AMINO ACID AND PROTEIN AS DRUG SUBSTANCES

*Asam aminoAsam amino adalah suatu senyawa kristalin yang tidak berwarna, larut dalam air, but sparingly dalam pelarut organik. Merupakan senyawa amfoterikAsam amino diklafikasikan sebagaiAsam amino alifatik netral (glisin) CH2(NH2)-COOHAsam amino alifatik asam (asam aspartat) HOOC-CH2-CH(NH2)-COOHAsam amino alifatik basa (lisin) H2N-(CH2)4-CH(NH2)-COOHAsam amino aromatik (fenilalanin) C6H5-CH2-CH(NH2)-COOH Asam amino heterosiklik (triptofan)

6. Asam amino bersulfur (alifatik,netral, metionin)CH3-S(CH2)2CH(NH2)-COOH 26 asam amino telah dapatdiketahui/diisolasi dari protein secara hidrolisis dengan asam, alkali, dan enzim. Dan ada 170 jenis asam amino telah dapat diperoleh dari alam dalam bentuk bebas atau terikat dengan senyawa lain.

NPC-S2-2012

*Sintesis asam aminoAminasi asam-asam a-halogenRCH2-COOH RCHBr-COOH RCHNH2-COOH + NH4BrSintesis SteckerRCHO RCH(OH)CN RCH(NH2)CN RCH(NH2)COOHReduktive aminasiRCOCOOH + NH3 R-C(=NH)COOH RCH(NH2)COOH + H2OSintesis Erlenmeyer azlakton

Diastereoisomer Metoda ini didasarkan pada pembentukan garam diastereoisomer dari campuran rasemik dengan basa optis aktif (kuinin, striknin, brusin, kinkonin, kloramfenikol)Br22NH3KCNH+NH3(+) B + (dl) A (+) B. (+)A + (+)B. (-) ANPC-S2-2012

*Analisis asam amino Analisis GC dan analisis KLT10 mg (simplisia)Suspensikan dalam 20 ml amilalkohol, dijenuhkan dgn as. hidrobromat anhidrat, dipanaskan pada penangas minyak (165 oC) hingga bagian alkohol hilang dan diuapkanEster asam amino hidrobromidaLar. berbentuk sirup+ kan dengan 8 ml asam asetat anhidrat, dibiarkan pada suhu 25 oC selama 5 menit, kemudian diuapkanTambahkan 1-5 ml amilalkohol atau benzenNPC-S2-2012PreparatSiap untuk analisis (KLT, GC)

*NPC-S2-2012*Rf asam amino (analisis KLT) (air-EtOH-as. asetat = 1 : 5 : 0,5). Spray Ninhidrin Asam aminowarna RfAsam aspartatviolet 0,11Glisinpink 0,31A-alaninpink 0,45Isoleucinpink 0,70Tirosinpink 0,73Retensi waktu (Rt) analisis GC untuk asam amino

As aminoRt (m)As aminoRt (m)A-alanin14Glisin 24Valin 16b-alanin26as. a-butirat17As-b-butirat28Isoleucin 19Prolin29Norvalin20As.g-butirat32Leucin22Ornitrin33norleucin23treonin35

As aminoRt (m)Serin39Fenilalanin60As. aspartat84As. glutamat102Tirosin 132

*PeptidaPeptida adalah poliamida asam amino yang mempunyai rumus sebagaiSenyawa peptida mempunyai aktivitas biologis dan banyak terdapat di alam, diantaranya tripeptidaglutamin (dalam khamir).ProteinSemua kehidupan mengandung sejumlah besar protein yg berbeda.Perbedaannya adalah susunan/urutan asam amino, kandungan non asam amino,Bobot molekul, korformasinya.Contoh : albuminglobulinglutelinhiston, dll NPC-S2-2012

INTRODUCTION

Protein [Greek: protos = first] refers to a class of rather complex nitrogenous compounds that are synthesized by all living organisms, and yield respective array of amino acids when hydrolyzed.Importantly, proteins essentially provide the amino acids required for the growth and subsequent repair of impaired animal tissue.Composition of Proteins: Proteins, are composed of a host of vital elements, such as: C, H, O, N, P, S, and Fe, which ultimately make up the greater segment of the animal and plant tissue. In fact, the amino acids do represent the basic structure of proteins. Generally, foods invariably contain protein with varying numbers and types of amino acids. However, one may recognize a complete protein as one that predominantly contains all the essential amino acids viz., arginine, histidine, isoleucine, lysine, leucine, meltionine, phenylalanine, threonine, tryptophan, and valine. In human beings, they are indeed an absolute must for the maintenance of body weight and required growth.

PROTEIN : INTRODUCTIONSources: Interestingly, the various known and important sources of proteins are, namely: cheese, milk, eggs, meat, fish, and certain vegetables viz., soybeans are recognized as the best sources.Nevertheless, the proteins are invariably found in both animal and vegetable sources of food. It has been duly observed that there are many incomplete proteins which are found in vegetables; and they do contain some of the so-called essential amino acids. Thus, a vegetarian diet may judiciously make up for this by combining various vegetable groups which complement each other in their basic amino acid groups. This ultimately provides the body with complete protein.The major animal proteins are as enumerated under in Table-12.3:

Functionality of ProteinsThe ingested proteins serve as an important source of amino acids essentially required to synthesize the bodys own proteins, that are quite essential not only for the growth of new tissue but also the repair of damaged tissue. Importantly, proteins constitute an important segment of all the cell membranes. It is proved beyond any reasonable doubt that the excess of amino acids derived from the diet may be conveniently converted to rather simpler carbohydrates, and ultimately get oxidized to produce adenosine triphosphate (ATP) as well as heat. Thus, 1 g of protein gives rise to 4 kcal of heat.

BRIEF DESCRIPTION OF PROTEINS USED AS DRUGS

There are quite a few unique and remarkable proteins that are essentially present in the humans which attribute highly efficacious and critical functions in vivo. Some proteins that belong to this category, used normally as therapeutic agents (drugs), have been described briefly in the sections that follows:

Complement Protein (Complement Factor C-3)[Latin; Complere = to Complete]Complement protein refers to a group of proteins in the blood which play a vital role in the bodys immune defense mechanisms via a cascade of interactions. However, the components of complements are labeled C1 through C9. Nevertheless, C3 and C5 are the most important of these.Complement invariably acts by killing directly the organisms; by opsonizing an antigen, thereby stimulating phagocytosis; and by stimulating inflammation and the B-cell mediated immune response.Importantly, all complement proteins usually lie inactive in the blood unless and until activated by either the classic or the recognized alternative pathways.

Complement Protein (Complement Factor C-3)An observed abnormality or deficiency in complement protein strategically refers to an autosomal recessive trait. The lack of factor C3 enhances susceptibility to common microbial infections, whereas deficits in C5 through C9 are invariably associated with enhanced incidence of autoimmune ailments, such as:glomerulonephritis, andsystemic lupus erythematosusIt has been duly observed that the lack of any of the more than 25 proteins intimately involved in the complement system may affect the bodys defense mechanism adversely.

Gelatin [Latin: Gelatina = Gelatin]Gelatin refers to a derived protein duly obtained by the hydrolysis of collagen strategically located in the connective tissues of the skin, bones, and joints of animals. It is used profusely in food products e.g., fruit jellys; in the preparations of pharmaceutical dosage forms e.g., soft-gelatine capsules for Vitamin E, garlic pearls etc., and hard-gelatine capsules for chloramphenicol, tetracycline, acetamenophen (paracetamol) Tylenol (R) in US; and also as a medium for the culture of certain microorganisms.Gelatin is also employed as a vehicle for some highly specific pharmaceutical injections e.g., Pitkins menstrumwhich comprises of heparin, gelatin, dextrose, acetic acid and water.Gelatin is also used for the treatment of brittle finger nails, and non-mycotic defects of the nails in humans.

Gelatin [Latin: Gelatina = Gelatin]Types of Gelatin: Gelatin is normally available in two distinct forms, namely:(a) Absorbable Gelatin Sponge: It is a sterile, white, tough, and finely porous spongy, waterinsoluble, and absorbable substance. Even though it is water-insoluble but it is adequately absorbed in body fluids. Nevertheless, it usually takes upto not less than 30 folds its equivalent weight of water.It has been observed that 9 g of absorbable gelatin sponge takes upto 405 g (i.e., 45 times) of well-agitated oxalated whole blood. The various uses of absorbable gelatin sponge are as follows:as an effective haemostatic,as a localized anticoagulant, andwhen placed upon a surgical incision after being duly moistened with sterile NaCl solution, it gets slowly absorbed within a span of 4-6 weeks.

Gelatin [Latin: Gelatina = Gelatin](b) Absorbable Gelatin Film: Absorbable gelatin film refers to a light amber coloured, sterile, non-antigenic thin film invariably produced from a especially prepared gelatin-formaldehyde solution by careful drying followed by subsequent sterilization.Absorbable gelatin film is largely employed in the form of saline-soaked rubber-like thin sheets chiefly in surgical repair of such observed defects in membranes, such as: dura and pleura matter, where it grossly serves as a mechanical means of protection, replacement matrix, and temporary supportive structural wall.

Collagen [Synonym: Ossien]: (Greek: kolla = glue, + gennan = to produce)Collagen refers to a strong, fibrous insoluble protein found in the connective tissue, including the dermis, tendous, ligaments, deep fascia, bone, and cartilage. Collagen is the protein typical of dental tissues (except the enamel of teeth), thereby forming the matrix of dentin, cementum, and alveolar bone proper. Collagen fibers also form the periodontal ligament, that eventually attaches the teeth to their respective bony sockets of the lower and upper jaws that are strategically located in the central core of the triple helical molecule of the collagen. However, collagen may be easily differentiated from other accompanying fibrous proteins e.g., elastin,* reticulin etc. Interestingly, collagen is duly characterized by the presence of a host of vital amino acids, such as: glycine, hydroxyproline, hydroxylysine, proline, and tyrosine; whereas, elastin essentially comprises of absolutely non-polar amino acids, for instance: isoleucine, leucine, and valine. Nevertheless, one may come across a plethora of collagen variants which solely depends upon the presence of the amino-acid sequence. When collagen is carefully boiled with water it gets duly converted into gelatin. Collagen finds its typical applications in the preparation of photographic emulsions, sutures, and also as a gel in food casings.

Casein [Latin: caseus = cheese]Casein designates the principal protein in milk. It is present in milk curds. It essentially provides all the amino acids that are necessary for the growth and development in humans. When milk is subjected to coagulation by rennin or acid, casein becomes one of the principle ingredients of cheese.Casein actually represents the phosphoprotein with a composition of 0.85% P and 0.75% S.Characteristic Featuresare as follows:(i) % N = 15 to 16(ii) Sulphated Ash (%) = NMT** 1.5(iii) Loss on Drying (%) = NMT 6(iv) Isoclectric Point = 4.7(v) Specific Gravity = 1.25 to 1.31(vi) Molecular Weight = 75 K*** to 370 K

Casein [Latin: caseus = cheese]Casein Variants: Casein has several known variants, such as:(a) Lactalbumin: Lactalbumin refers to the albumin of milk and cheese; and it is a soluble simple protein. It is present in relatively higher concentration in human milk in comparison to the cows milk. When milk is heated, the latalbumin aptly coagulates and appears as a film on the surface of the milk.(b) Lactoglobulin: Lactoglobulin refers to a protein found most abundantly in milk. Both casein and lactoglobulin are the most common proteins invariably seen in the cows milk.(c) Acid Casein: The warm skimmed milk when acidified with a diluted mineral acid, the whey usually gets separated. The solid curd is duly separated by any suitable means, residual solid mass is now washed thoroughly, dried and pulverized to obtain acid casein powder.(d) Rennet Casein: The skimmed milk is adequately treated with an enzyme, rennet extract, whereby the product is first separated carefully, and subsequently purified to obtain the Rennet Casein.

Casein [Latin: caseus = cheese]Casein (or soluble casein) is usually recommended as a dietary supplement for protein in both pre and post-operative care of the patients.Casein is also employed as a base in the proper standardization of the proteolytic enzymes.Casein is also exploited as an emulsifying agent.Casein is used in sizing of paper and textile.Casein is employed as an unique adhesive agent for the preparation of large-scale casein paints and casein plastics.

Lectins [Synonyms: Agglutinins; Affinitins; Phasins; Protectins;]Lectin refers to one of the several plant proteins that specifically stimulate the lymphocytes to undergo proleferation.Examples: Phytohemagglutinin; Concanavalin.Alternatively, lectins are proteins or glucoproteins without having an immune origin. Lectins may be isolated from various natural sources, such as: bark, fungi, fresh-eggs, roots, microorganisms, body fluids of lower-vertebrates, invertebrates, sea-weed and sponges, and the mammalian cell membranes.

Lectins [Synonyms: Agglutinins; Affinitins; Phasins; Protectins;]Importantly, the lectins are not used directly as a medicine, but they do have the following usages elsewhere, namely:For determining blood-groups; and for carrying out erythrocytic polyagglutination investigative studies.For performing histochemical studies related to either normal and pathological status.For establishing structural elucidation studies of the carbohydrate bearing molecules.For carrying out the mitogenic stimulation of lymphocytes.As tools for studying cell-surface properties in cancer research.Natural Sources of Lectins: a few typical natural sources of lectins are as given under:abrin: Abrus precaturius;concanavalin A: Conovalia ensioformis;green marine algae: Codium fragile;red kidney bean: Phaseolus vulgaris; andhorse gram: Dolichos biflorus.

YeastYeast invariably refers to any of several unicellular fungi of the genus Saccharomyces, that particularly reproduce by budding. They are capable of fermenting carbohydrates. Yeasts, especially candida albicans, may cause systemic infections as well as vaginitis. It has been generally observed that yeast infections are frequently present in patients with malignant lymphomas, AIDS, severe diabetes mellitus, and several other conditions causing immunocompromise.Types of Yeast: There are in fact two types of yeast, namely:

Yeast(a) Brewers Yeast: Brewers Yeast refers to the specific yeast obtained duly during the brewing of beer. However, it may also be used in its dried form as a good source of Vitamin B. (b) Dried Yeast: Dried Yeast designates the particular dried yeast cells obtained from the strains of Saccharomyces cerevisiae. It is mostly used as a viable source of proteins and Vitamins (especially Vitamin B Complex).Interestingly, as to date approximately 350 yeast variants have been duly isolated, purified, characterized for their specific uses. In a rather broader sense, the yeasts have been duly classified according to their actual usages to which they are based upon their typical morphological characteristic features, such as:For making Wines : Winers Yeast;For making Bakery Products : Bakers Yeast;For making Lager Beers : Brewers Yeast;For making Alcohol : Distillers Yeast;From Malt Wort, MolassesFor making Drugs : Brewers Yeast; Bakers Yeast;

YeastSources of Yeast: The natural habitats of this specific type of microorganism (i.e., yeast) are namely: fruit juice, bread, fermented media, and fermenting media. On a commercial scale the yeast is usually produced by making use of citrus fruits, molasses, grain wort, malt wort, molasses wort etc.Chemical Composition of Yeast: Yeast usually contains a wide range of vital and important chemical constituents, namely: nitrogenous ingredients (i.e., proteins); vitamins (e.g., thiamine, riboflavine, folic acid, pantothenic acid, biotine etc.); enzymes (e.g., diastase, maltase, zymase etc.); and glycogens and minerals (ash).Applications of Yeast: Following are some of the vital applications of yeast in various food, beverage, and pharmaceutical industries:(1) Beverage Industries viz., Bears, Wines, Alcoholic Beverages such as: Gins, Whiskies, Rums, Vodkas etc.,(2) Food Industries viz., Bakery Products, Biscuit Industries;(3) Pharma. Industries viz., antibiotis, papain etc.

Thaumatin [Synonym: Talin;]Thaumatin is the sweet-tasting basic protein duly extracted from the fruits of the tropical plant, Thaumatococcus danielli Benth., Marantaceae, found extensily in West Africa from Sierre Leone to Zaire, in Sudan and Uganda. It is mostly composed of five distinct forms viz., thaumatins I, II, III, b, and c. However, thaumatins I and II predominate invariably. Importantly, all of them are almost 100,000 times sweeter than sucrose, and do have molecular weights around 22,000.

Thaumatin [Synonym: Talin;]Characteristic Features: The various characteristic features of thaumatin are as enumerated under:(i) It has increasingly sweet taste with a licorice-like distinct after taste.(ii) It is strongly cationic having isoelectric point greater than or equivalent to 11.7. It exhibits UVmax: 278 nm at pH 5.6; and 283,290 nm at pH 13.0.(iv) It is about 750-1600 times sweeter than sucrose on a weight basis; and 30,000 to 100,000 times on a mole basis.(v) Its threshold values are very close to 104%.(vi) The proteins usually lose sweetness upon heating, whereby the disulphide-bridges undergo strategical cleavages in their basic structures.(vii) Thaumatin also loses its sweetness at pHs