Lecture №18

1. Lecture №18 Heterocyclic antibiotics: aminoglycosides, macrolides. Polypeptide antibiotics, polyenesandantitumorantibiotics. Fluoroquinolonesasantibioticdrugs. prepared ass. Kozachok S.S.

2. Aminoglycosidesdivided on the following groups: • Strepyomycin (separately, NH2- group is substituted); • Aminoglycosides(kanamycin, neomycin, gentamycin, monomycin, amikacin); • Macrolides (erythromycin, oleandomycin, roxitromycin, claritromicin, dirythromycin, spiramycin, azitromycin); • Ansamycin(rifampicin)

3. Strepyomycinsulfate (Streptomycini sulfas) Streptomycin sulfate* (UP) Bis-[N,N’-bis(aminoiminomethyl)-4-O-[5-deoxy-2-O-[2-deoxy-2-methylamino)-α-L-glucopyranosyl]-3-С-formyl- α-L -lyxofuranosyl]-D-streptamine] trisulfate

4. Streptomycin, discovered in 1944 by the American scientist Waxman. • Obtaining. Microbiological synthesis from Streptomyces griseus actinomycetes. • Streptomycin glycoside consists ofthe aglycone- streptydin (1,3-diguanidino-2,4,5,6-tetraoxicyclohexane) andsugar part-disaccharide streptobiozamine(N-methyl-L-glucosamineandL-streptose).

5. CHARACTERISTICS • White powder. Hygroscopic . Very soluble in water, practically insoluble in ethanol and ether. • Streptomycin has a basic property according to the appearance of the nitrogen containing group (twoguanidins andoneN-methylamino)that’s why easy creates salts. • In a weak acidic medium streptomycin is stable, but in strong acidic and especially in basic medium it easy hydrolyzes on a streptydin and streptobiozamine, which decomposes onN-methyl-L-glucosamine andL-streptos.

6. Identification • TLC. • Maltol testis caused by the ability of the streptose in a basic medium to convert on a maltol according to the dehydratation and isomerization. At the iron (III) ions interaction in an acidic medium maltol forms the violet products: Maltol (α-methyl-β-oxy-γ-piron)

7. Sakaguchi reaction on the remnants of guanidines. Formation of a violet-red colour, which appearances in a basic medium under the action of -naphthol and concentrated sodium hypochlorite. • Molish reaction on carbohydrates (streptos). After acidic hydrolysis the substance does not give reaction with -naphthol and concentrated sodium hypochlorite in a basic solution. Yellow colour. • It gives the reaction of sulfate.

8. Un pharmacopoeia reaction : • Escaping of ammonia by heating of the substance with sodium hydroxide (guanidine residues); • Weber Reagent (it is oxidized by sodium nitroprussideNa[Fe(CN)5NO]+K3[Fe(CN)6]+NaOH) – red colour (guanidine residues); • Brown colour with basic potassium tetraiodomerrcurate (Nernst reagent) and red sediment with copper-tartrate reagent– Feling reagent; “silver mirror” reaction (on aldehyde group); • Condensation with phenols in the presence of concentrated sulfuric acid – aurine dye; • On the aldehyde group of streptos (with a hydrazine, a semicarbazide – white sediments).

10. Aurine dye red colour

11. Assay • Microbiological method (UP). • Photocolorymetry which is based on the Maltol test. • Cerimetry. Titrant – Cerium sulfate. Indicator – iron (III) chloride. Em = М. m./2

12. Storage In the dry place. Application Broad-spectrum antibiotic. Antituberculosis drug. At the treatment of pneumonia, peritonitis, , gonorrhea, brucellosis. Produced in bottles, sealed with rubber stoppers with crimped aluminum caps on 0,25, 0,5 and 1,0 of the active substance in terms of streptomycin-base. Inject inner muscular 0,5-1,0 gon a day. Side effects. Kidney-and oto-toxic, respiratory depression, is rapidly developing resistance (2-3 days).

13. Aminoglycosides The general formula Characteristic. According to the physical properties aminoglycosides antibiotics are powders of white, yellow or creams colors, hydroscopic. Freely soluble in water and practically insoluble or low soluble in the most organic solvents. Opticallyactive.

14. Kanamycinmonosulfate (UP) (Kanamycini monosulfas)

15. Composition of Kanamycin is aglucone 2-deoxystreptomine (meso-1,3-diamino-4,5,6-trioxycyclohexan) and two saccharates - 3-amino-3-deoxy-D-glucoseand 6-аmino-6-deoxy-D-glucose. • Drug is obtained by the fermentation ofStreptomyces kanamycetis at 27-29 оС and рН=7, the environment that contains starch and soy flour. From the environment Kanamycin A, B and C is extracted by an ionchangers.The list toxic Kanamycin A has, that’s why it composes the mass of the drug Kanamycin monosulfate (94 %). • The semi-synthetic analog of Kanamycin is amikacin sulfate. Its main difference from kanamycin is a present of 4-аmino-L-2-oxybtyl radical. Except aglucone 2- deoxy-D-streptamine amikacin has 2-deoxy-L-streptamine.

16. Amykacinsulfate(Amyсаcinisulfas)

17. Neomycinlike Kanamycin is a mixture of two stereoisomers В and С. Aglucone – 2-deoxystreptamine ties three saccharides. • The drug is obtained from the liquid Streptomyces fradiae culture by the carboxylic cation exchange resins. In 1949 neomycin was the first obtained by Waxman and le Shatelye. • Monomycin sulfate drug is a mixture of monomycins that are produced by Actinomyces circulatus var. monomicini. The difference of the monomycin chemical structure from neomycin is a present in 6’ position – СН2ОН group (paramosa) exept – CH2NH2. • Gentamycin is similar to kanamycin. It is a metabolizing product ofMicromonospora purpurea.It is a mixture of:

18. Neomycin sulfate(Neomycini sulfas)

19. Monomycin sulfate(Monomycinisulfas)

20. Gentamycin sulfate(Gentamycinisulfas)

21. Aminoglycoside’s identification • Physical constants: melting temperature, UV- and ProtonNuclearMagnetic Resonanse-spectroscopy, TLC, Liquid chromatography. • Reaction on the aliphatic amino-group of Kanamycinmonosulfate – at the heating with a ninhydrin solution observed violet color. • Melting temperature of Kanamycinpicrate. • They give reaction of sulfate. • Bial’s reaction (pentose opening). For the identification of Kanamycinmonosulfate and neomycin sulfate using a color reaction with a alcohol solution of an orcyn (methylresorcinol) andconcentratedhydrochloricacidatthepresentofiron (III) chloride. Solution gets green color.

23. After acidic hydrolysis kanamycin and amikacin give the reaction of the reducing saccharides (Feling reagent. Nernst, Tolens). • Amykacin according to the amide group forms color complexes with the hard metal’s salts (reaction with cobalt nitrate after sodium hydroxide neutralization– violet). • Amikacin at the heating with concentrated mineral acids forms from the saccharides 5-aminomethylfurfural, which gives the colour reaction with antron:

24. Assay • Microbiological method (UP). • Polarimetry (gentamycin sulfate). • Photocolorimetry. • Absorption spectroscopy inUV- and visible region. Storage Protection from light. Application Broad spectrum antibiotics. Forthetreatmentofthegastrointestinaltractdiseases, tuberculosisandinfectiousskindiseases, sepsis, urinarytractinfections.

25. Kanamycinmonosulfate – bottleon 0,5 and 1,0 g. Inner muscular injection 5-7 days. H.d.d. – 2 g Gentamycin sulfate – Solution for injection.: exports producing. 40 or 80 mg 2,0 ml №10; national (Ukrainian) producing 4% 1,0 or 2,0 ml №10; cream 0,1% 15 г; eye oinment 0,3% 5,0 g. Average day’s dose – 3 mg/kg, 2-3 times, inner muscular injection. Amykacin – КМ – bottleon 0,1 and 0,25 g. Average day’s dose 15 mg/kg by 2 times inner muscular injections. Tergynan– vaginaltablets(ternidazole, neomycin sulfate, nystatin, prednisolone). Side effects – Kidney-and oto-toxicaction, allergyand others.

26. Lincomycins In medical practical using Lincomycin hydrochloride and clindamycin Lincomycin hydrochloride(Lyncomycini hydrochloridum) (UP) Monohydratemethyl-6,8-dideoxy- 6-[(2S,4R)-1-ethyl-4-propyl- pyrrolidinyl-2-carboxamido]- 1-thio-D-erythro-α-D-galacto- octopyranosidehydrochloride

27. Lincomycin hydrochloride–used at sepsis, staphylococ infections, acute and chronic osteomyelitis, purulent skin infections, otitis. Externally at purulent diseases of skin and soft tissue. Intravenous, inner muscular injection. Once dose - 0,5 g, day dose – 1,5 g. Capsules on 0,25 g №20; solution for injection 30% 1,0 or 2,0 № 10. Contraindications– severe liver and kidney disease, myasthenia gravis. Clindamycin (Dalacin) – broad-spectrum ANT. Capsule on 150 and 300 mg № 16; solution for injection 150 mg/ml 2,0 or 4,0 № 1; vaginal cream and suppositories.

28. Macrolides antibiotics They contain a lactone ring with 12-17 carbon atoms, it ties with amino carbohydrates (for the amino glycosides type) and neutral sugars. Reserve antibiotics. Modern classification • 14 – members: natural (erythromycin, oleandomycin); prodrug(estherand erythromycin salts, oleandomycinesthers); semi-synthetic (roxithromycin, clarithromycin, dirithromycin, josamycin, its active metabolite is 14-hydroxi clarithromycin). • 15 – members: semi-synthetic (additional nitrogen atom – azalides - azithromycin (sumamed) • 16 – members: natural (spiramycin (rovamycin), midecamycin (macropen), semi-synthetic (midecamycinacetate (miocamycin).

29. It is now known about 100 macrolide antibiotics, the general formula: In medical practice the most often used: Erythromycinphosphate (altrocin –S); Spiramycin (rovamycin, rovalen); Midecamycin(macropen); Roxithromycin (roxide, rulid, renicin, rovenal); Josamycin(wilprafen ); Clarithromycin (fromilid, klabax , clacid ); Azithromycin (Azitrox, Azithro, Zithromax , sumamed ).

30. Erythromycin(Erythromycinum)

31. Erythromycin is produced from a strain of the actinomycete Saccharopolyspora erythraea.. • Erythromycin consists of aglucone erythrolide (14-members lactone) and two sugars: cladynos (3-methyl-3-methoxy-2,6-didedeoxyhexopyranos) anddeamine (pycrocin, 3- dimethylamino-4,6- didedeoxyhexopyranos). Cladynossplits off at the interaction with eryehromycinwith 0,5 М НСl solution at the cold, and deamine – at the interaction with eryehromycinwith 6 М НСl solution. • Erythromycin – white crystalline substance bitter on taste, melting at 190-193 оС, soluble in water, freely soluble in ethanol, acetoneand chloroform. With concentrated H2SO4gives red-brown color, and with concentrated HCl – orange colorthat transfers into red after puple. • 1mgof Erythromycin equals 1000 U.A.

32. OleandomycinphosphateOleandomycini phosphas

33. Azithromycin (UP) (Azithromycinum) • Identification: IR-spectroscopy • Assay: Liquid chromotography

34. Erythromycin is available in enteric-coated tablets, slow-release capsules, oral suspensions, ophthalmic solutions, ointments, gels, and injections. • Erythromycin tablets on 100000 and 250000 U.A. are used for the treatment of pneumonia, scarlatina, angina,anthrax etc. For the local using at the infection wounds,burns, trophic exulcerates, trachoma, bedsores there is recommended an ointment Unguentum erythromycini, its 1 g contains 10000 U.A. • Other antibiotics-macrolides are produced as oral medicine forms. • Macrolide antibiotics are used by prescription, determine the sensitivity to this group and compared with a sensitivity to other groups as well as macrolides quickly develop resistance. If antibiotics- macrolides are not used continuously, the sensitivity of the microorganism to them restored.

35. Ansamycin’s antibiotics • Atthecoreofthestructureisaromaticring, coupledwithmacrocyclicaliphaticchain, called Anza-chain. Aliphaticchaindoesnotcontainthelactonebondsthataretypicalformacrolide’santibiotics, itattachestothecorewiththeamidenitrogenatom. • Thefollowingantibioticsbelong to ansamycin’s: rifampicin, streptovaricin, tolipomicin, galomicin, naphtomicinetc. • Rifampicin and its semi-synthetic analogs are used as medicines [3-(4-methyl-1-piperazinyliminomethyl)- rifampicin ], rifabutin, rifampitin and the combine drugs. • Ansamycin’s antibioticshave broud-spectrumactionand a highefficiency.Prescribedinthecaseswhereotherantibioticsareineffective. Usedforthetreatmentofallformsoftuberculosis, diseasesofgastrointestinaltractandpyogenicinfectionsonthedosesof 0,3-0,45 g. • MicroorganismsveryquickdeveloptheresistancetoRifampicin.

36. Rifampicin’s general formula and radicals

37. Polyenes antibiotics • Polyenes antibioticshave an antifungalactivity. Theyare mixturesofsubstancesthatareverycloseinstructure. Moleculeofeachcomponentconsistsoftheaglyconehavingmacrocyclicstructure, andaminosugars, connectedbyaglycosidicbond. Polyenestructureoftheaglyconehas6-7doublebondsand 35-40 carbonatoms. • In medical practice the following medicines used: nystatin, AmphotericinВ, levorin, trihmicin, candocidin, mycoheptin • , griseofulvin, amphoglucamin, etc. • Application. Forthetreatmentofcandidiasis, dermatomycoses, trichomoniasis, fungaldiseases. • The modern antifungal medicines – imidazole derivatives: Ketoconazole(1-2%), Oxiconazole(mifungar), Intraconazole(orungal); allylamine: Terbinafine(lamisil, terbisil, exifine); triazol derivatives : Fluconazole(diflucan, micosyst).

38. NystatinNystatinumtablets, ointment , suppository • Griseofulvin Griseofulvinum tablets, liniment

39. Nystatin (Nystatinum) (UP) • Identification: UV-, ІR-spectroscopy; Substance+ concentrated hydrochloric acid – brown color; Substance+ concentrated sulfuric acid – brown color that transfers into a violet; Liquid chromatography. • Assay: Microbiological method • For a substance that is designed to produce medicines for oral use must be withstanded the test on abnormal toxicity.

40. Amphotericin ВAmphotericinum Bamphomin, amphotret, fungizome, fungizone • Yellow or yellow-orange powder. Hygroscopic . Sensitive to the action of light and temperature. It is easy inactivated in acid and base medium. • Application - systemic and deep mycoses (blastomycosis, criptococoz), mold mycosis, chronic and granulomatous forms of candidiasis. Intravenous injection, inhalation and topical (cream). Used only in hospital settings: a highly toxic drug, capable to cumulation, kidney throtoxic, causesthe reducing of potassium quantity in a blood.

41. Polypeptides antibiotics • Polypeptides antibiotics according to their amino acids composition, chemical structure are different from other peptides (proteins). Amino acids associatein a cycle. • In medical practice the following medicines used: GramicidinS, Polymyxin M . • Storage. Protection from light. • Application. According to a high toxicty of polypeptides the are applied outside, at the internal using they cause hemolysisof erythrocytes. At the heavy septic and gastroenteric diseases (as enemas), when other antibiotics are uneffective, for washing of purulentwounds, ulcers, bedsores, rinse of throat. • GramicidinS has the spermocidic action (as paste). • Gramidin (tablet fro resorb) – GramicidinS1,5 mgand lidocaineh/ch 10 mg. • Sofradex (drops eye/ears) – gramicidin0,05 mg, neomycin 5,0 mg, Dexamethason0,5 mg.

42. GramicidinS(Gramicidin S) • GramicidinSisproducedfromabacterial strainoftheBacillus brevis, which are in the soil. The first it was obtained in USA.

43. Antineoplasticantibiotics • In 1940 actinomycinwas obtained by the americanscientist Waxman, and in 1952 there was determined the antineoplastic action of the substance. • Antitumorantibioticsthatareusedinmedicalpracticecanbedividedintoderivatives: tetracycline(doxorubicinhydrochloride); Aurelic acid (olivomycin); Anthracycline(rubomycin); Quinoline-5,8-dion (Вruneomycin). • For the analysis of these drugs physical, physical-chemical and chemical methods are used. • Storage. In a wellstopperedcontainerin a dryplace, protectedfromlightatroomtemperature.

44. Doxorubicinhydrochloride(UP)Doxorubicini hydrochloridumAdriblastin • Crystalline orange-red powder. Hygroscopic. • Identifiction: ІR-spectroscopy, gives the reaction of chloride ions. • Assay: Liquid chromatography

45. Olivomycin А

46. Rubomycinhydrochloride Вruneomycin

47. Olivomycin А– yellow powder with a green tint. Hygroscopic. Freely soluble in water, alcohol, practically insoluble in chloroform, ether. Storage as a toxic substance at below 20 оС. Using intravenous injection or local (oinmant). • Вruneomycin(streptonigrin) – brown crystallinepowder. Practically insoluble in water, alcohol, soluble in DMFA, pyridine. Storage as a toxic substance at below 20 оСin a dark place. Using intravenous injection as a sodium salt, hypodermic injection – necrosis. • Rubomycinh/ch – red powder. Hygroscopic. Freely soluble in water, alcohol, slightly soluble in chloroform. The color of medicine in acidic medium– red, in basic – blue. Storage as a toxic substance at below 20 оС. Using intravenous injection, inner muscular injection or hypodermic injection – necrosis.

48. 6 – Fluoroquinolones Antibiotics of a 4-quinoline derivatives, which in the 6-th position contain Fluorine atom, and in the 7-th position -piperazine cycle. Their precursors such as derivatives of quinoline and naphtiridin, are started to use in 60 years of ХХ century. The first representativeof this class is nalidyxone acid (nevigramon,negram) and its analogs is a pipemіdine acid (pipemidine, palin, urosept, pimidel). Thesedrugsarecharacterizedprimarilybytheactivityto gram-negativemicroorganisms, therapiddevelopmentofresistancetotheseantibioticscausingtheirusingonlyforinfectiousdiseasesoftheurinarytract. Nalidyxone acid (belongs to naphtiridines):

49. Fluoroquinolonesbegantobeintroducedintomedicalpracticein 80 yearsofthelastcentury. Introductionoffluorineatomsinthemoleculeofquinolinederivativesandnaphtiridinledsto a groupofdrugswithfundamentallynewpharmacologicalproperties. Fluoroquinolonesare a highlyactiveantimicrobialcompounds, whichareusedatsevereinfectionsofthedifferentetiologyandlocalization (respiratorytractinfections, skinandsofttissues, bonesandjoints, gastrointestinaltract, postoperativeinfections, otherchronicinflammatoryprocesses).Themechanismoffluoroquinolonesaction: theyaffectonthemetabolismofbacterial DNA byinhibitingtheenzymeinbacterialcells, DNA gyrase, whichcontrolsthestructureandfunctionof DNA. Inhibitionof DNA gyraseleadstodestructionofthebacteria (bactericidaleffect). Antibacterialactivityofquinolonesisalsoconnectedwiththeeffecton RNA synthesisofbacteriaandbacterialproteins, thestabilityofmembranesandothervitalprocessesofthebacterialcells.

50. Quinolones have a high effect on aerobic gram-negative bacteria and have little effect on anaerobic bacteria. They are rapidly absorbed from the gastrointestinal tract and are effective at the inside administration.            Quinolones inhibit the oxidazing enzymes of liver and can increase the effects of drugs (increase the side effects of theophylline).            The quinolones activity increases when administered in their molecules of two (lomefloxacin) or three (hemafloxacin) fluorine atoms.            Replacement of the ethyl radical in norfloxacin on cyclopropyl leads to the increasing of the substances activity (ciprofloxacin) by 3-8 times.            Some drugs of this group after extensive clinical using have been banned because of serious side effects.