Lecture № 4

LectureLecture № №44General characteristic of General characteristic of antibiotics, their types. antibiotics, their types. The main stages of the industrial production of antibiotics and the creation of new drugs in this

series. The identification methods, the purity tests and quantitative determination of determination of

antibioticsantibiotics. Using in medical practice, their side effects. Antibiotics of the alicyclic series.

ass. Kozachok S.S.ass. Kozachok S.S.

PlannerPlannerThe principle of chemotherapyThe principle of chemotherapy Determination of antibioticsDetermination of antibioticsClassification of antibioticsClassification of antibiotics. . Chemical classificationChemical classification..Obtaining methodsObtaining methods: :

а) а) microbial synthesis;microbial synthesis;bb) ) chemicalchemical synthesissynthesis; ; cc)) Combining of the microbial and chemical synthesis Combining of the microbial and chemical synthesis

Analysis methods of antibioticsAnalysis methods of antibiotics..Determination the antibiotics units of actionDetermination the antibiotics units of action The side effects of antibioticsThe side effects of antibiotics..Antibiotics of alicyclic structureAntibiotics of alicyclic structure ((tetracyclinetetracycline))

Chemotherapeutic agents –Chemotherapeutic agents –drugs wich have a selective activity against to

certain infectious and parasitic diseases (sulphonylamides, antibiotics, antibacterial

agents of the various chemical structure, antitubercular, antisyphilis, antiviral,

antiparasitic, antihelminthic).

Depending on the source and method of the producing of chemotherapeutic agents can be divided into 2 groups:Antibiotics - substances of natural origin, or the substaces are obtained by the modifying of natural substances;Original definition of antibiotics was material derived from microorganisms that suppress growth and sometimes kill the microorganism. Synthetic material was added later, when they were able to make synthetic antibiotics.Synthetic chemotherapeutic agents (sulfanilamide, nitrofurans, quinolones, nitroimidazoles, etc.)

The principle of chemotherapySelectivity of the drug’s action against to certain Selectivity of the drug’s action against to certain microorganisms (the certain spectrum of the microorganisms (the certain spectrum of the antimicrobial action)antimicrobial action) Low toxicity to humans and animals.Low toxicity to humans and animals.

Antibiosis is the base of antibiotic’s action.is the base of antibiotic’s action.Antibiosis is the phenomenon of the antagonism of is the phenomenon of the antagonism of

microorganisms, the first was opened by Louis microorganisms, the first was opened by Louis Pasteur in the 80 years of the XIX century.Pasteur in the 80 years of the XIX century.         For the first time the term "         For the first time the term "antibiotic" was " was offered by SA Waxman (USA). offered by SA Waxman (USA). The word The word "antibiotics" comes from the Greek anti ("against") "antibiotics" comes from the Greek anti ("against") and bios ("life"). Antibiotics are drugs that either and bios ("life"). Antibiotics are drugs that either destroy bacteria or prevent their reproduction. destroy bacteria or prevent their reproduction.

AntibioticsAntibiotics– – chemotherapeutic agents of a biological origin that have the ability to selectively kill or inhibit the growth of pathogenic agent (bacteria, viruses, fungi, protozoa microorganisms) or delay the development of malignant tumor. Antibiotics are used in medical practice, are Antibiotics are used in medical practice, are produced by actinomycetes (radiant produced by actinomycetes (radiant mushrooms), fungis and some bacteriasmushrooms), fungis and some bacterias ..

Antibiotics that are obtained from a plant origin called phytoncyds. Antibiotics are the representatives of the secondary metabolites of living cells. Primary metabolites are called the substance, that are necessary for cell growth (amino acids, carbohydrates, nucleotides, vitamins, etc.). Secondary metabolites are low molecular weight compounds (antibiotics, alkaloids, pigments ments, etc.) produced in the cells after the growing phase.

Antibiotics have the following functions in the living producing cells:1. Means of attack and defense ("chemical weapons" cells);2. Detoxification from the harmful metabolites;control of some reactions in metabolism;3. Participation in the process of cell differentiation;4. Reserve nutrients

Classification of antibioticsClassification of antibiotics

MainMain andand reservereserveddBy the types of By the types of producerproducer According to the biological action typeAccording to the biological action type

According to the chemical structure.According to the chemical structure.

Classification of antibiotics according to the chemical structure

1. β-lactamsAntibiotics, which are present in the molecule β- lactam cycle.

bicyclic: derivatives of 6-aminopenicillic- acid (penicillins), derivatives of 7 - aminocephalosporinic acid (cephalosporines) andetc.; monocyclic: monobactams and others;

2. Macrolides and azalides are theantibiotics, which contain a lactone ring with 14 - 16 atoms in

their molecule. In azalides (azitromycin) the atom of nitrogen is present in a cycle.

14-members macrolides (erythromycin, oleandomycin, claritromycin, roxitromycin and other)15- members azalides (azitromycin);16- members macrolides (spiramycin and other)

3. AminoglycosidesAntibiotics which have the cyclohexan structure with OH- and NH2- or guanidino-derivatives and glycoside derivatives with one or certain OH-groups.

Derivatives of D-streptydine (streptomycin);Derivatives of 2-desoxy-D-streptamine (Kanamycin, gentamicin, amikacin sulfate (amikin) and others)

4. TetracyclinesAntibiotics, which have the partially hydrated nucleus of tetracene.

Tetracyclines, oxytetracyclines, doxycyclines and others.5. AmphenicolesChloramphenicol (levomycetin) have practical using, which according to its structure belongs to nitrophenylalkylamines.

6. Antibiotics of other structureLincosamides (lincomycin, clindamycin);Ansamycin (rifampicin) – antibiotics contain the aromatic nucleus in their molecules (usuallynaphthalene), which in two positions contains an aliphatic chain, consisting of 15 - 20 carbon’s atomsAnthracycline (rubomycin, doxorubomycin, karminomycin hydrochloride (carubicin))– glycosides in which the aglycone is a substituted tetrahydronaphtacenquinon; Polyenes antibiotics (nystatin, levorin) –macrolides, which molecules contain a system of conjugated double bonds; Glycopeptide (vancomycin) and other.

Bactericidal drugs kill the target bacterium or fungus and are more effective. You would use these types of drugs to treat endocarditis, meningitis, osteomyelitis, and other invasive bacterial infections. Hosts with low immune systems should also be treated with bactericidal drugs. These drugs include β- lactams, aminoglycosides, and fluoroquinolones.

Bacteriostatic drugs inhibit growth and can be helpful cuz they let the normal host defenses to destroy the microorganisms. You would use these to treat infections such as cystitis. These drugs include tetracycline, sulfonamides, clindamycin, and chloramphenicol. Some organisms require 2 agents for killing such as entercocci.

Chemical classificationChemical classification of antibioticsof antibiotics 1)1) Acyclic structureAcyclic structure ( (tetracyclinetetracycline))2)2) Aromatic structureAromatic structure ( (levomicetine and its derivativeslevomicetine and its derivatives).).3)3) Heterocyclic structure (penicillins, cephalosporins).Heterocyclic structure (penicillins, cephalosporins).4)4) Glycosides:Glycosides:

        a) streptomycin;        a) streptomycin;        b) the aminoglycosides (canamycin, neomycine, gentamicine,         b) the aminoglycosides (canamycin, neomycine, gentamicine, monomycin);monomycin);        c) macrolides (erythromycin, oleandomitsine);        c) macrolides (erythromycin, oleandomitsine);        d) anzamitsiny (rifampicin).        d) anzamitsiny (rifampicin).

5)5) Polyenes (nystatin, amphotericin).Polyenes (nystatin, amphotericin). 6)6) Polypeptide structures (gramicidin, polymyxin)Polypeptide structures (gramicidin, polymyxin)7)7) AntineoplasmicAntineoplasmic:: a) aureolic acid derivativesa) aureolic acid derivatives;; bb) anthracyclines;) anthracyclines; cc) ) derivatives of quinolinederivatives of quinoline-5,8--5,8-diondion;; dd) actinomycin) actinomycin

We can classify antibiotics by their mode of action.We can classify antibiotics by their mode of action.

Antibiotics that influence onAntibiotics that influence on Gram +ve cell wall Gram +ve cell wall of of bacteriums.bacteriums. Penicillins violate the synthesis of the bacterial Penicillins violate the synthesis of the bacterial cell wall - showing a bactericidal effect; macrolides violate cell wall - showing a bactericidal effect; macrolides violate the intracellular protein synthesis on the ribosome’s level - the intracellular protein synthesis on the ribosome’s level - bacteriostatic action.bacteriostatic action. Antibiotics that influence onAntibiotics that influence on Gram Gram --ve cell wall ve cell wall of of bacteriums bacteriums ((polymyxinpolymyxin violate the penetration of the violate the penetration of the cytoplasm membrane (bactericidal effect).cytoplasm membrane (bactericidal effect). Broad-spectrum antibiotics. Chloramphenicol and Broad-spectrum antibiotics. Chloramphenicol and tetracycline (bacteriostatic effect on the level of ribosomes), tetracycline (bacteriostatic effect on the level of ribosomes), cephalosporins (bactericidal action - inhibit synthesis of cephalosporins (bactericidal action - inhibit synthesis of bacterial cell walls). Streptomycin, neomycin, kanamycin bacterial cell walls). Streptomycin, neomycin, kanamycin (bacteriostatic action - inhibit protein synthesis at the (bacteriostatic action - inhibit protein synthesis at the ribosome’s level). Rifampicin violate the synthesis of RNA.ribosome’s level). Rifampicin violate the synthesis of RNA.

Methods of antibiotics obtainingMethods of antibiotics obtainingMicrobiological synthesis is based on the mold Microbiological synthesis is based on the mold (Penicillium) or the radiant mushrooms (Penicillium) or the radiant mushrooms (Streptomyces). There are obtained by this method (Streptomyces). There are obtained by this method such antibiotics as: tetracyclines, the natural such antibiotics as: tetracyclines, the natural penicillins, glycosides, macrolides.penicillins, glycosides, macrolides.

The main stages of microbiological synthesis are:The main stages of microbiological synthesis are:a) selection of a highly producing strains;a) selection of a highly producing strains;b) selection the medium (pH, metal salts, b) selection the medium (pH, metal salts, carbohydrates, fats);carbohydrates, fats);c) the process (biosynthesis) of fermentation;c) the process (biosynthesis) of fermentation;d) isolation and purification of antibiotic d) isolation and purification of antibiotic (crystallization, chromatography, extraction, (crystallization, chromatography, extraction, resedimentation).resedimentation).

1). Modified method of a diffusion in an in an agar2). 2). UreasedUreased 3). 3). FFermentativeermentative4). 4). RadioimmunoassayRadioimmunoassay

Chemical synthesis (levomycetin).A combination of microbiological and chemical methods. Obtaining semi-synthetic antibiotics based on the transformation of the molecules of natural antibiotics (semisynthetic tetracyclines, penicillins, cephalosporins, etc. ).

Modern methods of determining the biological activity of antibiotics:

QuantitativeQuantitative determination of antibiotics determination of antibiotics by a by a microbiological methodmicrobiological method

Activity of antibiotics is determined by a Activity of antibiotics is determined by a comparing of the suppression growth degree of the comparing of the suppression growth degree of the test antibiotic on the sensitive (test cultures) m /o test antibiotic on the sensitive (test cultures) m /o (microorganism) with a standard sample of (microorganism) with a standard sample of antibiotic with known concentrations. Quantitative antibiotic with known concentrations. Quantitative determination is carried out by the determination is carried out by the diffuse or diffuse or turbidimetric methodsturbidimetric methods. Calculation of the biological . Calculation of the biological activity is performed according to a standard curve activity is performed according to a standard curve that was constructed on the basis of the definitions that was constructed on the basis of the definitions of the five concentrations of standard sample of the of the five concentrations of standard sample of the antibiotic. Biological activity is expressed in units of antibiotic. Biological activity is expressed in units of action (UA).action (UA).

U.AU.A.. – – the minimum molar amount of the antibiotic, the minimum molar amount of the antibiotic, which inhibits the development of a test microorganism in which inhibits the development of a test microorganism in a certain volume of the medium. The number of grams of a certain volume of the medium. The number of grams of active ingredient in U.A. is variety for different active ingredient in U.A. is variety for different antibiotics!antibiotics!

The average value of the activity, which was found by a The average value of the activity, which was found by a biological method, somewhat lower than the theoretical biological method, somewhat lower than the theoretical activity. In the pharmacopoeial article is the theoretical activity. In the pharmacopoeial article is the theoretical value of the activity and the lower permissible boundary value of the activity and the lower permissible boundary of the activities of the investigated antibiotic U.A. / mg.of the activities of the investigated antibiotic U.A. / mg.

It was established that 1 mg of streptomycin equivalents to It was established that 1 mg of streptomycin equivalents to 1000 UA. Respectively, 1 UA of streptomycin equivalents 1000 UA. Respectively, 1 UA of streptomycin equivalents to 1 microgram of a pure streptomycin basis. For other to 1 microgram of a pure streptomycin basis. For other antibiotics UA differs from 1 microgram of the substance. antibiotics UA differs from 1 microgram of the substance. For example, for benzylpenicillin 1 UA equivalents to = For example, for benzylpenicillin 1 UA equivalents to = 0.6 mcg, that 1 mg of the antibiotic contains 1,667 UA.0.6 mcg, that 1 mg of the antibiotic contains 1,667 UA.

SIDE EFFECTS OF ANTIBIOTICSSIDE EFFECTS OF ANTIBIOTICS((according to Charkevychaccording to Charkevych))

SperinfectionSperinfection ( (allall))AAllergy (llergy (allall))Dispepsia phenomenon, pain feelingDispepsia phenomenon, pain feelingLocal irritating action – at inner muscular injection; phlebitis, Local irritating action – at inner muscular injection; phlebitis, thrombophlebitis at intravenous injectionthrombophlebitis at intravenous injectionEffect on a liverEffect on a liver ( (tetracyclinetetracycline))Effect on a kidneys (cephalorydin, neomycin, polymyxin)Effect on a kidneys (cephalorydin, neomycin, polymyxin) Hemodyscrasia (chloramphenicol)Hemodyscrasia (chloramphenicol)Impairment of the function of vision and hearing (chloramphenicol Impairment of the function of vision and hearing (chloramphenicol in high doses)in high doses) Impaired function of a pair of VIII cranial nerve (aminoglycosides)Impaired function of a pair of VIII cranial nerve (aminoglycosides) Mycosis (women - gynecologic diseases, male - a disease of the Mycosis (women - gynecologic diseases, male - a disease of the hearing aid)hearing aid) Deafness, impaired tooth enamel, deposition in the bones - Deafness, impaired tooth enamel, deposition in the bones - tetracyclines.tetracyclines.

Toxicity investigation of antibioticsToxicity investigation of antibioticsХ Х PharmacopoeiaPharmacopoeia

1.1. Quantity of miceQuantity of mice - 5 - 5 2.2. Mass of miceMass of mice – 18-20 – 18-20 gg3.3. injectioninjection – – intravenousintravenous , ,

hypodermichypodermic, , in an in an abdomenabdomen. . Observation time according to the Observation time according to the requirements specific requirements specific pharmacopoeial monographpharmacopoeial monograph

4.4. If, within the prescribed time there If, within the prescribed time there is no lost in mice, the medicine is no lost in mice, the medicine keeps up the investigation, in case keeps up the investigation, in case of the death of a one or more of the death of a one or more mouse, the investigation repeat on mouse, the investigation repeat on the 10 mice with the weigh of 10-the 10 mice with the weigh of 10-20 g. If during the secondary 20 g. If during the secondary investigation , there is no lost in investigation , there is no lost in mice 10 mice, the drug holds the mice 10 mice, the drug holds the research . In the case of the death research . In the case of the death of a one mouse the medicine is of a one mouse the medicine is defected.defected.

SPhUSPhU ( (anomal toxicityanomal toxicity))1.1. Quantity of miceQuantity of mice - 5 - 5 2.2. Mass of miceMass of mice - 17-22 г- 17-22 г3.3. injectioninjection – – intravenous. Solution is intravenous. Solution is

injected frominjected from 15 15secsec tilltill 30 30secsec, , if if there is no some remarksthere is no some remarks. . The The observation time is 24 hoursobservation time is 24 hours, , or a or a period of time according to the period of time according to the requirements of a separate article.requirements of a separate article.

4.4. Sample of the test is good, if none of Sample of the test is good, if none of the mice did not die within the the mice did not die within the specified time according to a separate specified time according to a separate article. In the case of the death of an article. In the case of the death of an animal test is repeated again. The animal test is repeated again. The sample test keeps up the investigation sample test keeps up the investigation when the animals of the second group when the animals of the second group did not died.did not died.

Investigation of the antibiotic’sInvestigation of the antibiotic’s pyrogenic pyrogenicityity Х Х PharmacopoeiaPharmacopoeia

1.1. Quantity of Quantity of rabbitrabbitss - 3 - 3 2.2. Mass of Mass of rabbitrabbits s – 1,5-2,5 – 1,5-2,5 kgkg3.3. injectioninjection– – in ear veinin ear vein, , quantity of quantity of

solution is solution is 10 10 mlml onon 1 1 kg of weighkg of weigh. . Injection time isInjection time is 2 2 minutesminutes..

4.4. The solution is a pyrogen-free if The solution is a pyrogen-free if after the injection no one of the after the injection no one of the rabbits do not have the rise in rabbits do not have the rise in temperature more than temperature more than 0,60,6СС comparing with the initial comparing with the initial temperature of the rabbitstemperature of the rabbits and in the and in the amount does not exceed than amount does not exceed than 1,41,4СС. . If one or two rabbits, temperature If one or two rabbits, temperature increases more than increases more than 1,4 1,4 СС, , the investigation is repeated on 5 repeated on 5 rabbits, the solution is pyrogen-free, rabbits, the solution is pyrogen-free, if there is no more than 3 of 8 if there is no more than 3 of 8 rebbits have the rise in temperature rebbits have the rise in temperature on on 0,6 0,6 00CC and the total amount of and the total amount of the rise in temperature does not the rise in temperature does not exceed than exceed than 3,73,7СС..

SPhUSPhU 1.1. Quantity of Quantity of rabbitrabbitss - 3 - 3 2.2. Mass of Mass of rabbitrabbits not less thans not less than 1,5 1,5 kgkg3.3. injectioninjection - - in ear veinin ear vein, , Injection Injection

time istime is 44 minutesminutes, , injection volume injection volume is 0,5 ml-10 ml on 1kg of massis 0,5 ml-10 ml on 1kg of mass

4.4. The investigation is realizedThe investigation is realized on 3 on 3 rabbits. The sample keeps up the rabbits. The sample keeps up the investigation, if the total increasing investigation, if the total increasing in temperature does not exceedin temperature does not exceed than than 1,151,15С,С, if the temperature rising isif the temperature rising is 1,151,15СС, , but not more thanbut not more than 2,652,65СС. . The investigation is repeatedThe investigation is repeated. . Then Then the the corresponding valuescorresponding values areare 2,82,8СС; ; andand 4,34,3СС. . If the amount If the amount temperature rising is more than temperature rising is more than 4,34,3СС. . The sample is defected.The sample is defected.

If you need the research is conducted If you need the research is conducted by another 3 or 6 rabbits.\by another 3 or 6 rabbits.\

Investigation of antibiotics sterilityInvestigation of antibiotics sterilityTo select the medium and test the suitability of the procedure’s To select the medium and test the suitability of the procedure’s

methodic. This test is performed using the membrane filtration methodic. This test is performed using the membrane filtration method or the method of a direct seeding. Regardless of the method or the method of a direct seeding. Regardless of the procedure’s method, realeze a corresponding negative control procedure’s method, realeze a corresponding negative control experiment using the samples, sterility of which has been proved experiment using the samples, sterility of which has been proved earlier. The inoculation are periodically looked through during earlier. The inoculation are periodically looked through during and after the incubation period, visually denote the presence of and after the incubation period, visually denote the presence of microorganism’s growth.microorganism’s growth.

If the test sample causes the turbidity of the medium, that makes If the test sample causes the turbidity of the medium, that makes impossible of the visual record, then after 14 days from the impossible of the visual record, then after 14 days from the beginning of the incubation from each of the vessels there is beginning of the incubation from each of the vessels there is carried a certain amount of the medium in the vessels with the carried a certain amount of the medium in the vessels with the same medium.same medium.

Continue the incubation of the initial and repeating inoculations. Continue the incubation of the initial and repeating inoculations. The total incubation time is 14 + 7 days from the beginning of The total incubation time is 14 + 7 days from the beginning of test. The medicine is sterile, if at the visual record tere is no test. The medicine is sterile, if at the visual record tere is no microorganism growth.microorganism growth.

Investigation of the depressor substanceInvestigation of the depressor substance This test is performed with the aim of exclusion the risk of This test is performed with the aim of exclusion the risk of

blood pressure reducing (depressor reactions) after drug’s blood pressure reducing (depressor reactions) after drug’s administration, if in medicine’s composition there was found administration, if in medicine’s composition there was found the depressor substances (histamine, serotonin, bradykinin) at the depressor substances (histamine, serotonin, bradykinin) at the prodaction.the prodaction.

The investigation is realized on a cat, its mass is not more than 2 The investigation is realized on a cat, its mass is not more than 2 kg. The cat was kg. The cat was narcotizenarcotized by a d by a chloralose, barbiturates or chloralose, barbiturates or urethane to maintain a constant blood pressure.urethane to maintain a constant blood pressure. Through the Through the carotid artery, the animal is attached to the instrument carotid artery, the animal is attached to the instrument recording the blood pressure. In the femoral vein through the recording the blood pressure. In the femoral vein through the cannula is injected by a histamine and the test drug solutions.cannula is injected by a histamine and the test drug solutions.

The test is an inefficacious if the reaction on 1.5 ml of histamine The test is an inefficacious if the reaction on 1.5 ml of histamine does not exceed the response to 1 ml. The sample fails the test does not exceed the response to 1 ml. The sample fails the test if the average response to its injection is higher than the if the average response to its injection is higher than the average responses to 1 ml of histamine per kg of a weight average responses to 1 ml of histamine per kg of a weight body, or if any one of its dose causes a greater depressor body, or if any one of its dose causes a greater depressor response than the final dose of a histamine solution.response than the final dose of a histamine solution.

Acyclic antibiotics and their semi-Acyclic antibiotics and their semi-synthetic analogs analogs ((tetracyclines)tetracyclines)

Antibiotics, which have the partially hydrated nucleus of tetracene ((naphthacenenaphthacene):):

This antibiotics are obtained from the microorganisms of This antibiotics are obtained from the microorganisms of Streptomyces aurofaciens i Streptomyces rimosusStreptomyces aurofaciens i Streptomyces rimosus..

In medical practice there is using the natural and semi- In medical practice there is using the natural and semi- synthetic tetracyclines analogs tetracyclines analogs..

Natural tetracyclinesTetracycline (Tetracyclinum)

(4S,4aS,5aS,6S,12aS)-4-(Dimethylamino)-3,6,10,12,12a-pentahydroxy-6-methyl-1,11-dioxo-1,4,4a,5,5a,6,11,12a-octahydrotetracene-2-carboxamide.

In medical practice tetracycline is used in the form of a base or a hydrochloride (Tetracyclini hydrochloridum) (SPhU 1.2).

OH

C

OHOHOH

OHH3CN

O O

NH2

CH3H3C

O

12

34

4a5

5a67

8

910 11 12

11a 12a

Natural tetracyclinesNatural tetracyclines: : tetracyclinetetracycline, , tetracycline htetracycline h//chch, , oxytetracyclineoxytetracycline dihydratedihydrate, , oxytetracyclineoxytetracycline hh//chch, ,

Chlor(o)tetracyclineChlor(o)tetracycline hh//chch ( (biomicynbiomicyn)) Chlor(o)tetracyclineChlor(o)tetracycline

Oxytetracycline Oxytetracycline (terramicyn)(terramicyn)

TetracyclineTetracycline

Physical propertiesPhysical properties Tetracyclines are an yTetracyclines are an yellow or an light-yellow

crystalline powder, odorless, bitter on test. Their solutions in chloric acid rotate the polarized plane of a Their solutions in chloric acid rotate the polarized plane of a

beam to the leftbeam to the left.. Soluble in water and their salts are a freely soluble in waterSoluble in water and their salts are a freely soluble in water. .

All substances of tetracyclines group are hard soluble in All substances of tetracyclines group are hard soluble in alcohol, but according to the amphoteric properties they are alcohol, but according to the amphoteric properties they are a freely soluble in diluted strong acids and basesa freely soluble in diluted strong acids and bases. . Basic Basic properties are caused of the properties are caused of the Dimethylamino Dimethylamino group present in the 4-th position, , and acidicand acidic - - phenolic hydroxyl in 10-th position and the enol in 3-d and 12-th.

The presence of in tetracycline molecule of two systems of The presence of in tetracycline molecule of two systems of conjugated double bonds at positions 1, 11 cause the conjugated double bonds at positions 1, 11 cause the characteristic absorption in the UV range.characteristic absorption in the UV range.

IdentificationIdentification 1. 1. The reaction the a concentrated sulfuric acidThe reaction the a concentrated sulfuric acid – – forming forming

the anhydrotetracycline’s derivativesthe anhydrotetracycline’s derivatives, , which have a specific which have a specific colourcolour ( (tetracyclinetetracycline h/chh/ch – – violet-redviolet-red, , oxytetracyclineoxytetracycline h/chh/ch – – redred), ), the colour transfers in a yellow at the adding of waterthe colour transfers in a yellow at the adding of water::

2.2. ТТLC method LC method compareing with a standard pharmacopoeial compareing with a standard pharmacopoeial sample.sample.

3.3. Hydrochloride’s of tetracyclines give the reaction of chlorides.Hydrochloride’s of tetracyclines give the reaction of chlorides.4.4. Determine an specific Determine an specific ooptical rotation..5.5. UVUV--spectrometryspectrometry. . Determine the maximum and minimum Determine the maximum and minimum

absorption and calculate a specific absorption rate.absorption and calculate a specific absorption rate. 6.6. Formation of the coloured complex salts in an alcohol medium Formation of the coloured complex salts in an alcohol medium

withwith FeClFeCl33- - brownbrown andand red-brownred-brown ( (present of the present of the phenolic hydroxyl in the 10-th position).).

7.7. Formation of the coloured complexes with the salts of copper Formation of the coloured complexes with the salts of copper (II), zinc(II), zinc..

8.8. Formation in certain conditions, the fluorescent products.Formation in certain conditions, the fluorescent products.9.9. In an express analysis of tetracycline drugs there is used color In an express analysis of tetracycline drugs there is used color

reactions with sodium nitroprusside, n-dimethylaminobenzaldehide, reactions with sodium nitroprusside, n-dimethylaminobenzaldehide, Nessler reagent, diazoreagent.Nessler reagent, diazoreagent.

Formation of the azo dye dues to the present of Formation of the azo dye dues to the present of phenolic phenolic hydroxylhydroxyl::

Quantitative determinationQuantitative determinationLiquid chromatographyLiquid chromatography. . Quantity is calculated Quantity is calculated in percentin percent..Tetracycline’s antibiotics are quantified by the Tetracycline’s antibiotics are quantified by the biological method of diffusion in an agar. In this biological method of diffusion in an agar. In this case, 1 mcg = 1 UA, then 1.0 g of material = case, 1 mcg = 1 UA, then 1.0 g of material = 1000000 UA.1000000 UA.Spectrophotometry in the UV region, Spectrophotometry in the UV region, photocolorimetry and fluorimetry.photocolorimetry and fluorimetry.

StorageStorage Protected from light.

At the storage of tetracycline’s antibiotics, observing a change in color - darkening according to the formation of impurities of angidrotetratcycline and 4-epianhydrotratcycline and the products of their transformation. These substances are more toxic than the initial drugs.

Aqueous solutions of the tetracyclines salts gradually becomes turbidity due to settle down a base.

In weakly acidic medium tetracyclines hydrochlorid solutions are relatively stable, but in an acidic and a base medium, they are easily destroyed. For example, in an alkaline medium formed isotetracycline’s derivatives:

tetracycline isotetracycline

ApplicationApplication Broad-spectrum antibiotics at pneumonia, dysentery, gonorrhea, typhoid and other infectious diseases.

Administration inside in the form of tablets, capsules, suspensions, rarely inner muscular injections. Externally - in the form of ointments for burns treating, phlegmon and eye diseases. H.d. - 0,5 g; H.d.d. - 2,0 g. Oletetrin – tetracycline + oleandomycin.Vitacycline – tetracycline + vitamins С, В1, В2.oxitetramicyn h/ch is included in the ointments composition “Hyoxyson”, “Оxycort”, aerosol “Oxycyclosol”.

Semi-synthetic tetracyclinesMethacycline h/ch

RondomycinGeneric name (Methacyclini hydrochloridum)

4-Dimethylamino-1,4,4а,5,5а,6,11,12а-octahydro-3,5,10,12,12а-pentaoxy-6-methylen-1,11 –dioxo-naphthacene-2-carboxamide

hydrochloride

OH

C

OHOHOH

N

O O

NH2

CH3H3C

O

*HCl

OHCH2

Doxycycline hyclate (Doxycyclini hyclas) (SPhU), Doxybene, Doxycycline hydrochloride

H/cl hemiethanol hemi-hydrate (4S,4aR,5S,5aR,6R,12aS)- 4-(dimethylamino)-3,5,10,12,12а-pentahydroxy-6-methyl-1,11-dioxo-1,4,4а,5,5а,6,11,12а-octahydrotetracene-2-carboxamide

Doxcycline monohydrite - Vibramycin, Doxy-М, Unidox solutab

OH

C

OHOHOH

N

O O

NH2

CH3H3C

O

* HCl * 1/2 C2H5OH * 1/2 H2O

OHCH3

Doxycycline hyclateDoxycycline hyclate

PPropertiesroperties.. Yellow, crystalline powder. . HHygroscopicygroscopic. . A freely soluble in waterA freely soluble in water, , methanolmethanol, , a a slightly soluble in aslightly soluble in a 96 % 96 % ethanol alcohol ethanol alcohol, , a practically a practically insoluble in an etherinsoluble in an ether..

IdentificationIdentification::1.1. TLC-methodTLC-method..2.2. With a concentrated sulfuric acid appearance yellow With a concentrated sulfuric acid appearance yellow

colour.colour.3.3. It gives reaction of chlorideIt gives reaction of chloride..

Quantitative determinationQuantitative determination.. 1. 1. Liquid chromatographyLiquid chromatography..

ApllicationApllication Doxycycline h/ch is highly effective for upper respiratory tract

infection (bronchitis, pleurisy, pneumonia), has a prolonged effect (1-2 times take a day). Methacycline h/ch is better absorbed after oral administration (capsule), lasts longer in the blood and is effective in gonorrhea treating. Antibiotics of this group cause cross-resistance, but microorganisms get accustomed to tetracycline’s raw less than to penicillin.

Tetracycline side effect Forming the complexes with Ca2+, Fe3+ ions that’s why

tetracyclines can accumulate in bones, tooth enamel. Do not recommend taking tetracyclines for children and pregnant women.

When receiving these drugs should not eat dairy products, iron supplements and antacids medicines containing an aluminum, magnesium and calcium salts.

Thanx for attentionThanx for attention