PREFORMULATION

INTRODUCTION The performulation is the first step in the rational development of a dosage form of a drug substance alone and when combined with excipients. The overall object of the preformulation is to generate useful information to the formulator to design an optimum drug delivery system. The formal preformulation study should start at the point after biological screening. When a decision is made for further development of the compound in clinical trials. Before embarking on a formal programme, the preformulation scientist must consider the following : 1. 2. 3. 4. 5. Available physicochemical data (including chemical structure, different salt available). Anticipated dose. Supply situation and development schedule. Availability of stability indicating assay. Nature of the information the formulator should have or would like to have.

Preformulation may be described as a stage of development during which the physical pharmacist characterises the physical chemical properties of the drug substance in question which are considered important in the formulation of a stable, effective and safe dosage form. Such parameters as crystal size and shape, pH solubility profile, pH stability profile, polymorphism, partitioning effect, and dissolution behavior are evaluated. During this evaluation possible interactions with various inert ingredients intended for use in the final dosage from are also considered. Some consequences of poor preformulation work are 1. 2. 3. 4. 5. Possible use of unsatisfactory salt form Poor stability of the active ingredient Testing compound of marginal activity Increased development costs Increased development time

When preformulation studies are completed the data are complied and transferred to the development pharmacist, who in turn utilities his information to plan his development work on finished dosage form. GOALS OF PREFORMULATION 1. 2. 3. 4. To establish the physicochemical parameter of new drug substances. To establish the kinetic rate profile. To establish physical characteristics. To establish compatibility with the common excipients.

Once the drug is synthesized and tested for pharmacological activity and if found promising following steps are taken

1.

Sufficient quantity is synthesized to a. Perform initial toxicity studies b. To do analytical work c. To do initial Preformulation studies. Actual formulation is done Formulation is subjected to Phase 2 and 3 clinical trials, during this period final formula is finalized. After completion of all the above process, NDA is submitted to the concerned authority After approval of the NDA, production can be started.

2. 3.

5.

6.

The physico-chemical properties of the drug molecule can affect the structure and stability of the formulation and alter the bioavailability. So due consideration should be given to this, before the dosage form design, i.e. during the preformulation. It is also important to have an understanding of the physical description of a drug substance before dosage form development. ORGANOLEPTIC PROPERTIES A typical preformulation should begin with the description of the drug substance, i.e. the colour, odour, taste, of the new drug must be recorded. It is kept as reference for comparing with the other batches during production. The product should be physically attractive in nature, if the taste is found to be unpalatable consideration ought to be given for the use of less soluble chemical form. The odour and colour may be suppressed by using appropriate flavors or other excipients or by coating of final product. But these organoleptic agents must be screened for their influence on the stability and bioavailability of the active drug. Terminology that was suggested to describe organoleptic properties are Colour Off white Cream yellow Tan Shiny Odour pungent fruity aromatic sulfurous Odourless Taste acidic bitter bland sweet tasteless

Colour Colour of early batches of new drug should be recorded using descriptive terminology. This is useful in establishing appropriate specifications for later production. When colour attributes are undesirable or variable, dye should be incorporated or coating of final product . ODOUR & TASTE In tasting a new drug, caution must be exerted. If taste is unpalatable, less soluble chemical form of drug should be used. Odour and taste may be suppressed by using flavors, or by coating a final product. Flavors and excipients should be screened for their influence on stability and bioavailability of drug. Purity Preformulation have some perception of purity of drug substance. An impurity like metal contamination at the level of few parts per million in which certain classes of compounds have deleterious effect. Off-colour materials upon recrystallization become white in many instances Appearance is another area where slight impurity can have a large effect

TECHNIQUES USED
1. Thin layer chromatography (TLC) 2. High-pressure liquid chromatography(HPLC)

PARTICLE SIZE SHAPE AND SURFACE AREA

Chemicals and physicals properties of drug substance are affected by their particles size distribution effect also biopharmaceutical behaviour. Eg:Bioavailability of griseofulvin and phenacetin directly related to particles size distribution of these drugs. poorly soluble drugs showing a dissolution rate limiting step in absorption process will be more readily bioaviability when given in finely subdivided state. Very fine materials are difficult to handle, but can be over by heating solid solution of material of interest in carrier ,such as water-soluble polymer size also playas role in homogeneity of final tablet when big difficulty in sizes exit between the active component and recipients mutual sieving effects can occur making thorough mixing difficult this effect is more when diluents and active raw materials are of significant different sizes. Size can also be a factor in stability fine materials are relatively more open to attack atmosphere or heat, light, humidity because of these significant roles, it is important to get a desired particles size grinding can be done to new drugs having particles that are above 100micrometre in diameter. If materials consists of particles of size 30 micrometer or less then grinding is an necessary, except if material exists in needly where grinding may improve flow and handling properties and if material is poorly water soluble increases dissolution rate. Grinding reduces coarse material preferably 10 to 40 micrometer range. Drawbacks Several drawbacks but of lesser importance 1- When grinding is done material may be lost. 2- Static electric build up occurs,making material difficult to handling. 3- Undue grinding can destroy solutes and there by changes of important character of drug substance.

Techniques Common techniques for measuring fine particles of various sizes Techniques Microscopic Sieve Sedimentation. Eleutrition Centrifugal Permeability Ligthscattering Techniques for derterminig particles o Several tools are employed to monitor particle size most rapid technique is microscopy very useful in estimating the range of size and shape. o Not suited for rapid, quantitative size determination because it requires counting of large number of particles when quantitative information is needed. o The range of sizes observable by microscopy is from about 0.3micrometer. o For optical microscopy, material observed by suspending in non dissolving fluid and using polarizing lenses to detect a change to an amorphous state after grinding. o For materials that range from 40micrometer ,sieving or screening is good, although shape has a influence on results most pharmaceutical powders range from 1 to 120micrometer to en compass these ranges variety of instruments has been developed their based on light scattering ,light blockage ,and blockage of electrical conductivity path[coulter counter] o Most of these instruments major the number of particles but distribution is readily converted to weight and size distributions o A number of classical techniques based on sedimentation methods utilizing such as Anderson pipette or recording balances they continuously collect settling suspensions. Disadvantage They tedious in nature mathematical expressions can be used in characterizing average size these referred to average volume or weights geometric mean diameters and relationships reflecting shapes such as ratio of area to volume or weight factor . FINE PARTICLE CHARACTERIZATION Bulk flow, formulation homogeneity, dissolution, and chemical reactivity and hence absorption is often related to size and shape of the drug particles. The new drug particle particle size 1-100 >50 >1 1-50 <50 >1 0.5-50

should be tested during preformulation with the smallest particle size as it maximize the drugs surface area for interaction The following method can be used for determination of particle size: 1. 2. 3. 4. Optical microscopy Sieve analysis Andreasen pipette Coulter counter

Fine particles have the following effect: 1. Dissolution: The rate of dissolution of small particles is usually faster than that of larger ones because rate of dissolution depend on specific surface area dA / dt = KS (Cs C) Where A is amount of drug in solution K- is intrinsic dissolution rate constant. S- is surface area. Cs- is the saturation of the concentrated solution of the drug. C- is the drug concentration at time `t. 1. Solubility: Solubility usually increases with the fire particles and therefore solubility affects the dissolution rate and in turn bio-availability were affected by particle size. 3. Flow properties: Reduction of particle sizes to extremely small sizes may cause the small

particles to cover by the air which is entrapped so that the drug particle surface may prevent the wetting of the drug by surrounding fluid. 4. Degradation of the drug: Particle size reduction may be deleterious for some Drugs substance. Increasing surface area by milling or other methods may lead to rapid degradation of a compound. Drug substance may also undergo polymorphic transformation during the milling. 5. Gastrointestinal absorption: The GI absorption of a poorly soluble drug may be by affected the particle size distribution. Smaller particles should increase dissolution rate and thus bring about more rapid gastrointestinal absorption. Eg: Sulphadiazine suspension having 1-3 in size was absorbed more rapidly than suspension containing larger particles. Surface area plays an important role in drug absorption. Eg. Griseofulvin has surface area not less than 13000 to 17000 cm2 \gm

If less, decreased absorption. Bephenium hydronaphthoate I.P. limit is 7000cm2/gm CRYSTALLINITY AND POLYMORPHISM: Crystal habit and internal structure of a drug can affect bulk and physicochemical properties, which ranges from flowability to chemical stability. Habit is the description of outer appearance of a crystal. Whereas the internal structure is the molecular arrangement within the solid. Changes with internal structure usually alter crystal habit. Eg. Conversion of sodium salt to its free acid form produce both; a change in internal structure and crystal habit. Depending on the internal structure a compound can be classified as 1.Crystalline 2.Amorphous

Crystals are characterized by repetitious spacing of constituent atoms or molecules in a three dimensional array. In case of amorphous forms atoms or molecules are randomly placed as in a liquid. The amorphous forms are usually of higher thermodynamic energy than the crystalline form. Upon storage, the amorphous forms tend to convert to more stable crystalline forms. So due consideration should be given for these changes. Eg: The amorphous forms of Novobiocin was found to be well absorbed, however when formulated into a suspension, convert into more stable crystalline form and results in poor absorption. A polymeric form may contain stiochiometric or non stiochiometric solvents. If the solvent is water, the complex is called a hydrate. The term hemihydrates, monohydrate, dihydrate etc describe hydrate forms with molar equivalence of water. A compound not containing any water molecule within its crystal structure is termed as anhydrous. Anhydrous forms are more soluble; Conversion of anhydrous form to hydrous form within dosage form may reduce the dissolution rate and extent of absorption. Identification of possible hydrate compounds is important since their aqueous solublities can be significantly less than anhydrous form. Conversion of anhydrous form to hydrate with in dosage form may reduce dissolution and extent of absorption of drug. Eg: Anhydrous form of Ampilcillin produced higher blood concentration than the trihydrate form of Ampilcillin.

Non stoichiometric adducts such as inclusions or clatharates involve entrapped solvent molecules with in crystal lattice. This adduct is undesirable because of its lack of reproducibility and should be avoided for development.

POLYMORPHISM: Polymorphism is the ability of the compound to crystallize as more than one distinct crystalline species with different internal structure. Different crystalline forms are called Polymorphs. Polymorphs may be of two types :(a) Enantiotropic (b) Monotropic When the change from one form to another is reversible it is called as enantiotropic If transition take place in only one direction is called as monotropic. Chemical stability and solubility changes due to polymorphism can have impact on drugs bio-availability. Most of the pharmaceutical products exhibit polymorphism Eg. Sulfonamides and steroids. Eg: Cortisone acetate is available in 5 different polymorphic forms. Among this 4 are unstable, if they are used in the preparation of suspension in presence of water, it changes to stable form, accompanied by caking of the crystals. So we should select stable form for the preparation of suspension. The amorphous form of Novabiocin was found to be well absorbed. However when formulated into a suspension a reversion of metastable state to more stable crystalline form occurs resulting in poor absorption. In case of Calcium pantothenate the preferred form is the crystalline form. In the preparation of multivitamin formulation, calcium pantothenate is granulated with other ingredients. If the amorphous form is used during granulation, polymorphic transformation may occur and it makes the granulating mass stick, making further process impossible. The parameters routinely investigated are: - Number of polymorphs that exist, relative degree of stability of various polymorphs, presence of a glassy state, stabilization of metastable forms, temperature stability ranges for each polymorph, solubilities etc.

Polymorphic forms are physically more stable then others. They have high melting point, least aqueous solubility & lowest energy. Remaining polymorphs called Metastable forms i.e. they have low melting point, high aqueous solubility, & high energy. Therefore metastable forms are preferred in formulation. Eg. Chloramphenicol exist as A, B, C; B form is preferred Riboflavin has I, II, III forms; III rd form shows 20 times more water solubility than I. Therefore dissolution of different solid forms of drug is

Amorphous > metastable > stables Polymorphic stability is also needed to predict long term physical stability of dosage forms. Eg. Capping like cracking in tablets of anhydrous crystalline carbochromen hydrochloride upon storage under high humidity conditions. This was determined to be due to transformation of the anhydrous form into a dihydrate form Some problems in development that may result from inadequate investigation of polymorphic drug forms are: 1. Crystal growth in suspensions and creams, resulting in a product with poor uniformity, appearance and/or biological availability. Eg. Parenteral cortisone acetate suspensions cake if prepared with the wrong polymorphic form. 2. 3. Precipitation of less soluble polymorphic form in liquid dosage forms. Poor bioavailability from a less soluble polymorph, eg, metastable fluprednisolone implants has a higher absorption rate than the stable form. 4. Crystal transitions resulting from milling or wet granulation, producing changes in the physical and biological characteristics of the dosage form. 5. Poor chemical stability, eg. amorphous penicillin is less stable than the crystal salt.

Crystal habit may influence such properties as: 1. Suspension stability and syringability: plate shaped crystals will flow through a needle or orifice much more readily than needle shaped crystals. 2. Tableting properties - - These may be altered by crystal packing during compression. Eg. Particle size of lactose has a considerable influence upon tablet strength. 3. Dissolution Dissimilar crystalline habits may have different dissolution rates depending on their surface area. Cubic and spherical particles dissolve equally from all sides whereas other habits change their shape factor during dissolution, altering the dissolution rate. Various techniques are available for the investigation of the solid state .These include: 1) 2) 3) 4) 5) 6) 7) 8) 9) Microscopy (Including hot stage microscopy) Infrared spectrophotometry Single-crystal X-ray X-ray powder diffraction Thermal analaysis Dilatometry. Proton magnetic resonanace (PMR) Nuclear magnetic resonance spectroscopy (NMR) Scanning electron microscopy (SEM)

HYGROSCOPICITY: The amount of moisture absorbed by the fixed weight of anhydrous sample in equilibrium with the moisture in the air at a given temperature is referred as equilibrium moisture content. It may influence the flow and compression characteristics of powder and the hardness of the final tablet and granulation process. Many of the drug substances exhibit a tendency of absorbing moisture. So these hygroscopic compounds should be stored in a well closed container. And also during production of dosage from using these compounds the humidity should be maintained at a controlled manner. Eg. During capsule filling 30-50% relative humidity is maintained. During preformulation the moisture content range should be specified. If the granules have more moisture content it lead to poor flow and excess hardness of the tablet. If the granules contain less moisture, the compressed tablet may face problem of less hardness, and more friability. In such cases good packing like Eg. Strip or Blister packing is essential. It is better to add silica gel packs in the bulk container of tablet or capsules. Deliquescent substances absorb water to dissolve completely. Eg: NaCl. This hygroscopicity influence many important parameters like, chemical stability, flowability, and compatibility. Test to find out hygroscopicity: 2. Open containers: Bulk drug is placed in open containers with a thin powder bed to assure maximum atmospheric exposure. These samples are then exposed to a range of controlled relative humidity. Moisture up-take should be monitored at time points representative of handling (0 to 24 hrs) and storage (0 to 12 weeks). 2. Analytical method: - Gravimetry, Karl fisher titration gas chromatography etc. are used to find the hygroscopicity.

TYPES: (1) Non Hygroscopic - If stored at RH < 90%, No moisture content increase If stored at RH > 90% for one week, moisture content increase Up to < 20% (2) Slightly hygroscopic - If stored at RH < 80%, No moisture content increase If stored at RH > 80% for one week, moisture content increase Up to < 40%

(3) Moderately Hygroscopic - If stored at RH < 60%, moisture content does not increase above 5 % If stored at RH >80% for one week, moisture content increase Up to < 50%.

(4) Very Hygroscopic - If stored at RH as low as 40-50 % moisture content may Increase If stored at above 90% for one week, moisture content may Exceed 30 % . BULK DENSITY Bulk density of a compound varies with the method of crystallization, milling, or formulation. Bulk density is of importance in capsule filling and in selection of appropriate size of the empty capsule and also in tablet granules flows in to the die. Usually bulk density is of great importance when one considers size of a high dose capsule product or homogeneity of a low dose formulation in which there are large differences in drug & excipient densities. Types of densities 1. True density: It is the ratio of weight to volume. It is determined by liquid displacement method. (Helium displacement method) 2. Granule density: The powder volume includes volume of the particles with the intraparticle voids and this is determined by mercury displacement method. 3. Bulk density; It is the weight of the powder divided by the volume of the particle; it includes intra and inter particle voids. Measured by tapping method.

IONIZATION CONSTANT (PKa): Many drugs are either weakly acidic or basic compounds. In solution depending on the pH values, they exist as ionized or unionized species. The unionized species are liquid soluble and hence more readily absorbed. The gastrointestinal absorption of weakly acidic or basic drugs is dependent on many factors. They are fraction of drugs in unionized form, pH at the site of absorption, ionization constant, lipid solubility of the unionized species. The relative concentration of ionized and unionized form of weakly acidic or basic drug in a solution can be readily calculated using Henderson Hasselbach equation. pH = PKa+ log ionized form Unionized form pH = PKa+ log unionized form Ionized form for acidic drug

for basic drug

The stomach conditions are acidic in nature ranging in pH from 1-3. Whereas the pH of intestinal fluids ranges from 5-8. Hence weakly acidic drugs are preferentially absorbed from the stomach, whereas basic drugs predominately absorbed from the intestine.

Eg: Aspirin PKa- 3.5

% absorbed (pH range) 3.6 4.3 4.7-5.0 7.2-7.1 40% 27% -----

6.0-7.8 -----

Several methods are available for the determination of ionization constant. If unionized and ionized form of the drug in the solution exhibit significantly different, from U.V or visible spectra the absorbency data can be used for determination of ionization constant. Other method, which can be used, is Potentiometric Titrations, which offer maximum sensitivity for compounds with pKa value in 3-10 range. But is often hindered by precipitation of unionized form during titration since high concentration is usually required to obtain a significant titration curve. To prevent precipitation, a co solvent such as methanol or DMSO can be incorporated to maintained sufficient solubility for unionized species. PARTITION CO-EFFICIENT When a material be it a drug or anything else is placed in an environment consisting of organic and aqueous phase. It gets distributed into two phases. The relative quantities that get distributed are expressed in the form of ratio known as partition co-efficient. P 0/w = C oil/ C water Partition co efficient of the drugs are useful in the study of its absorption, distribution, metabolism, elimination. If the lipid solubility of the drug is more it is having more penetration power through the biological membrane. Aqueous solubility is essential for the absorption at the site. In most cases a drug substances with a pka of 3 will be move rapidly absorbed in the stomach. On the other hand a drug with pka of 8 will be absorbed more rapidly in the more alkaline intestines. During preformulation study, the partition co-efficient of the all the excipients likely to be added in to the formulation should considered. In the body the neutral fraction of the drug partitioned in to non-polar lipid membrane. The fraction of the remaining drug rapidly equilibrates so that non ionized drug again formed and available for the partition. This dynamic process takes place until entire drug partitions through the membrane. PERMEABILITY A preformulation evaluation should include studies to assess the passage of drug molecules across biological membrane. The data obtained from the basic physical and chemical studies may give the information about absorption difficulties. Experimental techniques are available to give a more accurate assessment of absorption, one of that kind is, Everted sac method : In this method a piece of intestine is removed from an intact animal (preferentially rat) everted and filled with a solution of the drug substance and the degree and rate of passage of drug through this sac is determined. By this method both active and passive transport can

be evaluated. One of the disadvantage of this technique is the tissue is not supplied with blood as that in the intact animal. DISSOLUTION It is the process of conversion or change from drug particles into solution form, so that body can absorb the drug. The time taken for this process is called as dissolution.The absorption of solid dosage form administered through orally can be represented by following chart. Solid drug in solution Drug in solution in GI fluids Drug in systemic circulation Kd Ka Where Kd is dissolution rate constant and Ka is absorption constant. Dissolution of a drug particle is controlled by several physico chemical properties like crystal habit, particle size, solubility, surface area and wetting properties. Dissolution test can be used to identify potential problem areas. The dissolution rate of a drug substance can described by Noyes whitney equation. dc/ dt = D. A (Cs-C) / h.v -------------------- (1)

where D is diffusion co-efficient A - Surface area of the drug exposed to dissolution media h - Thickness of the diffusion layer at solid-liquid interface v- The volume of the dissolution media Cs- Concentration of a saturated solution of the solute in the dissolution media C- Concentration drug in solution at time t dc/dt- Dissolution rate INTRINSIC DISSOLUTION : This is the dissolution from the constant area. A constant surface is obtained by compressing the powder into a disc of known area with a die punch apparatus. This differs from dissolution of conventional dosage form which is known as total dissolution (mg/min). where the exposed surface area is uncontrolled as disintegration, de-aggregation, occurs as the dissolution proceed. In the intrinsic dissolution case the dissolving substance area is kept constant and CsC, the equation can be rearranged and written as W / A = k.t Where k = D.Cs / h W = weight of drug dissolved in time t A plot of w versus t gives a straight line with the slope equal to the intrinsic dissolution rate constant k, usually expressed in mg/cm2/min. PARTICULATE DISSOLUTION The particulate dissolution is another method of studying dissolution of solids. A weighed amount of sample from a particulate sieve fraction is introduced in to the dissolution medium. A constant speed propeller provides agitation. Particulate dissolution is used to study the influence of surface area, particle size, and mixing with excipients on dissolution. In case of dissolution of Phenacetin granules, prepared using different sieve fraction of the drug powder (different particle size) As expected the rate of dissolution increased with a

decrease in the particle size. Dissolution rate data when combined with solubility, partition co-efficient and Pka results, provide some insight into the potential in vivo absorption characteristics of a drug entity. Dissolution and its correlation with biological activity is studied by measuring the pure drugs dissolution rate in 0.1 N Hcl, water and pH 7.4 buffer. The dissolution rate may be affected by chemical form, crystal form, particle size and surface properties of the drug. (a) Chemical form : The acid, base and different salt forms may give significant difference in dissolution rate, Eg. the dissolution rate of the sodium salt of sulphathiazole in O.1 N Hcl is 5500 times faster than the free base. (b) Crystal form : Metastable polymorphs such as sulfathiazole II have a greater dissolution rate compared to the stable sulfathiazole I. (c) Particle size : Reduction of particles size produces a greater surface area available for liquid contact. (d) Surface properties of the drug : The high surface energy of a micronized powder may result in poor wetability and agglomerates in the dissolution media. Under these circumstances, the dissolution rate decreases, because the total surface area is unavailable to the fluid. The dissolution rate may be determined by one of the two general techniques : 1. Suspension method : The prescreened drug powder is added to the dissolution fluid without any exact control of surface area. Samples of dissolution media are assayed at appropriate time intervals and amount dissolved vs time is charted on a graph. For metastable polymorphs or solvated compounds, dissolution data provide additional information, such as peak and equilibrium solubility, initial rate of dissolution and relative stability of the metastable form. 2. Constant surface method : Surface area and surface electrical charges dissolution variables are eliminated constant surface area is achieved by compressing the powder into a disc of known area with a die-punch apparatus and a carver press. q