BY PHASE


Steps on Drug Production for Small Molecules
  • Drug Discovery

    Searching for a potential molecule to become a new drug.
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  • Pre-Clinical Trial

    Utilizing laboratory and animal testing to determine if the drug is safe for human testing.
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  • Clinical Trial Approval

    Applying for Investigational New Drug (IND) to ensure safety of clinical trial.
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  • Clinical Trials

    Determining the safety and efficacy of the drug on clinical trial volunteers.
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  • Marketing Approval

    Applying for New Drug Application (NDA) or biologics license application for marketing .
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  • Post-Marketing Surveillance

    Monitoring the long term effects of the drug on a greater population.
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Drug Discovery

Drug Discovery
Lead Identification Lead Optimization Lead Development
The choice of antibody platform, appropriate in vivo and in vitro standards and the pharmacodynamics and pharmacokinetic models should have been first considered.
It utilizes natural, synthetic and /or semi synthetic sources for potential selection of candidate molecule.
Structure-Activity Relationships are performed to improve potency and selectivity of the candidate molecule.
Improvement of biochemical biophysical, physicochemical and druglike properties like expression, solubility, stability, aggregation and glycosylation.
Optimized lead compound through the stages necessary to allow it to be tested in animal and human trials.
Involvement of pharmacokinetic studies, toxicology testing, and formulation development.

Identifying Bioactive Molecules

Biologically active small molecules have often been discovered by testing a single compound at a time, but such an approach is obviously highly inefficient and cost intensive in terms of both reagents and personnel time. As a result, HTS technologies have been developed to screen large numbers of compounds simultaneously, typically through the miniaturization and automation of assay protocols. What constitutes high throughput will vary depending on technical considerations and the screener’s economic situation, but generally, the number of compounds involved may range from tens of thousands to several million. High-throughput screening assays can be divided into two main classes: “pure protein” and “cell-based”. Pure protein screens generally have optical assay readouts that monitor enzymatic or binding activity. For instance, fluorescence polarization or FRET techniques are commonly used to screen for compounds that affect binding between protein partners. In pure protein assays, every compound screened should have equal access to the target. However, the membrane permeability characteristics of any active molecules that are identified may subsequently pose a major concern if the target is intracellular.

Target Identification

When biologically active small molecules are identified through chemical screens, particularly forward genetic screens, a substantial amount of work is often required to identify the molecular target. Ignorance of the target does not preclude clinical or research use of the molecule; indeed, many clinical agents have been used effectively even when their targets were not known (e.g. aspirin, nitroglycerin, fumagillin, epoxomycin).

Hypothesis-driven Target Identification

For well-characterized cellular pathways, it is sometimes possible to deduce the target of a small molecule by comparing data from across the field. For instance, characteristic phenotypes induced by the small molecule may permit the assignment of the target to a previously identified complementation group. Subsequent hypothesis-driven testing of potential targets can then be undertaken.

With affinity labeling, molecules may be radioactively or chemically labeled; they may be synthetically modified with reactive groups to promote covalent attachment to the target or tagged with specific moieties to facilitate detection. Standard protein fractionation and detection techniques can then be used to identify proteins from crude extracts that are specifically labeled by the molecule. Examples of drugs whose cellular targets were determined by such methods include acetylcholine, the anti-angiogenic agent fumagillin and the antifungal lipopetide echinocandin . Affinity purification of putative targets from cellular extracts is accomplished with molecules immobilized on a solid support used either as a column matrix or as a bead slurry. Small molecules for which cellular targets were identified through affinity purification include the immunosuppressant FK506, the kinase inhibitor purvalanol and the anti-inflammatory compound SB 203580.

Additional studies involving affinity chromatography were instrumental in further characterizing the biochemistry of FK506 and purifying the target of the structurally related compound rapamycin. A natural technological development of affinity-based target identification involves probing proteome chips with small molecules.

Genomic Methods of Target Identification

A. Yeast three-hybrid system

The yeast two-hybrid system has been widely used to study protein–protein interactions and the approach was adapted to create a yeast three-hybrid system for identifying protein–ligand interactions. The method relies on the use of a hybrid “bait” ligand consisting of the query molecule linked to a known ligand. A protein fusion between the known ligand’s receptor and a DNA  binding domain serves as the “hook”, while the “fish” is a protein fusion between a transactivation domain and the target protein. If the target protein interacts with the query molecule, the “fish” and “hook” will be brought together by the “bait” and transcription from a reporter gene is activated. Hence it should be possible to identify the target of the query molecule by cloning a library into the “fish” domain and screening or selecting for activity of the reporter.

A three-hybrid approach was recently used to identify targets of various kinase inhibitors. For each inhibitor, a hybrid ligand was synthesized by attachment to methotrexate and the hook protein was a LexA–DHFR fusion. A three-hybrid screen of a human cDNA library with a purvalanol B–methotrexate ligand identified several (but not all) known purvalanol targets and also several new candidate targets, and all identified targets were kinases. Affinity chromatography and enzymatic assays confirmed 12 of 16 novel candidate targets identified in the cDNA screens.

B. Induced haploinsufficiency

One genomic approach to target identification relies on the premise that altering the gene dosage of the target will affect sensitivity to the small molecule. For example, inactivating one copy of the target gene in a diploid organism would in many cases be expected to increase sensitivity. This method of identifying drug targets through induced haploinsufficiency was established by Giaever et al., who constructed and screened a set of 233 heterozygous yeast deletion mutants for those demonstrating increased sensitivity to known drugs. Each mutant was chromosomally tagged with a unique oligonucleotide and the mutants were pooled and grown in the presence of tunicamycin, at a level of drug that is sublethal for wild type. The relative number of each mutant in the pool was monitored at various time points by polymerase chain reaction (PCR) amplification and fluorescence labeling of all tags in the pool, followed by hybridization of the PCR-generated probes to an oligonucleotide microarray. The fluorescence intensity generated at each spot on the array permitted quantitation of the relative abundance of each corresponding heterozygote in the pool. Mutants unaffected by tunicamycin showed no reduction in signal over time, whereas tunicamycin-sensitive heterozygotes showed varying decreases in signal. In this study, one known and two new loci were identified and confirmed as involved in tunicamycin resistance.

C. Expression profiling

Expression profiling was recently employed in identifying the targets of a class of small-molecule antagonists of FK506 in yeast cells subjected to salt stress. These molecules (termed SFKs for suppressors of FK506) were identified in a chemical genetic screen for molecules that rescue yeast growth in the presence of high salt and FK506 [34]. Expression profiling results obtained with SFK2-treated yeast suggested that the Ald6 p pathway was a target of SFK2 and haploinsufficency screening supported this hypothesis. Overexpression of ALD6 was found to suppress the effects of SFK2–SFK4 on growth and the ability of SFKs to inhibit Ald6 p in vitro was subsequently demonstrated. In addition to using DNA microarray technologies in conjunction with haploinsufficiency studies, it is likely that gene expression profiling will be used increasingly as a primary means of identifying the targets of bioactive small molecules. Many research groups have used expression profiling to identify characteristic patterns of gene expression (“fingerprints”) that are associated with certain disease states or biological pathways. If the patterns are sufficiently unique, then it is sometimes possible to assign previously uncharacterized mutants to specific cellular pathways or complementation groups based on their expression profiles. Analogously, by profiling cells grown with and without a small-molecule modulator, it may be possible to identify genes or pathways that are affected by the small molecule.

LEAD OPTIMIZATION

In analog design, molecular modifications of the lead compound can involve one or more of the following strategies:

  1. Identification of the active part (pharmacophore)
  2. Functional group optimization
  3. Structure activity relationship studies
  4. Bioisosteric replacement
  5. Design rigid analogs
  6. Homologation of alkyl chains or alteration of chain branching, design of aromatic ring position isomers, alteration of ring size, and substitution of an aromatic ring for a saturated one or the converse.
  7. Alteration of stereochemistry, or design of geometric isomers or stereoisomers.
  8. Design of fragments of the lead molecule that contain the pharmacophoric group (bond disconnection).
  9. Alteration of interatomic distances within the pharmacophoric group or in other parts of the molecule.

IDENTIFICATION OF THE ACTIVE PART (PHARMACOPHORE)

Any drug molecule consists of both, essential and nonessential parts. Essential part is important in governing pharmacodynamics (drug receptor interactions) property while non-essential part influences pharmacokinetic features. The relevant groups on a molecule that interact with a receptor are known as bioactive functional groups. They are responsible for the activity. The schematic representation of nature of such bioactive functional groups along with their interatomic distances is known as pharmacophore.

Once such pharmacophore is identified, structural modification can be done to improve pharmacokinetic properties of the drug. For example, the presence of a phenyl ring, asymmetric carbon, ethylene bridge and tertiary nitrogen are found to be minimum structural requirement for a narcotic analgesic to become active. Similarly the presence of two anionic sites and one cationic site must be present in cholinergic agent.

FUNCTIONAL GROUP OPTIMIZATION

The activity of a drug can be correlated to its structure in terms of the contribution of its functional group, to the lipophilicity, electronic and steric features of the drug skeleton. Hence by selecting proper functional group, one can govern the drug distribution pattern and can avoid the occurrence of side-effects.

STRUCTURE ACTIVITY RELATIONSHIPS

The physiological action of a molecule is a function of its chemical constitution. This observation is the basis of interpretation of activity in terms of the structural features of a drug molecule. Generalized conclusion is then can be made after examining a sufficient number of drug analogs.

BIOISOTERIC REPLACEMENT

The concept of bioisosterism is derived from Langmuir’s observation that certain physical properties of chemically different substances are strikingly similar. These similarities were rationalized on the basis that carbon monoxide and nitrogen both have 14 orbital electrons and similarly, diazomethane and ketone both have 22 orbital electrons.

Bioisosteres are substituents or groups that have chemical or physical similarities and which produce broadly similar biological properties. Bioisosteres have been classified as either classical or nonclassical. Classical bioisosteres are those which have similar steric and electronic features and have the same number of atoms as the substituent moiety for which they are used as replacement. Non classical bioisosteres do not obey the strict steric and electronic definition of classical isosteres and they don not have the same number of atoms as the substituent moiety for which they are used as a replacement. These isosteres are capable of maintaining similar biological activity be mimicking the spatial arrangement, electronic properties, or some other physicochemical property of the molecule of functional group that is critical for the retention of the biological activity.

DESIGN OF RIGID ANALOGS

Imposition of some degree of molecular rigidity on a flexible organic molecule (e.g., by incorporation of elements of the flexible molecule into a rigid ring system or by introduction of a carbon-carbon double or triple bond) may result in potent, biologically active agents that show a higher degree of specificity of pharmacological effect. The three-dimensional geometry of the pharmacophore can be determined and the key functional groups are held in one steric disposition, or in the case of a semirigid structure, the key functional groups are constrained to a limited range of steric dispositions and interatomic distances. By the rigid analog strategy, it is possible to approximate “frozen” conformations of a flexible lead molecule that, if an enhanced pharmacological effect results, may assists in defining and understanding structure activity parameters.

HOMOLOGATTION OF ALKYL CHAIN OR ALTERATION OF CHAIN BRANCHING, CHANGES IN RING SIZE, AND RING POSITION ISOMERS.

Change in size or branching of an alkyl chain on a bioactive molecule may have profound and sometimes unpredictable effects on physical and pharmacological properties. Alteration of the size and shape of an alkyl substituent can affect the conformational preference of a flexible molecule and may alter the spatial relationships of the components of the pharmacophore, which may be reflected in the ability of the molecule to achieve complementarily with its receptor or with the catalytic surface of a metabolizing enzyme. The alkyl group itself may represent a binding site with the receptor through hydrophobic interactions, and alteration of the chain may alter its binding capacity. Position isomers of substituents even alkyl groups on an aromatic ring may possess different pharmacological properties. In addition to their ability to affect electron distribution over an aromatic ring system, position isomers may differ in their complementarily to receptors, and the position of a substituent on a ring may influence the spatial occupancy of the ring system.

ALTERATION OF STEREOCHEMISTRY AND DESIGN OF STERIOISOMERS AND GEOMETRIC ISOMERS.

In the case of chiral molecules, one enantiomer would be expected to demonstrate pharmacological activity and the other enantiomer should be expected to be pharmacologically inert, is not valid.

At high doses (R)-enantiomer selectively stimulated presynaptic dopaminergic receptor sites, while at lower doses, it selectively stimulated postsynaptic receptor sites. In contrast, the (S)-enantiomer stimulated presynaptic dopamine receptors and at the same dose level, it blocked postsynaptic dopamine receptors. Thus, this enantiomer exhibits a bifunctional mode of dopaminergic attenuation: that of presynaptic agonism and postsynaptic antagonism.

FRAGMENT OF THE LEAD MOLECULE

Design of fragments of a lead molecule is based on the premise that some lead molecules, especially polycyclic natural products, may be much more structurally complex than is necessary for optimal pharmacologic effect. A bond disconnection strategy may be employed, in which bonds in the polycyclic structure are broken or removed or destroy one or more of the rings.

VARIATION IN INTERATOMIC DISTANCES.

Alteration of distances between portions of the pharmacophore of a molecule or even between other portions of the molecule may [produce profound qualitative and quantitative changes in pharmacological actions.

LEAD DEVELOPMENT

The screening cascade is designed to decrease the number of compounds examined at each level in order to ensure that compounds with flaws are removed as early as possible. The cascade, also referred to as a screening tree, begins with in vitro profiling and then transitions into in vivo studies designed to determine a compound’s pharmacokinetic profile and demonstrate efficacy in an appropriate animal model.

At the top of the cascade, compounds are screened for activity against the biological target and a threshold of interest is generally set to determine if compounds are active enough to warrant further investigation. Potency is, of course, an important issue, as dosing requirements are lower for compounds that are more potent. All other things being equal, compounds with higher potency can be dosed at lower levels, decreasing the likelihood of side effects. A compound with target potency of 5nM in theory could be provided to a patient at a significantly lower dose than a compound serving the same function but with a potency of 5μM.

Once a compound has satisfied the potency criteria, selectivity and physicochemical properties criteria are typically examined. Nature has developed exquisite systems to accomplish very specific tasks with highly selective systems, but many of these systems overlap structurally, and this can have a significant impact on the biological properties of a given test compound. Thus, the next biological screening step in a typical screening cascade is often an assessment of a compound’s potency at biological systems that are closely related to the target of interest. The Kv1.5 channel, for example, is a voltage-gated potassium channel that has been the target of research programs atrial arrhythmia, and many compounds have been identified that can block this channel with a high level of potency.82 There are, however, over 70 other voltage-gated potassium channels with varying degrees of similarity to the Kv1.5 channel, and undesired activity at any of these related channels could create unwanted side effects in human or animal studies. For example, the Kv1.5 channel is closely related to the hERG channel. Blockade of the hERG channel has been linked to torsade de pointes and sudden cardiac death,83 so any compound moving forward in this area would need to be counterscreened for hERG activity in order to ensure that advancing compounds do not present a risk of sudden cardiac death in a clinical setting. This is a rather extreme example of the importance of proper selectivity, but it should be clear that failure to achieve proper target selectivity in this area represents a significant barrier to moving a program forward.

Similarly, there are over 500 known kinase enzymes,84 and any drug discovery program designed to target a single kinase, or even a family of kinases, has an associated risk of identifying compounds that are active at multiple members of this large family of related enzymes. In order to mitigate this risk, kinase programs routinely screen test compounds against panels of related kinases in order to understand the risks associated with off-target activity.

In general, compounds that are potent at the target of interest, but are also potent at a variety of other targets (“promiscuous compounds”), do not move forward in a drug discovery program, as the risk of undesired (or unpredicted) side effects is too high. The level of selectivity required, however, is dependent on the program, the nature of the potential side effect presented by off-target activity (off-target activity leading to excessive hair growth might be tolerable, whereas sudden cardiac death through poor hERG selectivity is not), the target patient population, duration of treatment (some side effects only appear upon extended exposure to a drug), and a variety of other factors. Overall, target selectivity is a major factor to consider.

An active and suitably selective compound, however, is not necessarily a good drug candidate. Physicochemical properties also play a major role in determining whether or not a compound is suitable for further investigation. In vitro screens designed to predict absorption, distribution, metabolism, and excretion (in vitro ADME) are generally performed early in a program in order to ensure that candidates reaching the drug development pipeline are “druglike” in nature. Compounds that have poor aqueous solubility, for instance, are often difficult to develop as drugs. In order for a drug to exert an influence on a biological target, it must be soluble in biological fluid at a level consistent with its potency. Thus, the level of solubility required for a given compound is directly linked to its potency. As potency increases, the requisite solubility decreases, as less drug is required to provide the intended effect. Solubility also has a direct impact on absorption, as a compound must be soluble in biological fluids in order for it to successfully pass through a biological membrane and reach its intended target.

The ability of a compound to penetrate cellular membrane (its permeability) can also be a determining factor in the success or failure of a given candidate compound. If a compound is potent, selective, and soluble, but unable to pass through a biological membrane, it may not be able to reach the target of interest and fail to demonstrate the desired efficacy. Orally active drugs must be absorbed in the gastrointestinal tract, and drugs that target intracellular system must also pass through the cell membrane in order to reach their intended targets. Extracellular targets, of course, do not face this added issue, but CNS drug candidates face the added complexity of required permeability through the blood–brain barrier (BBB). Additionally, there are efflux pumps (e.g., P-glycoproteins (Pgps))85 designed to remove xenobiotic material that can limit permeability, preventing efficacy. The inability of a compound to penetrate a cell membrane represents a significant issue that could prevent further investigation of the candidate compound.

Metabolic and chemical stability are also important considerations. If all of the previously mentioned criteria are met, and a compound is able to enter the body, but is immediately metabolized, efficacy studies will fail to show the desired results. However, the relative rate of metabolism that can be allowed for a successful compound depends on the goals of the project. If, for example, the goal is to develop a new antibacterial agent, then high-metabolic stability will likely be desired so that the potential drug candidate will be available in the circulation long enough to kill the invading organism. If, however, the goal is to develop a new surgical anesthetic, metabolic stability may be less of an issue, as it may be desirable for drug efficacy to fade rapidly upon termination of dosing regimens.

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Pre-Clinical Trial

Pre-Clinical
Pharmacokinetics
Formulation
Toxicology
Synthesis Scale up/Kilo Labs

Pharmacokinetics

The study of drug absorption, distribution, metabolism and excretion of drugs in the body.

PKA Analysis

In drug discovery, initial PK studies are most commonly conducted in rodents and/or the species used for the assessment of in vivo efficacy. Subsequently, experiments in a large animal species such as dog or monkey are performed to better describe the disposition of a compound (in multiple species), support toxicology studies, and generate data useful in predicting human PK parameters. It should be noted that low aqueous solubility of discovery molecules often necessitates significant formulation work prior to dosing. The route of compound administration is generally dictated by that which will best ensure clinical and commercial success for a given indication (e.g., an oral agent for diabetes) or the stage of the project along the discovery path (e.g. intravenous dosing to support proof-of-principle studies). Throughput in PK studies can be improved by resorting to cassette dosing of multiple compounds in a single animal, although its potential limitations (e.g., enzyme inhibition leading to exaggerated exposure or “false positives”) must be considered. The bioanalytical or assay phase typically involves sample extraction with organic solvents and LC/MS or LC/MS/MS separation and detection of analytes.

NON COMPARTMENTAL (MODEL INDEPENDENT) PK

Pharmacokinetic parameter estimation based on noncompartmental analysis relies on empirical determination of AUC and AUMC. Therefore, no assumptions are made based on the shape of the blood or plasma drug concentration versus time curve, as is the case with compartmental methods (described below). Noncompartmental methods thus furnish important PK data such as AUC, CL, F, t1/2, and Vss in a simple and rapid manner. However, note that in calculating CL, an important assumption must still be made: that the drug undergoes first-order (unsaturable) elimination. In other words, that clearance is constant and the rate of drug elimination from blood or plasma is linearly proportional to its concentration. An important limitation of noncompartmental PK analysis is that it lacks the ability to predict PK when there are alterations in a dosing regimen, which compartmental PK methods afford.

COMPARTMENTAL PK ANALYSIS

In drug concentration versus time profiles, levels are often observed to decline in a mono– or multiexponential fashion. In compartmental PK analysis, these data are fit to an exponential equation that is derived by assuming that the body consists of one or more compartments through which the administered drug distributes. At least one of the compartments must be a sampling compartment (typically the central compartment). Rate constants describe drug transfer between compartments and a set of differential equations is written to describe the concentrations in each compartment versus time. The equation describing the change in concentration in the sampled compartment is fit to the blood or plasma drug concentration versus time data and PK parameters in the model are estimated using nonlinear regression. In general, the number of compartments used in the model is equal to the number of identifiable phases when the in-transformed concentrations are plotted versus time. Compartmental approaches allow some level of “physiological” interpretation of what the body does to the drug (i.e. should be consistent with physiologically reasonable pathways of drug elimination)


Formulation

It is the process in which different chemical substances such as active ingredient and excipients will combine together to produce a dosage form.

List of equipment/facilities/reagents:
  1. Airflow Containment (Pharmacon Downflow Booths, Ceiling Laminar Airflow Units, Laminar Floor Horizontal Trolley/ Laminar Flow Vertical Trolley, Enterprise Laminar Flow straddle Units, ESCO Garment storage cabinet, Ventilated Balance Enclosure)
  2. Isolation Containment (Aseptic Containment Isolator, Containment Barrier Isolator, General Processing Platform Isolator, Isoclean Healthcare Platform Isolator, Streamline Compounding Isolator, Weighing and Dispensing Containment Isolator.
  3. Cross Contamination Facility Integrated Barrier (Biopass Pass Through, Infinity Air Shower Pass Box, Cleanroom Air showers, Infinity Cleanroom Transfer Hatch, Infinity Pass Boxes, Soft Capsule Soft Wall Cleanroom, Dynamic Passboxes and Dynamic Floor Label hatches.
  4. Barrier Isolation System (CradlePro-Iso, Cytotoxic Safety cabinets)
  1. Sample Storage and protection solution (Lab Refrigerators and Freezers, Voyager – Remote monitoring programming, datalogging software, ESCO PROtect Wireless Monitoring System)
  2. Laboratory Thermostatic Products (Irefrigerated incubator, Laboratory Ovens)
  3. SubliMate Freeze Dryer or lyophilizers
  4. Stability Isolators

Lyophilized Dosage Forms

Lyophilized dosage forms are frequently used to mitigate stability issues. Development of lyophilized formulations requires selection of suitable excipients such as bulking agents that will deliver a pharmaceutically elegant and physically and chemically stable dosage form. If pH modification is required, the buffer selection may need to be modified to account for volatility of the components and compatibility of concentrated ingredients.

Oral Dosage Form Development

The general approach towards oral formulation development involves first assessing simple formulations and increasing the complexity, if needed, until sufficient performance is obtained. Wolfe Laboratories then optimizes dosage form composition and adjusts analytical methods.In collaboration with the sponsor, Wolfe Laboratories conducts a proof-of-concept (POC) PK study using the prototype formulations. Upon evaluation of the PK data, the team determines whether the results justify refinement of the prototype formulations. The dosage form is evaluated for its storage stability, and excipient component ratios are optimized for optimal in vitro performance. The performance of new prototype formulations are verified in vivo, after which process optimization ensues.

The simplest oral dosage form utilizes powder in capsule (PIC) whenever possible. However, when there is insufficient bioavailability, more complex formulation approaches may be warranted. The specific program design considers pharmaceutical and biopharmaceutical factors such as: aqueous, organic and lipid solubility, crystallinity, pKa, logP, food effects, permeability and efflux, non-enzymatic degradation in the GI tract, and metabolism. Consequently, the specific formulation design approach depends upon the attributes of a given molecule.

Considerations for Liquid Oral Formulations

Liquid dosage forms require that sufficient solubility and stability are maintained to support the dosing studies. If preformulation studies revealed sufficient solubilities, then developing a liquid dosage form is relatively straightforward, but if the solubility is insufficient, suspension dosage forms may be required. The formulation may also require taste masking and other excipients.

Considerations for Solid Oral Formulations

The success of solid dosage formulations depends on achieving the sufficient dissolution rate and stability of the physical form of the compound. Polymorphs can result in different solubility and dissolution properties, and thus adversely impact the bioavailability. The dissolution rate is dependent upon the intrinsic dissolution rate, the volume into which the drug must be dissolved, and the surface area. For poorly soluble drugs, it may limit the extent of absorption and therefore must be well understood. For the purposes of informing rational formulation development, the choice of the biorelevant media is also important.

Enabling Oral Formulations

Self-emulsifying drug delivery systems

Self-emulsifying drug delivery systems (SEDDS) generally contain oil, surfactant, co-surfactant and a co-solvent. Wolfe Laboratories evaluates compound performance in various blends to develop prototype and optimized formulations.

Amorphous Dosage Forms

Thorough characterization of solid-state properties and physical form stability reduces the risk in product development. In situations where poor oral bioavailability is caused at least in part by limited dissolution of the starting material, and if the starting material is crystalline, altering the solid-state from crystalline to amorphous state may lead to improved solubility by reducing the energy barrier to dissolution. The amorphous dosage forms can be formulated as solid dispersions, liquids or suspensions. A more complex approach involves development of a lipid-solid dispersion. Additives can increase or decrease the melting temperature for ease of preparation and affect the properties of the solidified material. At the manufacturing stage, the solidified melts can be pulverized into powder or melt extruded and then pulverized into powder, which can be compressed into tablets. Alternatively, liquid melts can be loaded into capsules and cooled.


Toxicology

The evaluation of safety of potential drug candidates. This step is accomplished by using relevant animal models and validated procedures. It encompasses acute, sub acute or chronic toxicity studies.

List of equipment/facilities/reagents:

  1. PCR Thermal Cyclers (Conventional Thermal Cyclers, Real-time PCR systems)
  2. PCR Sample Handling (Microplate shakers, PCR cabinets)
  3. VIVA Universal Animal Containment Workstations
  4. VIVA Dual Access Animal Containment Work Stations
  5. VIVA Bedding Disposal Animal Containment Work Stations
  1. Target Safety Review
    1. Identify potential on-target toxicities.
    2. Define the early discovery experiments required to evaluate potential target-related safety issues.
    3. Inform the design of subsequent regulatory studies.
  2. Bespoke in vivo
    1. Confirm any expected on-target/off-target, tissue specific toxicities.
    2. Compare potential lead candidates, and make an informed candidate selection.
    3. Inform design of early Good Laboratory Practices (GLP) toxicology studies.

Typically, these programs have two stages. In the first, the potential leads are dosed in a single dose, ascending dose toxicokinetic study in rats. The aims of this study are two-fold:

  1. Identify acute toxicities.
  2. Define dose and exposure relationships, thus compounds can be compared in repeat-dose study on the basis of comparable exposure rather than dose.
  1. Bespoke in vitro

Issue specific, mechanistic in vitro assays

  1. Range of assays available
  2. New assays established on request
  1. Supporting DMPK

Rat CYP Induction – used to predict issues with autoinduction (where the compound induces its own metabolism in repeat-dose preclinical studies, leading to a lowering of exposure), to explain certain liver histopathological changes and to alert for non-genotoxic carcinogenicity in the liver and thyroid.

This assay is defined to identify compounds that interact with transcription factors involved in the induction of CYP1A, CYP2B, CYP3A and CYP4A isoforms. These are, respectively, the aryl hydrocarbon receptor (Ahr), the constitutive androstane receptor (CAR), the pregnane X receptor (PXR) and peroxisome proliferator-activated receptor alpha (PPARα). The most relevant test system is to freshly prepared rat hepatocytes.

In vitro metabolism – used to support the selection of the most appropriate preclinical studies, and to aid in the design and interpretation of those studies, by comparing rates of metabolism and identifying metabolites formed in different species. MIST (Metabolites in Safety Testing) guidelines encourage the identification of differences in drug metabolism between animals used in nonclinical safety assessments and humans as early as possible during the drug development process. This is important as, potentially, a full toxicity study on a metabolite may be required if that metabolite is not present in a preclinical species at sufficient levels. It is important to use in vitro studies to predict metabolites across species and inform the choice of preclinical safety species.

  1. Downstream Investigative Toxicology

Problem resolution can identify mechanisms of toxicity, and feedback into discovery, allowing the design of molecules with reduced or removed toxicological activities.


Synthesis Scale Up / Kilo Labs

Kilo labs are designed to yield quantities ranging from 100 grams to under 10 kilogram of materials with typical batch sizes of 2–3 kilograms. Reactor sizes usually range from 20–100 liters and are made from glass. The kilo lab is the first step out of laboratory, hence, equipment are typically closer to a laboratory than in a manufacturing plant.

List of equipment/facilities/reagents:

  1. Airflow Containment (Pharmacon Downflow Booths, Ceiling Laminar Airflow Units, Laminar Floor Horizontal Trolley/ Laminar Flow Vertical Trolley, Enterprise Laminar Flow straddle Units, ESCO Garment storage cabinet, Ventilated Balance Enclosure)
  2. Isolation Containment (Aseptic Containment Isolator, Containment Barrier Isolator, General Processing Platform Isolator, Isoclean Healthcare Platform Isolator, Streamline Compounding Isolator, Weighing and Dispensing Containment Isolator.
  3. Cross Contamination Facility Integrated Barrier (Biopass Pass Through, Infinity Air Shower Pass Box, Cleanroom Air showers, Infinity Cleanroom Transfer Hatch, Infinity Pass Boxes, Soft Capsule Soft Wall Cleanroom, Dynamic Passboxes and Dynamic Floor Label hatches.
  4. Barrier Isolation System (CradlePro-Iso, Cytotoxic Safety cabinets)
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Clinical Trial Approval

Clinical Trial Approval

Investigational New Drug (IND) is the general application to administer a new drug to humans. Studies in human can only start upon the approval of IND by the Food and Drug Administration (FDA) and an institutional review board (IRB).

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Clinical Trials

Clinical Trials
Phase I
Phase II
Phase III
The main goal is to assess the safety of the candidate drug in humans. It is usually conducted in a small number of healthy volunteer (100 or less). Pharmacokinetic, pharmacodynamics, and safe dosing range are evaluated. This phase is necessary to determine if the candidate drug will move on to the next phase.
The effectiveness of the drug candidate is evaluated in this phase using 100-500 patient volunteers with the condition under study. Optimal dose strength, short term side effects and risk are also analyzed. This phase is necessary to determine if the candidate drug will move on to a larger phase III trial.
Demonstrate safety and efficacy in a large group of patient usually 1000-5000 patients or more across numerous clinical trial sites around the world. Planning for the full scale production of medicine is conducted on this stage. Drugs at this stage must be produced in a CGMP facility and is the final phase in which the process can be changed before commercial production.

Phase I

During Phase 1 studies, researchers test a new drug in normal volunteers (healthy people). In most cases, 20 to 80 healthy volunteers or people with the disease/condition participate in Phase 1. However, if a new drug is intended for use in cancer patients, researchers conduct Phase 1 studies in patients with that type of cancer.

Phase 1 studies are closely monitored and gather information about how a drug interacts with the human body. Researchers adjust dosing schemes based on animal data to find out how much of a drug the body can tolerate and what its acute side effects are.

As a Phase 1 trial continues, researchers answer research questions related to how it works in the body, the side effects associated with increased dosage, and early information about how effective it is to determine how best to administer the drug to limit risks and maximize possible benefits. This is important to the design of Phase 2 studies.


Phase II

In Phase 2 studies, researchers administer the drug to a group of patients with the disease or condition for which the drug is being developed. Typically involving a few hundred patients, these studies aren't large enough to show whether the drug will be beneficial.

Instead, Phase 2 studies provide researchers with additional safety data. Researchers use these data to refine research questions, develop research methods, and design new Phase 3 research protocols.


Phase III

Researchers design Phase 3 studies to demonstrate whether or not a product offers a treatment benefit to a specific population. Sometimes known as pivotal studies, these studies involve 300 to 3,000 participants.

Phase 3 studies provide most of the safety data. In previous studies, it is possible that less common side effects might have gone undetected. Because these studies are larger and longer in duration, the results are more likely to show long-term or rare side effects.

Drugs at this stage must be produced in a CGMP facility and is the final phase in which the process can be changed before commercial production.

For biosimilars or biologics a change in bioprocess method such as changing bioreactors from roller bottles to TideCell may require a bridging study or redoing phase 3

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Marketing Approval

Marketing Approval

The New Drug Application (NDA)/ Biologics License Application (BLA) is submitted to FDA for marketing approval. These application contains results and data analysis from the previous clinical trials.

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Post-Marketing Surveillance

Commercial Marketing

Production plants of active pharmaceutical ingredients, and drug products are primarily batch operated multi purpose manufacturing plants. At this facilities, commercial supplies of API intermediates, APIs and drug products are manufactured before being packaged, labeled, and distributed.

List of equipment/facilities/reagents:
  1. Airflow Containment (Pharmacon Downflow Booths, Ceiling Laminar Airflow Units, Laminar Floor Horizontal Trolley/ Laminar Flow Vertical Trolley, Enterprise Laminar Flow straddle Units, ESCO Garment storage cabinet, Ventilated Balance Enclosure)
  2. Isolation Containment (Aseptic Containment Isolator, Containment Barrier Isolator, General Processing Platform Isolator, Isoclean Healthcare Platform Isolator, Streamline Compounding Isolator, Weighing and Dispensing Containment Isolator.
  3. Cross Contamination Facility Integrated Barrier (Biopass Pass Through, Infinity Air Shower Pass Box, Cleanroom Air showers, Infinity Cleanroom Transfer Hatch, Infinity Pass Boxes, Soft Capsule Soft Wall Cleanroom, Dynamic Passboxes and Dynamic Floor Label hatches.
  4. Barrier Isolation System (CradlePro-Iso, Cytotoxic Safety cabinets)
  1. ASEPTiCell
  2. Vial Washing
  3. Automatic Trayloaders
  4. Depyrogenation Tunnel
  5. Filling/Stopper Inserting Machines
  6. Capping Machines
  7. Automatic Loading/Unloading Equipment
  8. External Vial Washer

Primary production is the processing of raw materials to create active pharmaceutical ingredients (APIs) and ancillary substances used in pharmaceutical formulations. The final API in the pharmaceutical that produces the therapeutic effect, should meet pharmacopoeial requirements. Primary manufacturing may involve either chemical or biological processes requiring different types of production facilities, technologies, skills, and knowledge. The manufacture of active ingredients is the most expensive aspect of pharmaceutical production because of the necessary investment in capital equipment, process development, and quality assurance systems.

Pharmaceutical Ingredients: These are portions of any drug considered active with which the reactions and results vary (depending on the drug’s administered dosage). Most drugs are made of more than one kind of pharmaceutical ingredients.

Components:

API or Active Pharmaceutical Ingredients -- the main ingredient known to have a biologic/ therapeutic activity.

Excipient or ancillary substance – the inert or inactive substance combined with a drug.

Secondary production is the large-scale processing of finished dosage forms (like tablets, capsules, parenteral, syrups etc.) from raw materials or intermediate products, from both local and imported sources. Production of sterile preparations (examples are as follow: injections, antibiotics, and intravenous fluids) and non-sterile preparations (oral solids, liquids, and topical drugs) can be carried out with either locally produced or imported packing materials. Just like primary production, this stage must be accomplished to clear-cut specifications. It necessitates modern, high-speed, precision equipment to produce drugs in their final form, in large quantities and at very low unit costs, meeting international GMP standards.

Tertiary production includes packaging and labelling finished products from primary and secondary sources into bulk packs, smaller dispensing packets, bottles, or course-of-therapy units for individual use. The initial quality of the pharmaceutical product established in the earlier phases of production must be maintained in the tertiary and final step, so ensuring high quality standards through rigorous operational procedures is important. This type of production can be developed first in many countries as a positive contribution that also builds industrial skill and experience. Tertiary production also addresses specific local needs for certain formulations, labeling, and packaging.


Phase 4

Post marketing surveillance is done to continually assess the safety of the drug and may include incidence, severity of rare adverse reaction, cost effectiveness analysis, comparative trials, and quality of life studies.