' Corpora non agunt nisi fixata ‘ „a drug will not work unless it is bound ” (Ehrlich)

1. 'Corpora non agunt nisi fixata‘ „a drug will not work unless it is bound” (Ehrlich)

2. Pharmacodynamics Mechanism of drug action Dependent and structure – independent drug action Definition and classification of receptors Quantitative aspects of drug receptor interaction

3. Dependent and structure – independent drug action • Drug effects are produced by altering the normal functions of cells and tissues in the body via one of four general mechanisms: • Interaction with receptors (1) • Alteration of the activity of enzymes(2) • Antimetabolite action (3) • Nonspecific chemical or physical interactions (4)

4. Interaction with receptors (1) Drug-Receptor Interactions

5. Conformation and Chemistry of Drugs and Receptors • Why is it that one drug affects cardiac function and another alters water and ion balance in the kidney? • Why does ciprofloxacin effectively kill bacteria but rarely harms a patient?

6. Conformation and Chemistry of Drugs and Receptors • These questions can be answered by first examining the interaction between a drug and its specific molecular target.

7. Conformation and Chemistry of Drugs and Receptors • Most drugs achieve their therapeutic effects by interacting selectively with target molecules that play important roles in physiologic or pathophysiologic functioning. • In many cases, selectivity of drug binding to receptors also determines the adverse effects of a drug.

8. Conformation and Chemistry of Drugs and Receptors • In general, drugs are molecules that interact with specific molecular components of an organism tocause biochemical and physiologic changes within that organism. • Drug receptors are macromolecules that, upon binding to a drug, mediate those biochemical and physiologic changes.

9. Conformation and Chemistry of Drugs and Receptors • The site on the receptor at which the drug binds is called its binding site. • Each drug-binding site has unique chemical characteristics that are determined by the specific properties of the amino acids that make up the site. • The three-dimensional structure of the drug determine the orientation of the drug, with respect to the receptor, and govern how tightly these molecules bind to one another.

10. Conformation and Chemistry of Drugs and Receptors • Drug–receptor binding is the result of multiple chemical interactions between the two molecules, some of which are weak (such as van der Waals' forces) and some of which are extremely strong (such as covalent bonding). • The sum total of these interactions provides the specificity of the drug–receptor interaction. • The favorability of a drug–receptor interaction is referred to as the affinity of the drug for its binding site on the receptor.

11. Conformation and Chemistry of Drugs and Receptors • A typical drug–receptor interaction may consist of 10 or more van der Waals' interactions and a few hydrogen bonds; ionic interactions. • For example, imatinib forms many van der Waals' interactions and hydrogen bonds with the ATP-binding site of the BCR-Abl tyrosine kinase. The sum total of these forces creates a strong (high affinity) interaction between this drug and its receptor.

12. Conformation and Chemistry of Drugs and Receptors • Covalent interactions between a drug and its receptor are relatively rare. • Covalent bond is often irreversible, and the drug and receptor form an inactive complex. • To regain activity, the cell must synthesize a new receptor molecule to replace the inactivated protein; and the drug molecule, which is also part of the inactive complex, is not available to inhibit other receptor molecules. • Drugs that modify their target receptors (often enzymes) through this mechanism are sometimes called suicide substrates.

13. Impact of Drug Binding on the Receptor • How does drug binding produce a biochemical and/or physiologic change in the organism? • In the case of receptors with enzymatic activity, the binding site of the drug is often the active site at which an enzymatic transformation is catalyzed. • Therefore, the catalytic activity of the enzyme is inhibited by drugs that prevent substrate binding to the site or that covalently modify the site.

14. Membrane Effects on Drug–Receptor Interactions • Drugs that are highly water-soluble are often less able to pass through the plasma membrane and bind to target in the cytoplasm. In contrast, certain hydrophilic drugs that are able to pass through transmembrane channels or use other transport mechanisms can gain ready access to cytoplasmic receptors. • Drugs that are highly lipophilic (such as many steroid hormones) are able to pass through the plasma membrane, without special channels or transporters.

15. Membrane Effects on Drug–Receptor Interactions • Many cell surface protein receptors have extracellular domains that are linked to intracellular effector molecules through receptor domains that span the plasma membrane. • Changing the shape of the extracellular domain can alter the conformation of the intracellular domains of the receptor, resulting in a change in receptor function. • In other cases, drugs can cross link the extracellular domains of two receptor molecules, forming a dimeric receptor complex that activates effector molecules inside the cell.

16. Membrane Effects on Drug–Receptor Interactions • All of these factors • drug and receptor structure, • the chemical forces influencing drug–receptor interaction, • drug solubility in water and in the plasma membrane, and • the function of the receptor in its cellular environment • confer significant specificity on the interactions between drugs and their target receptors.

17. Molecular and Cellular Determinants of Drug Selectivity • The ideal drug would interact only with a molecular target that causes the desired therapeutic effectbut not with molecular targets that cause unwanted adverse effects. Although no such drug has yet been discovered (i.e., all drugs currently in clinical use have the potential to cause adverse effects as well as therapeutic effects). • Selectivity of drug action can be conferred by at least two classes of mechanisms, including • (1) the cell-type specificity of receptor subtypes and • (2) the cell-type specificity of receptor-effector coupling.

18. Molecular and Cellular Determinants of Drug Selectivity • Drugs that target DNA synthesis, are likely to cause significant toxic side effects; this is the case with many currently available chemotherapeutics for the treatment of cancer. • Other drugs that target cell-type restricted processes, such as acid generation in the stomach, may have fewer adverse effects. • Imatinib is an extremely selective drug because the BCR-Abl protein is not expressed in normal (noncancerous) cells. • In general, the more restricted the cell-type distribution of the receptor targeted by a particular drug, the more selective the drug is likely to be.

19. Molecular and Cellular Determinants of Drug Selectivity • Voltage-gated calcium channels are expressed in the heart, cardiac pacemaker cells are relatively more sensitive to the effects of calcium channel blocking agents than are cardiac ventricular muscle cells. • In general, the more the receptor–effector coupling mechanisms differ among the various cell types that express a particular molecular target for a drug, the more selective the drug is likely to be.

20. Molecular and Cellular Determinants of Drug Selectivity • Agonistsare molecules that binding to their targets, cause a change in the activity of those targets. • Full agonists bind to and activate their targets to the maximal extent possible. For example, acetylcholine binds to the nicotinic acetylcholine receptor and induces a conformational change in the receptor-associated ion channel from a nonconducting to a fully conducting state. • Partial agonists produce a submaximal response upon binding to their targets. • Inverse agonists cause constitutively active targets to become inactive.

21. Molecular and Cellular Determinants of Drug Selectivity • Antagonistsinhibit the ability of their targets to be activated (or inactivated) by physiologic or pharmacologic agonists. • Drugs that directly block the binding site of a physiologic agonist are called competitive antagonists; • drugs that bind to other sites on the target molecule, and prevent the conformational change required for receptor activation (or inactivation), may be either • noncompetitive (allosteric) or • /uncompetitive antagonists (they require receptor activation by an agonist before they can bind to a separate allosteric binding site)/.

22. Major Types of Drug Receptors • Given the great diversity of drug molecules, it might seem likely that the interactions between drugs and their molecular targets would be equally diverse. This is only partly true. • In fact, most of the currently understood drug–receptor interactions can be classified into major groups.

23. (1) Interaction with receptors (Major Types of Drug Receptors) -Ligand-gated ion channels (1/1) -G-protein–coupled receptors (1/2) -Receptor-activated tyrosine kinases (1/3) -Intracellular nuclear receptors (1/4)

24. (1) Interaction with receptors (Major Types of Drug Receptors) • A. Drugs can bind to ion channels, causing an alteration in the channel's conductance. • B. Heptahelical receptors spanning the plasma membrane are functionally coupled to intracellular G proteins. • C. Drugs can bind to the extracellular domain of a transmembrane receptor and cause a change in signaling within the cell by activating or inhibiting an enzymatic intracellular domain of the same receptor molecule. • D. Drugs can diffuse through the plasma membrane and bind to cytoplasmic or nuclear receptors.

25. (1/1) Ligand-gated ion channels Acetylcholine interacts with a nicotinic receptor that is a nonspecific Na+/K+ transmembrane ion channel.

26. Ligand-gated nicotinic acetylcholine receptor • A. The plasma membrane acetylcholine (ACh) receptor is composed of five subunits—two α subunits, a β subunit, a γ subunit, and a δ subunit. • B. In the absence of ACh, the receptor gate is closed, and cations [most importantly, sodium ions (Na+)] are unable to traverse the channel. • C. When ACh is bound to both α subunits, the channel opens, and sodium can pass down its concentration gradient into the cell.

27. Ligand-gated nicotinic acetylcholine receptor Nicotinic receptors are localizedat -the motor endplate of the myoneural (neuromuscular) junctions of somatic nerves and skeletal muscle (NM), -autonomic ganglia (NG), including the adrenal medulla, and -certain areas in the brain.

28. Ligand-gated ion channels Skeletal muscle - ACh interacts with nicotinic receptors - open channels - permit passage of ions, mostly Na+ - Na+ current - membrane depolarization - resulting in the release of Ca2+ - muscle contraction - hydrolysis of ACh by AChE results in muscle cell repolarization

29. Transmembrane Ion Channels • nicotinic ACh receptor - only two states, open or closed • many ion channels are able to assume other states: • voltage-gated sodium channels are able to become refractory or inactivated. In this state, the channel's permeability cannot be altered for a certain period of time. During the inactivation (refractory) period, the channel cannot be reactivated for a number of milliseconds, even if the membrane potential returns to a voltage that normally stimulates the channel to open. • This state-dependent binding is important in the mechanism of action of some local anesthetic.

30. Transmembrane Ion Channels • Two important classes of drugs that act by altering the conductance of ion channels are the local anesthetics and the benzodiazepines. • Local anesthetics block the conductance of sodiumions through voltage-gated sodium channels in neurons that transmit pain information from the periphery to the central nervous system, preventing action potential propagation and pain perception (nociception). • Benzodiazepines inhibit neurotransmission in the central nervous system by potentiating the ability of the neurotransmitter gamma aminobutyric acid (GABA) to increase the conductance of chloride ions across neuronal membranes, preventing action potential propagation and pain perception (nociception).

31. (2/1) G-protein–coupled receptors The biologic activity of the receptors is mediated via interaction with a number of G (GTP binding)-proteins. Gαs(Gαstimulatory)-coupled receptors (2/1/1) Gαi (Ginhibitory)-coupled receptors (2/1/2) Gq (and G11)-coupled receptors (2/1/3)

32. G-protein–coupled receptors Epinephrin (adrenalin) binds its receptor, that associates with an heterotrimeric G protein. The G protein associates with adenylate cyclase that converts ATP to cAMP, spreading the signal.

33. (2/1/1) Gαs-coupled receptors β-adrenoceptor, which when activated by ligand binding (e.g., epinephrine) exchanges GDP for GTP. This facilitates the migration of Gαs (Gαstimulatory) and its interaction with adenylyl cyclase (AC). Gαs-bound AC catalyzes the production of cAMP from ATP; cAMP activates protein kinase A, which subsequently acts to phosphorylate and activate a number of effector proteins. The γ dimer may also activate some effectors. Hydrolysis of the GTP bound to the Gα to GDP terminates the signal.

34. Gαs-coupled receptors Histamine (H2)-receptors: - H2-receptors are found in the brain, heart, vascular smooth muscles, leukocytes, and parietal cells - response of H2-receptors is coupled via Gαs - activation of H2-receptors increases gastric acid production, causes vasodilation, and generally relaxes smooth muscles

35. Gαs-coupled receptors • β-Adrenoceptors • - located in postjunctional effector cells • - β1-receptors (primarily excitatory) • - β2-receptors (primarily inhibitory) • β1-receptors mediate increased contractility (cardiac muscle) and heart rate (SA node of heart), fat cell lipolysis • β2-receptors mediate vasodilation and intestinal, bronchial, and uterine smooth muscle relaxation

36. (2/1/2) Gαi (Ginhibitory)-coupled receptors Ligand binding (e.g., somatostatin), to Gαi (Gαinhibitory)-coupled receptors, similarly exchanges GTP for GDP, but Gαi inhibits adenylyl cyclase, leading to reduced cAMP production.

37. Gαi (Ginhibitory)-coupled receptors α2-Adrenoceptors - α2-receptors are located primarily in prejunctional adrenergic nerve terminals - prejunctional inhibition of release of norepinephrine and other neurotransmitters (α2) - α2-Receptorsactivate (Gi) (like muscarinic M2-cholinoceptors)

38. (2/1/3) Gq (and G11)-coupled receptors Gq (and G11) interact with ligand (e.g., serotonin)-activated receptors and increase the activity of phospholipase C (PLC). PLC cleaves the membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PIP2) to diacylglycerol (DAG) and inositol 1,4,5-triphosphate (IP3). DAG activates protein kinase C, which can subsequently phosphorylate and activate a number of cellular proteins; IP3 causes the release of Ca2+ from the endoplasmic reticulum into the cytoplasm, where it can activate many cellular processes.

39. Gq (and G11)-coupled receptors Histamine (H1)-receptors: - H1-receptors are found in the brain, heart, bronchi, gastrointestinal tract, vascular smooth muscles, and leukocytes. - H1-receptors activation causes an increase in diacylglycerol and intracellular Ca2. - Activation of H1-receptors in the brain increases wakefulness. - Activation of H1-receptors in vessels causes vasodilation and an increase in permeability.

40. Gq (and G11)-coupled receptors α1-Adrenoceptors - α1-receptors are located in postjunctional effector cells, vascular smooth muscle (mainly excitatory) - α-adrenoceptors mediate vasoconstriction (α1), gastrointestinal relaxation (α1), mydriasis (α1) - α1-receptorsactivate (Gq) (like muscarinic M1 and M3 cholinoceptors)

41. (1/3) Receptor-activated tyrosine kinases Many growth-related signals (e.g., insulin) are mediated via membrane receptors that possess intrinsic tyrosine kinase activity. Ligand binding causes conformational changes in the receptor. The liganded receptors then autophosphorylate tyrosine, and activated.

42. (1/4) Intracellular nuclear receptors Ligands (e.g., cortisol) for nuclear receptors are lipophilic and can diffuse rapidly through the plasma membrane. In the absence of ligand, nuclear receptors are inactive because of their interaction with chaperone proteins such as heat-shock proteins like HSP-90. Binding of ligand promotes structural changes in the receptor that facilitate dissociation of chaperones, entry of receptors into the nucleus, hetero- or homodimerization of receptors, and high-affinity interaction with the DNA of target genes.

43. Cellular Regulation of Drug–Receptor Interactions • Mechanisms that mediate drug-induced activation or inhibition are important because they prevent overstimulation that could lead to cellular damage or adversely affect the organism as a whole. Many drugs show diminishing effects over time; this phenomenon is calledtachyphylaxis. In pharmacologic terms, the receptor and the cell become desensitized to the action of the drug. • Mechanisms of desensitization can be divided into two types: • homologous, in which the effects of agonists at only one type of receptor are diminished; and • heterologous, in which the effects of agonists at two or more types of receptors are coordinately diminished.

44. Cellular Regulation of Drug–Receptor Interactions • β-adrenergic receptor desensitization is mediated by epinephrine-induced phosphorylationof the cytoplasmic tail of the receptor. • This phosphorylation promotes the binding of β-arrestin to the receptor; in turn, β-arrestin inhibits the receptor's ability to stimulate the G protein Gs. With lower levels of activated Gs present, adenylyl cyclase produces less cAMP. In this manner, repeated cycles of ligand–receptor binding result in smaller and smaller cellular effects. • Other molecular mechanisms have even more profound effects, completely turning off the receptor to stimulation by ligand. The latter phenomenon, referred to as inactivation, may also result from phosphorylation of the receptor; in this case, the phosphorylation completely blocks the signaling activity of the receptor or causes removal of the receptor from the cell surface.

45. β-Adrenergic receptor regulation • A. Repeated or persistent stimulation of the receptor by agonist results in phosphorylation of amino acids at the C-terminus of the receptor, thereby decreasing adenylyl cyclase (effector) activity. • B. Binding of β-arrestin also leads to receptor sequestration. The receptor can then be recycled and reinserted into the plasma membrane. • C. Prolonged receptor occupation by agonist can lead to receptor down-regulation and eventual receptor degradation. Cells can also reduce the number of surface receptors.

46. Cellular Regulation of Drug–Receptor Interactions • Another mechanism that can affect the cellular response caused by drug–receptor binding is called refractoriness. Receptors that assume a refractory state following activation require a period of time to pass before they can be stimulated again. As noted previously, voltage-gated sodium channels, which mediate the firing of neuronal action potentials, are subject to refractory periods. Following channel opening induced by membrane depolarization, the voltage-gated sodium channel spontaneously closes and cannot be reopened for some period of time (called the refractory period). This inherent property of the channel determines the maximum rate at which neurons can be stimulated and transmit information.

47. Cellular Regulation of Drug–Receptor Interactions • The effect of drug–receptor binding can also be influenced by drug-induced changes in the number of receptors on or in a cell. One example of a molecular mechanism by which receptor number can be altered is called down-regulation. In this phenomenon, prolonged receptor stimulation by ligand induces the cell to endocytose and sequester receptors in endocytic vesicles, resulting in cellular desensitization. When the stimulus that caused the receptor sequestration subsides, the receptors can be recycled to the cell surface and thereby rendered functional again. • Cells also have the ability to alter the level of synthesis of receptors and thereby to regulate the number of receptors available for drug binding.

48. idáig Mechanisms of Receptor Regulation

49. Drug effects (2) Alteration of the activity of enzymes Neostigmine and physostigmine are indirect-acting parasympathomimetic agents inhibit AChE and increase ACh levels at both muscarinic and nicotinic cholinoceptors.

50. Drug effects (3) Antimetabolite action • Antimetabolites are S-phase—specific drugs that are structural analogues of essential metabolites and that interfere with DNA synthesis. CytarabineFluorouracilMercaptopurineThioguanine