General characteristic of hormones as General characteristic of hormones as medicines, their classification, the medicines, their classification, the

obtaining methods, the identification obtaining methods, the identification methods, amethods, assays, storage conditions and

application. Thyroid, adrenal medulla, pancreas Thyroid, adrenal medulla, pancreas

hormones.hormones. prepared Kozachok S.S.prepared Kozachok S.S.

LectureLecture № №0909

Hormones are biologically active substances that are Hormones are biologically active substances that are produced by glands of internal secretion in very produced by glands of internal secretion in very small quantities, and regulate all the vital processes small quantities, and regulate all the vital processes occurring in the body. Only sex hormones affect occurring in the body. Only sex hormones affect more than 120 body functions.more than 120 body functions. The formation process of hormones is regulated by the The formation process of hormones is regulated by the hypothalamus it is a brain region which is under the optic hypothalamus it is a brain region which is under the optic tubercles.tubercles. Nowadays there are about 50 natural hormones. As drugs Nowadays there are about 50 natural hormones. As drugs there are used galena preparations and individual hormones, there are used galena preparations and individual hormones, which are derived from endocrine glands of a beef cattle as which are derived from endocrine glands of a beef cattle as well as synthetic analogs of natural hormones.well as synthetic analogs of natural hormones.Hormones have long been classified according to their place Hormones have long been classified according to their place of origin, taking into account their physiological effect: thyroid of origin, taking into account their physiological effect: thyroid hormones, pancreatic hormones, adrenal hormones, etc.hormones, pancreatic hormones, adrenal hormones, etc.

Over the past decade in the chemistry of hormones Over the past decade in the chemistry of hormones there there have been have been many new studies. many new studies. It was sIt was synthesized the hypophysis hormones, ynthesized the hypophysis hormones, several hypothalamic hormones. Some of them (rifatiroin, several hypothalamic hormones. Some of them (rifatiroin, somatostatin) are recommended for clinical use. somatostatin) are recommended for clinical use. Discover Discover morphinemorphine--similar similar hormones (enkephalins, endorphins), hormoneshormones (enkephalins, endorphins), hormones of of memory memory and sleep. This is one of the ways to create new analgesics that are and sleep. This is one of the ways to create new analgesics that are not addictive. not addictive.

Create the new methods of synthesis of neurohormones with peptide Create the new methods of synthesis of neurohormones with peptide structure, and many of their structural analogs. It turned out that structure, and many of their structural analogs. It turned out that some of them may affect on the heart, showing mediator or some of them may affect on the heart, showing mediator or modulator of the function, which allows using them for modulator of the function, which allows using them for atherosclerosis and myocardial infarction.atherosclerosis and myocardial infarction. Established the chemical structure and the synthesis of insulin. Established the chemical structure and the synthesis of insulin. However, the chemical synthesis of this hormone has only However, the chemical synthesis of this hormone has only theoretical interest, since the yield of the final product is very small theoretical interest, since the yield of the final product is very small and the cost of performing the synthesis of significant. Therefore, and the cost of performing the synthesis of significant. Therefore, insulin is extracted based on animal products or by genetic insulin is extracted based on animal products or by genetic engineering.engineering.

Recent advances in the chemistry of hormones

TTissue hormonesissue hormones or kininsor kinins have the i have the importanmportancece role role for for the body's vital functions. They are formed not in the the body's vital functions. They are formed not in the endocrine glands, endocrine glands, butbut at various points in the body, where at various points in the body, where their influence is currently required. Kinins have a peptide their influence is currently required. Kinins have a peptide structure. They regulate the basic biochemical and structure. They regulate the basic biochemical and physiological processes in the body and affect physiological processes in the body and affect on on many of many of its functions. This gave impetus to the use of some of its functions. This gave impetus to the use of some of kinins in human and veterinary medicine.kinins in human and veterinary medicine.Modern studies of drugs of animal origin are carried out in Modern studies of drugs of animal origin are carried out in several directions. several directions. TThe components of organs and tissueshe components of organs and tissues cause the greatest interest in studyingcause the greatest interest in studying, which in chemical , which in chemical structure are proteins, glycosaminoglycans and nucleic structure are proteins, glycosaminoglycans and nucleic acids. They have a wide range of actions. Obtained drugs acids. They have a wide range of actions. Obtained drugs from the thymus gland, prostate, trachea, blood vessels from the thymus gland, prostate, trachea, blood vessels and other organs of the animal materials. and other organs of the animal materials.

Chemical classification of hormonesChemical classification of hormones1.1. Hormones that have the structure of amino-acid, Hormones that have the structure of amino-acid, amino alcohols, amino alcohols,

polypeptides, proteins and their derivativespolypeptides, proteins and their derivatives ::a)a) thyroid hormones and their synthetic analoguesthyroid hormones and their synthetic analogues;;b)b) parathyroid hormoneparathyroid hormone;;c)c) pituitary hormonespituitary hormones;;d)d) Hormones of the pancreasHormones of the pancreas;;e)e) Hormones of adrenal medullaHormones of adrenal medulla ..2. Hormones that have the steroid structure2. Hormones that have the steroid structure ::a)a) Cortical layer of the adrenal hormones (corticosteroids) and Cortical layer of the adrenal hormones (corticosteroids) and

their semisynthetic analogstheir semisynthetic analogs;;b)b) progestin (lutoidni) hormones and their semisynthetic progestin (lutoidni) hormones and their semisynthetic

analogsanalogs;;c)c) androgenic hormones and semisynthetic anabolic drugs;androgenic hormones and semisynthetic anabolic drugs;d)d) estrogenic hormones and their synthetic analogs of non-estrogenic hormones and their synthetic analogs of non-

steroidal structure.steroidal structure.

Thyroid hormonesThyroid hormones Thyronine derivatives of the thyroid gland have biological

activity::

In 1914, Mr. Kendall is siparated out from the thyroid gland of a cattle thyronine tetraiodidne derivative (chemical structure was established in 1927). This substance called thyroxine (3,5,3',5'-tetraiodothyronine):

The presence of an asymmetric carbon atom determines the The presence of an asymmetric carbon atom determines the presence of two optical isomers, of which L-thyroxine presence of two optical isomers, of which L-thyroxine which is 3 times more active than D-isomer.which is 3 times more active than D-isomer.

In the 1952-1955 years , it was proved that the hormonal activity of the In the 1952-1955 years , it was proved that the hormonal activity of the thyroid gland the thyroxine does not only have, but other iodo thyroid gland the thyroxine does not only have, but other iodo derivatives of the thyronine: 3,5,3'-triiodothyronine and 3,3'- of the thyronine: 3,5,3'-triiodothyronine and 3,3'-diiodothironinediiodothironine

Biosynthesis of these hormones in the body is produced by the tyrosine and iodine, which enter to the body with food and water.              In medical practice, there is using thyroidin, synthetic L-thyroxine and triiodothyronine as sodium salts.

ThyroidinThyroidinThyreoidin contains hormones ofThyreoidin contains hormones of LL- - thyroxinthyroxin andand LL-3,5,3-3,5,3’’--triiodo thyronine. . Obtaining by the drying and grinding the thyroid Obtaining by the drying and grinding the thyroid glands of a cattleglands of a cattle. . Yellowish-white powder with faint odor that is a Yellowish-white powder with faint odor that is a characteristic of the dried animal tissuescharacteristic of the dried animal tissues. . Not soluble in water and other solvents, soluble in Not soluble in water and other solvents, soluble in alkalis and carbonates (acid properties).alkalis and carbonates (acid properties). Store in a well-sealed jars of dark glassStore in a well-sealed jars of dark glass..Apply at the hypofunction of Apply at the hypofunction of thyroid glandthyroid gland, which leads , which leads to the hypothyroidism, myxedema, cretinism, obesity to the hypothyroidism, myxedema, cretinism, obesity or endemic goiter. H.s.d.-0, 3 g; H.d.d.-1, 0 g.or endemic goiter. H.s.d.-0, 3 g; H.d.d.-1, 0 g.

Identification of Identification of ThyroidinThyroidin 1.1. Protein is determined by the formation of yellow Protein is determined by the formation of yellow

color after boiling the drug in alkaline solution. With color after boiling the drug in alkaline solution. With further addition of diluted sulfuric acid solution there further addition of diluted sulfuric acid solution there is decolorizing and droping out the white colloidal is decolorizing and droping out the white colloidal precipitate.precipitate.

2.2. To determine the organically bound of iodine the To determine the organically bound of iodine the preparation is destroyed by the calcination with a preparation is destroyed by the calcination with a mixture of KNOmixture of KNO33 and Na and Na22COCO33. Forming iodides are . Forming iodides are extracted with water and identified by the oxidation extracted with water and identified by the oxidation of chloramines in the HCl environment. Iodine of chloramines in the HCl environment. Iodine liberated, the chloroform layer is coloured in red-liberated, the chloroform layer is coloured in red-violet color.violet color.

3.3. Thyroidin is burned with oxygen in the flask. As an Thyroidin is burned with oxygen in the flask. As an absorption substance using a starch solution that absorption substance using a starch solution that contains 0.2% sulfamic acid. Iodine forms during contains 0.2% sulfamic acid. Iodine forms during combustion, it paints an absorbing layer in blue.combustion, it paints an absorbing layer in blue.

4.4. Gives reaction of the Gives reaction of the phenolic hydroxyl withphenolic hydroxyl with FeClFeCl33..

Quantitative determinationQuantitative determination IIn thyroidin there is determined the content of n thyroidin there is determined the content of

the organically iodine bound.the organically iodine bound.Spending the mineralization by perhydrol in the Spending the mineralization by perhydrol in the presence of concentrated Hpresence of concentrated H22SOSO44 (USPh (USPh X).Forming the iodides and partial oxidation of X).Forming the iodides and partial oxidation of iodides to iodates. Iodides is oxidized to iodates by iodides to iodates. Iodides is oxidized to iodates by the solution of KMnOthe solution of KMnO44::

The excess ofThe excess of KMnOKMnO44 is removed by NaNOis removed by NaNO22 solution:solution:

Possible excess of the nitrates can be destroyed by an ureaPossible excess of the nitrates can be destroyed by an urea::

In the solution, there is only one oxidant - iodate acid in an In the solution, there is only one oxidant - iodate acid in an amount equivalents to the iodine content in thyroidin precise amount equivalents to the iodine content in thyroidin precise mass. To add a solution of KI and iodine, that was separated, mass. To add a solution of KI and iodine, that was separated, it titrated by Nait titrated by Na22SS22OO33 solution solution ::

TThyroidin hyroidin containscontains 0,17-0,23 % 0,17-0,23 % of the iodineof the iodine. Е. Емм=1/6А=1/6Амм(І).(І).

2.2. Mineralization is carried out in a flask Mineralization is carried out in a flask combustion with oxygen (SP XI):combustion with oxygen (SP XI):

RСНRСН22І + ОІ + О22 → І→ І2 2 + СО+ СО22 + Н + Н22ОО Evolving iodine is absorbed by Evolving iodine is absorbed by NаОН:NаОН:

ІІ2 2 + 2NаОН → NaІO+ 2NаОН → NaІO + NaІ + Н+ NaІ + Н22ОО For the oxidation of the forming iodides in the flask to For the oxidation of the forming iodides in the flask to

put acetate bromide solution till yellow colourput acetate bromide solution till yellow colour::2Br2Br++(СH(СH33СOO)СOO)- - + + NaІ + НNaІ + Н22О → BrО → Br22 + NaІO + NaІO + 2CH+ 2CH33СОOHСОOH

NaІONaІO + NaІ + + NaІ + 55BrBr22 + + 55НН22О → О → 22NaІONaІO33 + 10H+ 10HBr Br

Then to add the concentrated Then to add the concentrated formic acid solution till formic acid solution till the colourlessing of the solutionthe colourlessing of the solution: :

НСООН + НСООН + BrBr22 → 2НBr + СО → 2НBr + СО22

Bromine residual traces is pumped.Bromine residual traces is pumped.

At the At the thyroidin combustion also forming nitrogen oxides, thyroidin combustion also forming nitrogen oxides, which dissolves in a solution of NaOH, converting to nitrite which dissolves in a solution of NaOH, converting to nitrite ions. For the delation of them to add to the reaction mixture ions. For the delation of them to add to the reaction mixture 3% solution of sulfamic acid3% solution of sulfamic acid : :

HH22N-SON-SO22OH + HNOOH + HNO2 2 → → HH22SOSO44 ++ N N22 + Н + Н22ОО

Therefore, remove all oxidants except iodate equivalent to Therefore, remove all oxidants except iodate equivalent to the content of iodine bound in the sample. After that, in the the content of iodine bound in the sample. After that, in the flask to add 1 g of KI, which interacts with iodate, allocates flask to add 1 g of KI, which interacts with iodate, allocates an equivalent amount of iodine. It is titrated by 0.005 M an equivalent amount of iodine. It is titrated by 0.005 M NaNa22SS22OO33 (indicator - starch): (indicator - starch):

22NaІONaІO33 + 10+ 10КІ + 6КІ + 6HH22SOSO4 4 → 6І→ 6І2 2 + + NaNa22SSОО44 + + 5K5K22SOSO44 + 6 + 6НН22ОО

II22 + 2 + 2 NaNa22SS22OO3 3 → NaІ + Na→ NaІ + Na22SS44OO66

Parallel to do a control experiment (without burning).Parallel to do a control experiment (without burning). ЕЕмм=1/6А=1/6Амм(І).(І).

Levothyroxine sodium (SPU) Levothyroxinum natricum

Sodium (2S)-2-amino-3-[4-(4-hydroxy-3,5- diiodidophenoxy)-3,5- diiodophenyl] propanoate

hydrate The synthetic levogyrate isomer of thyroxine (L-thyroxine). Properties. Fine crystalline powder, almost white or slightly

brownish-yellow color. Very slightly soluble in the water, slightly soluble in 96% ethanol, practically insoluble in the ether. Soluble in dilute alkali.

I

HO

I

O

I

I

CH2 CH C

O

ONa

NH2

H2O

Liothyronine Sodium (SPU)Liothyroninum natriсum

Sodium (2S)-2-amino-3-[4-(4-hydroxy-3-iodophenoxy)-3,5- diiodophenyl] propanoate

Synthetic analogue of triiodothyronine to 5 times stronger than L-thyroxine.

Properties. The white or slightly colored powder. Very slightly soluble in the water, slightly soluble in 96% ethanol. Soluble in dilute alkali.

I

HO O

I

I

CH2 CH C

O

ONa

NH2

Identification of solium saltsIdentification of solium saltsLevothyroxineLevothyroxine

1.1. Specific optical rotationSpecific optical rotation 2.2. IR spectroscopyIR spectroscopy 3.3. TLCTLC

LiothyronineLiothyronine1.1. Specific optical rotationSpecific optical rotation 2.2. IR spectroscopyIR spectroscopy 3.3. UVUV - - spectroscopyspectroscopy

4.4. To the substance to add some drops of the concentratedTo the substance to add some drops of the concentrated HH22SOSO44 and and heatheat – – forming the steam of the violet colorforming the steam of the violet color ( (iodineiodine).).

5.5. After After mineralizationmineralization ((under the action of the diluted Hunder the action of the diluted H22SOSO44 ) it gives reaction of sodium.

Assay 1. Both drugs were determined by liquid chromatography.2. Combustion in a flask with oxygen, using for the absorption

NaOH. Than Iodometry (see thyroidin).3. The method is based on the dehalogen at the heating with

zinc powder in alkaline medium:

Then to do argentometric titration educated iodide ions (indicator - sodium eosinate). Еm=1/4М.м.

4КІ + 4AgNO3 → 4AgI + 4KNO3

HO

I

I

O

I

CH2

HC

NH2

COOH

I

HO O CH2

HC

NH2

COOK

+ 2 Zn + 9 KOH

+ 4 KI + 2 K2[Zn(OH)4] + H2O

Storager – in the airtight containers. In the dark place, at 2-8 0С.Remedy of the synthetic analogs of thyreodine: L - thyroxine (levothyroxine) (Pharmak) 50 mg and 100 mg № 50. Triiodothyronine (Borschagovsky Chemical Plant ) - liotironine Novotiral (levothyroxine + liotironine) Tiratrikol (Triakan) - 3,5,3 '- triiodotiacetate acid - a physiological metabolite of liotironine. Iodtirox - levothyroxine + potassium chloride Antithyroid drugs (for treatment of toxic goiter) - merkazolil (tiamazol) Metizol.

Hormones of adrenal medulla (catecholamines and Hormones of adrenal medulla (catecholamines and their synthetic analogues)their synthetic analogues)

Adrenal glands - paired glands, which are composed of two layers: cortical and medullary (inner). Medulla layer produces the hormone of adrenaline and noradrenaline accompanying to it, and the cortical layer - about 40 different hormones, which are called corticosteroids. Adrenaline was first identified by a russian scientist Cybulski M.O. at the end of the XIX century, and in 1903 established its structure, which is confirmed by synthesis. Raw material for production of adrenaline and corticosteroids is a adrenal (gland) of a beef cattle. So from 45 kg of the adrenal glands receive only 9.5 grams of pure adrenaline. Therefore, in the present preparations of adrenal hormones are obtained only synthetically ways.

Despite there are several Despite there are several hydrophilic groups in the hydrophilic groups in the molecule, the adrenaline molecule, the adrenaline practically insolubles in practically insolubles in water according to the water according to the formation of betaine (inner formation of betaine (inner salt, zwitter-ion).salt, zwitter-ion).From aqueous solutions of salts From aqueous solutions of salts (hydrochloride, hydrotartrate) it (hydrochloride, hydrotartrate) it can be settled down the base of can be settled down the base of adrenaline with ammonia, adrenaline with ammonia, filtered, dried and used for the filtered, dried and used for the determining of the specific determining of the specific rotation.rotation.

In medical practice, there are using of adrenaline (epinephrine) hydrochloride, epinephrine tartrate, noradrenaline hydrotartrate and their synthetic analogs - mezaton (phenylephrine h/ch) izadrin (isoprenaline h/ch) berotek (fenoterol h/br), salbutamol, verapamil h/ch. Properties. Adrenaline and noradrenaline hydrotartratis - white or white with a yellowish white crystalline substance, odorless. It easily solubles in water, practically insoluble in ether and chloroform, practically insoluble in ethanol. Like other phenols, these compounds are soluble in alkaline solutions, can be oxidized. Under the influence of light and oxygen they form a colored oxidation products. They have amphoteric properties. As diatom phenol it is easily oxidized and is a good reducing agent. Natural adrenaline - levogyrate, synthetic - racemate.

Adrenaline tartrate(Adrenalinі tartras)(SPhU 1.1) Epinephrine bitartrate*

(1R)-1 -(3,4-dihydroxyphenyl)-2-(methylamino) ethanol hydrogen (2R,ЗR)-2,3-dihydroxy butandioate

CH

CH2

NH

CH3

OHHO

HO

HC

CH

COOHHO

OHHOOC*

Noradrenaline hydrotartrate(Noradrenalini hydrotartras)Levarterenol bitartrate*

CH

CH2

NH2

OHHO

HO

HC

CH

COOHHO

OHHOOC* * H2O

(-)-1-(3,4-Dioxyphenyl)-2-amino ethanol hydrotartrate

Synthesis scheme of remedies of Synthesis scheme of remedies of adrenaline and noradrenalineadrenaline and noradrenaline

The gottenThe gotten racemate according the synthesis are separated racemate according the synthesis are separated by the tartratic acid, using different solubility of the by the tartratic acid, using different solubility of the hydrotartrates in alcohol. Levogyrate tartrate isomer is not hydrotartrates in alcohol. Levogyrate tartrate isomer is not soluble in alcohol, dexter levogyrate - soluble.soluble in alcohol, dexter levogyrate - soluble.

Identification 1. The specific rotation after transfering to the chloride salt, ultraviolet

and infrared spectroscopy. 2. To the aqueous solution of a substance to add diethoxy

tetrahydrofuran in the concentrated acetate acid and heated. To a cooled solution to add dimethyl amino benzaldehyde solution in a mixture of hydrochloric and acetate acids. The obtaining and the control solution have the same yellow color.

3. Reaction on tartrate: a substance is heated with potassium bromide and resorcin in the presence of concentrated sulfuric acid, appearance a dark blue color. After cooling to add water, color changes to red:

HC

CH

HO COOH

HOOC OH

OH

OH

C

COOH

O H

H2SO4

KBr

C

COOH

HO

OHOH

O

C COOH

O

H

CH

COOH

HO OH

OHOH

H2SO3 H2O ;

[O]

++ +2

2 +.- H2O

4. Oxidized by 0,1М iodine solution in the buffer solutions with рН 3,56 and 6,5. Adrenaline an this condition forms adrenochrome, which gives dark-red color of solution (рН 3,56) (SPU) or red-violet (рН 6,5) color.

Noradrenaline forms noradrenochrome (red-violet color) only in the solution with рН 6,5.

5. Not pharmacopeial reaction:a) Due to the presence of two phenolic hydroxyl epinephrine and

norepinephrine with a solution of FeCl3 form an emerald-green color, which transfers by the adding drops of ammonia solution to a cherry-red, then an orange-red colors.

b) On the tartrate ion reaction with a potassium salt. c) With Tollens reagent (silver is reduced from the amino

solution). d) With Felling reagent (orange-red sediment Cu2O).e) Color reaction with 1,2-dinitrobenzene - blue-violet color.

CHHO

HO

OHNH CH3 O

OH

NO2

NO2 H2

NHNO2

OH

KOH

CHO

O

OHNH CH3

NN+

KOOK

O-

+ H2

f) In alkaline solution of adrenaline is oxidized by atmospheric oxygen to form adreno lutain - there is a yellow-green fluorescence

g) At the heating with NaOH or KOH adrenaline and noradrenalin have hydroamino splitting to form 3,4-dioxyacetophen and methylamine or ammonia:

Purity testsPurity tests By the UV- spectrometry it is determined in adrenaline

tartrate adrenalon in norepinephrine hydrotartratis - noradrenalon; Noradrenaline in adrenaline tartrate is determined by TLC.

Assay of adrenaline and noradrenaline Assay of adrenaline and noradrenaline hydrotatrates (SPhU)hydrotatrates (SPhU)

1. Acidimetry in nonaqueous medium. Titrate in the in nonaqueous medium. Titrate in the present of the concentrated acetic acidpresent of the concentrated acetic acid, the indicator the indicator -- crystal violet (Е=М.м).

2.2. The content of drugs in solutions for injections it is The content of drugs in solutions for injections it is determined by photocolorimetry (color reaction with determined by photocolorimetry (color reaction with Iron-citrate reagent, 1,2-dinitrobenzene).Iron-citrate reagent, 1,2-dinitrobenzene).

3. UV- spectrometry..4.4. AlkalimetryAlkalimetry. Е. Емм = М.м. = М.м./2/2..

StorageStorageIn the dark place in airtight container. Easily oxidized In the dark place in airtight container. Easily oxidized when exposed to light and oxygen, and therefore when exposed to light and oxygen, and therefore stabilizstabilizinging the solutions for injection of adrenaline the solutions for injection of adrenaline and noradrenaline hydrotartratis and noradrenaline hydrotartratis by the by the addadding of theing of the reductant - 0,1% sodium metabisulfite (disulfitreductant - 0,1% sodium metabisulfite (disulfitee) ) NaNa22SS22OO55.. – ApplicationApplication

Adrenaline and noradrenaline hydrotartratis used as Adrenaline and noradrenaline hydrotartratis used as adrenomimetic (vasoconstrictor) adrenomimetic (vasoconstrictor) remedyremedy. The. They are y are prescribed prescribed aan the collapse, a sharp decrease in blood pressure as a n the collapse, a sharp decrease in blood pressure as a result of injuries, poisonings, during surgery to reduce result of injuries, poisonings, during surgery to reduce bleeding and blood loss. bleeding and blood loss. Adrenaline hydrotartrate is injected hypodermic as 0.18% solution of 0,1-0,5 ml, norepinephrine hydrotartratis - intravenous 0.2% solution. In oculus medical practice (dilates the pupil) using 0,1% solution of adrenaline.

Phenylephrine hydrochloride (Phenylephrini hydrocloridum) (SPhU)Mesaton (Mesatonum)

(1R)-1-(3-Hydroxyphenyl)-2-((methylamino) ethanol hydrochloride Synthetic analogue of the adrenaline. CHARACTERS. Appearance White or almost white, crystalline powder. Solubility Slightly soluble in water, sparingly soluble in methanol, slightly

soluble in ethanol (96 per cent). It dissolves in dilute mineral acids and in dilute solutions of alkali hydroxides.

CH

CH2

NH

CH3 * HCl

OHHO

IdentificationIdentification of of Phenylephrine Phenylephrine hhydrochlorideydrochloride

1. Specific optical rotation, , melting pointmelting point, І, ІRR--spectroscopyspectroscopy..2.2. Gives reaction of chlorideGives reaction of chloride..3.3. WithWith CuSOCuSO44 at present of at present of NaOHNaOH mezaton forms the complex of a mezaton forms the complex of a

violet colorviolet color, , which which , unlike ephedrine, insoluble in ether., unlike ephedrine, insoluble in ether.

4.4. Not pharmacopeial reactionNot pharmacopeial reaction: : withwith FeClFeCl33 ((on the phenol on the phenol hydroxylhydroxyl) – ) – violet colorviolet color..

ASSAYASSAY1.1. ((SPhUSPhU). ). AlkalimetryAlkalimetry. . Substance is dissolved in a mixture of 0.1 M Substance is dissolved in a mixture of 0.1 M

HCl and 96% ethanol and titrated by 0.1 M ethanolic solution of HCl and 96% ethanol and titrated by 0.1 M ethanolic solution of NaOH potentiometrically. At the calculation to taken the volume of NaOH potentiometrically. At the calculation to taken the volume of titrant between the two potential jumps on the titration curve. Em = titrant between the two potential jumps on the titration curve. Em = M.m. M.m.

2.2. Return bromatometryReturn bromatometry. . IndicatorIndicator – – starchstarch. . Parallel to do control testParallel to do control test. . ЕЕм м = 1/6 М.м.= 1/6 М.м.

2. Acidimetry in nonaqueous mediumin nonaqueous medium, a direct titrationa direct titration at the present of mercuryat the present of mercury (II) acetate. Е. Ем м = М.м.= М.м.

3.3. AlkalimetryAlkalimetry. Е. Ем м = М.м.= М.м.

4.4. ArgentometryArgentometry. Е. Ем м = М.м.= М.м.

5.5. MercurimetryMercurimetry. Е. Ем м = М.м.= М.м.

ApplicationApplication Alpha-adrenoceptor agonist, Adrenomimetic

(vasoconstrictor) agent.         Release. Solution for injection 1% 1,0 № 10: mezaton-, Mesafeton-Darnisa. Nazol-baby - a drop

of 0,125% 15 ml. Combination preparations: Farmacitron (10 mg), Coldrex (10mg), KOLDFLU,

rinza, TeraFlu, AntiFlu, Anticataral.

Isadrine (Isadrinum) hydrochloride*

1-(3,4-dioxyphenyl)-2-isopropilaminoethanol h/ch Synthetic analogue of adrenalіne . Properties. White crystalline powder, freely solubles in water.

Melting point 167-169 °C. Under the action of atmospheric oxygen and light, easily oxidized and becomes pink.           Adrenomimetic tool. Unlike adrenaline, greatly expands the bronchial tubes, slightly vasoconstrictor. Using at bronchial asthma. Administered by inhalation of 1 ml of 0.5% solution.

HOHC C

H2

NH

HO

OH

CH

CH3

CH3

* HCl

Identification of Identification of isadrineisadrine1.1. IR-IR- andand UVUV--spectroscopyspectroscopy..2.2. Chloride’s reactionChloride’s reaction..3.3. WithWith FeClFeCl33 – – green colorgreen color, , at the addingat the adding NaHCONaHCO33 transfers to a transfers to a

dark bluedark blue, а, аnd than to a rednd than to a red..4.4. The formation of the isopropyladrenochromeThe formation of the isopropyladrenochrome. . The drug is The drug is

dissolved in the two buffer solutions, whith pH 3.5 and 6, 5. dissolved in the two buffer solutions, whith pH 3.5 and 6, 5. When mixing these solutions with 0.05 M iodine solution When mixing these solutions with 0.05 M iodine solution and bleaching of the iodine excess after 5 minutes. the first and bleaching of the iodine excess after 5 minutes. the first solution becomes intensely red and the second one - a red-solution becomes intensely red and the second one - a red-violet (unlike noradrenaline).violet (unlike noradrenaline).

5.5. With Tollens and Felling reagentsWith Tollens and Felling reagents..6.6. WithWith HNOHNO22 – – dirty-violet colordirty-violet color..7.7. Azoconection reaction (on phenolyc radical) - cherry-red Azoconection reaction (on phenolyc radical) - cherry-red

colorationcoloration..8.8. With a basic CuSOWith a basic CuSO44 solution – the complex has dark green color solution – the complex has dark green color..

ASSAYASSAY KKjeldahl methodjeldahl method. Е. Ем м = М.м.= М.м. Acidimetry in nonaqueous mediumin nonaqueous medium. Е. Ем м = М.м= М.м

Hormones of the pancreas. Hormones of the pancreas. InsulinInsulin

Insulin is a hormone that is produced by Langerhans islets of the Insulin is a hormone that is produced by Langerhans islets of the pancreas (from Lat. insula - island).pancreas (from Lat. insula - island). It has the huge influense on the carbonate exchanging reaction in our It has the huge influense on the carbonate exchanging reaction in our body. Using inside insulin the level of suger in blood extremely body. Using inside insulin the level of suger in blood extremely reduces. Insulin is used for the treatment of diabetes.reduces. Insulin is used for the treatment of diabetes. Insulin is obtained by the extraction from the pancreas of beef cattle, Insulin is obtained by the extraction from the pancreas of beef cattle, or by genetic engineering. This crystalline powder, readily soluble in or by genetic engineering. This crystalline powder, readily soluble in water. Ebel was first extracted insulin in 1925. water. Ebel was first extracted insulin in 1925. Pharmaceutical agent is insulin injection (Insulinum pro Pharmaceutical agent is insulin injection (Insulinum pro injectionibus), which is prepared by dissolving the crystalline powder injectionibus), which is prepared by dissolving the crystalline powder of insulin in water, acidified with HCl. For the preservation of insulin of insulin in water, acidified with HCl. For the preservation of insulin was added to a solution of 0.3% solution trikrezola.was added to a solution of 0.3% solution trikrezola.Remedy is an insulin for injection (Insulinum pro injectionibus), Remedy is an insulin for injection (Insulinum pro injectionibus), which is prepared by dissolving the crystalline powder of insulin in which is prepared by dissolving the crystalline powder of insulin in water, acidified with HCl. For the preservation of insulin there was water, acidified with HCl. For the preservation of insulin there was added to a solution of 0.3% tricresol solution.added to a solution of 0.3% tricresol solution.

The drug is a clear, colorless or slightly yellowish The drug is a clear, colorless or slightly yellowish liquid acid reaction (pH 2,5-3,5), with the smell of liquid acid reaction (pH 2,5-3,5), with the smell of preservative.preservative.According to the chemical structure insulin is a According to the chemical structure insulin is a macromolecular compounds of protein nature. The empirical macromolecular compounds of protein nature. The empirical formula of its is formula of its is СС254254HH337337NN6565SS66. Molecular weight - 5733.5.. Molecular weight - 5733.5.In 1945-1956 Sanger for the first time established the In 1945-1956 Sanger for the first time established the chemical structure of insulin. At the oxidation of the formic acid chemical structure of insulin. At the oxidation of the formic acid HCOOOH it is divided into two monomeric peptides - A HCOOOH it is divided into two monomeric peptides - A peptide composed of 21 aminoacids, peptide B - with 30 peptide composed of 21 aminoacids, peptide B - with 30 aminoacids. aminoacids. Placement of the aminoacids in the insulin molecule doesn’t Placement of the aminoacids in the insulin molecule doesn’t have the order. This protein has an unique structure.have the order. This protein has an unique structure.Animals insulin have the same structure of the peptide B, but Animals insulin have the same structure of the peptide B, but slightly different structure of peptide A. Since pig’s insulin has slightly different structure of peptide A. Since pig’s insulin has in 8-th position threonine (instead of alanine), and in position in 8-th position threonine (instead of alanine), and in position 10-th isoleucine (instead of valine).10-th isoleucine (instead of valine).

For identification to the remedy to add by drops the diluted NaOH solution – forming the flakes sediment, which dissolves at the acidifiing.Insulin is easily destroyed by digestive enzymes, so it does not take per os.It is injected hypodermic, inner muscular. 1 МО = 0,045 mg of the crystalline insulin. Insulin syringes 100 AU/ml and 40 AU/ml.Released in sterile vials on 5 and 10 ml, sealed with rubber caps. 10 ml of liquid = 20 or 40 MO. The drug remains active for 1.5 years when stored in a cool place.Insulin is a short action (Farmasulin H humanity, Himulin regular humanity, Humudar R humanity, pork) and the midle actopn (Farmasulin HL humanity, Himulin NPH, Humodar B, B-insulin, pig.).

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