists of the production of a deÞ ned fragment of DNA and its propagation and ampliÞ cation in a suitable host cell. Drugs developed by recombinant technology or genetic engineering are known as biologics, biopharmaceuticals, recombinant DNA expressed products, bioengineered, or genetically engineered drugs. A current list of various products developed by recombinant technology includes erythropoietin, coagulation modulators, enzymes, hormones, interferons, interleukins, granulocyte colony-stimulating factors, anti-rheumatoid drugs, and various other agents like TNF, becaplermin, hepatitis-B vaccine, antibodies etc. This article provides general as well as recent pharmacological information on different aspects of recombinant drugs that may be useful for their better understanding by users and health care professionals. Key words: Biologics, erythropoietin, interferon, interleukins, insulin, thrombolytic enzymes DOI: 10.4103/0975-1483.55747 INTRODUCTION fragment of DNA and its propagation and ampliÞ cation in a suitable host cell. Recombinant technology was only Drugs developed using living organisms with the help of possible after the discovery of restriction endonucleases, biotechnology or genetic engineering are known as biologics, the enzymes used as cutters for a desired segment[3] of biopharmaceuticals, recombinant DNA expressed genes known as recognition sequences. There are various products, bioengineered, or genetically engineered drugs. other enzymes that have great value in recombinant The recombinant technique was developed by Cohen in technology such as DNA polymerases, ligases, kinases, 1973.[1] The general procedure starts from the identiÞ cation alkaline phosphatases, and nucleases.[4] For thousands of of the gene responsible for the production of the desired years, living microorganisms have been used to produce product. The gene is isolated from human cells and gases, leaven bread, ferment alcoholic liquids, and for other inserted into other carrier or vector cells like bacteria desired by-products, but their use in recombinant DNA (Escherichia coli) or yeast (Saccharomyces cerevisiae, Hansenulla technology is often described as a relatively new science.[5] polymorpha, Pichia pastoris) that proliferate and produce large Recombinant technology, thus, is a new method for the amounts of the desired product.[2] The central theme of development of drugs and other life-saving products that recombinant DNA technology is the process of “gene involves the blending of discoveries in molecular cloning” which consists of the production of a deÞ ned biology, DNA alteration, gene splicing, immunology, J Young Pharm Vol 1 / No 2 141 Kishore and Krishan. J Young Pharm. 2009;1(2):141-150 and immunopharmacology. Now a days, various human CLASSIFICATION OF RECOMBINANT PRODUCTS diseases are treated with the help of drugs developed by recombinant technology, such as erythropoietin, The following products or biologics are obtained by coagulation modulators, enzymes, hormones, interferons, recombinant or genetic engineering technology: interleukins, granulocyte colony-stimulating factors, anti-rheumatoid drugs, and various other agents like (I) Recombinant erythropoietin Tumor necrosis factor (TNF), becaplermin, hepatitis-B Epoetin-α vaccine, antibodies etc. This article provides general as Epoetin-β well as recent pharmacological information on different Epoetin-ω aspects like a brief description of recombinant products, Darbepoetin-α pharmacological actions, mechanism of action, adverse (II) Recombinant blood coagulation modulators effects, doses, contraindications, interactions and (a) Coagulants therapeutic applications. The pharmacological information Factor-VII A of bio-engineered products may be useful for better Factor-VIII understanding by users and professionals in the medical Factor-IX and health-related sciences. (b) Anticoagulants Lepirudin Desirudin MANUFACTURING PROCESS FOR RECOMBINANT Drotecogin alfa PRODUCTS (III) Recombinant enzymes (a) Thrombolytic enzymes The following steps are involved in the production of [4] Alteplase recombinant products: Anistreplase Duteplase Extraction of the DNA from a donor organism and the Reteplase plasmid from bacteria Saruplase ↓ Streptase or streptokinase Cleaved with restriction endonucleases Tenecteplase ↓ Tissue plasminogen activator Joined to form a recombinant DNA molecule or vector Urokinase ↓ (b) Therapeutic enzymes Insert this vector into host cell or expression system Agalsidase beta (E. coli or Yeast) Aeromonas aminopeptidase ↓ Alpha-glactosidase-A Multiplication of recombinant DNA molecules Dornase alpha Imiglucerase or glucocerebrosidase or ↓ alglucerase Formation of clone Laronidase or alpha-L-iduronidase ↓ Galsulfase or N-acetylgalactosamine-4-sulfatase Scale-up (harvesting or fermentation) of cell to produce (IV) Recombinant hormones recombinant drugs (a) Insulin and its analogs ↓ Human insulin PuriÞ cation (centrifugation or Þ ltration) Insulin aspart ↓ Insulin glargine Preformulation Insulin lispro Insulin glulisine ↓ Insulin-like growth factor-I Preclinical studies (animal testing) Insulin-like growth factor-II ↓ (b) Growth hormone and its analogs Clinical studies (human testing) Growth hormone ↓ Growth hormone (ovine) Final product Growth hormone binding protein 142 J Young Pharm Vol 1 / No 2 Pharmacology of recombinant drugs Growth hormone-20K Recombinant albumin Somatrem Fc puriÞ ed recombinant protein Somatropin Becaplermin Sermorelin Beta-nerve growth factor (c) Gonadotropins Bone morphogenetic protein-2 Recombinant follitropin-α Brain-derived neurotrophic factor Recombinant follitropin-β Ciliary neurotrophic factor Recombinant choriogonadotropin Chymosin Follicle-stimulating hormone Endostatin human Luteinizing hormone Enteropeptidase/Enterokinase (d) Other hormones Epidermal growth factors Calcitonin Fibroblast growth factor Glucagon Hepatitis-B vaccine (V) Recombinant interferons Hepatocyte growth factor Interferon alfa-2a Inß uenza A, nucleoprotein, recombinant protein Interferon alfa-2b Lung surfactant protein Interferon beta-1a Relaxin Interferon beta-1b TNF Interferon-gamma-1a Recombinant vaccines Interferon-gamma-1b α-antitrypsin Interferons-ω Thrombopoietin Consensus interferon PHARMACOLOGY OF INDIVIDUAL PRODUCTS (VI) Recombinant interleukins Interleukin-1 alpha Recombinant erythropoietin Interleukin-1 beta [6] Interleukin-2 (aldesleukin) Erythropoietin is a 165 amino acid glycoprotein hormone Interleukin-3 that is synthesized naturally in the kidney and the liver but Interleukin-4 is now prepared by recombinant technology. It stimulates Interleukin-10 erythrocyte formation and is available as epoetin-α, Interleukin-11 (oprelvekin) epoetin-β, and darbepoetin-α. Mechanism of action: It Interleukin-13 stimulates proliferation and differentiation of erythrocytes. Interleukin-15 Dose: 5-100 units/ kg/ s.c. or i.v./three times a week. Interleukin-17 Adverse effects: Allergic reaction, deÞ ciency of iron and folic acid, disorientation, encephalopathy, ß u-like symptoms, Interleukin-18 headache, mild hypertension, seizures, and thrombosis. (VII) Recombinant granulocyte colony stimulating Uses: It is preferred in cases of anemia due to AIDS, cáncer factors chemotherapy, chronic renal failure, and anemia in premature Filgrastim babies. It is also useful in hematopoietic disorders and is used Lenograstim to increase the beneÞ ts of autologous blood transfusión. Molgramostim Sargramostim Recombinant blood coagulation modulators Regrasmostim PegÞ lgrastim Coagulants[7] (VIII) Recombinant anti-rheumatoid agents Factor-VII A is a unique hemostatic agent containing Inß iximab 406 amino acids[8,9] that is supposed to activate the extrinsic Adalimumab pathway of the coagulation cascade.[10] Dose: 150- 500 μg/ kg Basiliximab of body weight. Adverse effects: Factor VIIa may cause Daclizumab adverse effects such as pyrexia, hemorrhage, injection Anakinra site reactions, arthralgia, headache, hypertension, nausea, Etanercept vomiting, pain, edema, and rashes. Uses: It is preferred (IX) Miscellaneous agents in cases of hemophilia-A and B, and in patients with Recombinant antibodies congenital and acquired bleeding abnormalities. J Young Pharm Vol 1 / No 2 143 Kishore and Krishan. J Young Pharm. 2009;1(2):141-150 Factor-VIII is an endogenous glycoprotein coagulating in bleeding, defective homeostasis, trauma, surgical factor that is necessary for blood clotting and hemostasis. procedures, stroke, acute pericarditis, hypoglycemia and It is a cofactor essential for factor IX to activate factor X in hyperglycemia. Dose: 100 mg/i.v. infusion. Adverse effects: the intrinsic pathway. It is a 1438 amino acid peptide that is Alteplase may cause adverse effects like nausea, vomiting, produced in CHO cells. DeÞ ciency of factor VIII causes fever, arrhythmias, allergy, hypotension, intracranial a common inherited bleeding disorder known as hemorrhage, and GIT bleeding. Uses: It is preferred in [11,12] hemophilia-A.