LectureLecture № №55Antibiotics of the aromatic row: Antibiotics of the aromatic row:

synthesis, stereoisomery, methods of synthesis, stereoisomery, methods of analysis. Antibiotics of the heterocyclic analysis. Antibiotics of the heterocyclic

structure: structure: ββ-lactam antibiotics.-lactam antibiotics.

ass. Medvid I.I,ass. Medvid I.I,

Antibiotics of the aromatic row Chloramphenicol (Chloramphenicolum) (SPhU)

Laevomycetin (Laevomycetinum)

2,2-dichloro-N-[(1′R,2′R)-2′-hydroxy-1′-(hydroxymethyl)-2′-(4"-nitrophenyl)ethyl]acetamide

D-(-)-threo-1 -p-Nitrophenyl-2-dichloroacetylamino-propanediol-1,3

Laevomycetin – fine white with a grey or yellowish-green tinge crystalline substance, odorless. Bitter taste. Slightly soluble in water, soluble in alcohol. Solution of the substance in ethanol rotates the plane of polarization on the right, in ethylacetone – on the left.

O2NHC C

HCH2OH

HN C CHCl2

O

OH

1 2

1'2'4''

Leavomycetin stearate

(Laevomycetini stearas)

D-(-)-threo-1-p-Nitrophenyl-2-dichloroacetylaminopropanediol-1,3-3 –atearate

Laevomycetin stearate - white with a yellowish or yellowish-green tinge crystalline substance, odorless. Laevomycetin stearatehas no taste. Practically insoluble in water, difficult soluble in alcohol, in all solvents forms a cloudy solutions.

O2NHC C

HCH2O

HN C CHCl2

O

OH

C C17H35

O

Leavomycetin succinate soluble (Laevomycetini succinas solubile)

D-(-)-threo-1 -p-Nitrophenyl-2-dichloroacetylaminopropanediol-1,3-3-sodium succinate

Laevomycetin succinate soluble -white or slightly yellowish porous mass with low specific smell. Laevomycetin succinate soluble – has a bitter taste. Very easily soluble in water, slightly soluble in alcohol, hygroscopic.

O2NHC C

HCH2O

HN C CHCl2

O

OH

C (CH2)2

O

COONa

LeavomycetinLeavomycetin molecule has molecule has 2 2 asymmetric carbon atoms, and asymmetric carbon atoms, and therefore it is possible the existence of four isomerstherefore it is possible the existence of four isomers: : D- and L-D- and L-

threothreo, , D- and L-erythroD- and L-erythro-, -, which differ by the spatial which differ by the spatial arrangement of functional groupsarrangement of functional groups::

LaevomycetinLaevomycetin isis a leftrotatorya leftrotatory threothreo--isomer ofisomer of the Dthe D rowrow.. Mixture of Mixture of DD(-) (-) andand LL(+) (+) threothreo--isomers isomers ofof laevomycetin islaevomycetin is called racemic called racemic mixturemixture, , opticallyoptically inactive compound known asinactive compound known as synthomycinsynthomycin ( (has has 50% of the laevomycetin physiological activity50% of the laevomycetin physiological activity). ). ErythroErythro--formsforms is not is not used in medicine because it is a toxicused in medicine because it is a toxic compoundscompounds..

Obtaining ofObtaining of laevomycetinlaevomycetinLaevomycetinLaevomycetin first was isolated in first was isolated in 1947 1947 yearyear from the from the cultural liquid of cultural liquid of actinomycetesactinomycetes, , in in 1949 1949 year was year was established the chemical structureestablished the chemical structure..

The drug is extractedThe drug is extracted by the growing of actinomycetes by the growing of actinomycetes Streptomyces venezuelae on the mediumStreptomyces venezuelae on the medium ( (broth, glycerol, broth, glycerol, molasses and mineral saltsmolasses and mineral salts) ) atat 23-27 23-27 °°СС and strong and strong aeration for 89 hoursaeration for 89 hours. . After the filtering of the mycelium After the filtering of the mycelium fungus antibiotic is extracted and purified by the fungus antibiotic is extracted and purified by the chromatographicchromatographic..

LaevomycetinLaevomycetin - - the first antibiotic, which began to obtain the first antibiotic, which began to obtain by the chemical synthesisby the chemical synthesis, , whereas the majority antibiotic whereas the majority antibiotic was obtained by biosynthesiswas obtained by biosynthesis..

Synthesis of laevomycetinSynthesis of laevomycetin

Extracted drug by the interaction withExtracted drug by the interaction with NaOHNaOH is transformedis transformed in a racemic in a racemic base of laevomycetinebase of laevomycetine and divided into optical isomers by using of and divided into optical isomers by using of the different solubilitythe different solubility of salts, formed withof salts, formed with dextrorotatory tartaric dextrorotatory tartaric acid, in methanolacid, in methanol. . However, only salt of DHowever, only salt of D--isomer is soluble in isomer is soluble in methanolmethanol. . The basis from solution is sedimented by the ammonia andThe basis from solution is sedimented by the ammonia and acylated by the methyl ester of dichloroacetic acid Clacylated by the methyl ester of dichloroacetic acid Cl22CHCOOCHCHCOOCH3.3.

IdentificationIdentification 1.1. By the physico-chemical constantsBy the physico-chemical constants: : melting pointmelting point, , IRIR- - andand

UVUV--spectroscopyspectroscopy, , TLCTLC, , specific rotationspecific rotation..

2.2. To the alcoholic solution of laevomycetin add calcium To the alcoholic solution of laevomycetin add calcium chloride and zinc powder and heatchloride and zinc powder and heat. . Filtrate the obtained Filtrate the obtained hot solutionhot solution, , after cooling add benzoylchlorideafter cooling add benzoylchloride. . Then add Then add iron iron (ІІІ) (ІІІ) chloride solution and chloroform, shake the chloride solution and chloroform, shake the solutionsolution; ; aqueous layer should be pained in the color from aqueous layer should be pained in the color from light violet-red to purplelight violet-red to purple..

3.3. Reaction on chloride-ion after the mineralization of the Reaction on chloride-ion after the mineralization of the substance by the anhydrous sodium carbonate and substance by the anhydrous sodium carbonate and dissolution of the residue in the diluted nitrate aciddissolution of the residue in the diluted nitrate acid..

4.4. Hydrolysis in acidic or basic mediumsHydrolysis in acidic or basic mediums with the with the following identification of the formed productsfollowing identification of the formed products. . ThusThus, , at at the heating of laevomycetin with sodium hydroxide at the heating of laevomycetin with sodium hydroxide at first yellow color appearsfirst yellow color appears,, that transfers to the red- that transfers to the red-orangeorange ( (as a result of formation ofas a result of formation of acy-nitroformacy-nitroform), ), after after the following heating brick-red precipitate formed and the following heating brick-red precipitate formed and smell of ammoniasmell of ammonia appearedappeared::

5.5. Hydroxame reaction on laevomycetin estersHydroxame reaction on laevomycetin esters..

6.6. Laevomycetin stearate at the heating with Laevomycetin stearate at the heating with concentrated chloride acid hydrolyzed concentrated chloride acid hydrolyzed – – stearic stearic acid is formedacid is formed, , which floats to the surface in the which floats to the surface in the form of oily drops which harden when cooledform of oily drops which harden when cooled::

7.7. By the reaction of azodyes red color By the reaction of azodyes red color formationformation, , after after the reduction of nitro-group to amino-group with the reduction of nitro-group to amino-group with following following diazotationdiazotation and azojoiningand azojoining::

8.8. In the express-analysisIn the express-analysis use reaction of use reaction of laevomycetin with copper laevomycetin with copper ((IIII) ) sulfate in alkali sulfate in alkali mediummedium in the presence ofin the presence of nn--butanolbutanol – – alcoholic alcoholic layer painted in blue-violet color due to the layer painted in blue-violet color due to the formation of complex saltformation of complex salt, , which supposedly has which supposedly has the following structurethe following structure::

AssayAssay 1.1. SPhUSPhU - - SpectrophotometrySpectrophotometry. . Laevomycetine content in the Laevomycetine content in the

substance Content of the laevomycetin in the substance is substance Content of the laevomycetin in the substance is calculated by the specific absorptioncalculated by the specific absorption..

2.2. NitritometryNitritometry after the previous reductionafter the previous reduction of nitro-group to of nitro-group to amino-group by the zinc dustamino-group by the zinc dust in acidic mediumin acidic medium. Е = М.. Е = М.mm..

3.3. By liquid chromatographyBy liquid chromatography..

4.4. Cooper-metryCooper-metry,, direct titration.. The method is based on The method is based on the formation of soluble laevomycetin complex the formation of soluble laevomycetin complex compounds with copper (II) sulfate in alkaline medium compounds with copper (II) sulfate in alkaline medium (look identification)(look identification). . Titrant Titrant -- 0,01 М 0,01 М solution ofsolution of coopercooper ((IIII) ) sulfatesulfate, , indicatorindicator - - murexidemurexide. . Titration goes from the Titration goes from the purple to brownish-red color, uniform with color of the purple to brownish-red color, uniform with color of the “blind” sample“blind” sample. Е = 2 М.. Е = 2 М.mm..

5.5. Cooper-iodometryCooper-iodometry, , direct titrationdirect titration by the substituteby the substitute.. To the laevomycetin in alkali medium add cooper To the laevomycetin in alkali medium add cooper ((IIII) ) sulfatesulfate. . Precipitate of cooperPrecipitate of cooper (ІІ) (ІІ) hydroxidehydroxide is filtratedis filtrated, , in in the filtrate soluble cooper-laevomycetin complex is the filtrate soluble cooper-laevomycetin complex is destroyed by the action of sulfate acid with formation of destroyed by the action of sulfate acid with formation of equivalent amount of cooper equivalent amount of cooper ((IIII) ) sulfatesulfate, , which is which is determined by iodometrydetermined by iodometry, , indicatorindicator - - starchstarch. . In parallel In parallel control experiment is conductedcontrol experiment is conducted. Е = 2 М.. Е = 2 М.mm..

CuSO4

6.6. Precipitate titrationPrecipitate titration ( (argentometryargentometry oror mercurymetrymercurymetry). ). Methods based on Methods based on the the oxidationoxidation of laevomycetin by of laevomycetin by НН22ОО22 in alkali mediumin alkali medium, , as a result as a result в 2 в 2 molecules ofmolecules of NaCl NaCl formed, which are determined by argentometric method formed, which are determined by argentometric method (Folgard’s method) or mercurymetry(Folgard’s method) or mercurymetry with with diphenylcarbazone as indicatordiphenylcarbazone as indicator.. Е = Е = 1/1/2 М.2 М.mm..

7.7. PhotocolorimetryPhotocolorimetry by the formation of azodye after the by the formation of azodye after the reduction of of nitro-group to amino-group with followingreduction of of nitro-group to amino-group with following diazotation and azojoiningdiazotation and azojoining..

8.8. IodometryIodometry. . The method is based on the oxidation of the The method is based on the oxidation of the products of alkaline hydrolysis of laevomycetinproducts of alkaline hydrolysis of laevomycetin. . Experimentally determined Experimentally determined Е = 1/6 М.Е = 1/6 М.mm..

9.9. BromatometryBromatometry, , reverse titrationreverse titration. Е = 1/4 М.. Е = 1/4 М.mm.. 10.10. Acidimetry in non-aqueous medium after the acidic Acidimetry in non-aqueous medium after the acidic

hydrolysishydrolysis..

STORAGESTORAGEIn airtight containersIn airtight containers, , in the protected from light placein the protected from light place..

USAGEUSAGELaevomycetine belongs to the broad-spectrum antibiotics, used in the treatment of dysentery, pneumonia, whooping cough, typhoid and other infectious diseases. Course of treatment is 8-10 days. In pediatric practice use use llaevomycetine stearate, which hasn't bitter taste. Laevomycetine succinatecan be used for injections.

Issue: tablets, alcoholic solution, eye drops (0,25%), ointments Levosin, Levomekol.

Side effects. Disorders of the function of hematopoietic organs, so, at the treatment by laevomycetin blood test is required. Can cause an overgrowth .

Antibiotics heterocyclic structure

Penicillins (pennames) in the core of the penicillin's molecule is 6-amino-

penicillinic acid (6-APA), which consists of the condensed thiazolidine (А) and β-lactam (В) cycles:

N

S

O

H2N

CH3

CH3

COOH

12

3

456

7

Obtaining Obtaining of the natural penicillinsof the natural penicillinsFor the obtaining of penicillin growth mouldy fungusFor the obtaining of penicillin growth mouldy fungus Penicillium Penicillium

notatum on the mediumnotatum on the medium, , which consists ofwhich consists of corn extractcorn extract, , lactoselactose andand different mineral saltsdifferent mineral salts. . To increase the outlet of benzylpenicillinTo increase the outlet of benzylpenicillin, , to to the medium addthe medium add 0,02 – 0,08 % 0,02 – 0,08 % of phenylacetic acid of phenylacetic acid СС66НН55СНСН22СООН СООН

or its amideor its amide. . FermentationFermentation is conducted approximately for is conducted approximately for 70 70 hourshours atat 23-24 23-24 ооС, рН = 6-6,5 С, рН = 6-6,5 and strong aerationand strong aeration (1 (1 l of airl of air onon 1 1 l of l of environmentenvironment perper 1 1 minmin.). .). After the end of fermentation the obtained After the end of fermentation the obtained product is transformed by the scheme represented on the next slideproduct is transformed by the scheme represented on the next slide..

To the separation of the different types of penicillin and To the separation of the different types of penicillin and following purification of the penicillin sodium saltfollowing purification of the penicillin sodium salt, , this salt is this salt is transformed into the penicillin salt with organic bases or purified by transformed into the penicillin salt with organic bases or purified by chromatographic methodchromatographic method. . Water solution of benzylpenicillin Water solution of benzylpenicillin sodium salt is evaporated to the dry state at the temperature sodium salt is evaporated to the dry state at the temperature – 40– 40ооС С in the vacuumin the vacuum (0,1-0,2 (0,1-0,2 mmmm of mercuryof mercury columncolumn): ): iceice, , without without meltingmelting, , transformed in pairtransformed in pair, а, а preparation preparation remained dryremained dry ( (liophillicliophillic dryingdrying).).

For the manifestation of biological activity of great importance isFor the manifestation of biological activity of great importance is β-β-lactamlactam cyclecycle, , which is very unstable in acidic and alkaline which is very unstable in acidic and alkaline environmentsenvironments. . Penicillin is of the early era of antibiotics and Penicillin is of the early era of antibiotics and because of its wide usage nowbecause of its wide usage now 80-90 %80-90 % strains of staphylococci strains of staphylococci developed resistance to itdeveloped resistance to it, , producing the enzyme penicillinase, producing the enzyme penicillinase, which split the which split the ββ-lactam cycle-lactam cycle. . Account this and the fact that Account this and the fact that penicillins are the least toxic in the comparison with other penicillins are the least toxic in the comparison with other antibioticsantibiotics, , conduct a searchconduct a search of semi-synthetic penicillins on the of semi-synthetic penicillins on the basis of basis of 6-6-APAAPA..

Semi-synthetic penicillinsSemi-synthetic penicillins are an are an acylderivativesacylderivatives ofof 6- 6-APAAPA. .

General formula of penicillin’s medicines

Three generations of penicillins:• Natural (benzylpenicillin Na, К or Novocain salt,

phenoxymethylpenicillin, Bicillin (1,3,5))• Semi-synthetic penicillins (oxacillin, ampicillin,

amoxicillin, dicloxacillin, carbenicillin and others)• Semi-synthetic antibiotics + inhibitor of -lactamases

(amoxiclav (augmentine), timentin).

N

S

O

NH

CH3

CH3

COOR1

CR

O

Benzylpenicillin potassium (sodium) salt Benzylpenicillinum kalicum (natricum) SPhU

• Sodium (2S,5R,6R) – 3,3 – dimethyl – 7 – oxo – 6 [(phenylacetyl)amino] – 4 – thio – 1 –

azabicyclo[3.2.0]heptane – 2 – carboxylate• Sodium salt of 6 – phenylacetylaminopenicilanic acid

N

S

OCOOK(Na)

HH

H

HN

CH3

CH3

O

1 2

3

456

7

Benzylpenicillin Novocain salt Benzylpenicillinum novocainum

• Novocain salt of 6 –phenylacetylaminopenicilanic acid

N

S

OCOO

-

HH

H

HN

CH3

CH3

O

H2N C O

O

CH2

CH2

N+

C2H5

C2H5H

*H2O

*

*

Phenoxymethylpenicillin (ospen) Phenoxymethylpenicillinum

6 – phenylacetylaminopenicilinic acid

• Bicillin - 1 (N,N’ – dibenzylethyldiammonium salt of benzylpenicillin

• Bicillin - 3 (a mixture of equal parts of potassium (sodium) benzylpenicillin salt, benzylpenicillin and bicillin-1Novocain salt)

• Bicillin – 5 (a mixture of 1part of benzylpenicillin Novocain salt and 4 parts of bicillin – 1)

N

S

OCOOH

HH

H

HN

CH3

CH3

O

O

Bicillin-1(Вicillinum-1) Benzathine Benzylpenicillin

(Benzathine Benzylpenicillinum)

N,N′ – dibenzylethylenediammonium salt of benzylpenicillin

N

S

OCOO

-

HH

H

HN

CH3

CH3

O

CH2

NH2

CH2

CH2

NH2

*

* CH2

+ +

2

Oxacillin sodium salt Oxacillinum natrium

Sodium salt of 3-phenyl-5-methyl-4-isoxazolylpenicillin monohydrate

Ampiox (ampicillin and oxacillin sodium salts)

N

S

OCOONa

HH

H

HN

CH3

CH3

OO

N

CH3

*H2O

Ampicillin trihydrate (SPhU) Ampicillinum trihydricum

(2S,5R,6R)-6-[[(2′R)-2′-amino-2′-phenylacetyl]amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-

carboxylic acid trihydrate Also pharmacopoeial preparation is Ampicillin sodium

salt.

N

S

OCOOH

HH

H

HN

CH3

CH3

O

NH2

*3H2O1

2

3

456

7

1'2'

Amoxicillin trihydrate (SPhU) Amoxicillinum trihydricum

amoxyl, gramox, ospamox, hiconcil, Phemoxin-solutab

2S,5R,6R)-6-[[(2′R)-2′-amino-2′-(4′′′-hydroxyphenyl)-acetyl]amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid

trihydrateVampiox (amoxicillin trihydrate + cloxacillin + Lactobacillus)

N

S

OCOOH

HH

H

HN

CH3

CH3

O

NH2

HO

1 – 1 – character of the radical which dererminescharacter of the radical which dererminesthe level of bounding with proteinsthe level of bounding with proteins;;2 – 2 – Substitute inSubstitute in оо--position ofposition ofthe phenylthe phenyl radicalradical influencesinfluenceson theon the stability to the penicillinasestability to the penicillinase;;3 – 3 – character of the boundingcharacter of the bounding of the phenylof the phenylradical with methyl groupradical with methyl groupdeterminesdetermines the acidic stability of the penicillinthe acidic stability of the penicillin;;4 – 4 – substitutesubstitute of the Hydrogen atom in the methyl group of the Hydrogen atom in the methyl group в в determines the determines the

spectrum of penicillin actionspectrum of penicillin action;;5 – 5 – splitting of thesplitting of the ββ--lactamlactam boundbound conducts to the disappearance of conducts to the disappearance of

antibiotic properties and appearance of the allergic actionantibiotic properties and appearance of the allergic action;;6 – 6 – substitutesubstitute in the carboxylic group in the carboxylic group gives the possibility to obtain the gives the possibility to obtain the

penicillin salty formspenicillin salty forms;;PP – – penicillinasepenicillinase splitssplits ββ--lactamlactam cyclecycle;;А – А – amylaseamylase splitssplits amideamide boundbound..

Dependence between the chemical structureDependence between the chemical structure and biological action of penicillinsand biological action of penicillins

CHARACTERS Medical drugs of the natural and semi-Medical drugs of the natural and semi-synthetic penicillins synthetic penicillins --white crystalline white crystalline substance, odorless, bitter tastesubstance, odorless, bitter taste. . Sodium and Sodium and potassium benzylpenicillin saltspotassium benzylpenicillin salts are are hygroscopic and easily soluble in waterhygroscopic and easily soluble in water. . Benzylpenicillin Novocain saltBenzylpenicillin Novocain salt, , phenoxymethylpenicillin and ampicillin few phenoxymethylpenicillin and ampicillin few soluble in watersoluble in water..

Water or alcohol solutions penicillins rotate Water or alcohol solutions penicillins rotate plane polarized beam to the rightplane polarized beam to the right..

IdentificationIdentification By the By the physico-chemical constantsphysico-chemical constants: : IRIR- - andand UVUV--SpectroscopySpectroscopy, , TLCTLC..

Reaction with formaldehydeReaction with formaldehyde in the presence of in the presence of concentrated sulfate acid concentrated sulfate acid ((Marki reagentMarki reagent). ). Reaction is Reaction is distinguisheddistinguished, , because each penicillin produces at these because each penicillin produces at these conditions characteristic colorconditions characteristic color ((benzylpenicillins benzylpenicillins --reddish-brown color, amoxicillin - dark yellow, etcreddish-brown color, amoxicillin - dark yellow, etc.)..).

SubstancesSubstances give reactions on potassium, sodium and give reactions on potassium, sodium and NovocainNovocain..

Unpharmacopoeial reactionsUnpharmacopoeial reactions: :

а) а) reaction reaction of copperof copper (II(II) () (greengreen) ) or ironor iron ( (IIIIII) () (red colorred color) ) penicilloinhydroxamates formation after the penicilloinhydroxamates formation after the hydroxylaminolysis hydroxylaminolysis ((ββ--lactam cyclelactam cycle):):

bb)) reaction with chromotropic acidreaction with chromotropic acid in the presence of in the presence of concentrated sulfate acidconcentrated sulfate acid,, which which is distinguishedis distinguished, , because because each penicillin produces at these conditions characteristic each penicillin produces at these conditions characteristic colorcolor ((benzylpenicillins benzylpenicillins -- brown color, amoxicillin – violet brown color, amoxicillin – violet etsets.);.);

cc)) determination of the determination of the organically bound sulfur (dry and wet organically bound sulfur (dry and wet pyrolysispyrolysis););

dd) ) determine the melting temperature of the determine the melting temperature of the NN--ethylpiperidine ethylpiperidine salt of benzylpenicillin salt of benzylpenicillin ((for natural penicillinsfor natural penicillins););

ee)) reaction on reaction on aliphatic aminoaliphatic amino--group (group (aampicillin, amoxicillinmpicillin, amoxicillin) ) –– at the at the heatheatinging with a with a ninhydrin ninhydrin solution observed purple solution observed purple color color ..

ff) ) Vitali – MorenVitali – Moren’s reaction’s reaction..

Reaction ofReaction of thethe azo-dye of amoxicillin formationazo-dye of amoxicillin formation ( (redred colorcolor))

PenicillinsPenicillins due to the Sulfur atom have the reducing due to the Sulfur atom have the reducing properties and can reduce silver from the Tollense reagentproperties and can reduce silver from the Tollense reagent, , mercury from the Nesler reagent ets..mercury from the Nesler reagent ets..

AssayAssayBy By the the liquid chromatography (SPliquid chromatography (SPhhU)U)..

Microbiological methods of Microbiological methods of the the diffusion in agar diffusion in agar ((reproducingreproducing of the resultsof the results - 5-10 %). - 5-10 %).

Spectrophotometric determination Spectrophotometric determination of the of the semi-synthetic semi-synthetic penicillinspenicillins..

Chemical methods in two phasesChemical methods in two phases::

а)а) determination of the penicillins amountdetermination of the penicillins amount;;

bb)) identify the content of the relevant drugidentify the content of the relevant drug..

The amount of penicillins for the medical drugs of natural The amount of penicillins for the medical drugs of natural penicillins determinepenicillins determine by iodometric methodby iodometric method, , the essence the essence of which is that the products of alkaline hydrolysis of of which is that the products of alkaline hydrolysis of penicillin canpenicillin can be oxidized by iodine in the presence of be oxidized by iodine in the presence of acetic buffer with acetic buffer with рН = 4,5. Е = М.рН = 4,5. Е = М.mm/8/8

For determination of the benzylpenicillinFor determination of the benzylpenicillin content use content use gravimetric method by the reaction of Ngravimetric method by the reaction of N--ethylpiperidine salt ethylpiperidine salt formationformation::

Amount ofAmount of penicillins in the semi-synthetic compounds penicillins in the semi-synthetic compounds determine by the alkalimetry, reverse titrationdetermine by the alkalimetry, reverse titration, , with control with control experimentexperiment, , indicatorindicator – –phenolphthaleinphenolphthalein. Е = М.. Е = М.mm..

Test on purityTest on purityMeasure the optical density of solutions of natural penicillins at a Measure the optical density of solutions of natural penicillins at a

wavelength ofwavelength of 264, 280 264, 280 andand 325 325 nmnm. . Specific impurities determined Specific impurities determined by the liquid chromatography, residual amounts of the organic by the liquid chromatography, residual amounts of the organic solvents - gas chromatographysolvents - gas chromatography. . Pyrogens, abnormal toxicity and Pyrogens, abnormal toxicity and sterility are also determinedsterility are also determined..

StorageStorage In airtight containersIn airtight containers, , in dry, dark place at the room temperaturein dry, dark place at the room temperature..

UsageUsage Natural penicillins affect on the gram-positive bacteria and used to Natural penicillins affect on the gram-positive bacteria and used to

treat pneumonia, gonorrhea, syphilis, septic infections, diphtheria, treat pneumonia, gonorrhea, syphilis, septic infections, diphtheria, scarlet feverscarlet fever. . They can not be received They can not be received per os,per os, because in acidic because in acidic environments inactivation occurs (semi-synthetic penicillins and environments inactivation occurs (semi-synthetic penicillins and phenoxymethylpenicillin are stable in acid solutionsphenoxymethylpenicillin are stable in acid solutions). ). Penicillins Penicillins destroy under the action of penicillinase, semi-synthetic analogs destroy under the action of penicillinase, semi-synthetic analogs resistant to it and have a broader spectrum of activityresistant to it and have a broader spectrum of activity ..

Side effectsSide effects.. Safest Safest. . Penicillins may cause gastro-intestinal Penicillins may cause gastro-intestinal disorders, very rare - allergic reactionsdisorders, very rare - allergic reactions..

Cephalosporins (cephems) In the structure of cephalosporin is condensed system

consisting of -lactam and dihydrothiazine cycles. Cephalosporin are derivatives of 7-aminocephalosporanic acid (7-ACA) (cefalotin, cephalopirin, cefuroxime, sodium cefotaxime) and 7-amino-desacetoxycephalosporanic acid (7-АDCA) (cefalexin monohydrate, cefazolin sodium, sodium ceftriaxon, cefixime, cefaloridine).

The general formula of cephalosporin group of medicines:

• Mouldy fungi Cephalosporium sort and actinomycetes produce natural antibiotic - cephalosporin C, which did not found application in medicine because of low activity. Cephalosporin C is a source of obtaining of the semi-synthetic cephalosporins.

• In the medical practice use the modern semi-synthetic cephalosporins of four generations (classification by Kharkevich).

N

SR1HN

COOH

CH2R2O

Cefazolin sodium (SPhU)

(Cefazolinum natricum), kefzol , reflin

Sodium (6R,7R)-3-[[(5′-methy-1′,3′,4′-thiadiazol-2′-yl) sulphanyl]methyl]-8-oxo-7-[(1"Н-tetrazol-1"-yl-

acetyl)amino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate

N

S

COONaH2CO

NH

CCH2

O

H H

S

NN

N

N

N

S

N

CH31

2

3

4

5

67

8

1'

2'

3' 4'

5'

1''

2''3''

4'' 5''

N

S

COONaH2CO

NH

CCH2

O

H H

S

NN

N

N

N

S

N

CH31

2

3

4

5

67

8

1'

2'

3' 4'

5'

1''

2''3''

4'' 5''

Cefalexin monohydrate (SPhU)(Cefalexinum monohydricum)

ospexin, lexin

(6R,7R)-7-[[(2′R)-2′-amino-2′-phenylacetyl]amino]-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-

carboxylic acid monohydrate

N

S

COOH

CH3O

NH

CC

NH2

H

O

H H

*H2O

12

3

4

5

67

8

1'2'

Cefixime (SPhU) (Cefiximum), cefix, loprax

(6R,7R)-7-[[(2′R)-2′-(2′′-aminothiazol-4"-yl)-2′-[(carboxymethoxy)imino]acetyl]amino]-3-ethenyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid trihydrate

N

S

COOHHCO

NH

CC

O

H H

CH2

N

O

H2C COOH

N

S

NH21

2

3

4

5

67

8

4''

3''2''

1'' 5''

* 3 H2O

1'2'

Cefotaxime sodium (SPhU) (Cefotaximum natricum)

cefabol, cafantral

Sodium (6R,7R)-3-[(acetyloxy)methyl]-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate.

N

S

COONaH2CO

NH

CC

O

H H

O

N

O

CH3

C CH3

ON

S

NH21

2

3

4

5

67

8

4''

3''2''

1'' 5''

1'2'

Ceftriaxone sodium (SPhU) (Ceftriaxonum natricum)

Disodium (6R,7R)-7-[[(2′R)-2′-(2′′-aminothiazol-4"-yl)-2′-[(methoxyimino)acetyl]amino]-3-[[(2′′′-methyl-6′′′-oxido-5′′′-oxo-2′′′,5′′′-dihydro-1′′′,2′′′,4′′′-triazin-3′′′-

іл)sulphanyl]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate

N

S

COONaH2CO

NH

CC

O

H H

S

N

O

CH3

N

S

NH21

2

3

4

5

67

8

4''

3''2''

1'' 5''

1'2'

N

NN

ONa

CH3

O

1'''

2'''3'''

4'''5'''

6'''

* 3,5 H2O

Ways of introducti

on

Generations

І ІІ ІІІ ІV

Parenteral (i/v; i/m)

Cefazolin (kefzol, reflin)Cefalotin (ceporin)CefapirinCefaloridinCafradin

Cefuroxime (zinacef)Cafamandole (cafambol)CefoxitineCefonocideCefprozylCefmetazoleCefotetanCeforanid

CeftriaxoneCefotaxime (cefabol, cefantral)CeftrizoximeCefoperazone(hepacef-KMP, cefobit)Ceftazidime (forum, ceftum)CeftizoximeMoxalactal

Cefepime (maxipime)Cefoxitine

Enteral (per os)

Cefalexin (ospecsin, lexin)Cefadroxil (duracef, cefangin-KMP)Cefradine

Cefaclor (biclor-KMP)Cefuroxime (zinnate)LoracarbefCefprozyl

Cefixime (cefix, loprax)Cefpodroxime (cefodox)ProxetylCeftibutene (cedex)Cefetamethe pivoxyl

IdentificationIdentification By the physico-chemical constants: IR and UV By the physico-chemical constants: IR and UV spectrophotometry, thin layer chromatographyspectrophotometry, thin layer chromatography..The reaction with Marki reagentThe reaction with Marki reagent. . Reaction is distinguished Reaction is distinguished because each of them forms a characteristic color because each of them forms a characteristic color ((cafalexincafalexin - -light yellow color, which becomes dark light yellow color, which becomes dark yellow; ceftriaxon sodium salt - greenish-yellow, which yellow; ceftriaxon sodium salt - greenish-yellow, which turns into yellow, cefotaxime sodium salt - bright yellow, turns into yellow, cefotaxime sodium salt - bright yellow, which turns into brownwhich turns into brown).).The presence of theThe presence of the β- β-lactam cyclelactam cycle causes the formation causes the formation of cooper of cooper ((IIII) ) or ironor iron ((IIIIII)) hydroxamateshydroxamates..With the mixture of acidsWith the mixture of acids HH22SOSO44 and HNOand HNO33 cefalexin cefalexin becomes yellowbecomes yellow, , cefalotincefalotin - -olive-green, which transforms olive-green, which transforms into a red-browninto a red-brown ..Sodium salts give an appropriate reactions on sodium Sodium salts give an appropriate reactions on sodium cationcation..

CHARACTERS Medicinal substances - white, sometimes yellowish Medicinal substances - white, sometimes yellowish

powderpowderss. . Sparingly soluble in water (except for sodium Sparingly soluble in water (except for sodium salts of cefalexin ceftriaxone, cefatoxime), hard soluble in salts of cefalexin ceftriaxone, cefatoxime), hard soluble in alcohol. Some of them have a characteristic smell and alcohol. Some of them have a characteristic smell and sensitive to lightsensitive to light. . Optically active substances of the Optically active substances of the cephalosporin droup rotate the plane polarization to the cephalosporin droup rotate the plane polarization to the rightright..

AssayAssayBy liquid chromatography By liquid chromatography ((SPUSPU).).Chemical methods (like penicillinChemical methods (like penicillinss).).Biological methodsBiological methods..Physico-chemical methods (spectrophotometry, Physico-chemical methods (spectrophotometry, photocolorimetryphotocolorimetry). ).

AssayAssay cefalexincefalexin

Acidimetry in non-aqueous environmentAcidimetry in non-aqueous environment. . Equivalent Equivalent point is determined by potentiometric methodpoint is determined by potentiometric method..

StorageStorage In airtight containersIn airtight containers, в, вin dryin dry, , pprotected from lightrotected from light place place at at

the the room temperatureroom temperature..

UsageUsageCephalosporins have a Cephalosporins have a more more broader spectrum than broader spectrum than penicillinpenicillinss, and less toxicity, and less toxicity..The difference in the chemical structure of penicillins The difference in the chemical structure of penicillins and cephalosporinsand cephalosporins leads to the resistance to leads to the resistance to staphylococcus penicillinase and greater resistance to staphylococcus penicillinase and greater resistance to acidsacids. . Therefore prescribe cephalosporins are Therefore prescribe cephalosporins are prescribed to the treatment of penicillin stable prescribed to the treatment of penicillin stable infectionsinfections..Medications of cephalosporin’s froup used in the Medications of cephalosporin’s froup used in the treatment of acute and chronic respiratory diseases, treatment of acute and chronic respiratory diseases, urinary tract, genitals, with postoperative and other urinary tract, genitals, with postoperative and other infectionsinfections..

Other antibiotics of heterocyclic Other antibiotics of heterocyclic structurestructure

The structure that connects The structure that connects ββ-lactam and-lactam and oxazolidine cycles oxazolidine cycles ––clavulanic clavulanic acidacid::

3-(2-3-(2-HydroxyethylideneHydroxyethylidene)-7-)-7-oxooxo-4--4-oxaoxa-1--1-azabicycloazabicyclo[3.2.0]-[3.2.0]-heptaneheptane-2--2-carboxylic acidcarboxylic acid

It is used as an inhibitor of It is used as an inhibitor of ββ-lactamase of gram-positive -lactamase of gram-positive and gram-negative bacteria with penicillin and and gram-negative bacteria with penicillin and cephalosporins, increasing their effect.cephalosporins, increasing their effect.

Also it is opened a new antibiotics of heterocyclic structureAlso it is opened a new antibiotics of heterocyclic structure:: cephamycincephamycin, , thienamycin and othersthienamycin and others..

Thank youThank you for for attentionattention ! !